Name: assessing hepatic metabolic changes during progressive colonization of germ-free mouse by 1h nmr spectroscopy
Uploaded: 2011-12-15T13:00:00
Description: Watch this Scientific Journal Video about Assessing Hepatic Metabolic Changes During Progressive Colonization of Germ-free Mouse by 1H NMR Spectroscopy at JoVE.com

Tutti gli spettri NMR utilizzati come esempi illustrativi sono derivati ​​da uno studio precedentemente pubblicato con il 7 che è stato sostenuto finanziariamente dalla Nestlé.

1. Colonizzazione di germi senza animali e la raccolta dei campioni <ol><li> Rimuovere sterile animali da isolatori e la casa in una stanza di allevamento in gabbie tradizionali dotati di filtro davanti alla animali convenzionali, che servirà come controlli (Figura 1). </li><li> Mescolare mezzo della cucciolata (3 giorni) preso dalla gabbia di controllo convenzionale con la lettiera del germe senza animali. Tenere sempre 1 / 3 della lettiera sporca convenzionale ogni volta che è necessario rinnovarlo al fine di mant...

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In questo protocollo, abbiamo descritto una procedura di progressiva colonizzazione in un ambiente aperto di approfondire l&#39;impatto della flora intestinale sul metabolismo epatico valutate di 1 profiling H NMR HR MAS di intatto biopsia. Vari metodi di colonizzazione sono state descritte in letteratura. I metodi più comuni per colonizzare gli animali con un microbiota definiti sono sonda gastrica o l&#39;acqua potabile contaminata 19,20. Inoculazione fecale può essere utilizzato anche come des...

Tabella di reagenti e attrezzature specifiche: <table border="1"><tbody><tr><th> Nome del reagente </th><th> Azienda </th><th> Numero di catalogo </th><th> Commenti </th></tr><tr><td> 2,5 millimetri microprovette </td><td> New Era </td><td> NE-H5/2.5-V-Br </td><td></td></tr><tr><td> 1,7 millimetri tubo capillare </td><td> Sigma-Aldrich </td><td> NORS175001 </td><td></td></tr><tr><td> Adattatore capillare </td><td> New Era </td><td> NE-325-5/1.7 </td><td></td></tr><tr><td> Estrazione asta </td><td> New Era </td><td> NE-341-5 </td><td></td></tr><tr><td> HR-MAS BL4 rotore con 50 microlitri sferica Teflon spacer kit </td><td> Bruker </td><td> HZ07213 </td><td></td></tr><tr><td> Kit di strumenti per 50 inserti microlitri </td><td> Bruker </td><td> B2950 </td><td></td></tr><tr><td> Advance III 600MHz NMR </td><td> Bruker </td><td></td><td></td></tr><tr><td> 1 H HR MAS NMR solido sonda </td><td> Bruker </td><td></td><td></td></tr><tr><td> Ossido di deuterio 99,9% </td><td> Sigma-Aldrich </td><td> 530867-1L </td><td></td></tr><tr><td> 3 - (trimetilsilil) propionico acido-d 4 (TSP) </td><td> Sigma-Aldrich </td><td> 269913 </td><td></td></tr></tbody></table>

assessing hepatic metabolic changes during progressive colonization of germ-free mouse by 1h nmr spectroscopy

