La Medic ina nel l ’era del Microbiota

Come le recenti conoscenze sul microbiota sconvolgono i concetti

della medicina e della nutrizione.

Paolo Mainardi

Funzioni fisiologiche, es.:

• Pressione sanguigna (Yang T, Zubcevic J. Gut-Brain Axis in Regulation of Blood Pressure.

Front Physiol. 2017 Oct 25;8:845).

• Colesterolemia (Gérard P. Metabolism of cholesterol and bile acids by the gut microbiota.

Pathogens. 2013 Dec 30;3(1):14-24.)

• Funzioni cardiache (Luedde M, Winkler T, Heinsen FA, Rühlemann MC, Spehlmann ME,

Bajrovic A, Lieb W, Franke A, Ott SJ, Frey N. Heart failure is associated with depletion of

coreintestinal microbiota. ESC Heart Fail. 2017 Aug;4(3):282-290)

• Funzioni renali (Jiang S, Xie S, Lv D, Wang P, He H, Zhang T, Zhou Y, Lin Q, Zhou H, Jiang J,Nie

J, Hou F, Chen Y. Alteration of the gut microbiota in Chinese population with chronic kidney disease.

Sci Rep. 2017 Jun 6;7(1):287)0

• Sistema Neuroendocrino (Farzi A, Fröhlich EE, Holzer P. Gut Microbiota and the

Neuroendocrine System. Neurotherapeutics. 2018 Jan;15(1):5-22)

• …

Allo stesso modo con cui noi teniamo pulita e efficiente la nostra casa

Microbiota intestinale:

Ago della bilancia tra salute e malattia.

Feng Q, Chen W-D and Wang Y-D (2018) Gut Microbiota: An Integral Moderator in Health and Disease. Front. Microbiol. 9:151

2018: 4573 articles on pubmed

Paolo Mainardi

Taste receptors to produce anticipatory response to foods or pathogens.

• They recognize food

• Memorized it

• To produce fast response

✓ The main role of digestive system is to

protect gut ecosystem

✓ By the food we can talk with microbiota

✓ Food information goes beyond thenutritional aspect

Let the food to be the medicine and the medicine to be the food.

Hippocrates

La loro scoperta rivoluziona i concetti della nutrizione.

When are stimulated by bitter compounds, such as

nicotine or quinine, the cilia waved back and forth

vigorously, helping to clear the airways of irritating

compounds

Taste receptors response to bitter-tasting compunds in airway

Bitter receptors are also found on cilia on

human airway epithelium cells

“they stop breathing,

they cough and sneeze,”

This response is to stop the irritant from being inhaled deep into the lungs

Microbiota alters taste receptors

• Germ-free mice had altered TRs for fat on tongues and on intestine.(Duca FA et al. 2012 Increased oral detection, but decreased intestinal signaling for fats in mice lacking gut microbiota. PLoS ONE 7:

e39748)

• Germ-free mice preferred more sweets, have more sweet TRs in gut vs normal

mice.(Swartz T et al. 2012 Up-regulation of intestinal type 1 taste receptor 3 and sodium glucose luminal transporter-1 expression and

increased sucrose intake on mice lacking microbiota. Br J Nutr 107:621)

Microbiota alters taste receptors

• Germ-free mice had altered TRs for fat on tongues and on intestine.(Duca FA et al. 2012 Increased oral detection, but decreased intestinal signaling for fats in mice lacking gut microbiota. PLoS ONE 7:

e39748)

• Germ-free mice preferred more sweets, have more sweet TRs in gut vs normal

mice.(Swartz T et al. 2012 Up-regulation of intestinal type 1 taste receptor 3 and sodium glucose luminal transporter-1 expression and

increased sucrose intake on mice lacking microbiota. Br J Nutr 107:621)

• Gastric bypass surgery modifies TR, change microbiota and alters satiety and

food preferences.(Miras AD, Ie Roux CW. 2013 Mechanisms underlying weight loss after bariatic surgery. Nat Rev Gastroenterol Hepatol 10:)575-84

Microbes could influence food preferences by altering receptor expression or transduction

Microbiota alters taste receptors

• Germ-free mice had altered TRs for fat on tongues and on intestine.(Duca FA et al. 2012 Increased oral detection, but decreased intestinal signaling for fats in mice lacking gut microbiota. PLoS ONE 7:

e39748)

