la ricerca: trasferibilità del modello...
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![Page 1: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/1.jpg)
Salvatore Cuzzocrea
Dipartimento di Scienze Biologiche ed Ambientali
La ricerca trasferibilitagrave del modello
animale
Donrsquot ask what your country can do for you Ask instead what
you can do for your countryrdquo ndash John F Kennedy
Donrsquot ask what a mice can do for
you Ask instead what you can
take from a mice studyrdquo
Traslational Research depends on your perspective
Traslational Research
Cell culture
Animal research
Clinical research
Should animals be used as
research subjects
Rats
I giovedigrave della cultura scientificaI giovedigrave della cultura scientificaldquoEtica e sperimentazione animale nella ricerca biomedicardquoldquoEtica e sperimentazione animale nella ricerca biomedicardquo
Genomica e modelli animali
di malat t ie umane
Mariateresa Mancuso
BI OTEC- MEDMice CD1
Mice C57BL6
Why mice
Easy to reproduce
Easy to handle
Genetically similar to humans
GENES 35000 vs 30000
80 of human genes
have a corresponding gene
in mice
Human Brain Mouse Brain
Mice VS Humans
ldquoBasicrdquo vs ldquoAppliedrdquo Research
Applied Research Taking the information
discovered in basic research and
investigating how to use it to treat and
prevent sicknesses
Example A researcher uses the
information about turning genes off and on
to find a drug that is used to turn off genes
that cause diseases and disorders in
humans
Segment of DNA
Many such segments
act as genes
Our animal models to study
Major Depressive Disorder (MDD)
Alzheimer disease (AD)
Parkinson disease (PD)
Major Depressive Disorder (MDD)
altered mood altered cognitive
functions
Severe forms of depression affect 2ndash5 of the US population and up to
20 of the population suffer from milder forms of the illness
Depression and Aging
Thielke Aging and Mental Health 2009
Incidence
Persistence
Endocannabinoid signalling
Boosting endocannabinoid tone might be
a useful alternative therapeutic approach
for depressive disorders
Neuroprotection (Alzheimer and
Huntington diseases Depression and
Schizophrenia)
Reduced
functionality might
be considered a
predisposing factor
for major
depression
PEA levels in rat brain following acute and chronic drug
administration
bull Naturally occurring fatty acid amide
bull Lipophilic features
bull Suitable for oral use
bull Present in many animal and plants commonly used for human nutrition
(Food for Special Medical Purposes)
bull In mammals it has been found in many tissues and body fluids
PEA
The levels of endogenous PEA in normal muscle and adipose tissues (004 - 600 pmolmg) are similar to those observed in the CNS and PNS and something higher compared to the gastrointestinal tract (005-150 pmolmg) skin (03- 069 pmolmg) eye (010 - 064 pmolmg) and kidney (25 ndash 71 pmolmg)
PEA levels in normal tissues
PEA
neurons microglia
astrocytes
mast cells
epidermal cells
macrophages
bull Produced ldquo on demand rdquo (enzymatically precursors) by neuro-immuno-inflammatory cells (eg mast cells microglia) exposed to potential damage from membrane
Analgesic
Anti-inflammatory
N-palmitoylethanolamide (PEA)
ldquoAliamides may represent the major breakthrough in inflammation we have been waiting forrdquo Jack D 1996 DNampP
Autacoid Local Injury Antagonism
Mini-Reviews in Medicinal Chemistry 2013 Vol 13 No 1
flavonoids
Pharmacological activity
Micronized coultraPEALut association
ratio of 101 in mass
was obtained by a co-ultramicronization process in an jet-mill equipment
Could ndash PEALut - treatment reduce the
behavioral responses associated with
depression
Ideg Hypothesis
Control group
Corticosterone-treated group
Corticosterone + PEA 1mgkg ip
Corticosterone + PEA 10mgkg ip
Corticosterone + luteolin 1mgkg ip
Corticosterone + luteolin 3mgkg ip
Corticosterone + luteolin 10mgkg ip
Corticosterone + PEA and lut (single
treatment combination ) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA luteolinand PEA and luteolin combination
Behavior (NSF)
Start of chronic
PEA and Luteolin treatment
Corticosterone
I Exp
Experimental design
Control group
Corticosterone-treated group
Corticosterone + PEA luteolin (co-ultramicronized) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA+luteolin compound
Behavior (NSF OF FST EPM)
Neurogenesis neuroplasticity neurotrophic and apoptotic
studies
Start of chronic
PEA+Luteolin treatment
Corticosterone
II Exp
Experimental design
Results-1 Anxiety-like behavior
deg
degdeg
Luteolin
deg
PEA
PEA and Lut
PEA + Lut
Results-1 Depression-like behavior - PEALut
FST
Could ndash PEALut - treatment increase
neurogenesis and neuroplasticity
IIdeg Hypothesis
Results-2 Neurogenesis - PEALut
A
50μm 50μm 50μm
Cort- Cort+ PEALut
Results-2 Neurogenesis - PEALut
Cort-
Cort+
PEA+Lut
50μm
50μm
50μm
Results-2 Neuroplasticity ndash PEALut
10microm
10microm
10microm
PEA+Lut
Cort+
Cort-
I
Adult Neurogenesis in Humans
β-actin
BDNF
Cort- Cort+ PEA+Lut
14kD
Results-2 Neurotrophic factor ndash PEALut
20microm 20microm 20microm
Cort- Cort+ PEA+Lut
IIIdeg Hypothesis
Could ldquoPEALutrdquo treatment modify
apoptotic protein expression
deg
Bax
β-actin
Cort- Cort+ PEA+Lu
t 23kD
Results-3 Apoptotic protein expression
β-actin
Bcl-2 29kD
PEA+Lut Cort+ Cort-
IVdeg Hypothesis
Could PEALut treatment reduce
neuroinflammation associated with Alzheimer
diseases
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 2: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/2.jpg)
Donrsquot ask what your country can do for you Ask instead what
you can do for your countryrdquo ndash John F Kennedy
Donrsquot ask what a mice can do for
you Ask instead what you can
take from a mice studyrdquo
Traslational Research depends on your perspective
Traslational Research
Cell culture
Animal research
Clinical research
Should animals be used as
research subjects
Rats
I giovedigrave della cultura scientificaI giovedigrave della cultura scientificaldquoEtica e sperimentazione animale nella ricerca biomedicardquoldquoEtica e sperimentazione animale nella ricerca biomedicardquo
Genomica e modelli animali
di malat t ie umane
Mariateresa Mancuso
BI OTEC- MEDMice CD1
Mice C57BL6
Why mice
Easy to reproduce
Easy to handle
Genetically similar to humans
GENES 35000 vs 30000
80 of human genes
have a corresponding gene
in mice
Human Brain Mouse Brain
Mice VS Humans
ldquoBasicrdquo vs ldquoAppliedrdquo Research
Applied Research Taking the information
discovered in basic research and
investigating how to use it to treat and
prevent sicknesses
Example A researcher uses the
information about turning genes off and on
to find a drug that is used to turn off genes
that cause diseases and disorders in
humans
Segment of DNA
Many such segments
act as genes
Our animal models to study
Major Depressive Disorder (MDD)
Alzheimer disease (AD)
Parkinson disease (PD)
Major Depressive Disorder (MDD)
altered mood altered cognitive
functions
Severe forms of depression affect 2ndash5 of the US population and up to
20 of the population suffer from milder forms of the illness
Depression and Aging
Thielke Aging and Mental Health 2009
Incidence
Persistence
Endocannabinoid signalling
Boosting endocannabinoid tone might be
a useful alternative therapeutic approach
for depressive disorders
Neuroprotection (Alzheimer and
Huntington diseases Depression and
Schizophrenia)
Reduced
functionality might
be considered a
predisposing factor
for major
depression
PEA levels in rat brain following acute and chronic drug
administration
bull Naturally occurring fatty acid amide
bull Lipophilic features
bull Suitable for oral use
bull Present in many animal and plants commonly used for human nutrition
(Food for Special Medical Purposes)
bull In mammals it has been found in many tissues and body fluids
PEA
The levels of endogenous PEA in normal muscle and adipose tissues (004 - 600 pmolmg) are similar to those observed in the CNS and PNS and something higher compared to the gastrointestinal tract (005-150 pmolmg) skin (03- 069 pmolmg) eye (010 - 064 pmolmg) and kidney (25 ndash 71 pmolmg)
PEA levels in normal tissues
PEA
neurons microglia
astrocytes
mast cells
epidermal cells
macrophages
bull Produced ldquo on demand rdquo (enzymatically precursors) by neuro-immuno-inflammatory cells (eg mast cells microglia) exposed to potential damage from membrane
Analgesic
Anti-inflammatory
N-palmitoylethanolamide (PEA)
ldquoAliamides may represent the major breakthrough in inflammation we have been waiting forrdquo Jack D 1996 DNampP
Autacoid Local Injury Antagonism
Mini-Reviews in Medicinal Chemistry 2013 Vol 13 No 1
flavonoids
Pharmacological activity
Micronized coultraPEALut association
ratio of 101 in mass
was obtained by a co-ultramicronization process in an jet-mill equipment
Could ndash PEALut - treatment reduce the
behavioral responses associated with
depression
Ideg Hypothesis
Control group
Corticosterone-treated group
Corticosterone + PEA 1mgkg ip
Corticosterone + PEA 10mgkg ip
Corticosterone + luteolin 1mgkg ip
Corticosterone + luteolin 3mgkg ip
Corticosterone + luteolin 10mgkg ip
Corticosterone + PEA and lut (single
treatment combination ) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA luteolinand PEA and luteolin combination
Behavior (NSF)
Start of chronic
PEA and Luteolin treatment
Corticosterone
I Exp
Experimental design
Control group
Corticosterone-treated group
Corticosterone + PEA luteolin (co-ultramicronized) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA+luteolin compound
Behavior (NSF OF FST EPM)
Neurogenesis neuroplasticity neurotrophic and apoptotic
studies
Start of chronic
PEA+Luteolin treatment
Corticosterone
II Exp
Experimental design
Results-1 Anxiety-like behavior
deg
degdeg
Luteolin
deg
PEA
PEA and Lut
PEA + Lut
Results-1 Depression-like behavior - PEALut
FST
Could ndash PEALut - treatment increase
neurogenesis and neuroplasticity
IIdeg Hypothesis
Results-2 Neurogenesis - PEALut
A
50μm 50μm 50μm
Cort- Cort+ PEALut
Results-2 Neurogenesis - PEALut
Cort-
Cort+
PEA+Lut
50μm
50μm
50μm
Results-2 Neuroplasticity ndash PEALut
10microm
10microm
10microm
PEA+Lut
Cort+
Cort-
I
Adult Neurogenesis in Humans
β-actin
BDNF
Cort- Cort+ PEA+Lut
14kD
Results-2 Neurotrophic factor ndash PEALut
20microm 20microm 20microm
Cort- Cort+ PEA+Lut
IIIdeg Hypothesis
Could ldquoPEALutrdquo treatment modify
apoptotic protein expression
deg
Bax
β-actin
Cort- Cort+ PEA+Lu
t 23kD
Results-3 Apoptotic protein expression
β-actin
Bcl-2 29kD
PEA+Lut Cort+ Cort-
IVdeg Hypothesis
Could PEALut treatment reduce
neuroinflammation associated with Alzheimer
diseases
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 3: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/3.jpg)
Traslational Research depends on your perspective
Traslational Research
Cell culture
Animal research
Clinical research
Should animals be used as
research subjects
Rats
I giovedigrave della cultura scientificaI giovedigrave della cultura scientificaldquoEtica e sperimentazione animale nella ricerca biomedicardquoldquoEtica e sperimentazione animale nella ricerca biomedicardquo
Genomica e modelli animali
di malat t ie umane
Mariateresa Mancuso
BI OTEC- MEDMice CD1
Mice C57BL6
Why mice
Easy to reproduce
Easy to handle
Genetically similar to humans
GENES 35000 vs 30000
80 of human genes
have a corresponding gene
in mice
Human Brain Mouse Brain
Mice VS Humans
ldquoBasicrdquo vs ldquoAppliedrdquo Research
Applied Research Taking the information
discovered in basic research and
investigating how to use it to treat and
prevent sicknesses
Example A researcher uses the
information about turning genes off and on
to find a drug that is used to turn off genes
that cause diseases and disorders in
humans
Segment of DNA
Many such segments
act as genes
Our animal models to study
Major Depressive Disorder (MDD)
Alzheimer disease (AD)
Parkinson disease (PD)
Major Depressive Disorder (MDD)
altered mood altered cognitive
functions
Severe forms of depression affect 2ndash5 of the US population and up to
20 of the population suffer from milder forms of the illness
Depression and Aging
Thielke Aging and Mental Health 2009
Incidence
Persistence
