fast-simplified-method-for-high-through-put-isolation-mirna-from

Fast and Simplified Method for High Through-put Isolation of miRNA from Highly Purified High Density Lipoprotein

Small non-coding RNAs (miRNAs) have been implicated in a variety of human diseases including metabolic syndromes. They may be utilized as biomarkers for ...

Virginia Commonwealth University

Bile acids are well known for their effects on cholesterol homeostasis and lipid digestion. Since the discovery of bile acid receptors, of which there are farnesoid X receptor (FXR), a nuclear receptor, and the plasma membrane G-protein receptor, as well as Takeda G-protein coupled receptor clone 5, further roles have been elucidated for bile acids including glucose and lipid metabolism as well as inflammation. Additionally, treatment with bile acid receptor agonists has shown a decrease in the amount of atherosclerosis plaque formation and decreased portal vascular resistance and portal hypotension in animal models. Furthermore, rodent models have demonstrated antifibrotic activity using bile acid receptor agonists. Early human data using a FXR agonist, obeticholic acid, have shown promising results with improvement of histological activity and even a reduction of fibrosis. Human studies are ongoing and will provide further information on bile acid receptor agonist therapies. Thus, bile acids and their derivatives have the potential for management of liver diseases and potentially other disease states including diabetes and the metabolic syndrome.

Bile acids: emerging role in management of liver diseases.

Establishment of intestinal infection with Entamoeba histolytica depends on the mouse strain; C57BL/6 mice are highly resistant, and C3H/HeJ mice are relatively susceptible. We found that resistance to intestinal infection was independent of lymphocyte activity or H-2 haplotype and occurred in the first hours to days postchallenge according to in vivo imaging. At 18 h postchallenge, the ceca of resistant C57BL/6 mice were histologically unremarkable, in contrast to the severe inflammation observed in susceptible C3H/HeJ mice. Comparison of cecal gene expression in C3H/HeJ and C57BL/6 mice demonstrated that there was parasite-induced upregulation of proinflammatory and neutrophil chemotaxis transcripts and there was downregulation of transforming growth factor beta signaling molecules. Pretreatment with dexamethasone abrogated the partial resistance of C3H/HeJ or CBA mice through an innate, lymphocyte-independent mechanism, but it had no effect on the high-level resistance of C57BL/6 mice. Similarly, administration of a neutrophil-depleting anti-Gr-1 monoclonal antibody (RB6-8C5) decreased the partial resistance of CBA mice and led to severe pathology compared to control antibody-treated mice, but it had no effect on C57BL/6 resistance. These data indicate that there are discrete mechanisms of innate resistance to E. histolytica depending on the host background and, in contrast to other reports, imply that neutrophils are protective and not damaging in intestinal amebiasis.

Resistance to intestinal Entamoeba histolytica infection is conferred by innate immunity and Gr-1+ cells.

Few data exist on the relative importance of individual Cryptosporidium species in acquired immunodeficiency syndrome cryptosporidiosis. We characterized 127 inpatients infected with human immunodeficiency virus (HIV) in Tanzania for their CD4 cell count and by stool analysis, including Cryptosporidium immunofluorescence and polymerase chain reaction-restriction fragment length polymorphism. Cryptosporidium was detected in patients both with and without diarrheal symptoms (defined as > or = 3 liquid stools/day, 11 of 61 versus 11 of 66; P = not significant) and was a marker for low CD4 cell count (median = 124/microL versus 212/microL in Cryptosporidium-negative patients; P < 0.04). Cryptosporidium hominis was the predominant species in this region and was associated with a longer duration of symptoms, a higher rate of asymptomatic infection, and a lower CD4 cell count versus C. parvum-infected patients (P < 0.05). This study suggests there may be important differences in the natural history of Cryptosporidium infection in HIV-infected persons depending on parasite species.

Short report: asymptomatic Cryptosporidium hominis infection among human immunodeficiency virus-infected patients in Tanzania.