• Germ-free mice preferred more sweets, have more sweet TRs in gut vs normal

mice.(Swartz T et al. 2012 Up-regulation of intestinal type 1 taste receptor 3 and sodium glucose luminal transporter-1 expression and

increased sucrose intake on mice lacking microbiota. Br J Nutr 107:621)

• Gastric bypass surgery modifies TR, change microbiota and alters satiety and

food preferences.(Miras AD, Ie Roux CW. 2013 Mechanisms underlying weight loss after bariatic surgery. Nat Rev Gastroenterol Hepatol 10:)575-84

Microbes could influence food preferences by altering receptor expression or transduction

Allergie/Intolleranze= reazioni paradosso dei recettori del gusto

NON DIPENDONO DAL CIBO MA DA UNA RISPOSTA SBAGLIATA

Influenza selettiva del microbiota sulla dieta:

Wu GD, Chen J, Hoffmann C, et al. Linking Long-Term Dietary Patterns with Gut Microbial Enterotypes. Science (New York, N.y). 2011;334(6052):105-108. doi:10.1126/science.1208344.

CarboidratiPrevotella

FibreBifidobacteria

Alcuni grassiBacteroidetes

Topi privati dei TLR5 diventano obesi anche se a dieta standard.

Paolo Mainardi

Un’insalata, per favore

Per noi cheeseburger

e patatine

Controllano la trasformazione del cibo in energia

Queipo-Ortuño MI, Seoane LM, Murri M, et al. Gut Microbiota Composition in Male Rat Models under Different NutritionalStatus and Physical Activity and Its Association with Serum Leptin and Ghrelin Levels. Sanz Y, ed. PLoS ONE. 2013;8(5):e65465.

Gut Microbiota modifies Serum Leptin and Ghrelin Levels

Wu GD, Chen J, Hoffmann C, et al. Linking Long-Term Dietary Patterns with Gut Microbial Enterotypes. Science (New York, N.y). 2011;334(6052):105-108. doi:10.1126/science.1208344.

Attraverso i recettori del gusto:

Taste receptors response to carbohydrates and proteins

T1R3 receptors secrete ghrelin, encouraging eating

when important nutrients are available .

The sweet receptor on L cells produce the gastrointestinal

incretin hormone GLP-1, which stimulates insulin

production and sends a satiety signal to the brain

Jang,H.J.etal.Proc.NatlAcad.Sci.USA104, 15069–15074 (2007).

Margolskee, R. F. et al. Proc. Natl Acad. Sci. USA 104, 15075–15080 (2007).

Ci incoraggiano a mangiare se c’è cibo “buono” (Proteine e carboidrati),

ma se ingeriamo carboidrati indigeribili, questi, poi, ci danno senso di sazietà.

Taste receptors response bitter-tasting compunds

Initially, T1R3 receptors secrete ghrelin,

encouraging eating when important nutrients are

available .

This curbs the appetite by prolonging the sense of fullness

and satiety — perhaps to prevent the ingestion of toxic food.

Margolskee, R. F. et al. Proc. Natl Acad. Sci. USA 104, 15075–15080 (2007).

But after 30 minutes, food intake decreased, as

did gastric emptying, keeping the food in the

stomach

The Romans drank wine infused with bitter herbs to prime the appetite and preventover-eating.

Taste receptors response bitter-tasting compunds

Initially, T1R3 receptors secrete ghrelin,

encouraging eating when important nutrients are

available .

This curbs the appetite by prolonging the sense of fullness

and satiety — perhaps to prevent the ingestion of toxic food.

Margolskee, R. F. et al. Proc. Natl Acad. Sci. USA 104, 15075–15080 (2007).

But after 30 minutes, food intake decreased,

as did gastric emptying, keeping the food in

the stomach

The Romans drank wine infused with bitter herbs to prime the appetite and preventover-eating.

Ippocrate consigliava a chi proprio non aveva voglia di

mangiare un medicinale di sua invenzione, il vinum

ippocraticum, vino bianco dolce, speziato con macerati di

dittamo, assenzio e ruta.

Un vino che i romani, per migliorarne il sapore amarognolo,

mischiarono con rosmarino e salvia. E che chiamarono,

pertanto, vinum absinthiatum.