Endocannabinoid signalling
Boosting endocannabinoid tone might be
a useful alternative therapeutic approach
for depressive disorders
Neuroprotection (Alzheimer and
Huntington diseases Depression and
Schizophrenia)
Reduced
functionality might
be considered a
predisposing factor
for major
depression
PEA levels in rat brain following acute and chronic drug
administration
bull Naturally occurring fatty acid amide
bull Lipophilic features
bull Suitable for oral use
bull Present in many animal and plants commonly used for human nutrition
(Food for Special Medical Purposes)
bull In mammals it has been found in many tissues and body fluids
PEA
The levels of endogenous PEA in normal muscle and adipose tissues (004 - 600 pmolmg) are similar to those observed in the CNS and PNS and something higher compared to the gastrointestinal tract (005-150 pmolmg) skin (03- 069 pmolmg) eye (010 - 064 pmolmg) and kidney (25 ndash 71 pmolmg)
PEA levels in normal tissues
PEA
neurons microglia
astrocytes
mast cells
epidermal cells
macrophages
bull Produced ldquo on demand rdquo (enzymatically precursors) by neuro-immuno-inflammatory cells (eg mast cells microglia) exposed to potential damage from membrane
Analgesic
Anti-inflammatory
N-palmitoylethanolamide (PEA)
ldquoAliamides may represent the major breakthrough in inflammation we have been waiting forrdquo Jack D 1996 DNampP
Autacoid Local Injury Antagonism
Mini-Reviews in Medicinal Chemistry 2013 Vol 13 No 1
flavonoids
Pharmacological activity
Micronized coultraPEALut association
ratio of 101 in mass
was obtained by a co-ultramicronization process in an jet-mill equipment
Could ndash PEALut - treatment reduce the
behavioral responses associated with
depression
Ideg Hypothesis
Control group
Corticosterone-treated group
Corticosterone + PEA 1mgkg ip
Corticosterone + PEA 10mgkg ip
Corticosterone + luteolin 1mgkg ip
Corticosterone + luteolin 3mgkg ip
Corticosterone + luteolin 10mgkg ip
Corticosterone + PEA and lut (single
treatment combination ) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA luteolinand PEA and luteolin combination
Behavior (NSF)
Start of chronic
PEA and Luteolin treatment
Corticosterone
I Exp
Experimental design
Control group
Corticosterone-treated group
Corticosterone + PEA luteolin (co-ultramicronized) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA+luteolin compound
Behavior (NSF OF FST EPM)
Neurogenesis neuroplasticity neurotrophic and apoptotic
studies
Start of chronic
PEA+Luteolin treatment
Corticosterone
II Exp
Experimental design
Results-1 Anxiety-like behavior
deg
degdeg
Luteolin
deg
PEA
PEA and Lut
PEA + Lut
Results-1 Depression-like behavior - PEALut
FST
Could ndash PEALut - treatment increase
neurogenesis and neuroplasticity
IIdeg Hypothesis
Results-2 Neurogenesis - PEALut
A
50μm 50μm 50μm
Cort- Cort+ PEALut
Results-2 Neurogenesis - PEALut
Cort-
Cort+
PEA+Lut
50μm
50μm
50μm
Results-2 Neuroplasticity ndash PEALut
10microm
10microm
10microm
PEA+Lut
Cort+
Cort-
I
Adult Neurogenesis in Humans
β-actin
BDNF
Cort- Cort+ PEA+Lut
14kD
Results-2 Neurotrophic factor ndash PEALut
20microm 20microm 20microm
Cort- Cort+ PEA+Lut
IIIdeg Hypothesis
Could ldquoPEALutrdquo treatment modify
apoptotic protein expression
deg
Bax
β-actin
Cort- Cort+ PEA+Lu
t 23kD
Results-3 Apoptotic protein expression
β-actin
Bcl-2 29kD
PEA+Lut Cort+ Cort-
IVdeg Hypothesis
Could PEALut treatment reduce
neuroinflammation associated with Alzheimer
diseases
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 4: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/4.jpg)
Should animals be used as
research subjects
Rats
I giovedigrave della cultura scientificaI giovedigrave della cultura scientificaldquoEtica e sperimentazione animale nella ricerca biomedicardquoldquoEtica e sperimentazione animale nella ricerca biomedicardquo
Genomica e modelli animali
di malat t ie umane
Mariateresa Mancuso
BI OTEC- MEDMice CD1
Mice C57BL6
Why mice
Easy to reproduce
Easy to handle
Genetically similar to humans
GENES 35000 vs 30000
80 of human genes
have a corresponding gene
in mice
Human Brain Mouse Brain
Mice VS Humans
ldquoBasicrdquo vs ldquoAppliedrdquo Research
Applied Research Taking the information
discovered in basic research and
investigating how to use it to treat and
prevent sicknesses
Example A researcher uses the
information about turning genes off and on
to find a drug that is used to turn off genes
that cause diseases and disorders in
humans
Segment of DNA
Many such segments
act as genes
Our animal models to study
Major Depressive Disorder (MDD)
Alzheimer disease (AD)
Parkinson disease (PD)
Major Depressive Disorder (MDD)
altered mood altered cognitive
functions
Severe forms of depression affect 2ndash5 of the US population and up to
20 of the population suffer from milder forms of the illness
Depression and Aging
Thielke Aging and Mental Health 2009
Incidence
Persistence
Endocannabinoid signalling
Boosting endocannabinoid tone might be
a useful alternative therapeutic approach
for depressive disorders
Neuroprotection (Alzheimer and
Huntington diseases Depression and
Schizophrenia)
Reduced
functionality might
be considered a
predisposing factor
for major
depression
PEA levels in rat brain following acute and chronic drug
administration
bull Naturally occurring fatty acid amide
bull Lipophilic features
bull Suitable for oral use
bull Present in many animal and plants commonly used for human nutrition
(Food for Special Medical Purposes)
bull In mammals it has been found in many tissues and body fluids
PEA
The levels of endogenous PEA in normal muscle and adipose tissues (004 - 600 pmolmg) are similar to those observed in the CNS and PNS and something higher compared to the gastrointestinal tract (005-150 pmolmg) skin (03- 069 pmolmg) eye (010 - 064 pmolmg) and kidney (25 ndash 71 pmolmg)
PEA levels in normal tissues
PEA
neurons microglia
astrocytes
mast cells
epidermal cells
macrophages
bull Produced ldquo on demand rdquo (enzymatically precursors) by neuro-immuno-inflammatory cells (eg mast cells microglia) exposed to potential damage from membrane
Analgesic
Anti-inflammatory
N-palmitoylethanolamide (PEA)
ldquoAliamides may represent the major breakthrough in inflammation we have been waiting forrdquo Jack D 1996 DNampP
Autacoid Local Injury Antagonism
Mini-Reviews in Medicinal Chemistry 2013 Vol 13 No 1
flavonoids
Pharmacological activity
Micronized coultraPEALut association
ratio of 101 in mass
was obtained by a co-ultramicronization process in an jet-mill equipment
Could ndash PEALut - treatment reduce the
behavioral responses associated with
depression
Ideg Hypothesis
Control group
Corticosterone-treated group
Corticosterone + PEA 1mgkg ip
Corticosterone + PEA 10mgkg ip
Corticosterone + luteolin 1mgkg ip
Corticosterone + luteolin 3mgkg ip
Corticosterone + luteolin 10mgkg ip
Corticosterone + PEA and lut (single
treatment combination ) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA luteolinand PEA and luteolin combination
Behavior (NSF)
Start of chronic
PEA and Luteolin treatment
Corticosterone
I Exp
Experimental design
Control group
Corticosterone-treated group
Corticosterone + PEA luteolin (co-ultramicronized) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA+luteolin compound
Behavior (NSF OF FST EPM)
Neurogenesis neuroplasticity neurotrophic and apoptotic
studies
Start of chronic
PEA+Luteolin treatment
Corticosterone
II Exp
Experimental design
Results-1 Anxiety-like behavior
deg
degdeg
Luteolin
deg
PEA
PEA and Lut
PEA + Lut
Results-1 Depression-like behavior - PEALut
FST
Could ndash PEALut - treatment increase
neurogenesis and neuroplasticity
IIdeg Hypothesis
Results-2 Neurogenesis - PEALut
A
50μm 50μm 50μm
Cort- Cort+ PEALut
Results-2 Neurogenesis - PEALut
Cort-
Cort+
PEA+Lut
50μm
50μm
50μm
Results-2 Neuroplasticity ndash PEALut
10microm
10microm
10microm
PEA+Lut
Cort+
Cort-
I
Adult Neurogenesis in Humans
β-actin
BDNF
Cort- Cort+ PEA+Lut
14kD
Results-2 Neurotrophic factor ndash PEALut
20microm 20microm 20microm
Cort- Cort+ PEA+Lut
IIIdeg Hypothesis
Could ldquoPEALutrdquo treatment modify
apoptotic protein expression
deg
Bax
β-actin
Cort- Cort+ PEA+Lu
t 23kD
Results-3 Apoptotic protein expression
β-actin
Bcl-2 29kD
PEA+Lut Cort+ Cort-
IVdeg Hypothesis
Could PEALut treatment reduce
neuroinflammation associated with Alzheimer
diseases
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 5: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/5.jpg)
Why mice
Easy to reproduce
Easy to handle
Genetically similar to humans
GENES 35000 vs 30000
80 of human genes
have a corresponding gene
in mice
Human Brain Mouse Brain
Mice VS Humans
ldquoBasicrdquo vs ldquoAppliedrdquo Research
Applied Research Taking the information
discovered in basic research and
investigating how to use it to treat and
prevent sicknesses
Example A researcher uses the
information about turning genes off and on
to find a drug that is used to turn off genes
that cause diseases and disorders in
humans
Segment of DNA
Many such segments
act as genes
Our animal models to study
Major Depressive Disorder (MDD)
Alzheimer disease (AD)
Parkinson disease (PD)
Major Depressive Disorder (MDD)
altered mood altered cognitive
functions
Severe forms of depression affect 2ndash5 of the US population and up to
20 of the population suffer from milder forms of the illness
Depression and Aging
Thielke Aging and Mental Health 2009
Incidence
Persistence
Endocannabinoid signalling
Boosting endocannabinoid tone might be
a useful alternative therapeutic approach
for depressive disorders
Neuroprotection (Alzheimer and
Huntington diseases Depression and
Schizophrenia)
Reduced
functionality might
be considered a
predisposing factor
for major
depression
PEA levels in rat brain following acute and chronic drug
administration
bull Naturally occurring fatty acid amide
bull Lipophilic features
bull Suitable for oral use
bull Present in many animal and plants commonly used for human nutrition
(Food for Special Medical Purposes)
bull In mammals it has been found in many tissues and body fluids
PEA
The levels of endogenous PEA in normal muscle and adipose tissues (004 - 600 pmolmg) are similar to those observed in the CNS and PNS and something higher compared to the gastrointestinal tract (005-150 pmolmg) skin (03- 069 pmolmg) eye (010 - 064 pmolmg) and kidney (25 ndash 71 pmolmg)
PEA levels in normal tissues
PEA
neurons microglia
astrocytes
mast cells
epidermal cells
macrophages
bull Produced ldquo on demand rdquo (enzymatically precursors) by neuro-immuno-inflammatory cells (eg mast cells microglia) exposed to potential damage from membrane
Analgesic
Anti-inflammatory
N-palmitoylethanolamide (PEA)
ldquoAliamides may represent the major breakthrough in inflammation we have been waiting forrdquo Jack D 1996 DNampP
Autacoid Local Injury Antagonism
Mini-Reviews in Medicinal Chemistry 2013 Vol 13 No 1
flavonoids
Pharmacological activity
Micronized coultraPEALut association
ratio of 101 in mass
was obtained by a co-ultramicronization process in an jet-mill equipment
Could ndash PEALut - treatment reduce the
behavioral responses associated with
depression
Ideg Hypothesis
Control group
Corticosterone-treated group
Corticosterone + PEA 1mgkg ip
Corticosterone + PEA 10mgkg ip
Corticosterone + luteolin 1mgkg ip
Corticosterone + luteolin 3mgkg ip
Corticosterone + luteolin 10mgkg ip
Corticosterone + PEA and lut (single
treatment combination ) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA luteolinand PEA and luteolin combination
Behavior (NSF)
Start of chronic
PEA and Luteolin treatment
Corticosterone
I Exp
Experimental design
Control group
Corticosterone-treated group
Corticosterone + PEA luteolin (co-ultramicronized) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA+luteolin compound
Behavior (NSF OF FST EPM)
Neurogenesis neuroplasticity neurotrophic and apoptotic
studies
Start of chronic
PEA+Luteolin treatment
Corticosterone
II Exp
Experimental design
Results-1 Anxiety-like behavior
deg
degdeg
Luteolin
deg
PEA
PEA and Lut
PEA + Lut
Results-1 Depression-like behavior - PEALut
FST
Could ndash PEALut - treatment increase
neurogenesis and neuroplasticity