&#x963F;&#x7C73;&#x5DF4;&#x7684;&#x5168;&#x57FA;&#x56E0;&#x7EC4;&#x8F6C;&#x5F55;&#x5206;&#x6790;&#x6267;&#x884C;&#x5728;&#x9694;&#x79BB;&#x6765;&#x81EA;&#x516D;&#x4E2A;&#x611F;&#x67D3;&#x5C0F;&#x9F20;&#x7ED3;&#x80A0;&#x548C;&#x4F53;&#x5916;&#x57F9;&#x517B;&#x7684;&#x6ECB;&#x517B;&#x3002;9435 &#x5F00;&#x653E;&#x9605;&#x8BFB;&#x6846;&#x67B6; &#xFF08;ORFs&#xFF09; &#x548C; 9066 5&#39; &#x548C; 3&#39; &#x4FA7;&#x7FFC;&#x5730;&#x533A;&#x8BBE;&#x7F6E;&#x5E73;&#x53F0;&#x57FA;&#x56E0;&#x8868;&#x8FBE;&#x9635;&#x5217;&#x4E3A;&#x8FD9;&#x79CD;&#x5206;&#x6790;&#xFF0C;&#x5176;&#x4E2D;&#x5305;&#x62EC;&#x63A2;&#x9488;&#x8BBE;&#x8BA1;&#x597D;&#x5F97;&#x591A;&#x3002;&#x8A8A;&#x672C;&#x88AB;&#x68C0;&#x6D4B;&#x5230;&#x7684;&#x6240;&#x6709; ORFs &gt; 80%&#x3002;&#x5171; 523 &#x6210;&#x7EE9;&#x5355; (&#x6240;&#x6709; E.&#x7EC4;&#x7EC7;&#x5185;&#x963F;&#x7C73;&#x5DF4;&#x57FA;&#x56E0;&#x7684; 5.2%) &#x6709;&#x91CD;&#x5927;&#x53D8;&#x5316;&#x5728; amebae &#x5929; 1 &#x548C; 29 &#x80A0;&#x9053;&#x611F;&#x67D3;&#x540E;&#x5206;&#x79BB;&#xFF1A; 326 &#x548C;&#x9760;&#x5929; 1&#x3001; 29&#x3001; 109 &#x548C; 88 &#x4E24;&#x5929;&#x3002;&#x5B9A;&#x91CF;&#x5B9E;&#x65F6;&#x9006;&#x8F6C;&#x5F55;&#x805A;&#x5408;&#x9176;&#x94FE;&#x53CD;&#x5E94;&#x8BC1;&#x5B9E;&#x8FD9;&#x4E9B;&#x53D8;&#x5316; 11/12 &#x57FA;&#x56E0;&#x4F7F;&#x7528;&#x989D;&#x5916;&#x7684;&#x516D;&#x4E2A;&#x5C0F;&#x9F20;&#x4E2D;&#x5206;&#x79BB;&#x7684; mRNA &#x8FDB;&#x884C;&#x6D4B;&#x8BD5;&#x3002;&#x5BF9;&#x80A0;&#x9053;&#x73AF;&#x5883;&#x7684;&#x9002;&#x5E94;&#x88AB;&#x4F34;&#x968F;&#x589E;&#x52A0;&#x7EC6;&#x80DE;&#x4FE1;&#x53F7;&#x5305;&#x62EC;&#x8DE8;&#x819C;&#x6FC0;&#x9176;&#x3001; ras &#x548C; rho &#x5BB6;&#x65CF;&#x86CB;&#x767D;&#xFF0C;&#x9499;&#x7ED3;&#x5408;&#x86CB;&#x767D;&#x7684;&#x57FA;&#x56E0;&#x7684;&#x4E00;&#x4E2A;&#x5B50;&#x96C6;&#x3002;&#x57FA;&#x56E0;&#x7684; mRNA &#x4E30;&#x5EA6;&#x7684;&#x663E;&#x8457;&#x8DCC;&#x5E45;&#x53C2;&#x4E0E;&#x7CD6;&#x9175;&#x89E3;&#x548C;&#x8102;&#x80AA;&#x9176;&#x7684;&#x76F8;&#x5E94;&#x5347;&#x5E45;&#x5206;&#x522B;&#x4E3A;&#x4E0E;&#x80FD;&#x91CF;&#x4EE3;&#x8C22;&#x7684;&#x53D8;&#x5316;&#x76F8;&#x4E00;&#x81F4;&#x3002;&#x9632;&#x5FA1;&#x7EC6;&#x83CC;&#x5B58;&#x5728;&#x4E8E;&#x80A0;&#x9053; &#xFF08;&#x4F46;&#x7F3A;&#x4E4F;&#x4ECE;&#x4F53;&#x5916;&#x57F9;&#x517B;&#xFF09; &#x63D0;&#x51FA;&#xFF0C;&#x7C7B;&#x4F3C;&#x4E8E; AIG1 &#x690D;&#x7269;&#x6297;&#x83CC;&#x86CB;&#x767D;&#x7684;&#x57FA;&#x56E0;&#x8868;&#x8FBE;&#x6C34;&#x5E73;&#x7684;&#x53D8;&#x5316;&#x3002;&#x6C27;&#x89E3;&#x6BD2;&#x9014;&#x5F84;&#x8DCC;&#x5E45;&#x5728;&#x538C;&#x6C27;&#x7ED3;&#x80A0;&#x5185;&#x8154;&#x4E2D;&#x6309;&#x9884;&#x671F;&#x3002;&#x5DF2;&#x77E5;&#x7684;&#x6BD2;&#x529B;&#x56E0;&#x5B50;&#x7684;&#x6700;&#x663E;&#x8457;&#x7684;&#x53D8;&#x5316;&#x662F;&#x589E;&#x52A0; 20-35-&#x500D;&#x534A;&#x80F1;&#x6C28;&#x9178;&#x86CB;&#x767D;&#x9176;&#x56DB;&#x6837;&#x57FA;&#x56E0;&#x548C;&#x4E24;&#x4E2A;&#x5BB6;&#x5EAD;&#x6210;&#x5458;&#x7684; Gal/&#x591A;&#x51DD;&#x96C6;&#x7D20;&#x5149;&#x4E9A;&#x57FA;&#x7684;&#x8DCC;&#x5E45;&#x4E3A; 2-3 &#x500D;&#x3002;&#x63A7;&#x5236;&#x80A0;&#x9053; mRNA &#x4E30;&#x5EA6;&#x89C2;&#x6D4B;&#x5230;&#x53D8;&#x5316;&#x7684;&#x6F5C;&#x5728;&#x53EF;&#x80FD;&#x4E0E;&#x56DB;&#x4E2A;&#x76F8;&#x5173;&#x86CB;&#x767D;&#x8D28;&#x4E0E; DNA &#x7ED3;&#x5408;&#x57DF;&#x7684;&#x4E0B;&#x8C03; 6-16 &#x500D;&#x80A0;&#x9053;&#x73AF;&#x5883;&#x4E2D;&#x4F11;&#x606F;&#x3002;&#x6700;&#x540E;&#xFF0C;E.&#x7EC4;&#x7EC7;&#x5185;&#x963F;&#x7C73;&#x5DF4;&#x7684;&#x8F6C;&#x5F55;&#x8FC7;&#x7A0B;&#x7684;&#x7B2C;&#x4E00;&#x4E2A;&#x5168;&#x57FA;&#x56E0;&#x7EC4;&#x5206;&#x6790;&#x8868;&#x660E;&#x7EDD;&#x5927;&#x591A;&#x6570;&#x7684;&#x57FA;&#x56E0;&#x7684;&#x8F6C;&#x5F55;&#x4E2D;&#x6ECB;&#x517B;&#x4F53;&#xFF0C;&#x800C;&#x4E14;&#x6B64;&#x4E3B;&#x673A;&#x4E2D;&#x80A0;&#x6ECB;&#x517B;&#x6539;&#x52A8;&#x5206;&#x6CCC;&#x4EE3;&#x8C22;&#x3001; &#x6C27;&#x9632;&#x5FA1;&#x3001; &#x7EC6;&#x80DE;&#x4FE1;&#x53F7;&#x8F6C;&#x5BFC;&#x3001; &#x6BD2;&#x529B;&#x3001; &#x6297;&#x83CC;&#x6D3B;&#x6027;&#x53CA; DNA &#x7ED1;&#x5B9A;&#x4E2D;&#x53D7;&#x7275;&#x8FDE;&#x7684;&#x57FA;&#x56E0;&#x7684;&#x8868;&#x8FBE;&#x3002;