When are stimulated by bitter compounds,

such as nicotine or quinine, the cilia waved back and

forth vigorously, helping to clear the airways of

irritating compounds

Taste receptors response to bitter-tasting compunds in airway

Bitter receptors are also found on cilia on

human airway epithelium cells

“they stop breathing,

they cough and sneeze,”

This response is to stop the irritant from being inhaled deep into the lungs

Urinary metabolites in Chocolate indifferent and Chocolate desidering

Rezzi S, et al. Human metabolic phenotypes link directly to specific dietary preferences in healthy individuals. J Proteome Res. 2007 Nov;6(11):4469-77.

Microbiota dei golosi diverso da quello dei non golosi

Oggi non ha più senso:

- Parlare di calorie

- Dividere il cibo in grassi, carboidrati, e proteine.

• Quali grassi?

• Quali Proteine?

• Quali carboidrati?

Pasquaretta, C.; Gómez-Moracho, T.; Heeb, P.; Lihoreau, M. Exploring Interactions between the Gut Microbiota and Social Behaviorthrough Nutrition. Genes 2018, 9, 534.

- Comportamento alimentare

- Comportamento sociale

- Comportamento sessuale

Processi infiammatori per riparare i tessuti

Paolo Mainardi

Il microbiota controlla ANCHE la risposta infiammatoria

Andreasen AS, Krabbe KS, Krogh-Madsen R, Taudorf S, Pedersen BK, Møller K. Human Endotoxemia as a model of systemicinflammation. Curr Med Chem. 2008;15(17):1697-705.

CASCATA INFIAMMATORIA:

• Le pro-infiammatorie producono necrosi

• Le anti-infiammatorie ricostruiscono i tessuti

Nel sistema nervoso centrale:

• Le pro-infiammatorie inibiscono i processi di sinaptogenesi e neurogenesi,

• Le anti-infiammatorie attivano i processi di sinaptogenesi e neuogenesi

Microbiota intestinale e Neurosviluppo Fondazione Domus Angelorum - Onlus

Paolo Mainardi

Danno Tissutale

Vasta demolizione

Ricostruzione

Paolo Mainardi

Risposta infiammatoria acuta o cronica

Demoliscono Abili aricostruire

Inabili aricostruire

Hang Ch et al, Intestinal mucosa structure after TBI. World J Gastroenterol 2003,9 (12) 2776-2781

Prima trauma cranico

3 giorni dopo trauma cranico

3 giorni dopo Trauma Cranico collassa la membrana intestinale

Brain

GutIntestinal mucosa

Paolo Mainardi

IL-6 elevation in the brain is involved in the mediation of autism-like behaviors through impairments of neuroanatomical

structures and neuronal plasticity

Wei H, Alberts I, Li X. Brain IL-6 and autism. Neuroscience. 2013 Nov 12;252:320-5.

Brain

Gut

Paolo Mainardi

• Danni neuroanatomici strutturali

• Riduce la neuroplasticità

23

Paolo Mainardi

24

• Le citochine INABILI non consentono una

corretta rigenerazione cellulare.

(degenerazione)

Paolo Mainardi

25

• Le citochine INABILI non consentono una

corretta rigenerazione cellulare.

(degenerazione)

Paolo Mainardi

Patologie autoimmuni: (sistema immunitario impazzito?)

Linfociti T: prodotti casualmente, eliminati il 97% per apoptosi

Linfociti B: le citochine controllano la tolleranza.

Autoanticorpi= cacciatori nella caccia di selezione nei parchi naturali

26

• Le citochine INABILI non consentono una

corretta rigenerazione cellulare.

(degenerazione)

Paolo Mainardi

- Cytokines participate in neuronal development in brain functioning. Inappropriate activity can produce

different neurological symptomsGoines PE, Ashwood P. Cytokine dysregulation in autism spectrum disorders (ASD): possible role of the environment. Neurotoxicol Teratol.

2013 Mar-Apr;36:67-81.

• Citochine partecipano al neurosviluppo

Metabolic changes during pregnancy:

- Same nutritional intake

- Increased energy extraction by foods:

• the microbiota undergoes profound changes in T3 vs T1

• It is not due to nourish the fetus, but to realize his tissues by anti-

inflammatory citokines

No-problems

Houston, we have a problem!