IIdeg Hypothesis
Results-2 Neurogenesis - PEALut
A
50μm 50μm 50μm
Cort- Cort+ PEALut
Results-2 Neurogenesis - PEALut
Cort-
Cort+
PEA+Lut
50μm
50μm
50μm
Results-2 Neuroplasticity ndash PEALut
10microm
10microm
10microm
PEA+Lut
Cort+
Cort-
I
Adult Neurogenesis in Humans
β-actin
BDNF
Cort- Cort+ PEA+Lut
14kD
Results-2 Neurotrophic factor ndash PEALut
20microm 20microm 20microm
Cort- Cort+ PEA+Lut
IIIdeg Hypothesis
Could ldquoPEALutrdquo treatment modify
apoptotic protein expression
deg
Bax
β-actin
Cort- Cort+ PEA+Lu
t 23kD
Results-3 Apoptotic protein expression
β-actin
Bcl-2 29kD
PEA+Lut Cort+ Cort-
IVdeg Hypothesis
Could PEALut treatment reduce
neuroinflammation associated with Alzheimer
diseases
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 6: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/6.jpg)
Human Brain Mouse Brain
Mice VS Humans
ldquoBasicrdquo vs ldquoAppliedrdquo Research
Applied Research Taking the information
discovered in basic research and
investigating how to use it to treat and
prevent sicknesses
Example A researcher uses the
information about turning genes off and on
to find a drug that is used to turn off genes
that cause diseases and disorders in
humans
Segment of DNA
Many such segments
act as genes
Our animal models to study
Major Depressive Disorder (MDD)
Alzheimer disease (AD)
Parkinson disease (PD)
Major Depressive Disorder (MDD)
altered mood altered cognitive
functions
Severe forms of depression affect 2ndash5 of the US population and up to
20 of the population suffer from milder forms of the illness
Depression and Aging
Thielke Aging and Mental Health 2009
Incidence
Persistence
Endocannabinoid signalling
Boosting endocannabinoid tone might be
a useful alternative therapeutic approach
for depressive disorders
Neuroprotection (Alzheimer and
Huntington diseases Depression and
Schizophrenia)
Reduced
functionality might
be considered a
predisposing factor
for major
depression
PEA levels in rat brain following acute and chronic drug
administration
bull Naturally occurring fatty acid amide
bull Lipophilic features
bull Suitable for oral use
bull Present in many animal and plants commonly used for human nutrition
(Food for Special Medical Purposes)
bull In mammals it has been found in many tissues and body fluids
PEA
The levels of endogenous PEA in normal muscle and adipose tissues (004 - 600 pmolmg) are similar to those observed in the CNS and PNS and something higher compared to the gastrointestinal tract (005-150 pmolmg) skin (03- 069 pmolmg) eye (010 - 064 pmolmg) and kidney (25 ndash 71 pmolmg)
PEA levels in normal tissues
PEA
neurons microglia
astrocytes
mast cells
epidermal cells
macrophages
bull Produced ldquo on demand rdquo (enzymatically precursors) by neuro-immuno-inflammatory cells (eg mast cells microglia) exposed to potential damage from membrane
Analgesic
Anti-inflammatory
N-palmitoylethanolamide (PEA)
ldquoAliamides may represent the major breakthrough in inflammation we have been waiting forrdquo Jack D 1996 DNampP
Autacoid Local Injury Antagonism
Mini-Reviews in Medicinal Chemistry 2013 Vol 13 No 1
flavonoids
Pharmacological activity
Micronized coultraPEALut association
ratio of 101 in mass
was obtained by a co-ultramicronization process in an jet-mill equipment
Could ndash PEALut - treatment reduce the
behavioral responses associated with
depression
Ideg Hypothesis
Control group
Corticosterone-treated group
Corticosterone + PEA 1mgkg ip
Corticosterone + PEA 10mgkg ip
Corticosterone + luteolin 1mgkg ip
Corticosterone + luteolin 3mgkg ip
Corticosterone + luteolin 10mgkg ip
Corticosterone + PEA and lut (single
treatment combination ) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA luteolinand PEA and luteolin combination
Behavior (NSF)
Start of chronic
PEA and Luteolin treatment
Corticosterone
I Exp
Experimental design
Control group
Corticosterone-treated group
Corticosterone + PEA luteolin (co-ultramicronized) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA+luteolin compound
Behavior (NSF OF FST EPM)
Neurogenesis neuroplasticity neurotrophic and apoptotic
studies
Start of chronic
PEA+Luteolin treatment
Corticosterone
II Exp
Experimental design
Results-1 Anxiety-like behavior
deg
degdeg
Luteolin
deg
PEA
PEA and Lut
PEA + Lut
Results-1 Depression-like behavior - PEALut
FST
Could ndash PEALut - treatment increase
neurogenesis and neuroplasticity
IIdeg Hypothesis
Results-2 Neurogenesis - PEALut
A
50μm 50μm 50μm
Cort- Cort+ PEALut
Results-2 Neurogenesis - PEALut
Cort-
Cort+
PEA+Lut
50μm
50μm
50μm
Results-2 Neuroplasticity ndash PEALut
10microm
10microm
10microm
PEA+Lut
Cort+
Cort-
I
Adult Neurogenesis in Humans
β-actin
BDNF
Cort- Cort+ PEA+Lut
14kD
Results-2 Neurotrophic factor ndash PEALut
20microm 20microm 20microm
Cort- Cort+ PEA+Lut
IIIdeg Hypothesis
Could ldquoPEALutrdquo treatment modify
apoptotic protein expression
deg
Bax
β-actin
Cort- Cort+ PEA+Lu
t 23kD
Results-3 Apoptotic protein expression
β-actin
Bcl-2 29kD
PEA+Lut Cort+ Cort-
IVdeg Hypothesis
Could PEALut treatment reduce
neuroinflammation associated with Alzheimer
diseases
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 7: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/7.jpg)
ldquoBasicrdquo vs ldquoAppliedrdquo Research
Applied Research Taking the information
discovered in basic research and
investigating how to use it to treat and
prevent sicknesses
Example A researcher uses the
information about turning genes off and on
to find a drug that is used to turn off genes
that cause diseases and disorders in
humans
Segment of DNA
Many such segments
act as genes
Our animal models to study
Major Depressive Disorder (MDD)
Alzheimer disease (AD)
Parkinson disease (PD)
Major Depressive Disorder (MDD)
altered mood altered cognitive
functions
Severe forms of depression affect 2ndash5 of the US population and up to
20 of the population suffer from milder forms of the illness
Depression and Aging
Thielke Aging and Mental Health 2009
Incidence
Persistence
Endocannabinoid signalling
Boosting endocannabinoid tone might be
a useful alternative therapeutic approach
for depressive disorders
Neuroprotection (Alzheimer and
Huntington diseases Depression and
Schizophrenia)
Reduced
functionality might
be considered a
predisposing factor
for major
depression
PEA levels in rat brain following acute and chronic drug
administration
bull Naturally occurring fatty acid amide
bull Lipophilic features
bull Suitable for oral use
bull Present in many animal and plants commonly used for human nutrition
(Food for Special Medical Purposes)
bull In mammals it has been found in many tissues and body fluids
PEA
The levels of endogenous PEA in normal muscle and adipose tissues (004 - 600 pmolmg) are similar to those observed in the CNS and PNS and something higher compared to the gastrointestinal tract (005-150 pmolmg) skin (03- 069 pmolmg) eye (010 - 064 pmolmg) and kidney (25 ndash 71 pmolmg)
PEA levels in normal tissues
PEA
neurons microglia
astrocytes
mast cells
epidermal cells
macrophages
bull Produced ldquo on demand rdquo (enzymatically precursors) by neuro-immuno-inflammatory cells (eg mast cells microglia) exposed to potential damage from membrane
Analgesic
Anti-inflammatory
N-palmitoylethanolamide (PEA)
ldquoAliamides may represent the major breakthrough in inflammation we have been waiting forrdquo Jack D 1996 DNampP
Autacoid Local Injury Antagonism
Mini-Reviews in Medicinal Chemistry 2013 Vol 13 No 1
flavonoids
Pharmacological activity
Micronized coultraPEALut association
ratio of 101 in mass
was obtained by a co-ultramicronization process in an jet-mill equipment
Could ndash PEALut - treatment reduce the
behavioral responses associated with
depression
Ideg Hypothesis
Control group
Corticosterone-treated group
Corticosterone + PEA 1mgkg ip
Corticosterone + PEA 10mgkg ip
Corticosterone + luteolin 1mgkg ip
Corticosterone + luteolin 3mgkg ip
Corticosterone + luteolin 10mgkg ip
Corticosterone + PEA and lut (single
treatment combination ) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA luteolinand PEA and luteolin combination
Behavior (NSF)
Start of chronic
PEA and Luteolin treatment
Corticosterone
I Exp
Experimental design
Control group
Corticosterone-treated group
Corticosterone + PEA luteolin (co-ultramicronized) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA+luteolin compound
Behavior (NSF OF FST EPM)
Neurogenesis neuroplasticity neurotrophic and apoptotic
studies
Start of chronic
PEA+Luteolin treatment
Corticosterone
II Exp
Experimental design
Results-1 Anxiety-like behavior
deg
degdeg
Luteolin
deg
PEA
PEA and Lut
PEA + Lut
Results-1 Depression-like behavior - PEALut
FST
Could ndash PEALut - treatment increase
neurogenesis and neuroplasticity
IIdeg Hypothesis
Results-2 Neurogenesis - PEALut
A
50μm 50μm 50μm
Cort- Cort+ PEALut
Results-2 Neurogenesis - PEALut
Cort-
Cort+
PEA+Lut
50μm
50μm
50μm
Results-2 Neuroplasticity ndash PEALut
10microm
10microm
10microm
PEA+Lut
Cort+
Cort-
I
Adult Neurogenesis in Humans
β-actin
BDNF
Cort- Cort+ PEA+Lut
14kD
Results-2 Neurotrophic factor ndash PEALut
20microm 20microm 20microm
Cort- Cort+ PEA+Lut
IIIdeg Hypothesis
Could ldquoPEALutrdquo treatment modify
apoptotic protein expression
deg
Bax
β-actin
Cort- Cort+ PEA+Lu
t 23kD
Results-3 Apoptotic protein expression
β-actin
Bcl-2 29kD
PEA+Lut Cort+ Cort-
IVdeg Hypothesis
Could PEALut treatment reduce
neuroinflammation associated with Alzheimer
diseases
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 8: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/8.jpg)
Our animal models to study
Major Depressive Disorder (MDD)
Alzheimer disease (AD)
Parkinson disease (PD)
Major Depressive Disorder (MDD)
altered mood altered cognitive
functions
Severe forms of depression affect 2ndash5 of the US population and up to
20 of the population suffer from milder forms of the illness
Depression and Aging
Thielke Aging and Mental Health 2009
Incidence
Persistence
Endocannabinoid signalling
Boosting endocannabinoid tone might be
a useful alternative therapeutic approach
for depressive disorders
Neuroprotection (Alzheimer and
Huntington diseases Depression and
Schizophrenia)
Reduced
functionality might
be considered a
predisposing factor
for major
depression
PEA levels in rat brain following acute and chronic drug
administration
bull Naturally occurring fatty acid amide
bull Lipophilic features
bull Suitable for oral use
bull Present in many animal and plants commonly used for human nutrition
(Food for Special Medical Purposes)
bull In mammals it has been found in many tissues and body fluids
PEA
The levels of endogenous PEA in normal muscle and adipose tissues (004 - 600 pmolmg) are similar to those observed in the CNS and PNS and something higher compared to the gastrointestinal tract (005-150 pmolmg) skin (03- 069 pmolmg) eye (010 - 064 pmolmg) and kidney (25 ndash 71 pmolmg)
PEA levels in normal tissues
PEA
neurons microglia
astrocytes
mast cells
epidermal cells
macrophages
bull Produced ldquo on demand rdquo (enzymatically precursors) by neuro-immuno-inflammatory cells (eg mast cells microglia) exposed to potential damage from membrane
Analgesic
Anti-inflammatory
N-palmitoylethanolamide (PEA)
ldquoAliamides may represent the major breakthrough in inflammation we have been waiting forrdquo Jack D 1996 DNampP
Autacoid Local Injury Antagonism
Mini-Reviews in Medicinal Chemistry 2013 Vol 13 No 1
flavonoids
Pharmacological activity
Micronized coultraPEALut association
ratio of 101 in mass
was obtained by a co-ultramicronization process in an jet-mill equipment
Could ndash PEALut - treatment reduce the
behavioral responses associated with
depression
Ideg Hypothesis
Control group
Corticosterone-treated group
Corticosterone + PEA 1mgkg ip
Corticosterone + PEA 10mgkg ip
Corticosterone + luteolin 1mgkg ip
Corticosterone + luteolin 3mgkg ip
Corticosterone + luteolin 10mgkg ip
Corticosterone + PEA and lut (single
treatment combination ) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA luteolinand PEA and luteolin combination
Behavior (NSF)
Start of chronic
PEA and Luteolin treatment
Corticosterone
I Exp
Experimental design
Control group