&#x80A0;&#x9053;&#x5B9A;&#x690D;&#x548C;&#x5165;&#x4FB5;&#x5BF9;&#x963F;&#x7C73;&#x5DF4;&#x7EC4;&#x7EC7;&#x5185;&#x963F;&#x7C73;&#x5DF4;&#x8F6C;&#x5F55;&#x8FC7;&#x7A0B;&#x7684;&#x5F71;&#x54CD;&#x3002;

A genome-wide transcriptional analysis of Entamoeba histolytica was performed on trophozoites isolated from the colon of six infected mice and from in vitro culture. An Affymetrix platform gene expression array was designed for this analysis that included probe sets for 9435 open reading frames (ORFs) and 9066 5' and 3' flanking regions. Transcripts were detected for > 80% of all ORFs. A total of 523 transcripts (5.2% of all E. histolytica genes) were significantly changed in amebae isolated from the intestine on Days 1 and 29 after infection: 326 and 109 solely on Days 1 and 29, and 88 on both days. Quantitative real-time reverse transcriptase PCR confirmed these changes in 11/12 genes tested using mRNA isolated from an additional six mice. Adaptation to the intestinal environment was accompanied by increases in a subset of cell signaling genes including transmembrane kinases, ras and rho family GTPases, and calcium binding proteins. Significant decreases in mRNA abundance for genes involved in glycolysis and concomitant increases in lipases were consistent with a change in energy metabolism. Defense against bacteria present in the intestine (but lacking from in vitro culture) was suggested by alterations in mRNA levels of genes similar to the AIG1 plant antibacterial proteins. Decreases in oxygen detoxification pathways were observed as expected in the anaerobic colonic lumen. Of the known virulence factors the most remarkable changes were a 20-35-fold increase in a cysteine proteinase four-like gene, and a 2-3-fold decrease in two members of the Gal/GalNAc lectin light subunit family. Control of the observed changes in mRNA abundance in the intestine might potentially rest with four related proteins with DNA binding domains that were down-regulated 6-16-fold in the intestinal environment. In conclusion, the first genome-wide analysis of the transcriptome of E. histolytica demonstrated that the vast majority of genes are transcribed in trophozoites, and that in the host intestine trophozoites altered the expression of mRNAs for genes implicated in metabolism, oxygen defense, cell signaling, virulence, antibacterial activity, and DNA binding.

Impact of intestinal colonization and invasion on the Entamoeba histolytica transcriptome.