Stig Bengmark, Acute and ‘‘chronic’’ phase reaction - a mother of disease. Clinical Nutrition (2004) 23, 1256–1266

• Poi, nei primi 1000 giorni di vita, il microbiota si evolve aumenta la biodiversità dei

ceppi

• Nei primi giorni di vita fetale è stato campionato una piccola quantità di ceppi

batterici (simbionti permanenti)

Paolo Mainardi

Insediamento del microbiota, verso un aumento della biodiversità dei ceppi.

???

Jeremy E. Koenig et al. Succession of microbial consortia in the developing infant gut microbiome Proceedings of the National Academy ofSciences Mar 2011, 108 (Supplement 1) 4578-4585;

Febbre

RisoAntibiotic

o

Antibiotico

Antibioticocefalosporina

adulto

Più FORTE

Più DEBOLE

adulto anziano

Invecchiando si torna bambini.

“Fragilità dell’anziano”

Invecchiando si torna bambini.

Vu l n e ra b i l i tà F r a g i l i t à

?

Ridotta diversità ceppi:

• Ridotta capacità di riparare i danni

• Ridotta capacità a rigenerare i tessuti

• Ridotta capacità a scremare linfociti T

• Ridotta capacità a controllare organi

…

Tu questa la chiami vita?

Lo scopo principale del sistema digerente è mantenere ambiente confortevole ai

batteri residenti nel colon, non è quello di nutrirci!!!!!!

Nel corpo umano vivono 100 trilioni di forme di vita microbiche

Pat.

H+Mucin

Mucus

pH=7.0

pH=8.2

“good” Bacteria

“Bad” Bacteria

Gastric Juices, biliary salts

SCFAs:The nutrients of the cells of digestive systemof all mammals

First of all: Nourish

the cells of digestive

system

Protonic pumps

L’intestinonon

nasceimparato

Il distacco del neonato dalla mamma non avviene al taglio del cordone ombelicale.

Il colostro si va a sostituire nella duplice funzione di PROTEGGERE e nutrire

Inoltra INSEGNA al neonato a prodursi gli anticorpi e i fattori di crescita.

• Alla nascita è completamente permeabile e sterile• Prima maturazione con il colostro• Maggiore maturazione con lo svezzamento

L’intestinonon

nasceimparato

Azioni intestinali dell’alfa-lattoalbumina

• gastrite cronica• atrofia gastrica• …

Stomaco

Corpus AntrumDuodeno

Digiuno

Ileo

Colon

gel sol

pH 7pH 5-6

pH 1.2pH 5- 7 pH 1.2

pH 2

pH 7

H+

HCO3-

H2O + CO2

Demolisce cibo eAgenti patogeni

Pompa protonica

muco

Il pH controlla il rapporto tra i ceppi batterici.

Alpha-lactalbumin protects in a

dose-dependent manner from

gastric ulcers induced by alcool or

stress

Hung CR, Cheng JT, Shih CS. Gastric mucosal damage induced by arecoline seizure in rats. Life Sci. 2000 May 5; 66 (24): 2337-49.

- The amount of gastric ulcers depend in a

dose-dependent manner on the amount of

epileptogenic agent injected i.c.v.

Alpha-lactalbumin protects in a

dose-dependent manner from gastric

ulcers induced by alcool or stress

Alpha-lactalbumin increases IL-1ra/IL-1 ratio:

OFF

Reparativeprocesses

ON

Reparativeprocesses

It becomes REALLY INTERESTING to correlate the ability to act on intestinal

inflammation with to control epileptic seizures.

Alpha-lactalbumin increases IL-1ra/IL-1 ratio:

OFF

Reparativeprocesses

ON

Reparativeprocesses

• brings out chronic inflammation,

• Restore endogenous riparative processes

• MORE POWERFUL THAN ANY DRUGS

It becomes REALLY INTERESTING to correlate the ability to act on intestinal

inflammation with to control epileptic seizures.

➢ Bovine milk-derived α-lactalbumin inhibits colon inflammation andcarcinogenesis in azoxymethane and dextran sodium sulfate-treated mice. Yamaguchi M, Takai S, Hosono A, Seki T.. Biosci Biotechnol Biochem.

2014;78(4):672-9.