Corticosterone-treated group
Corticosterone + PEA luteolin (co-ultramicronized) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA+luteolin compound
Behavior (NSF OF FST EPM)
Neurogenesis neuroplasticity neurotrophic and apoptotic
studies
Start of chronic
PEA+Luteolin treatment
Corticosterone
II Exp
Experimental design
Results-1 Anxiety-like behavior
deg
degdeg
Luteolin
deg
PEA
PEA and Lut
PEA + Lut
Results-1 Depression-like behavior - PEALut
FST
Could ndash PEALut - treatment increase
neurogenesis and neuroplasticity
IIdeg Hypothesis
Results-2 Neurogenesis - PEALut
A
50μm 50μm 50μm
Cort- Cort+ PEALut
Results-2 Neurogenesis - PEALut
Cort-
Cort+
PEA+Lut
50μm
50μm
50μm
Results-2 Neuroplasticity ndash PEALut
10microm
10microm
10microm
PEA+Lut
Cort+
Cort-
I
Adult Neurogenesis in Humans
β-actin
BDNF
Cort- Cort+ PEA+Lut
14kD
Results-2 Neurotrophic factor ndash PEALut
20microm 20microm 20microm
Cort- Cort+ PEA+Lut
IIIdeg Hypothesis
Could ldquoPEALutrdquo treatment modify
apoptotic protein expression
deg
Bax
β-actin
Cort- Cort+ PEA+Lu
t 23kD
Results-3 Apoptotic protein expression
β-actin
Bcl-2 29kD
PEA+Lut Cort+ Cort-
IVdeg Hypothesis
Could PEALut treatment reduce
neuroinflammation associated with Alzheimer
diseases
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 9: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/9.jpg)
Major Depressive Disorder (MDD)
altered mood altered cognitive
functions
Severe forms of depression affect 2ndash5 of the US population and up to
20 of the population suffer from milder forms of the illness
Depression and Aging
Thielke Aging and Mental Health 2009
Incidence
Persistence
Endocannabinoid signalling
Boosting endocannabinoid tone might be
a useful alternative therapeutic approach
for depressive disorders
Neuroprotection (Alzheimer and
Huntington diseases Depression and
Schizophrenia)
Reduced
functionality might
be considered a
predisposing factor
for major
depression
PEA levels in rat brain following acute and chronic drug
administration
bull Naturally occurring fatty acid amide
bull Lipophilic features
bull Suitable for oral use
bull Present in many animal and plants commonly used for human nutrition
(Food for Special Medical Purposes)
bull In mammals it has been found in many tissues and body fluids
PEA
The levels of endogenous PEA in normal muscle and adipose tissues (004 - 600 pmolmg) are similar to those observed in the CNS and PNS and something higher compared to the gastrointestinal tract (005-150 pmolmg) skin (03- 069 pmolmg) eye (010 - 064 pmolmg) and kidney (25 ndash 71 pmolmg)
PEA levels in normal tissues
PEA
neurons microglia
astrocytes
mast cells
epidermal cells
macrophages
bull Produced ldquo on demand rdquo (enzymatically precursors) by neuro-immuno-inflammatory cells (eg mast cells microglia) exposed to potential damage from membrane
Analgesic
Anti-inflammatory
N-palmitoylethanolamide (PEA)
ldquoAliamides may represent the major breakthrough in inflammation we have been waiting forrdquo Jack D 1996 DNampP
Autacoid Local Injury Antagonism
Mini-Reviews in Medicinal Chemistry 2013 Vol 13 No 1
flavonoids
Pharmacological activity
Micronized coultraPEALut association
ratio of 101 in mass
was obtained by a co-ultramicronization process in an jet-mill equipment
Could ndash PEALut - treatment reduce the
behavioral responses associated with
depression
Ideg Hypothesis
Control group
Corticosterone-treated group
Corticosterone + PEA 1mgkg ip
Corticosterone + PEA 10mgkg ip
Corticosterone + luteolin 1mgkg ip
Corticosterone + luteolin 3mgkg ip
Corticosterone + luteolin 10mgkg ip
Corticosterone + PEA and lut (single
treatment combination ) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA luteolinand PEA and luteolin combination
Behavior (NSF)
Start of chronic
PEA and Luteolin treatment
Corticosterone
I Exp
Experimental design
Control group
Corticosterone-treated group
Corticosterone + PEA luteolin (co-ultramicronized) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA+luteolin compound
Behavior (NSF OF FST EPM)
Neurogenesis neuroplasticity neurotrophic and apoptotic
studies
Start of chronic
PEA+Luteolin treatment
Corticosterone
II Exp
Experimental design
Results-1 Anxiety-like behavior
deg
degdeg
Luteolin
deg
PEA
PEA and Lut
PEA + Lut
Results-1 Depression-like behavior - PEALut
FST
Could ndash PEALut - treatment increase
neurogenesis and neuroplasticity
IIdeg Hypothesis
Results-2 Neurogenesis - PEALut
A
50μm 50μm 50μm
Cort- Cort+ PEALut
Results-2 Neurogenesis - PEALut
Cort-
Cort+
PEA+Lut
50μm
50μm
50μm
Results-2 Neuroplasticity ndash PEALut
10microm
10microm
10microm
PEA+Lut
Cort+
Cort-
I
Adult Neurogenesis in Humans
β-actin
BDNF
Cort- Cort+ PEA+Lut
14kD
Results-2 Neurotrophic factor ndash PEALut
20microm 20microm 20microm
Cort- Cort+ PEA+Lut
IIIdeg Hypothesis
Could ldquoPEALutrdquo treatment modify
apoptotic protein expression
deg
Bax
β-actin
Cort- Cort+ PEA+Lu
t 23kD
Results-3 Apoptotic protein expression
β-actin
Bcl-2 29kD
PEA+Lut Cort+ Cort-
IVdeg Hypothesis
Could PEALut treatment reduce
neuroinflammation associated with Alzheimer
diseases
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 10: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/10.jpg)
Depression and Aging
Thielke Aging and Mental Health 2009
Incidence
Persistence
Endocannabinoid signalling
Boosting endocannabinoid tone might be
a useful alternative therapeutic approach
for depressive disorders
Neuroprotection (Alzheimer and
Huntington diseases Depression and
Schizophrenia)
Reduced
functionality might
be considered a
predisposing factor
for major
depression
PEA levels in rat brain following acute and chronic drug
administration
bull Naturally occurring fatty acid amide
bull Lipophilic features
bull Suitable for oral use
bull Present in many animal and plants commonly used for human nutrition
(Food for Special Medical Purposes)
bull In mammals it has been found in many tissues and body fluids
PEA
The levels of endogenous PEA in normal muscle and adipose tissues (004 - 600 pmolmg) are similar to those observed in the CNS and PNS and something higher compared to the gastrointestinal tract (005-150 pmolmg) skin (03- 069 pmolmg) eye (010 - 064 pmolmg) and kidney (25 ndash 71 pmolmg)
PEA levels in normal tissues
PEA
neurons microglia
astrocytes
mast cells
epidermal cells
macrophages
bull Produced ldquo on demand rdquo (enzymatically precursors) by neuro-immuno-inflammatory cells (eg mast cells microglia) exposed to potential damage from membrane
Analgesic
Anti-inflammatory
N-palmitoylethanolamide (PEA)
ldquoAliamides may represent the major breakthrough in inflammation we have been waiting forrdquo Jack D 1996 DNampP
Autacoid Local Injury Antagonism
Mini-Reviews in Medicinal Chemistry 2013 Vol 13 No 1
flavonoids
Pharmacological activity
Micronized coultraPEALut association
ratio of 101 in mass
was obtained by a co-ultramicronization process in an jet-mill equipment
Could ndash PEALut - treatment reduce the
behavioral responses associated with
depression
Ideg Hypothesis
Control group
Corticosterone-treated group
Corticosterone + PEA 1mgkg ip
Corticosterone + PEA 10mgkg ip
Corticosterone + luteolin 1mgkg ip
Corticosterone + luteolin 3mgkg ip
Corticosterone + luteolin 10mgkg ip
Corticosterone + PEA and lut (single
treatment combination ) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA luteolinand PEA and luteolin combination
Behavior (NSF)
Start of chronic
PEA and Luteolin treatment
Corticosterone
I Exp
Experimental design
Control group
Corticosterone-treated group
Corticosterone + PEA luteolin (co-ultramicronized) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA+luteolin compound
Behavior (NSF OF FST EPM)
Neurogenesis neuroplasticity neurotrophic and apoptotic
studies
Start of chronic
PEA+Luteolin treatment
Corticosterone
II Exp
Experimental design
Results-1 Anxiety-like behavior
deg
degdeg
Luteolin
deg
PEA
PEA and Lut
PEA + Lut
Results-1 Depression-like behavior - PEALut
FST
Could ndash PEALut - treatment increase
neurogenesis and neuroplasticity
IIdeg Hypothesis
Results-2 Neurogenesis - PEALut
A
50μm 50μm 50μm
Cort- Cort+ PEALut
Results-2 Neurogenesis - PEALut
Cort-
Cort+
PEA+Lut
50μm
50μm
50μm
Results-2 Neuroplasticity ndash PEALut
10microm
10microm
10microm
PEA+Lut
Cort+
Cort-
I
Adult Neurogenesis in Humans
β-actin
BDNF
Cort- Cort+ PEA+Lut
14kD
Results-2 Neurotrophic factor ndash PEALut
20microm 20microm 20microm
Cort- Cort+ PEA+Lut
IIIdeg Hypothesis
Could ldquoPEALutrdquo treatment modify
apoptotic protein expression
deg
Bax
β-actin
Cort- Cort+ PEA+Lu
t 23kD
Results-3 Apoptotic protein expression
β-actin
Bcl-2 29kD
PEA+Lut Cort+ Cort-
IVdeg Hypothesis
Could PEALut treatment reduce
neuroinflammation associated with Alzheimer
diseases
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 11: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/11.jpg)
Endocannabinoid signalling
Boosting endocannabinoid tone might be
a useful alternative therapeutic approach
for depressive disorders
Neuroprotection (Alzheimer and
Huntington diseases Depression and
Schizophrenia)
Reduced
functionality might
be considered a
predisposing factor
for major
depression
PEA levels in rat brain following acute and chronic drug
administration
bull Naturally occurring fatty acid amide
bull Lipophilic features
bull Suitable for oral use
bull Present in many animal and plants commonly used for human nutrition
(Food for Special Medical Purposes)
bull In mammals it has been found in many tissues and body fluids
PEA
The levels of endogenous PEA in normal muscle and adipose tissues (004 - 600 pmolmg) are similar to those observed in the CNS and PNS and something higher compared to the gastrointestinal tract (005-150 pmolmg) skin (03- 069 pmolmg) eye (010 - 064 pmolmg) and kidney (25 ndash 71 pmolmg)
PEA levels in normal tissues
PEA
neurons microglia
astrocytes
mast cells
epidermal cells
macrophages
bull Produced ldquo on demand rdquo (enzymatically precursors) by neuro-immuno-inflammatory cells (eg mast cells microglia) exposed to potential damage from membrane
Analgesic
Anti-inflammatory
N-palmitoylethanolamide (PEA)
ldquoAliamides may represent the major breakthrough in inflammation we have been waiting forrdquo Jack D 1996 DNampP
Autacoid Local Injury Antagonism
Mini-Reviews in Medicinal Chemistry 2013 Vol 13 No 1
flavonoids
Pharmacological activity
Micronized coultraPEALut association
ratio of 101 in mass
was obtained by a co-ultramicronization process in an jet-mill equipment
Could ndash PEALut - treatment reduce the
behavioral responses associated with
depression
Ideg Hypothesis
Control group
Corticosterone-treated group
Corticosterone + PEA 1mgkg ip
Corticosterone + PEA 10mgkg ip
Corticosterone + luteolin 1mgkg ip
Corticosterone + luteolin 3mgkg ip
Corticosterone + luteolin 10mgkg ip
Corticosterone + PEA and lut (single
treatment combination ) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA luteolinand PEA and luteolin combination
Behavior (NSF)
Start of chronic
PEA and Luteolin treatment
Corticosterone
I Exp
Experimental design
Control group
Corticosterone-treated group
Corticosterone + PEA luteolin (co-ultramicronized) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA+luteolin compound
Behavior (NSF OF FST EPM)
Neurogenesis neuroplasticity neurotrophic and apoptotic
studies
Start of chronic
PEA+Luteolin treatment
Corticosterone
II Exp
Experimental design
Results-1 Anxiety-like behavior
deg
degdeg
Luteolin
deg
PEA
PEA and Lut
PEA + Lut
Results-1 Depression-like behavior - PEALut
FST
Could ndash PEALut - treatment increase
neurogenesis and neuroplasticity
IIdeg Hypothesis
Results-2 Neurogenesis - PEALut
A
50μm 50μm 50μm
Cort- Cort+ PEALut
Results-2 Neurogenesis - PEALut
Cort-
Cort+
PEA+Lut
50μm
50μm
50μm
Results-2 Neuroplasticity ndash PEALut
10microm
10microm
10microm
PEA+Lut
Cort+
Cort-
I
Adult Neurogenesis in Humans
β-actin
BDNF
Cort- Cort+ PEA+Lut
14kD
Results-2 Neurotrophic factor ndash PEALut
20microm 20microm 20microm
Cort- Cort+ PEA+Lut
IIIdeg Hypothesis
Could ldquoPEALutrdquo treatment modify
apoptotic protein expression
deg
Bax
β-actin
Cort- Cort+ PEA+Lu
t 23kD
Results-3 Apoptotic protein expression
β-actin
Bcl-2 29kD