Une analyse transcriptionnelle de G&eacute;nome-large d&#39;Entamoeba histolytica a &eacute;t&eacute; r&eacute;alis&eacute;e sur les trophozo&iuml;tes isol&eacute;es du colon de six souris infect&eacute;es et de culture in vitro. Un alignement tableau plate-forme d&#39;expression des g&egrave;nes a &eacute;t&eacute; con&ccedil;u pour cette analyse qui comprenait la sonde d&eacute;finit pour 9435 open reading cadres (ORF) et 9066 5&#39; et 3&#39; des r&eacute;gions flanquantes. Relev&eacute;s de notes ont &eacute;t&eacute; d&eacute;tect&eacute;es pour &gt; 80 % de tous les ORF. Un total de 523 transcriptions (5,2 % de tous les g&egrave;nes d&#39;Entamoeba histolytica) ont &eacute;t&eacute; significativement modifi&eacute;es dans l&#39;amibiase isol&eacute;e de l&#39;intestin les jours 1 et 29 apr&egrave;s l&#39;infection : 326 et 109 uniquement sur des jours 1 et 29 et 88 sur les deux jours. Quantitative en temps r&eacute;el reverse transcriptase PCR a confirm&eacute; ces changements en 11/12 g&egrave;nes test&eacute;s &agrave; l&#39;aide de mRNA isol&eacute; d&#39;une souris six suppl&eacute;mentaires. Adaptation &agrave; l&#39;environnement intestinal a &eacute;t&eacute; accompagn&eacute;e d&#39;augmentations dans un sous-ensemble de g&egrave;nes, y compris les kinases transmembranaires, ras et famille rho GTPases et prot&eacute;ines liant le calcium de la signalisation cellulaire. Des diminutions significatives en abondance des ARNm de g&egrave;nes impliqu&eacute;s dans la glycolyse et augmentation concomitante des lipases &eacute;tait conformes &agrave; un changement dans le m&eacute;tabolisme &eacute;nerg&eacute;tique. D&eacute;fense contre les bact&eacute;ries pr&eacute;sentes dans l&#39;intestin (mais il manque de culture in vitro) a &eacute;t&eacute; sugg&eacute;r&eacute;e par les modifications des niveaux d&#39;ARNm de g&egrave;nes semblables aux prot&eacute;ines antibact&eacute;riennes AIG1 plante. Diminutions dans les voies de d&eacute;toxication de l&#39;oxyg&egrave;ne ont &eacute;t&eacute; observ&eacute;es comme pr&eacute;vu dans la lumi&egrave;re colique ana&eacute;robie. Les facteurs de virulence connus des changements les plus remarquables ont &eacute;t&eacute; une augmentation de 20 &agrave; 35 fois dans un g&egrave;ne de quatre type de cyst&eacute;ine prot&eacute;inase et une diminution de 2-3 fois dans les deux membres de la famille de lumi&egrave;re sous-unit&eacute; Gal/GalNAc lectine. Contr&ocirc;le des changements observ&eacute;s dans l&#39;abondance des ARNm dans l&#39;intestin pourrait potentiellement &ecirc;tre quatre prot&eacute;ines apparent&eacute;es avec les domaines de liaison de l&#39;ADN qui ont &eacute;t&eacute; diminu&eacute;es 6-16-pli dans le milieu intestinal. En conclusion, la premi&egrave;re analyse du transcriptome d&#39;e. histolytica genome-wide ont montr&eacute; que la grande majorit&eacute; des g&egrave;nes est transcrits en trophozo&iuml;tes, et que chez l&#39;h&ocirc;te trophozo&iuml;tes intestins modifiait l&#39;expression des ARNm de g&egrave;nes impliqu&eacute;s dans le m&eacute;tabolisme, d&eacute;fense de l&#39;oxyg&egrave;ne, signalisation cellulaire, virulence, activit&eacute; antibact&eacute;rienne et l&#39;attache d&#39;ADN.

Impact de la colonisation de l&#39;intestin et l&#39;invasion sur le transcriptome de histolytica d&#39;Entamoeba.

Eine genomweiten transkriptionelle Analyse von Entamoeba Histolytica erfolgte auf Trophozoiten isoliert aus dem Doppelpunkt von sechs infizierten M&auml;usen und in-vitro-Kultur. Ein Affymetrix, die Plattform-Gen-Ausdruck-Array f&uuml;r diese Analyse entwickelt wurde, die Sonde enthalten legt f&uuml;r 9435 Open lesen Frames (ORFs) und 9066 5&#39; und 3&#39; angrenzende Regionen. Abschriften wurden f&uuml;r alle ORFs &gt; 80 % erkannt. Insgesamt 523 Transkripte (5,2 % aller Gene in E. Histolytica) wurden in Amebae isoliert aus dem Darm auf Tage 1 und 29 nach Infektion wesentlich ge&auml;ndert: 326 und 109 ausschlie&szlig;lich am Tage 1 und 29 und 88 an beiden Tagen. Quantitative Echtzeit R&uuml;cktranscriptase PCR best&auml;tigt diese Ver&auml;nderungen im 11/12-Gene mit mRNA isoliert von einer zus&auml;tzlichen sechs M&auml;usen getestet. Anpassung an die intestinale Umwelt wurde durch Zuw&auml;chse in eine Teilmenge der Zelle Signalisierung Transmembran-Kinasen, Ras und Rho Familie GTPasen und Kalzium-bindende Proteine Gene begleitet. Erhebliche R&uuml;ckg&auml;nge in H&uuml;lle und F&uuml;lle der mRNA f&uuml;r Gene beteiligt Glykolyse und damit einhergehenden Anstieg der Lipasen entsprachen einer &Auml;nderung im Energiestoffwechsel. Verteidigung gegen Bakterien im Darm (aber fehlt von in-vitro-Kultur) schlug Ver&auml;nderungen in mRNA-Stufen von Genen, die &auml;hnlich wie die AIG1 Pflanze antibakterielle Proteine. Sauerstoff-Entgiftung-Bahnen wurde festgestellt wie erwartet in die anaerobe Kolon Lumen. Von der bekannten Virulenzfaktoren waren bemerkenswertesten &Auml;nderungen ein 20-35-fach in einem Cystein-Proteinase-vier-wie-gen, und 2-3-fach W&auml;hrungsver&auml;nderungen in zwei Mitglieder der Gal/GalNAc Lektin Licht Untereinheit Familie. Kontrolle &uuml;ber die beobachteten Ver&auml;nderungen in mRNA F&uuml;lle im Darm k&ouml;nnte potenziell mit vier Verwandte Proteine mit DNA-Bindung-Domains ruhen, die unten-geregelt 6-16-fach in der Darm-Umgebung waren. Abschlie&szlig;end zeigte die erste genomweiten Analyse das Transkriptom von E. Histolytica, dass die &uuml;berwiegende Mehrheit der Gene in Trophozoiten transkribiert werden, und im Host Darm Trophozoiten mRNAs f&uuml;r Gene, die im Stoffwechsel, Sauerstoff-Verteidigung, Zelle signalisieren, Virulenz, antibakterielle Wirkung und DNA-Bindung impliziert den Ausdruck ge&auml;ndert.