➢ Anti-inflammatory mechanisms of bioactive milk proteins in theintestine of newborns Chatterton DE, Nguyen DN, Bering SB, Sangild PT.. Int J

Biochem Cell Biol. 2013 Aug;45(8):1730-47.

L’alfa-lattoalbumina esplica azioni SPECIFICHE nel digerente dell’uomo,

in quanto selezionata dalla Natura ad attivarlo dopo la nascita

46

Favale E, Rubino V, Mainardi P, Lunardi G, Albano C. Anticonvulsant effect of fluoxetine in humans. Neurology. 1995Oct;45(10):1926-7.

Lunardi G, Mainardi P, Rubino V, Fracassi M, Pioli F, Cultrera S, Albano C. Tryptophan and epilepsy. Adv Exp Med Biol.1996;398:101-2.

Albano C, Cupello A, Mainardi P, Scarrone S, Favale E. Successful treatment of epilepsy with serotonin reuptake inhibitors:proposed mechanism. Neurochem Res. 2006 Apr;31(4):509-14.

Mainardi P, Leonardi A, Albano C. Potentiation of brain serotonin activity may inhibit seizures, especially in drug-resistantepilepsy. Med Hypotheses. 2008;70(4):876-9.

Mainardi P, Albano C. Is the antiepileptic effect of the ketogenic diet due to ketones? Med Hypotheses. 2008;70(3):536-9.

Striano P, D'Amato E, Pezzella M, Mainardi P, Zara F, Striano S. Sudden death in Unverricht-Lundborg patients: is serotoninthe key? Neurol Sci. 2010 Feb;31(1):115-6.

Citraro R, Scicchitano F, De Fazio S, Raggio R, Mainardi P, Perucca E, De Sarro G, Russo E. Preclinical activity profile of α-lactoalbumin, a whey protein rich in tryptophan, in rodent models of seizures and epilepsy. Epilepsy Res. 2011 Jun;95(1-2):60-9.

Errichiello L, Pezzella M, Santulli L, Striano S, Zara F, Minetti C, Mainardi P, Striano P. A proof-of-concept trial of the wheyprotein alfa-lactalbumin in chronic cortical myoclonus. Mov Disord. 2011 Dec;26(14):2573-5.

Russo E, Scicchitano F, Citraro R, Aiello R, Camastra C, Mainardi P, Chimirri S, Perucca E, Donato G, De Sarro G. Protectiveactivity of α-lactoalbumin (ALAC), a whey protein rich in tryptophan, in rodent models of epileptogenesis. Neuroscience.2012 Dec 13;226:282-8.

Dal ruolo della serotonina cerebrale, allo studio del sistema digerente…

47

Favale E, Rubino V, Mainardi P, Lunardi G, Albano C. Anticonvulsant effect of fluoxetine in humans. Neurology. 1995Oct;45(10):1926-7.

Lunardi G, Mainardi P, Rubino V, Fracassi M, Pioli F, Cultrera S, Albano C. Tryptophan and epilepsy. Adv Exp Med Biol.1996;398:101-2.

Albano C, Cupello A, Mainardi P, Scarrone S, Favale E. Successful treatment of epilepsy with serotonin reuptake inhibitors:proposed mechanism. Neurochem Res. 2006 Apr;31(4):509-14.

Mainardi P, Leonardi A, Albano C. Potentiation of brain serotonin activity may inhibit seizures, especially in drug-resistantepilepsy. Med Hypotheses. 2008;70(4):876-9.

Mainardi P, Albano C. Is the antiepileptic effect of the ketogenic diet due to ketones? Med Hypotheses. 2008;70(3):536-9.

Striano P, D'Amato E, Pezzella M, Mainardi P, Zara F, Striano S. Sudden death in Unverricht-Lundborg patients: is serotoninthe key? Neurol Sci. 2010 Feb;31(1):115-6.

Citraro R, Scicchitano F, De Fazio S, Raggio R, Mainardi P, Perucca E, De Sarro G, Russo E. Preclinical activity profile of α-lactoalbumin, a whey protein rich in tryptophan, in rodent models of seizures and epilepsy. Epilepsy Res. 2011 Jun;95(1-2):60-9.

Errichiello L, Pezzella M, Santulli L, Striano S, Zara F, Minetti C, Mainardi P, Striano P. A proof-of-concept trial of the wheyprotein alfa-lactalbumin in chronic cortical myoclonus. Mov Disord. 2011 Dec;26(14):2573-5.