E &#39;noto che i batteri intestinali contribuiscono significativamente alla omeostasi host, fornendo una serie di benefici come la protezione immunitaria e la sintesi della vitamina. Essi hanno inoltre fornire l&#39;host con una notevole quantità di sostanze nutritive, facendo di questo ecosistema un organo metabolico essenziale. Nel contesto di una crescente evidenza del legame tra la flora intestinale e la sindrome metabolica, comprendere l&#39;interazione metabolica tra l&#39;host e la sua flora intestinale sta diventando una sfida importante della biologia moderna. 1-4 Colonizzazione (noto anche come processo di normalizzazione), designa la creazione di micro-organismi in un antico animale privo di germi. Mentre è un processo naturale che si verificano al momento della nascita, è utilizzato anche in adulti privo di germi animali per controllare l&#39;ecosistema intestinale floreali e ulteriormente determinare il suo impatto sul metabolismo host. Una procedura comune per controllare il processo di colonizzazione è quello di utilizzare il metodo gavage con un singlE o una miscela di microrganismi. Questo metodo comporta una colonizzazione molto veloce e presenta lo svantaggio di essere estremamente stressante 5. E &#39;quindi utile per ridurre al minimo lo stress e per ottenere un processo di colonizzazione lento ad osservare gradualmente l&#39;impatto di stabilimento sul metabolismo batterico ospite. In questo manoscritto, si descrive una procedura per valutare la modifica del metabolismo epatico durante un processo di colonizzazione graduale utilizzando una tecnica non distruttiva profili metabolici. Noi proponiamo di monitorare la colonizzazione microbica intestinale, mediante la valutazione dell&#39;attività microbica intestinale metabolica riflette l&#39;escrezione urinaria di metaboliti microbici co-profilatura di 1 H NMR basati metabolica. Questo consente un apprezzamento della stabilità dell&#39;attività microbica intestinale al di là della sede stabile l&#39;ecosistema microbico intestinale solitamente valutata attraverso il monitoraggio dei batteri fecali da DGGE (gel elettroforesi denaturante gradiente). 6 Ilcolonizzazione si svolge in un ambiente convenzionale aperta ed è iniziata da una cucciolata sporca sporca da animali convenzionali, che servirà come controlli. Roditori sono animali coprofagi, questo garantisce una colonizzazione omogenea come descritto in precedenza 7. Profiling metabolico epatico viene misurato direttamente da una biopsia del fegato intatto con 1 H ad alta risoluzione Magic Angle Spinning spettroscopia NMR. Questo semi-quantitativa tecnica offre un modo rapido per valutare, senza danneggiare la struttura delle cellule, i metaboliti importanti come trigliceridi, glucosio e glicogeno al fine di ulteriormente valutare la complessa interazione tra il processo di colonizzazione e il metabolismo epatico 7-10. Questo metodo può essere applicato a qualsiasi tessuto bioptico 11,12.

Watch this Scientific Journal Video about Assessing Hepatic Metabolic Changes During Progressive Colonization of Germ-free Mouse by 1H NMR Spectroscopy at JoVE.com

Assessing Hepatic Metabolic Changes During Progressive Colonization of Germ-free Mouse by 1H NMR Spectroscopy

Una procedura di progressiva colonizzazione è descritto per valutare ulteriormente il suo impatto sul metabolismo epatico ospitante. La colonizzazione è monitorato non invasivo, valutando l&#39;escrezione urinaria di co-metaboliti microbici da NMR basati su profili metabolici, mentre il metabolismo epatico viene valutata ad alta risoluzione Angle Spinning Magic (MAS HR) profilazione NMR di intatto biopsia.

Valutare alterazioni metaboliche epatiche durante la colonizzazione progressiva di Germe senza mouse da 1 H spettroscopia NMR

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Fluorescence correlation spectroscopy (FCS) is a powerful technique for studying the diffusion of molecules within biological membranes with high spatial ...

Detection of Enterohemorrhagic Escherichia Coli Colonization in Murine Host by Non-invasive In Vivo Bioluminescence System

Detection of Enterohemorrhagic Escherichia Coli Colonization in Murine Host by Non-invasive In Vivo Bioluminescence System

Rilevamento della colonizzazione di Escherichia Coli enteroemorragica in murino Host di sistema Non invasivo In Vivo bioluminescenza

Enterohemorrhagic E. coli (EHEC) O157:H7, which is a foodborne pathogen that causesdiarrhea, hemorrhagic colitis (HS), and hemolytic uremic syndrome ...