Auswirkungen der intestinale Kolonisation und Invasion auf die Entamoeba Histolytica Transkriptom.

Su trofozoiti isolati dal colon di sei topi infetti e da coltura in vitro &egrave; stata eseguita un&#39;analisi trascrizionale genome-wide di Entamoeba histolytica. Una matrice di espressione genica platform &egrave; stato progettato per questa analisi che includeva la sonda di Affymetrix imposta per 9435 open reading frame (ORF) e 9066 5&#39; e 3&#39; delle regioni fiancheggianti. Le trascrizioni sono state rilevate &amp;gt; 80% del ORFs tutti. Un totale di 523 trascrizioni (5.2% di tutti i geni di E. histolytica) sono state significativamente modificate nel amebae isolato dall&#39;intestino su giorni 1 e 29 dopo l&#39;infezione: 326 e 109 esclusivamente su giorni 1 e 29 e 88 su entrambi i giorni. Quantitativo in tempo reale il transcriptase PCR ha confermato questi cambiamenti nei geni del 11/12 testati usando mRNA isolato da un ulteriore sei topi. Adattamento all&#39;ambiente intestinale &egrave; stato accompagnato da aumenti in un sottogruppo di geni del transmembrane chinasi, ras e famiglia rho GTPases e proteine leganti il calcio di segnalazione cellulare. Significative diminuzioni in abbondanza di mRNA di geni coinvolti nella glicolisi e concomitante aumento della lipasi era coerenti con un cambiamento nel metabolismo energetico. Difesa contro i batteri presenti nell&#39;intestino (ma mancano da coltura in vitro) &egrave; stato suggerito da alterazioni nei livelli di mRNA di geni simili alle proteine antibatteriche AIG1 pianta. Diminuzioni in percorsi di disintossicazione di ossigeno sono state osservate come previsto nel lume del colon anaerobico. I fattori di virulenza noti i cambiamenti pi&ugrave; notevoli sono stati un aumento di 20-35 volte in un gene di quattro-come cisteina proteinasi e una diminuzione di-2-3 volte in due membri della famiglia Gal/GalNAc lectina subunit&agrave; leggera. Controllo dei cambiamenti osservati in abbondanza di mRNA nell&#39;intestino potrebbe potenzialmente riposo con quattro proteine correlate con domini di legame del DNA che erano gi&ugrave;-regolato 6-16 volte nell&#39;ambiente intestinale. In conclusione, la prima analisi genome-wide transcriptome di E. histolytica ha dimostrato che la stragrande maggioranza dei geni &egrave; trascritti in trofozoiti, e che nell&#39;host di trofozoiti intestino alterato l&#39;espressione del mRNA di geni implicati nel metabolismo, difesa di ossigeno, segnalazione delle cellule, virulenza, attivit&agrave; antibatterica e grippaggio del DNA.

Impatto della colonizzazione intestinale e invasione su transcriptome Entamoeba histolytica.