PEA+Lut Cort+ Cort-
IVdeg Hypothesis
Could PEALut treatment reduce
neuroinflammation associated with Alzheimer
diseases
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 12: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/12.jpg)
PEA levels in rat brain following acute and chronic drug
administration
bull Naturally occurring fatty acid amide
bull Lipophilic features
bull Suitable for oral use
bull Present in many animal and plants commonly used for human nutrition
(Food for Special Medical Purposes)
bull In mammals it has been found in many tissues and body fluids
PEA
The levels of endogenous PEA in normal muscle and adipose tissues (004 - 600 pmolmg) are similar to those observed in the CNS and PNS and something higher compared to the gastrointestinal tract (005-150 pmolmg) skin (03- 069 pmolmg) eye (010 - 064 pmolmg) and kidney (25 ndash 71 pmolmg)
PEA levels in normal tissues
PEA
neurons microglia
astrocytes
mast cells
epidermal cells
macrophages
bull Produced ldquo on demand rdquo (enzymatically precursors) by neuro-immuno-inflammatory cells (eg mast cells microglia) exposed to potential damage from membrane
Analgesic
Anti-inflammatory
N-palmitoylethanolamide (PEA)
ldquoAliamides may represent the major breakthrough in inflammation we have been waiting forrdquo Jack D 1996 DNampP
Autacoid Local Injury Antagonism
Mini-Reviews in Medicinal Chemistry 2013 Vol 13 No 1
flavonoids
Pharmacological activity
Micronized coultraPEALut association
ratio of 101 in mass
was obtained by a co-ultramicronization process in an jet-mill equipment
Could ndash PEALut - treatment reduce the
behavioral responses associated with
depression
Ideg Hypothesis
Control group
Corticosterone-treated group
Corticosterone + PEA 1mgkg ip
Corticosterone + PEA 10mgkg ip
Corticosterone + luteolin 1mgkg ip
Corticosterone + luteolin 3mgkg ip
Corticosterone + luteolin 10mgkg ip
Corticosterone + PEA and lut (single
treatment combination ) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA luteolinand PEA and luteolin combination
Behavior (NSF)
Start of chronic
PEA and Luteolin treatment
Corticosterone
I Exp
Experimental design
Control group
Corticosterone-treated group
Corticosterone + PEA luteolin (co-ultramicronized) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA+luteolin compound
Behavior (NSF OF FST EPM)
Neurogenesis neuroplasticity neurotrophic and apoptotic
studies
Start of chronic
PEA+Luteolin treatment
Corticosterone
II Exp
Experimental design
Results-1 Anxiety-like behavior
deg
degdeg
Luteolin
deg
PEA
PEA and Lut
PEA + Lut
Results-1 Depression-like behavior - PEALut
FST
Could ndash PEALut - treatment increase
neurogenesis and neuroplasticity
IIdeg Hypothesis
Results-2 Neurogenesis - PEALut
A
50μm 50μm 50μm
Cort- Cort+ PEALut
Results-2 Neurogenesis - PEALut
Cort-
Cort+
PEA+Lut
50μm
50μm
50μm
Results-2 Neuroplasticity ndash PEALut
10microm
10microm
10microm
PEA+Lut
Cort+
Cort-
I
Adult Neurogenesis in Humans
β-actin
BDNF
Cort- Cort+ PEA+Lut
14kD
Results-2 Neurotrophic factor ndash PEALut
20microm 20microm 20microm
Cort- Cort+ PEA+Lut
IIIdeg Hypothesis
Could ldquoPEALutrdquo treatment modify
apoptotic protein expression
deg
Bax
β-actin
Cort- Cort+ PEA+Lu
t 23kD
Results-3 Apoptotic protein expression
β-actin
Bcl-2 29kD
PEA+Lut Cort+ Cort-
IVdeg Hypothesis
Could PEALut treatment reduce
neuroinflammation associated with Alzheimer
diseases
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 13: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/13.jpg)
bull Naturally occurring fatty acid amide
bull Lipophilic features
bull Suitable for oral use
bull Present in many animal and plants commonly used for human nutrition
(Food for Special Medical Purposes)
bull In mammals it has been found in many tissues and body fluids
PEA
The levels of endogenous PEA in normal muscle and adipose tissues (004 - 600 pmolmg) are similar to those observed in the CNS and PNS and something higher compared to the gastrointestinal tract (005-150 pmolmg) skin (03- 069 pmolmg) eye (010 - 064 pmolmg) and kidney (25 ndash 71 pmolmg)
PEA levels in normal tissues
PEA
neurons microglia
astrocytes
mast cells
epidermal cells
macrophages
bull Produced ldquo on demand rdquo (enzymatically precursors) by neuro-immuno-inflammatory cells (eg mast cells microglia) exposed to potential damage from membrane
Analgesic
Anti-inflammatory
N-palmitoylethanolamide (PEA)
ldquoAliamides may represent the major breakthrough in inflammation we have been waiting forrdquo Jack D 1996 DNampP
Autacoid Local Injury Antagonism
Mini-Reviews in Medicinal Chemistry 2013 Vol 13 No 1
flavonoids
Pharmacological activity
Micronized coultraPEALut association
ratio of 101 in mass
was obtained by a co-ultramicronization process in an jet-mill equipment
Could ndash PEALut - treatment reduce the
behavioral responses associated with
depression
Ideg Hypothesis
Control group
Corticosterone-treated group
Corticosterone + PEA 1mgkg ip
Corticosterone + PEA 10mgkg ip
Corticosterone + luteolin 1mgkg ip
Corticosterone + luteolin 3mgkg ip
Corticosterone + luteolin 10mgkg ip
Corticosterone + PEA and lut (single
treatment combination ) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA luteolinand PEA and luteolin combination
Behavior (NSF)
Start of chronic
PEA and Luteolin treatment
Corticosterone
I Exp
Experimental design
Control group
Corticosterone-treated group
Corticosterone + PEA luteolin (co-ultramicronized) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA+luteolin compound
Behavior (NSF OF FST EPM)
Neurogenesis neuroplasticity neurotrophic and apoptotic
studies
Start of chronic
PEA+Luteolin treatment
Corticosterone
II Exp
Experimental design
Results-1 Anxiety-like behavior
deg
degdeg
Luteolin
deg
PEA
PEA and Lut
PEA + Lut
Results-1 Depression-like behavior - PEALut
FST
Could ndash PEALut - treatment increase
neurogenesis and neuroplasticity
IIdeg Hypothesis
Results-2 Neurogenesis - PEALut
A
50μm 50μm 50μm
Cort- Cort+ PEALut
Results-2 Neurogenesis - PEALut
Cort-
Cort+
PEA+Lut
50μm
50μm
50μm
Results-2 Neuroplasticity ndash PEALut
10microm
10microm
10microm
PEA+Lut
Cort+
Cort-
I
Adult Neurogenesis in Humans
β-actin
BDNF
Cort- Cort+ PEA+Lut
14kD
Results-2 Neurotrophic factor ndash PEALut
20microm 20microm 20microm
Cort- Cort+ PEA+Lut
IIIdeg Hypothesis
Could ldquoPEALutrdquo treatment modify
apoptotic protein expression
deg
Bax
β-actin
Cort- Cort+ PEA+Lu
t 23kD
Results-3 Apoptotic protein expression
β-actin
Bcl-2 29kD
PEA+Lut Cort+ Cort-
IVdeg Hypothesis
Could PEALut treatment reduce
neuroinflammation associated with Alzheimer
diseases
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 14: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/14.jpg)
bull In mammals it has been found in many tissues and body fluids
PEA
The levels of endogenous PEA in normal muscle and adipose tissues (004 - 600 pmolmg) are similar to those observed in the CNS and PNS and something higher compared to the gastrointestinal tract (005-150 pmolmg) skin (03- 069 pmolmg) eye (010 - 064 pmolmg) and kidney (25 ndash 71 pmolmg)
PEA levels in normal tissues
PEA
neurons microglia
astrocytes
mast cells
epidermal cells
macrophages
bull Produced ldquo on demand rdquo (enzymatically precursors) by neuro-immuno-inflammatory cells (eg mast cells microglia) exposed to potential damage from membrane
Analgesic
Anti-inflammatory
N-palmitoylethanolamide (PEA)
ldquoAliamides may represent the major breakthrough in inflammation we have been waiting forrdquo Jack D 1996 DNampP
Autacoid Local Injury Antagonism
Mini-Reviews in Medicinal Chemistry 2013 Vol 13 No 1
flavonoids
Pharmacological activity
Micronized coultraPEALut association
ratio of 101 in mass
was obtained by a co-ultramicronization process in an jet-mill equipment
Could ndash PEALut - treatment reduce the
behavioral responses associated with
depression
Ideg Hypothesis
Control group
Corticosterone-treated group
Corticosterone + PEA 1mgkg ip
Corticosterone + PEA 10mgkg ip
Corticosterone + luteolin 1mgkg ip
Corticosterone + luteolin 3mgkg ip
Corticosterone + luteolin 10mgkg ip
Corticosterone + PEA and lut (single
treatment combination ) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA luteolinand PEA and luteolin combination
Behavior (NSF)
Start of chronic
PEA and Luteolin treatment
Corticosterone
I Exp
Experimental design
Control group
Corticosterone-treated group
Corticosterone + PEA luteolin (co-ultramicronized) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA+luteolin compound
Behavior (NSF OF FST EPM)
Neurogenesis neuroplasticity neurotrophic and apoptotic
studies
Start of chronic
PEA+Luteolin treatment
Corticosterone
II Exp
Experimental design
Results-1 Anxiety-like behavior
deg
degdeg
Luteolin
deg
PEA
PEA and Lut
PEA + Lut
Results-1 Depression-like behavior - PEALut
FST
Could ndash PEALut - treatment increase
neurogenesis and neuroplasticity
IIdeg Hypothesis
Results-2 Neurogenesis - PEALut
A
50μm 50μm 50μm
Cort- Cort+ PEALut
Results-2 Neurogenesis - PEALut
Cort-
Cort+
PEA+Lut
50μm
50μm
50μm
Results-2 Neuroplasticity ndash PEALut
10microm
10microm
10microm
PEA+Lut
Cort+
Cort-
I
Adult Neurogenesis in Humans
β-actin
BDNF
Cort- Cort+ PEA+Lut
14kD
Results-2 Neurotrophic factor ndash PEALut
20microm 20microm 20microm
Cort- Cort+ PEA+Lut
IIIdeg Hypothesis
Could ldquoPEALutrdquo treatment modify
apoptotic protein expression
deg
Bax
β-actin
Cort- Cort+ PEA+Lu
t 23kD
Results-3 Apoptotic protein expression
β-actin
Bcl-2 29kD
PEA+Lut Cort+ Cort-
IVdeg Hypothesis
Could PEALut treatment reduce
neuroinflammation associated with Alzheimer
diseases
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 15: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/15.jpg)
PEA
neurons microglia
astrocytes
mast cells
epidermal cells
macrophages
bull Produced ldquo on demand rdquo (enzymatically precursors) by neuro-immuno-inflammatory cells (eg mast cells microglia) exposed to potential damage from membrane
Analgesic
Anti-inflammatory
N-palmitoylethanolamide (PEA)
ldquoAliamides may represent the major breakthrough in inflammation we have been waiting forrdquo Jack D 1996 DNampP
Autacoid Local Injury Antagonism
Mini-Reviews in Medicinal Chemistry 2013 Vol 13 No 1
flavonoids
Pharmacological activity
Micronized coultraPEALut association
ratio of 101 in mass
was obtained by a co-ultramicronization process in an jet-mill equipment
Could ndash PEALut - treatment reduce the
behavioral responses associated with
depression
Ideg Hypothesis
Control group
Corticosterone-treated group
Corticosterone + PEA 1mgkg ip
Corticosterone + PEA 10mgkg ip
Corticosterone + luteolin 1mgkg ip
Corticosterone + luteolin 3mgkg ip
Corticosterone + luteolin 10mgkg ip
Corticosterone + PEA and lut (single
treatment combination ) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA luteolinand PEA and luteolin combination
Behavior (NSF)
Start of chronic
PEA and Luteolin treatment
Corticosterone
I Exp
Experimental design
Control group
Corticosterone-treated group
Corticosterone + PEA luteolin (co-ultramicronized) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA+luteolin compound
Behavior (NSF OF FST EPM)
Neurogenesis neuroplasticity neurotrophic and apoptotic
studies
Start of chronic
PEA+Luteolin treatment
Corticosterone
II Exp
Experimental design
Results-1 Anxiety-like behavior
deg
degdeg
Luteolin
deg
PEA
PEA and Lut
PEA + Lut
Results-1 Depression-like behavior - PEALut
FST
Could ndash PEALut - treatment increase
neurogenesis and neuroplasticity
IIdeg Hypothesis
Results-2 Neurogenesis - PEALut
A
50μm 50μm 50μm
Cort- Cort+ PEALut
Results-2 Neurogenesis - PEALut
Cort-
Cort+
PEA+Lut
50μm
50μm
50μm
Results-2 Neuroplasticity ndash PEALut
10microm
10microm
10microm
PEA+Lut
Cort+
Cort-
I
Adult Neurogenesis in Humans
β-actin
BDNF
Cort- Cort+ PEA+Lut
14kD
Results-2 Neurotrophic factor ndash PEALut
20microm 20microm 20microm
Cort- Cort+ PEA+Lut
IIIdeg Hypothesis
Could ldquoPEALutrdquo treatment modify
apoptotic protein expression
deg
Bax
β-actin
Cort- Cort+ PEA+Lu
t 23kD
Results-3 Apoptotic protein expression
β-actin
Bcl-2 29kD
PEA+Lut Cort+ Cort-
IVdeg Hypothesis
Could PEALut treatment reduce
neuroinflammation associated with Alzheimer
