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

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

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

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

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

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

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

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

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

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 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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

AEA levels in rat brain following acute and chronic drug

administration