6 &#x611F;&#x67D3;&#x30DE;&#x30A6;&#x30B9;&#x306E;&#x7D50;&#x8178;&#x304A;&#x3088;&#x3073;&#x4F53;&#x5916;&#x57F9;&#x990A;&#x304B;&#x3089;&#x5206;&#x96E2;&#x3055;&#x308C;&#x305F;&#x6804;&#x990A;&#x578B;&#x8D64;&#x75E2;&#x30A2;&#x30E1;&#x30FC;&#x30D0;&#x306E;&#x30B2;&#x30CE;&#x30E0;&#x306E;&#x8EE2;&#x5199;&#x89E3;&#x6790;&#x304C;&#x5B9F;&#x884C;&#x3055;&#x308C;&#x307E;&#x3057;&#x305F;&#x3002;&#x30D7;&#x30E9;&#x30C3;&#x30C8;&#x30D5;&#x30A9;&#x30FC;&#x30E0;&#x907A;&#x4F1D;&#x5B50;&#x767A;&#x73FE;&#x30A2;&#x30EC;&#x30A4; &#x30D7;&#x30ED;&#x30FC;&#x30D6;&#x304C;&#x542B;&#x307E;&#x308C;&#x3066;&#x3053;&#x306E;&#x5206;&#x6790;&#x7528;&#x306B;&#x8A2D;&#x8A08;&#x3055;&#x308C;&#x305F;&#x3001;&#x30A2;&#x30D5;&#x30A3;&#x30E1;&#x30C8;&#x30EA;&#x30AF;&#x30B9; &#x30FB; &#x30D5;&#x30EC;&#x30FC;&#x30E0; (ORFs) &#x3068; 9066 5&#39; 3&#39; &#x4E26;&#x3076;&#x9818;&#x57DF;&#x8AAD;&#x66F8; 9435 &#x30AA;&#x30FC;&#x30D7;&#x30F3;&#x3092;&#x8A2D;&#x5B9A;&#x3057;&#x307E;&#x3059;&#x3002;&#x6210;&#x7E3E;&#x8A3C;&#x660E;&#x66F8;&#x306F;&#x3059;&#x3079;&#x3066; ORFs &#x306E;&#x76EE; 80 &#xFF05; &#x304C;&#x691C;&#x51FA;&#x3055;&#x308C;&#x307E;&#x3057;&#x305F;&#x3002;523 &#x306E;&#x6210;&#x7E3E;&#x8A3C;&#x660E;&#x66F8; &#xFF08;&#x3059;&#x3079;&#x3066;&#x306E; E. &#x30A2;&#x30E1;&#x30FC;&#x30D0;&#x306E;&#x907A;&#x4F1D;&#x5B50;&#x306E; 5.2 &#xFF05;&#xFF09; &#x306E;&#x5408;&#x8A08;&#x611F;&#x67D3;&#x5F8C;&#x65E5; 1 &#x3068; 29 &#x306E;&#x8178;&#x7BA1;&#x304B;&#x3089;&#x5206;&#x96E2;&#x3055;&#x308C;&#x305F; amebae &#x3067;&#x5927;&#x5E45;&#x306B;&#x5909;&#x66F4;&#x3055;&#x308C;&#x305F;: 326 &#x3068;&#x3060;&#x3051;&#x3067; 1 &#x65E5;&#x3068; 29 &#x65E5; 109 88 &#x4E21;&#x65B9;&#x306E;&#x65E5;&#x3002;&#x30EA;&#x30A2;&#x30EB;&#x30BF;&#x30A4;&#x30E0;&#x306E;&#x5B9A;&#x91CF;&#x7684;&#x9006;&#x8EE2;&#x5199; PCR 11/12 &#x907A;&#x4F1D;&#x5B50;&#x306E; mRNA &#x3092;&#x8FFD;&#x52A0; 6 &#x30DE;&#x30A6;&#x30B9;&#x304B;&#x3089;&#x5206;&#x96E2;&#x3092;&#x4F7F;&#x7528;&#x3057;&#x3066;&#x30C6;&#x30B9;&#x30C8;&#x3067;&#x3053;&#x308C;&#x3089;&#x306E;&#x5909;&#x66F4;&#x3092;&#x78BA;&#x8A8D;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x8178;&#x5185;&#x306E;&#x74B0;&#x5883;&#x3078;&#x306E;&#x9069;&#x5FDC;&#x306F;&#x30AB;&#x30EB;&#x30B7;&#x30A6;&#x30E0;&#x7D50;&#x5408;&#x86CB;&#x767D;&#x8CEA;&#x306A;&#x3069; rho &#x30D5;&#x30A1;&#x30DF;&#x30EA;&#x30FC;&#x8F38;&#x9001;&#x3001;ras &#x3068;&#x819C;&#x8CAB;&#x901A;&#x30AD;&#x30CA;&#x30FC;&#x30BC;&#x907A;&#x4F1D;&#x5B50;&#x7D30;&#x80DE;&#x306E;&#x30B5;&#x30D6;&#x30BB;&#x30C3;&#x30C8;&#x306E;&#x5897;&#x52A0;&#x306B;&#x4F34;&#x308F;&#x308C;&#x307E;&#x3057;&#x305F;&#x3002;&#x6709;&#x610F;&#x306A;&#x6E1B;&#x5C11;&#x3092;&#x907A;&#x4F1D;&#x5B50;&#x306E; mRNA &#x8C4A;&#x5BCC;&#x89E3;&#x7CD6;&#x7CFB;&#x306E;&#x95A2;&#x4E0E;&#x3057;&#x3001;&#x30EA;&#x30D1;&#x30FC;&#x30BC;&#x306E;&#x5897;&#x5927;&#x3059;&#x308B;&#x30A8;&#x30CD;&#x30EB;&#x30AE;&#x30FC;&#x4EE3;&#x8B1D;&#x306E;&#x5909;&#x5316;&#x3068;&#x4E00;&#x81F4;&#x3057;&#x3066;&#x3044;&#x305F;&#x3002;&#x7D30;&#x83CC;&#x8178; (&#x3057;&#x304B;&#x3057;&#x4F53;&#x5916;&#x57F9;&#x990A;&#x304B;&#x3089;&#x6B20;&#x3051;&#x3066;&#x3044;&#x308B;) &#x306E;&#x5B58;&#x5728;&#x306B;&#x5BFE;&#x3059;&#x308B;&#x9632;&#x5FA1;&#x306F;&#x3001;aig 1 &#x690D;&#x7269;&#x306E;&#x6297;&#x83CC;&#x30BF;&#x30F3;&#x30D1;&#x30AF;&#x8CEA;&#x306B;&#x985E;&#x4F3C;&#x907A;&#x4F1D;&#x5B50;&#x306E; mRNA &#x306E;&#x30EC;&#x30D9;&#x30EB;&#x306E;&#x5909;&#x5316;&#x306B;&#x3088;&#x3063;&#x3066;&#x793A;&#x5506;&#x3055;&#x308C;&#x307E;&#x3057;&#x305F;&#x3002;&#x9178;&#x7D20;&#x89E3;&#x6BD2;&#x7D4C;&#x8DEF;&#x306E;&#x6E1B;&#x5C11;&#x3067;&#x5ACC;&#x6C17;&#x6027;&#x5927;&#x8178;&#x5185;&#x8154;&#x304C;&#x671F;&#x5F85;&#x3069;&#x304A;&#x308A;&#x306B;&#x89B3;&#x5BDF;&#x3055;&#x308C;&#x307E;&#x3057;&#x305F;&#x3002;&#x65E2;&#x77E5;&#x306E;&#x75C5;&#x539F;&#x56E0;&#x5B50;&#x306E;&#x6700;&#x3082;&#x9855;&#x8457;&#x306A;&#x5909;&#x5316; 20 35 &#x306E;&#x500D;&#x306F;&#x30B7;&#x30B9;&#x30C6;&#x30A4;&#x30F3;&#x306E;&#x30D7;&#x30ED;&#x30C6;&#x30A3;&#x30CA;&#x30FC;&#x30BC; 4 &#x306E;&#x3088;&#x3046;&#x306A;&#x907A;&#x4F1D;&#x5B50;&#x306E;&#x3068;&#x30AE;&#x30E3;&#x30EB;/GalNAc &#x30EC;&#x30AF;&#x30C1;&#x30F3;&#x5149;&#x30B5;&#x30D6;&#x30E6;&#x30CB;&#x30C3;&#x30C8;&#x306E;&#x5BB6;&#x65CF;&#x306E;&#x30E1;&#x30F3;&#x30D0;&#x30FC; 2 &#x4EBA;&#x306E; 2-3 &#x500D;&#x6E1B;&#x5C11;&#x3059;&#x308B;.MRNA &#x8C4A;&#x5BCC;&#x8178;&#x3067;&#x89B3;&#x5BDF;&#x3055;&#x308C;&#x305F;&#x5909;&#x5316;&#x306E;&#x5236;&#x5FA1;&#x306F;&#x3001;&#x6F5C;&#x5728;&#x7684;&#x30C0;&#x30A6;&#x30F3;&#x898F;&#x5236; 6-16-&#x500D;&#x8178;&#x5185;&#x74B0;&#x5883;&#x306B;&#x3057;&#x305F; DNA &#x7D50;&#x5408;&#x30C9;&#x30E1;&#x30A4;&#x30F3;&#x3092;&#x6301;&#x3064; 4 &#x3064;&#x306E;&#x95A2;&#x9023;&#x86CB;&#x767D;&#x8CEA;&#x3068;&#x6B8B;&#x308A;&#x53EF;&#x80FD;&#x6027;&#x304C;&#x3042;&#x308A;&#x307E;&#x3059;&#x3002;&#x7D50;&#x8AD6;&#x3068;&#x3057;&#x3066;&#x3001;E. &#x30A2;&#x30E1;&#x30FC;&#x30D0; &#x30C8;&#x30E9;&#x30F3;&#x30B9;&#x30AF;&#x30EA;&#x30D7;&#x30C8;&#x30FC;&#x30E0;&#x306E;&#x6700;&#x521D;&#x306E;&#x30B2;&#x30CE;&#x30E0;&#x89E3;&#x6790;&#x907A;&#x4F1D;&#x5B50;&#x306E;&#x5927;&#x534A;&#x304C;&#x6804;&#x990A;&#x578B;&#x5185;&#x8EE2;&#x5199;&#x3055;&#x308C;&#x3001;&#x30DB;&#x30B9;&#x30C8;&#x3067;&#x8178;&#x6804;&#x990A;&#x4EE3;&#x8B1D;&#x3001;&#x9178;&#x7D20;&#x9632;&#x885B;&#x3001;&#x30BB;&#x30EB;&#x30B7;&#x30B0;&#x30CA;&#x3001;&#x75C5;&#x539F;&#x3001;&#x6297;&#x83CC;&#x4F5C;&#x7528;&#x3001;DNA &#x7D50;&#x5408;&#x306B;&#x95A2;&#x4E0E;&#x3059;&#x308B;&#x907A;&#x4F1D;&#x5B50;&#x306E; Mrna &#x306E;&#x767A;&#x73FE;&#x3092;&#x5909;&#x66F4;&#x3053;&#x3068;&#x3092;&#x793A;&#x3057;&#x305F;&#x3002;