diseases
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 16: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/16.jpg)
Analgesic
Anti-inflammatory
N-palmitoylethanolamide (PEA)
ldquoAliamides may represent the major breakthrough in inflammation we have been waiting forrdquo Jack D 1996 DNampP
Autacoid Local Injury Antagonism
Mini-Reviews in Medicinal Chemistry 2013 Vol 13 No 1
flavonoids
Pharmacological activity
Micronized coultraPEALut association
ratio of 101 in mass
was obtained by a co-ultramicronization process in an jet-mill equipment
Could ndash PEALut - treatment reduce the
behavioral responses associated with
depression
Ideg Hypothesis
Control group
Corticosterone-treated group
Corticosterone + PEA 1mgkg ip
Corticosterone + PEA 10mgkg ip
Corticosterone + luteolin 1mgkg ip
Corticosterone + luteolin 3mgkg ip
Corticosterone + luteolin 10mgkg ip
Corticosterone + PEA and lut (single
treatment combination ) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA luteolinand PEA and luteolin combination
Behavior (NSF)
Start of chronic
PEA and Luteolin treatment
Corticosterone
I Exp
Experimental design
Control group
Corticosterone-treated group
Corticosterone + PEA luteolin (co-ultramicronized) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA+luteolin compound
Behavior (NSF OF FST EPM)
Neurogenesis neuroplasticity neurotrophic and apoptotic
studies
Start of chronic
PEA+Luteolin treatment
Corticosterone
II Exp
Experimental design
Results-1 Anxiety-like behavior
deg
degdeg
Luteolin
deg
PEA
PEA and Lut
PEA + Lut
Results-1 Depression-like behavior - PEALut
FST
Could ndash PEALut - treatment increase
neurogenesis and neuroplasticity
IIdeg Hypothesis
Results-2 Neurogenesis - PEALut
A
50μm 50μm 50μm
Cort- Cort+ PEALut
Results-2 Neurogenesis - PEALut
Cort-
Cort+
PEA+Lut
50μm
50μm
50μm
Results-2 Neuroplasticity ndash PEALut
10microm
10microm
10microm
PEA+Lut
Cort+
Cort-
I
Adult Neurogenesis in Humans
β-actin
BDNF
Cort- Cort+ PEA+Lut
14kD
Results-2 Neurotrophic factor ndash PEALut
20microm 20microm 20microm
Cort- Cort+ PEA+Lut
IIIdeg Hypothesis
Could ldquoPEALutrdquo treatment modify
apoptotic protein expression
deg
Bax
β-actin
Cort- Cort+ PEA+Lu
t 23kD
Results-3 Apoptotic protein expression
β-actin
Bcl-2 29kD
PEA+Lut Cort+ Cort-
IVdeg Hypothesis
Could PEALut treatment reduce
neuroinflammation associated with Alzheimer
diseases
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 17: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/17.jpg)
ldquoAliamides may represent the major breakthrough in inflammation we have been waiting forrdquo Jack D 1996 DNampP
Autacoid Local Injury Antagonism
Mini-Reviews in Medicinal Chemistry 2013 Vol 13 No 1
flavonoids
Pharmacological activity
Micronized coultraPEALut association
ratio of 101 in mass
was obtained by a co-ultramicronization process in an jet-mill equipment
Could ndash PEALut - treatment reduce the
behavioral responses associated with
depression
Ideg Hypothesis
Control group
Corticosterone-treated group
Corticosterone + PEA 1mgkg ip
Corticosterone + PEA 10mgkg ip
Corticosterone + luteolin 1mgkg ip
Corticosterone + luteolin 3mgkg ip
Corticosterone + luteolin 10mgkg ip
Corticosterone + PEA and lut (single
treatment combination ) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA luteolinand PEA and luteolin combination
Behavior (NSF)
Start of chronic
PEA and Luteolin treatment
Corticosterone
I Exp
Experimental design
Control group
Corticosterone-treated group
Corticosterone + PEA luteolin (co-ultramicronized) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA+luteolin compound
Behavior (NSF OF FST EPM)
Neurogenesis neuroplasticity neurotrophic and apoptotic
studies
Start of chronic
PEA+Luteolin treatment
Corticosterone
II Exp
Experimental design
Results-1 Anxiety-like behavior
deg
degdeg
Luteolin
deg
PEA
PEA and Lut
PEA + Lut
Results-1 Depression-like behavior - PEALut
FST
Could ndash PEALut - treatment increase
neurogenesis and neuroplasticity
IIdeg Hypothesis
Results-2 Neurogenesis - PEALut
A
50μm 50μm 50μm
Cort- Cort+ PEALut
Results-2 Neurogenesis - PEALut
Cort-
Cort+
PEA+Lut
50μm
50μm
50μm
Results-2 Neuroplasticity ndash PEALut
10microm
10microm
10microm
PEA+Lut
Cort+
Cort-
I
Adult Neurogenesis in Humans
β-actin
BDNF
Cort- Cort+ PEA+Lut
14kD
Results-2 Neurotrophic factor ndash PEALut
20microm 20microm 20microm
Cort- Cort+ PEA+Lut
IIIdeg Hypothesis
Could ldquoPEALutrdquo treatment modify
apoptotic protein expression
deg
Bax
β-actin
Cort- Cort+ PEA+Lu
t 23kD
Results-3 Apoptotic protein expression
β-actin
Bcl-2 29kD
PEA+Lut Cort+ Cort-
IVdeg Hypothesis
Could PEALut treatment reduce
neuroinflammation associated with Alzheimer
diseases
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 18: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/18.jpg)
Mini-Reviews in Medicinal Chemistry 2013 Vol 13 No 1
flavonoids
Pharmacological activity
Micronized coultraPEALut association
ratio of 101 in mass
was obtained by a co-ultramicronization process in an jet-mill equipment
Could ndash PEALut - treatment reduce the
behavioral responses associated with
depression
Ideg Hypothesis
Control group
Corticosterone-treated group
Corticosterone + PEA 1mgkg ip
Corticosterone + PEA 10mgkg ip
Corticosterone + luteolin 1mgkg ip
Corticosterone + luteolin 3mgkg ip
Corticosterone + luteolin 10mgkg ip
Corticosterone + PEA and lut (single
treatment combination ) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA luteolinand PEA and luteolin combination
Behavior (NSF)
Start of chronic
PEA and Luteolin treatment
Corticosterone
I Exp
Experimental design
Control group
Corticosterone-treated group
Corticosterone + PEA luteolin (co-ultramicronized) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA+luteolin compound
Behavior (NSF OF FST EPM)
Neurogenesis neuroplasticity neurotrophic and apoptotic
studies
Start of chronic
PEA+Luteolin treatment
Corticosterone
II Exp
Experimental design
Results-1 Anxiety-like behavior
deg
degdeg
Luteolin
deg
PEA
PEA and Lut
PEA + Lut
Results-1 Depression-like behavior - PEALut
FST
Could ndash PEALut - treatment increase
neurogenesis and neuroplasticity
IIdeg Hypothesis
Results-2 Neurogenesis - PEALut
A
50μm 50μm 50μm
Cort- Cort+ PEALut
Results-2 Neurogenesis - PEALut
Cort-
Cort+
PEA+Lut
50μm
50μm
50μm
Results-2 Neuroplasticity ndash PEALut
10microm
10microm
10microm
PEA+Lut
Cort+
Cort-
I
Adult Neurogenesis in Humans
β-actin
BDNF
Cort- Cort+ PEA+Lut
14kD
Results-2 Neurotrophic factor ndash PEALut
20microm 20microm 20microm
Cort- Cort+ PEA+Lut
IIIdeg Hypothesis
Could ldquoPEALutrdquo treatment modify
apoptotic protein expression
deg
Bax
β-actin
Cort- Cort+ PEA+Lu
t 23kD
Results-3 Apoptotic protein expression
β-actin
Bcl-2 29kD
PEA+Lut Cort+ Cort-
IVdeg Hypothesis
Could PEALut treatment reduce
neuroinflammation associated with Alzheimer
diseases
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 19: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/19.jpg)
flavonoids
Pharmacological activity
Micronized coultraPEALut association
ratio of 101 in mass
was obtained by a co-ultramicronization process in an jet-mill equipment
Could ndash PEALut - treatment reduce the
behavioral responses associated with
depression
Ideg Hypothesis
Control group
Corticosterone-treated group
Corticosterone + PEA 1mgkg ip
Corticosterone + PEA 10mgkg ip
Corticosterone + luteolin 1mgkg ip
Corticosterone + luteolin 3mgkg ip
Corticosterone + luteolin 10mgkg ip
Corticosterone + PEA and lut (single
treatment combination ) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA luteolinand PEA and luteolin combination
Behavior (NSF)
Start of chronic
PEA and Luteolin treatment
Corticosterone
I Exp
Experimental design
Control group
Corticosterone-treated group
Corticosterone + PEA luteolin (co-ultramicronized) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA+luteolin compound
Behavior (NSF OF FST EPM)
Neurogenesis neuroplasticity neurotrophic and apoptotic
studies
Start of chronic
PEA+Luteolin treatment
Corticosterone
II Exp
Experimental design
Results-1 Anxiety-like behavior
deg
degdeg
Luteolin
deg
PEA
PEA and Lut
PEA + Lut
Results-1 Depression-like behavior - PEALut
FST
Could ndash PEALut - treatment increase
neurogenesis and neuroplasticity
IIdeg Hypothesis
Results-2 Neurogenesis - PEALut
A
50μm 50μm 50μm
Cort- Cort+ PEALut
Results-2 Neurogenesis - PEALut
Cort-
Cort+
PEA+Lut
50μm
50μm
50μm
Results-2 Neuroplasticity ndash PEALut
10microm
10microm
10microm
PEA+Lut
Cort+
Cort-
I
Adult Neurogenesis in Humans
β-actin
BDNF
Cort- Cort+ PEA+Lut
14kD
Results-2 Neurotrophic factor ndash PEALut
20microm 20microm 20microm
Cort- Cort+ PEA+Lut
IIIdeg Hypothesis
Could ldquoPEALutrdquo treatment modify
apoptotic protein expression
deg
Bax
β-actin
Cort- Cort+ PEA+Lu
t 23kD
Results-3 Apoptotic protein expression
β-actin
Bcl-2 29kD
PEA+Lut Cort+ Cort-
IVdeg Hypothesis
Could PEALut treatment reduce
neuroinflammation associated with Alzheimer
diseases
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 20: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/20.jpg)
Pharmacological activity
Micronized coultraPEALut association
ratio of 101 in mass
was obtained by a co-ultramicronization process in an jet-mill equipment
Could ndash PEALut - treatment reduce the
behavioral responses associated with
depression
Ideg Hypothesis
Control group
Corticosterone-treated group
Corticosterone + PEA 1mgkg ip
Corticosterone + PEA 10mgkg ip
Corticosterone + luteolin 1mgkg ip
Corticosterone + luteolin 3mgkg ip
Corticosterone + luteolin 10mgkg ip
Corticosterone + PEA and lut (single
treatment combination ) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA luteolinand PEA and luteolin combination
Behavior (NSF)
Start of chronic
PEA and Luteolin treatment
Corticosterone
I Exp
Experimental design
Control group
Corticosterone-treated group
Corticosterone + PEA luteolin (co-ultramicronized) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA+luteolin compound
Behavior (NSF OF FST EPM)
Neurogenesis neuroplasticity neurotrophic and apoptotic
studies
Start of chronic
PEA+Luteolin treatment
Corticosterone
II Exp
Experimental design
Results-1 Anxiety-like behavior
deg
degdeg
Luteolin
deg
PEA
PEA and Lut
PEA + Lut
Results-1 Depression-like behavior - PEALut
FST
Could ndash PEALut - treatment increase
neurogenesis and neuroplasticity
IIdeg Hypothesis
Results-2 Neurogenesis - PEALut
A
50μm 50μm 50μm
Cort- Cort+ PEALut
Results-2 Neurogenesis - PEALut
Cort-
Cort+
PEA+Lut
50μm
50μm
50μm
Results-2 Neuroplasticity ndash PEALut
10microm
10microm
10microm
PEA+Lut
Cort+
Cort-
I
Adult Neurogenesis in Humans
β-actin
BDNF
Cort- Cort+ PEA+Lut
14kD
Results-2 Neurotrophic factor ndash PEALut
20microm 20microm 20microm
Cort- Cort+ PEA+Lut
IIIdeg Hypothesis
Could ldquoPEALutrdquo treatment modify
apoptotic protein expression
deg
Bax
β-actin
Cort- Cort+ PEA+Lu
t 23kD
Results-3 Apoptotic protein expression
β-actin
Bcl-2 29kD
PEA+Lut Cort+ Cort-
IVdeg Hypothesis
Could PEALut treatment reduce
neuroinflammation associated with Alzheimer
diseases
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 21: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/21.jpg)
Micronized coultraPEALut association
ratio of 101 in mass
was obtained by a co-ultramicronization process in an jet-mill equipment
Could ndash PEALut - treatment reduce the
behavioral responses associated with
depression
Ideg Hypothesis
Control group
Corticosterone-treated group
Corticosterone + PEA 1mgkg ip
Corticosterone + PEA 10mgkg ip
Corticosterone + luteolin 1mgkg ip
Corticosterone + luteolin 3mgkg ip
Corticosterone + luteolin 10mgkg ip
Corticosterone + PEA and lut (single
treatment combination ) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA luteolinand PEA and luteolin combination
Behavior (NSF)
Start of chronic
PEA and Luteolin treatment
Corticosterone
I Exp
Experimental design
Control group
Corticosterone-treated group
Corticosterone + PEA luteolin (co-ultramicronized) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA+luteolin compound
Behavior (NSF OF FST EPM)
Neurogenesis neuroplasticity neurotrophic and apoptotic
studies
Start of chronic
PEA+Luteolin treatment
Corticosterone