Russo E, Scicchitano F, Citraro R, Aiello R, Camastra C, Mainardi P, Chimirri S, Perucca E, Donato G, De Sarro G. Protectiveactivity of α-lactoalbumin (ALAC), a whey protein rich in tryptophan, in rodent models of epileptogenesis. Neuroscience.2012 Dec 13;226:282-8.

…alla rivalutazione delle diete del passato…

Mainardi P, Albano C.Is the antiepileptic effect of the ketogenic diet due to ketones?Med Hypotheses. 2008;70(3):536-9.

Dear Paolo,

I actually read your article a few weeks ago and agree. We have one of our researchers, dr

Adam Hartman, investiging KD in a mouse model and he agrees (as do I in my clinical

research) that ketosis is probably over-rates.

Erick Kossoff

L’efficacia della dieta chetogenica non è dovuta alla chetosi

ERICK KOSSOFF

49

La conoscenza del microbiota ci aiuta ad individuare una corretta dieta per la salute

Paolo MainardiCHIMICO

Pasquale StrianoNeurologo-Pediatra

Paolo CartaBIOLOGO

NUTRIZIONISTAEmilio RussoFarmacologo

La ricerca scientifica si basa sulla falsificabilità dei risultati. Karl Popper (1902-1994)

Prof.Emilio Perucca past prresident ILAE Prof. De SarroRettore Magna Grecia Catanzaro

Thanks to these results, ALAC entered in the NIH new drug program screening

Serplus is reported in a Thesis of Master of Nutrition on the

basis of results on autism.

With Dr Tosatto we are planning a clinical study at Dubai

Serplus in autism

The gut brain connection in pediatric migraine: an open label trial.M. Carotenuto et al.

52

Mainardi P, Carta P, Montinari M, Striano P et al. From the Ancient Diets to the RecentAcquisitions on the Role of Brain Inflammation in Epilepsy, Are there Any Links? J NeurolNeurophysiol 2015, 6:3

P.Mainardi, P.Carta, P.Striano. How Knowledge on Microbiota may be Helpful to Establishan Optimal Diet for Health Maintenance. Int J Nutrition, 2018..

Dalle antiche diete al ruolo dell’infiammazione

La conoscenza del microbiota ci aiuta ad individuare una corretta dieta per la salute

Intestinal inflammation is linked to anticonvulsant activity in a mouse model of epilepsy

Prof. Emilio Russo (erusso@unicz.it) (University of Catanzaro)

Contributing AuthorsProf. Carmen De Caro , Dr. Antonio Leo , Miss Valentina Nesci , Prof. Carla Ghelardini , Dr.Lorenzo Di Cesare Mannelli , Prof. Pasquale Striano , Dr. Carmen Avagliano , Antonio

Calignano , Dr. Paolo Mainardi , Dr. Rita Citraro , Prof. Giovambattista De Sarro

Nel 2018 a Vienna

Per la prima volta ben 2 sessioni su Gut Brain Axis al congresso europeo dell’ILAE

DSSDSS-ALAC

DSS-BuNa

DSS-VPA

CTR

DSS-MSZ

Intestinal inflammation is linked to anticonvulsant activity in a mouse model

E.Russo, P.Striano, P.Mainardi,…, submitted to Scientific Report (Nature)

FUTURO PROSSIMO:

Studi clinici su:

• Disturbi del sonno (Franzoni, presidente SINP)

• Disturbi del comportamento alimentare (Prof.Franzoni, Bologna, presidente SINP)

• S. Tourette –pandas (Dott. Pavone, Catania)

• Autismo (Dott. Viri, Novara)

• Epilessia (dott. Viri, Novara, Prof .Franzoni, Bologna, Prof. Striano, Genova)

• Cefalee (prof.Carotenuto, Napoli)

• Pre termine (prof.ssa Suppiej, Ferrara)

Verso una nuova medicina:

• Più potente di qualsiasi farmaco

Take – home message:

Cura il microbiota per consentirgli di prendersi cura di te.

• Capace di rispristinare i potenti meccanismi

endogeni di riparazione e di controllo.

2015

www.unamedicina.itwww.protocollorestart.it

info@kolfarma.it