II Exp
Experimental design
Results-1 Anxiety-like behavior
deg
degdeg
Luteolin
deg
PEA
PEA and Lut
PEA + Lut
Results-1 Depression-like behavior - PEALut
FST
Could ndash PEALut - treatment increase
neurogenesis and neuroplasticity
IIdeg Hypothesis
Results-2 Neurogenesis - PEALut
A
50μm 50μm 50μm
Cort- Cort+ PEALut
Results-2 Neurogenesis - PEALut
Cort-
Cort+
PEA+Lut
50μm
50μm
50μm
Results-2 Neuroplasticity ndash PEALut
10microm
10microm
10microm
PEA+Lut
Cort+
Cort-
I
Adult Neurogenesis in Humans
β-actin
BDNF
Cort- Cort+ PEA+Lut
14kD
Results-2 Neurotrophic factor ndash PEALut
20microm 20microm 20microm
Cort- Cort+ PEA+Lut
IIIdeg Hypothesis
Could ldquoPEALutrdquo treatment modify
apoptotic protein expression
deg
Bax
β-actin
Cort- Cort+ PEA+Lu
t 23kD
Results-3 Apoptotic protein expression
β-actin
Bcl-2 29kD
PEA+Lut Cort+ Cort-
IVdeg Hypothesis
Could PEALut treatment reduce
neuroinflammation associated with Alzheimer
diseases
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 22: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/22.jpg)
Could ndash PEALut - treatment reduce the
behavioral responses associated with
depression
Ideg Hypothesis
Control group
Corticosterone-treated group
Corticosterone + PEA 1mgkg ip
Corticosterone + PEA 10mgkg ip
Corticosterone + luteolin 1mgkg ip
Corticosterone + luteolin 3mgkg ip
Corticosterone + luteolin 10mgkg ip
Corticosterone + PEA and lut (single
treatment combination ) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA luteolinand PEA and luteolin combination
Behavior (NSF)
Start of chronic
PEA and Luteolin treatment
Corticosterone
I Exp
Experimental design
Control group
Corticosterone-treated group
Corticosterone + PEA luteolin (co-ultramicronized) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA+luteolin compound
Behavior (NSF OF FST EPM)
Neurogenesis neuroplasticity neurotrophic and apoptotic
studies
Start of chronic
PEA+Luteolin treatment
Corticosterone
II Exp
Experimental design
Results-1 Anxiety-like behavior
deg
degdeg
Luteolin
deg
PEA
PEA and Lut
PEA + Lut
Results-1 Depression-like behavior - PEALut
FST
Could ndash PEALut - treatment increase
neurogenesis and neuroplasticity
IIdeg Hypothesis
Results-2 Neurogenesis - PEALut
A
50μm 50μm 50μm
Cort- Cort+ PEALut
Results-2 Neurogenesis - PEALut
Cort-
Cort+
PEA+Lut
50μm
50μm
50μm
Results-2 Neuroplasticity ndash PEALut
10microm
10microm
10microm
PEA+Lut
Cort+
Cort-
I
Adult Neurogenesis in Humans
β-actin
BDNF
Cort- Cort+ PEA+Lut
14kD
Results-2 Neurotrophic factor ndash PEALut
20microm 20microm 20microm
Cort- Cort+ PEA+Lut
IIIdeg Hypothesis
Could ldquoPEALutrdquo treatment modify
apoptotic protein expression
deg
Bax
β-actin
Cort- Cort+ PEA+Lu
t 23kD
Results-3 Apoptotic protein expression
β-actin
Bcl-2 29kD
PEA+Lut Cort+ Cort-
IVdeg Hypothesis
Could PEALut treatment reduce
neuroinflammation associated with Alzheimer
diseases
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 23: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/23.jpg)
Control group
Corticosterone-treated group
Corticosterone + PEA 1mgkg ip
Corticosterone + PEA 10mgkg ip
Corticosterone + luteolin 1mgkg ip
Corticosterone + luteolin 3mgkg ip
Corticosterone + luteolin 10mgkg ip
Corticosterone + PEA and lut (single
treatment combination ) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA luteolinand PEA and luteolin combination
Behavior (NSF)
Start of chronic
PEA and Luteolin treatment
Corticosterone
I Exp
Experimental design
Control group
Corticosterone-treated group
Corticosterone + PEA luteolin (co-ultramicronized) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA+luteolin compound
Behavior (NSF OF FST EPM)
Neurogenesis neuroplasticity neurotrophic and apoptotic
studies
Start of chronic
PEA+Luteolin treatment
Corticosterone
II Exp
Experimental design
Results-1 Anxiety-like behavior
deg
degdeg
Luteolin
deg
PEA
PEA and Lut
PEA + Lut
Results-1 Depression-like behavior - PEALut
FST
Could ndash PEALut - treatment increase
neurogenesis and neuroplasticity
IIdeg Hypothesis
Results-2 Neurogenesis - PEALut
A
50μm 50μm 50μm
Cort- Cort+ PEALut
Results-2 Neurogenesis - PEALut
Cort-
Cort+
PEA+Lut
50μm
50μm
50μm
Results-2 Neuroplasticity ndash PEALut
10microm
10microm
10microm
PEA+Lut
Cort+
Cort-
I
Adult Neurogenesis in Humans
β-actin
BDNF
Cort- Cort+ PEA+Lut
14kD
Results-2 Neurotrophic factor ndash PEALut
20microm 20microm 20microm
Cort- Cort+ PEA+Lut
IIIdeg Hypothesis
Could ldquoPEALutrdquo treatment modify
apoptotic protein expression
deg
Bax
β-actin
Cort- Cort+ PEA+Lu
t 23kD
Results-3 Apoptotic protein expression
β-actin
Bcl-2 29kD
PEA+Lut Cort+ Cort-
IVdeg Hypothesis
Could PEALut treatment reduce
neuroinflammation associated with Alzheimer
diseases
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 24: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/24.jpg)
Control group
Corticosterone-treated group
Corticosterone + PEA luteolin (co-ultramicronized) 1mgkg ip
Start of chronic
corticosterone treatment
0 4 6
weeks
Corticosterone
Corticosterone + PEA+luteolin compound
Behavior (NSF OF FST EPM)
Neurogenesis neuroplasticity neurotrophic and apoptotic
studies
Start of chronic
PEA+Luteolin treatment
Corticosterone
II Exp
Experimental design
Results-1 Anxiety-like behavior
deg
degdeg
Luteolin
deg
PEA
PEA and Lut
PEA + Lut
Results-1 Depression-like behavior - PEALut
FST
Could ndash PEALut - treatment increase
neurogenesis and neuroplasticity
IIdeg Hypothesis
Results-2 Neurogenesis - PEALut
A
50μm 50μm 50μm
Cort- Cort+ PEALut
Results-2 Neurogenesis - PEALut
Cort-
Cort+
PEA+Lut
50μm
50μm
50μm
Results-2 Neuroplasticity ndash PEALut
10microm
10microm
10microm
PEA+Lut
Cort+
Cort-
I
Adult Neurogenesis in Humans
β-actin
BDNF
Cort- Cort+ PEA+Lut
14kD
Results-2 Neurotrophic factor ndash PEALut
20microm 20microm 20microm
Cort- Cort+ PEA+Lut
IIIdeg Hypothesis
Could ldquoPEALutrdquo treatment modify
apoptotic protein expression
deg
Bax
β-actin
Cort- Cort+ PEA+Lu
t 23kD
Results-3 Apoptotic protein expression
β-actin
Bcl-2 29kD
PEA+Lut Cort+ Cort-
IVdeg Hypothesis
Could PEALut treatment reduce
neuroinflammation associated with Alzheimer
diseases
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 25: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/25.jpg)
Results-1 Anxiety-like behavior
deg
degdeg
Luteolin
deg
PEA
PEA and Lut
PEA + Lut
Results-1 Depression-like behavior - PEALut
FST
Could ndash PEALut - treatment increase
neurogenesis and neuroplasticity
IIdeg Hypothesis
Results-2 Neurogenesis - PEALut
A
50μm 50μm 50μm
Cort- Cort+ PEALut
Results-2 Neurogenesis - PEALut
Cort-
Cort+
PEA+Lut
50μm
50μm
50μm
Results-2 Neuroplasticity ndash PEALut
10microm
10microm
10microm
PEA+Lut
Cort+
Cort-
I
Adult Neurogenesis in Humans
β-actin
BDNF
Cort- Cort+ PEA+Lut
14kD
Results-2 Neurotrophic factor ndash PEALut
20microm 20microm 20microm
Cort- Cort+ PEA+Lut
IIIdeg Hypothesis
Could ldquoPEALutrdquo treatment modify
apoptotic protein expression
deg
Bax
β-actin
Cort- Cort+ PEA+Lu
t 23kD
Results-3 Apoptotic protein expression
β-actin
Bcl-2 29kD
PEA+Lut Cort+ Cort-
IVdeg Hypothesis
Could PEALut treatment reduce
neuroinflammation associated with Alzheimer
diseases
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 26: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/26.jpg)
Results-1 Depression-like behavior - PEALut
FST
Could ndash PEALut - treatment increase
neurogenesis and neuroplasticity
IIdeg Hypothesis
Results-2 Neurogenesis - PEALut
A
50μm 50μm 50μm
Cort- Cort+ PEALut
Results-2 Neurogenesis - PEALut
Cort-
Cort+
PEA+Lut
50μm
50μm
50μm
Results-2 Neuroplasticity ndash PEALut
10microm
10microm
10microm
PEA+Lut
Cort+
Cort-
I
Adult Neurogenesis in Humans
β-actin
BDNF
Cort- Cort+ PEA+Lut
14kD
Results-2 Neurotrophic factor ndash PEALut
20microm 20microm 20microm
Cort- Cort+ PEA+Lut
IIIdeg Hypothesis
Could ldquoPEALutrdquo treatment modify
apoptotic protein expression
deg
Bax
β-actin
Cort- Cort+ PEA+Lu
t 23kD
Results-3 Apoptotic protein expression
β-actin
Bcl-2 29kD
PEA+Lut Cort+ Cort-
IVdeg Hypothesis
Could PEALut treatment reduce
neuroinflammation associated with Alzheimer
diseases
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 27: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/27.jpg)
Could ndash PEALut - treatment increase
neurogenesis and neuroplasticity
IIdeg Hypothesis
Results-2 Neurogenesis - PEALut
A
50μm 50μm 50μm
Cort- Cort+ PEALut
Results-2 Neurogenesis - PEALut
Cort-
Cort+
PEA+Lut
50μm
50μm
50μm
Results-2 Neuroplasticity ndash PEALut
10microm
10microm
10microm
PEA+Lut
Cort+
Cort-
I
Adult Neurogenesis in Humans
β-actin
BDNF
Cort- Cort+ PEA+Lut
14kD
Results-2 Neurotrophic factor ndash PEALut
20microm 20microm 20microm
Cort- Cort+ PEA+Lut
IIIdeg Hypothesis
Could ldquoPEALutrdquo treatment modify
apoptotic protein expression
deg
Bax
β-actin
Cort- Cort+ PEA+Lu
t 23kD
Results-3 Apoptotic protein expression
β-actin
Bcl-2 29kD
PEA+Lut Cort+ Cort-
IVdeg Hypothesis
Could PEALut treatment reduce
neuroinflammation associated with Alzheimer
diseases
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 28: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/28.jpg)
Results-2 Neurogenesis - PEALut
A
50μm 50μm 50μm
Cort- Cort+ PEALut
Results-2 Neurogenesis - PEALut
Cort-
Cort+
PEA+Lut
50μm
50μm
50μm
Results-2 Neuroplasticity ndash PEALut
10microm
10microm
10microm
PEA+Lut
Cort+
Cort-
I
Adult Neurogenesis in Humans
β-actin
BDNF
Cort- Cort+ PEA+Lut
14kD
Results-2 Neurotrophic factor ndash PEALut
20microm 20microm 20microm
Cort- Cort+ PEA+Lut
IIIdeg Hypothesis
Could ldquoPEALutrdquo treatment modify
apoptotic protein expression
deg
Bax
β-actin
Cort- Cort+ PEA+Lu
t 23kD
Results-3 Apoptotic protein expression
β-actin
Bcl-2 29kD
PEA+Lut Cort+ Cort-
IVdeg Hypothesis
Could PEALut treatment reduce
neuroinflammation associated with Alzheimer
diseases
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 29: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/29.jpg)
Results-2 Neurogenesis - PEALut
Cort-
Cort+
PEA+Lut
50μm
50μm
50μm
Results-2 Neuroplasticity ndash PEALut
10microm
10microm
10microm
PEA+Lut
Cort+
Cort-
I
Adult Neurogenesis in Humans
β-actin
BDNF
Cort- Cort+ PEA+Lut
14kD
Results-2 Neurotrophic factor ndash PEALut
20microm 20microm 20microm
Cort- Cort+ PEA+Lut
IIIdeg Hypothesis
Could ldquoPEALutrdquo treatment modify
apoptotic protein expression
deg
Bax
β-actin
Cort- Cort+ PEA+Lu
t 23kD
Results-3 Apoptotic protein expression
β-actin
Bcl-2 29kD
PEA+Lut Cort+ Cort-
IVdeg Hypothesis
Could PEALut treatment reduce
neuroinflammation associated with Alzheimer
diseases
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 30: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/30.jpg)
Results-2 Neuroplasticity ndash PEALut
10microm
10microm
10microm
PEA+Lut
Cort+
Cort-
I
Adult Neurogenesis in Humans
β-actin
BDNF
Cort- Cort+ PEA+Lut
14kD
Results-2 Neurotrophic factor ndash PEALut
20microm 20microm 20microm
Cort- Cort+ PEA+Lut
IIIdeg Hypothesis
Could ldquoPEALutrdquo treatment modify
apoptotic protein expression
deg
Bax
β-actin
Cort- Cort+ PEA+Lu
t 23kD
Results-3 Apoptotic protein expression
β-actin
Bcl-2 29kD
PEA+Lut Cort+ Cort-
IVdeg Hypothesis
Could PEALut treatment reduce
neuroinflammation associated with Alzheimer
diseases
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 31: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/31.jpg)
Adult Neurogenesis in Humans
β-actin
BDNF
Cort- Cort+ PEA+Lut
14kD
Results-2 Neurotrophic factor ndash PEALut
20microm 20microm 20microm
Cort- Cort+ PEA+Lut
IIIdeg Hypothesis
Could ldquoPEALutrdquo treatment modify
apoptotic protein expression
deg
Bax
β-actin
Cort- Cort+ PEA+Lu
t 23kD
Results-3 Apoptotic protein expression
β-actin
Bcl-2 29kD
PEA+Lut Cort+ Cort-
IVdeg Hypothesis
Could PEALut treatment reduce
neuroinflammation associated with Alzheimer
diseases
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 32: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/32.jpg)
β-actin
BDNF
Cort- Cort+ PEA+Lut
14kD
Results-2 Neurotrophic factor ndash PEALut
20microm 20microm 20microm
Cort- Cort+ PEA+Lut
IIIdeg Hypothesis
Could ldquoPEALutrdquo treatment modify
apoptotic protein expression
deg
Bax
β-actin
Cort- Cort+ PEA+Lu
t 23kD
Results-3 Apoptotic protein expression
β-actin
Bcl-2 29kD
PEA+Lut Cort+ Cort-
IVdeg Hypothesis
Could PEALut treatment reduce
neuroinflammation associated with Alzheimer
diseases
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 33: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/33.jpg)
IIIdeg Hypothesis
Could ldquoPEALutrdquo treatment modify
apoptotic protein expression
deg
Bax
β-actin
Cort- Cort+ PEA+Lu
t 23kD
Results-3 Apoptotic protein expression
β-actin
Bcl-2 29kD
PEA+Lut Cort+ Cort-
IVdeg Hypothesis
Could PEALut treatment reduce
neuroinflammation associated with Alzheimer
diseases
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 34: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/34.jpg)
deg
Bax
β-actin
Cort- Cort+ PEA+Lu
t 23kD
Results-3 Apoptotic protein expression
β-actin
Bcl-2 29kD
PEA+Lut Cort+ Cort-
IVdeg Hypothesis
Could PEALut treatment reduce
neuroinflammation associated with Alzheimer
diseases
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 35: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/35.jpg)
IVdeg Hypothesis
Could PEALut treatment reduce
neuroinflammation associated with Alzheimer
diseases
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 36: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/36.jpg)
Retinoic Acid 100nM
2 ml
2 h
Damage with
β-Amiloid1-42
(Aβ1-42)
(1μM)
3 d
bull Western
Blot
8x105 cells
2 d
Pre-treatment
with PEALut
(01 μM - 1μM - 10 μM )
1 d
Differentiation
with retinoic acid
100 nM
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 37: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/37.jpg)
Effect of co-ultra PEALut on viability of SH-
SY5Y
NF-kB pathway
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 38: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/38.jpg)
Incubation with appropriate media
2h 21days
PEALut
MTT assay Nitrite concentration Western Blot Analysis
Hippocampal slices
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 39: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/39.jpg)
A1-42+co-ultraPEALut
Ctr
1-42
A
+PEALut 0
009
gL
+PEALut 0
000
9 gL
+PEALut 0
000
09 g
L
0
2000
4000
6000
8000
Den
sit
om
etr
ic u
nit
s (
OD
mm
2)
IκB 37kDa
Ctr
β-actin 43 kDa
A 1-42
0009gL
(000009
gL)
A1-42+co-ultraPEALut
(00009
gL)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 40: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/40.jpg)
Ctr
1-42
A
+co-ultr
aPEALut 0
00009
gL
+ co-u
ltraP
EALut 00
009
gL
+ co-u
ltraP
EALut 00
09gL
0
2
4
6
8
10
NO
2-(
M)
A1-42+co-ultraPEALut
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 41: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/41.jpg)
Vdeg Hypothesis
Could PEALut treatment have potential
anti-inflammatory effects in an experimental
model of Parkinson disease
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 42: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/42.jpg)
1
7 days
2
hellip daily hellip
Experimental design
Administration of
Ten mice from each group were sacrificed at 7 days to collect samples for the future evaluation
MPTP (20 mgkg ip) 4 administration at 2-h intervals the total dose per mouse being 80 mgkg PEA Lut(1 mgkg ip) at 24h after MPTP administration and administred daily for 7 days Minociclina (30 mgkg ip) at 24h after MPTP administration and administred daily for 7 days
3
MPTP
4
2h 2h 2h 24h
PEA Minociclina
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 43: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/43.jpg)
0
2
4
6
8Sham
MPTP
MPTP+co-ultraPEALut
DATTyrosine Hydroxylase
o
f tot
al ti
ssue
are
a
TH and DAT expression
Sham MPTP MPTP + PEA_LUT
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 44: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/44.jpg)
Sha
m
MPTP
MPTP
+ co
-ultr
aPEALu
t
0
2000
4000
6000
8000
10000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Sham
TNF-α
MPTP
β-actin
MPTP +
co-ultraPEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
20000
Den
sito
met
ric
un
its
(OD
m
m2)
GFAP
β-actin
Sham MPTP MPTP +
co-ultraPEALut
Inflammatory studies
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 45: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/45.jpg)
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sit
om
etr
ic u
nit
s (
OD
m
m2)
Beclin-1
Sham MPTP
MPTP +
co-ultraPEALut
β-actin
p62
Sham MPTP
β-actin
MPTP +
co-ultraPEALut
Sham MPTP MPTP+ co-ultraPEALut0
10000
20000
30000
Den
sito
met
ric
un
its
(OD
m
m2)
Apoptotic studies
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 46: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/46.jpg)
B
A
C
D
E
Behavioral Studies
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 47: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/47.jpg)
1-Take home messagehellip
These results support the view that PEALut exerts an
antidepressant and neuroprotective effects being a useful novel
therapeutic strategy in neuroinflammatory diseases
Co-ultramicronized
PEALut
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 48: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/48.jpg)
divieto di
bull fare ricerca sugli xenotrapianti
bull fare ricerca sulle sostanze drsquoabuso
bull allevare cani gatti e primati non umani a fini scientifici pur
mantenendo la possibilitagrave di utilizzarli con la conseguente
necessitagrave di ricorrere ad allevamenti extranazionali
bull utilizzare animali nei corsi universitari delle facoltagrave
scientifiche legate alla ricerca biomedica ad esclusione delle
facoltagrave di Medicina veterinaria
In italiahellip
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 49: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/49.jpg)
OPIOIDS
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 50: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/50.jpg)
Biomed Res Int 2014 accepted
Animals were treated daily with 30 mg kg PEA sc or with vehicle The pain threshold was evaluated every day immediately before and 30 min after the injection ip of 10 mg kg morphine Each value represents the mean plusmn SEM of 12 rats per group performed in 2 different experimental sets Plt001 versus pretest values
Plantar test
Paw pressure test
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 51: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/51.jpg)
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Microglial (Iba1-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented microglia cell number increase induced by morphine in the dorsal horn
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 52: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/52.jpg)
Di Cesare Mannelli et al Biomed Res Int 2014 accepted
Astroglial (GFAP-positive) cell density in the dorsal horn of the spinal cord
30 mg kg PEA sc and 10 mg kg morphine ip were administered daily Immunohistochemical analysis were performed on days 6 and 11 Left Representative images of merged Iba1-labeled microglia cells (red) plus DAPI-labeled cell nuclei (blue) scale bar 50 microm Right quantitative analysis of cellular density was performed evaluating 6 animals for each group Each value represents the mean plusmnSEM of 6 rats per group performed in 2 different experimental sets Plt005 and Plt001 versus vehicle + vehicle Plt001 versus vehicle + morphine
PEA prevented astrocyte cell number increase induced by morphine in the dorsal horn
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 53: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/53.jpg)
The evidence of a delayed development of tolerance to the
antinociceptive effects of morphine in the presence of PEA
suggests a possible application of this endogenous compound in
opioid-based therapies
2-Take home messagehellip
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 54: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/54.jpg)
Le scoperte scientifiche dal laboratorio al letto
del paziente
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 55: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/55.jpg)
Traslational Research
Master Clinician Physician
Scientist
Clinical
Investigator
Bench
Pathophysiology
Clinical problems
Diseases
New infections
Genomics
Proteomics
TrangenicKnockouts
Stem cells
Structural
Biologyimaging
informatics
Clinical trials
Bedside Bedside
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 56: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/56.jpg)
What is Translational Research
Pubmed Search Translational Research - 33824 items
The evolution of chemoradiation for glioblastoma a modern success story
The importance of translational research in brain injury
Cell culture medium composition and translational adult bone marrow-derived stem cell
research
Insights from molecular investigations of traditional Chinese herbal stroke medicines
Role of proteomics in translational research in cervical cancer
A versatile tool for conditional gene expression and knockdown
Academic psychiatry and the pharmaceutical industry
Advances of Alzheimers disease research crossroad of basic and translational studies
RNA editing in plant organelles machinery physiological function and evolution
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 57: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/57.jpg)
The fact is animal research does occur and we
need to know our responsibilities
The Legal Obligations
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 58: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/58.jpg)
Endocannabinoid system and psychiatric
disorders
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 59: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/59.jpg)
It was known since 1980 that particular N-acylethanolamine-
related compounds accumulated in damaged tissues
N-palmitoyl-posphatidyl-
ethanolamine (NAPE)
NAPE-PLD
PALMITIC ACID
ETHANOLAMINE
Fatty acid amide hydrolase
(FAAH)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 60: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/60.jpg)
When animals can be used
1 When there are no other alternatives
2 When confirmation has been made that
research activities are not unnecessarily
duplicating previously conducted experiments
3 Experiments involving animals are relevant to
human or animal health will advance scientific
knowledge or will be for the good of society
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 61: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/61.jpg)
There are two types of Research
Basic and Applied
Basic Research discovering new facts about how things
work how they are made or what causes a biological event
to occur Basic research can explore a topic explain a topic
or describe a topic
For Example A researcher discovered that genes can be
turned off or on by small RNA molecules in the body This
study was conducted on worms It led to the Nobel Prize in
2006
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 62: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/62.jpg)
PALMITOYLETHANOLAMIDE AN ENDOGENOUS
PPAR-α AGONIST
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 63: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/63.jpg)
Results-1 Anxiety-like behavior - PEALut
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 64: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/64.jpg)
Results-1 Anxiety-like behavior -PEALut
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 65: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/65.jpg)
Sham
MPTP
MPTP
+ co
-ultr
aPEA
Lut
0
5000
10000
15000
Den
sito
met
ric
un
its
(OD
m
m2)
IκB-α
β-actin
Sha
m
MPTP
MPTP +
co-
ultraPEALut
Sham
MPTP
MPTP+co
-ultr
aPEA
Lut
0
10000
20000
30000
40000
Den
sito
met
ric
un
its
(OD
m
m2)
Sham MPTP
MPTP +
co-ultraPEALut
NF-κB p65
Laminin AC
Inflammatory studies
AEA levels in rat brain following acute and chronic drug
administration
![Page 66: La ricerca: trasferibilità del modello animaleassociazionegeriatri.it/wp-content/uploads/2015/04/15-30...La ricerca: trasferibilità del modello animale ... 30 mg/ kg PEA s.c. and](https://reader031.vdocuments.pub/reader031/viewer/2022021801/5b2617b87f8b9ad33d8b546d/html5/thumbnails/66.jpg)
AEA levels in rat brain following acute and chronic drug
administration