&#x8178;&#x7BA1;&#x5B9A;&#x7740;&#x3068;&#x8D64;&#x75E2;&#x30A2;&#x30E1;&#x30FC;&#x30D0; &#x30C8;&#x30E9;&#x30F3;&#x30B9;&#x30AF;&#x30EA;&#x30D7;&#x30C8;&#x30FC;&#x30E0;&#x306E;&#x4FB5;&#x5165;&#x306E;&#x5F71;&#x97FF;&#x3002;

Entamoeba histolytica&#xC758; &#xAC8C;&#xB188; &#xB113;&#xC740; transcriptional &#xBD84;&#xC11D; 6 &#xAC10;&#xC5FC; &#xB41C; &#xC950;&#xC758; &#xCF5C;&#xB860; &#xBC0F; &#xCCB4; &#xC678; &#xBB38;&#xD654;&#xC5D0;&#xC11C; &#xBD84;&#xB9AC; &#xB41C; trophozoites&#xC5D0;&#xC11C; &#xC218;&#xD589; &#xB418;&#xC5C8;&#xB2E4;. &#xD50C;&#xB7AB;&#xD3FC; &#xC9C4; &#xC2DD; &#xBC30;&#xC5F4; &#xD504;&#xB85C;&#xBE0C;&#xB97C; &#xD3EC;&#xD568; &#xD558;&#xB294;&#xC774; &#xBD84;&#xC11D;&#xC744; &#xC704;&#xD574; &#xC124;&#xACC4; &#xB418;&#xC5C8;&#xC2B5;&#xB2C8;&#xB2E4;&#xB294; Affymetrix 9435 &#xC624;&#xD508; &#xB3C5;&#xC11C; &#xD504;&#xB808;&#xC784; (ORFs) &#xBC0F; 9066 5&#39;&#xACFC; 3&#39; &#xC601;&#xC5ED; &#xCE21;&#xBA74;&#xC5D0; &#xB300; &#xD55C; &#xC124;&#xC815; &#xD569;&#xB2C8;&#xB2E4;. &#xC131;&#xC801; &#xC99D;&#xBA85;&#xC11C; &#xBAA8;&#xB4E0; Orfs&#xC758; &amp;gt; 80% &#xAC80;&#xCD9C; &#xB410;&#xB2E4;. &#xCD1D; 523 &#xC131;&#xC801;&#xD45C; (&#xBAA8;&#xB4E0; E. histolytica &#xC720;&#xC804;&#xC790;&#xC758; 5.2%)&#xC758; &#xAC10;&#xC5FC; &#xD6C4; &#xC77C; 1&#xACFC; 29&#xC5D0; &#xCC3D; &#xC790;&#xC5D0;&#xC11C; &#xBD84;&#xB9AC; &#xB41C; amebae&#xC5D0;&#xC11C; &#xD06C;&#xAC8C; &#xBC14;&#xB00C;&#xC5C8;&#xB2E4;: 326 &#xACE0; 109 &#xC77C; 1&#xACFC; 29 &#xC77C;&#xC5D0; &#xC720;&#xC77C; &#xD558; &#xAC8C; &#xB458; &#xB2E4; &#xC77C; 88. &#xC815;&#xB7C9; &#xC2E4;&#xC2DC;&#xAC04; &#xC5ED;&#xC804;&#xC0AC; PCR 11/12 &#xC720;&#xC804;&#xC790; mRNA &#xCD94;&#xAC00; 6 &#xC950;&#xC5D0;&#xC11C; &#xACA9;&#xB9AC;&#xB97C; &#xC0AC;&#xC6A9; &#xD558; &#xC5EC; &#xD14C;&#xC2A4;&#xD2B8;&#xC5D0;&#xC11C; &#xC774;&#xB7EC;&#xD55C; &#xBCC0;&#xD654;&#xB97C; &#xD655;&#xC778; &#xD588;&#xB2E4;. &#xC7A5; &#xD658;&#xACBD;&#xC5D0; &#xC801;&#xC751; &#xC99D;&#xAC00; &#xB9C9; &#xD6A1;&#xB2E8; kinases, ras &#xBC0F; rho &#xAC00;&#xC871; GTPases, &#xCE7C;&#xC298; &#xC758;&#xBB34;&#xC801;&#xC778; &#xB2E8;&#xBC31;&#xC9C8; &#xB4F1; &#xC720;&#xC804;&#xC790; &#xC2E0;&#xD638; &#xC140;&#xC758; &#xD558;&#xC704; &#xC9D1;&#xD569;&#xC5D0; &#xC758;&#xD574; &#xB3D9;&#xD589; &#xD588;&#xB2E4;. &#xD574;&#xB2F9; &#xBD84;&#xD574;&#xC5D0; &#xAD00;&#xC5EC; &#xD558;&#xB294; &#xC720;&#xC804;&#xC790;&#xC758; mRNA &#xD48D;&#xBD80;&#xC5D0; &#xC788;&#xB294; &#xC0C1;&#xB2F9;&#xD55C; &#xAC10;&#xC18C;&#xC640; lipases&#xC5D0; &#xC788;&#xB294; &#xC218; &#xBC18;&#xD558;&#xB294; &#xC99D;&#xAC00; &#xB41C; &#xC5D0;&#xB108;&#xC9C0; &#xBB3C;&#xC9C8; &#xB300;&#xC0AC;&#xC5D0; &#xBCC0;&#xD654;&#xB97C; &#xC77C;&#xCE58; &#xD569;&#xB2C8;&#xB2E4;. &#xBC15;&#xD14C;&#xB9AC;&#xC544; &#xC18C;&#xC7A5; (&#xD558;&#xC9C0;&#xB9CC; &#xCCB4; &#xC678; &#xBB38;&#xD654;&#xC5D0;&#xC11C; &#xBD80;&#xC871;)&#xC5D0; &#xB300; &#xD55C; &#xBC29;&#xC5B4;&#xB294; &#xBE44;&#xC2B7;&#xD55C; AIG1 &#xC2DD;&#xBB3C; &#xD56D;&#xADE0; &#xB2E8;&#xBC31;&#xC9C8; &#xC720;&#xC804;&#xC790;&#xC758; mRNA &#xC218;&#xC900;&#xC5D0;&#xC11C; &#xBCC0;&#xACBD; &#xD558; &#xC5EC; &#xC81C;&#xC548; &#xD588;&#xB2E4;. &#xD610; &#xAE30;&#xC131; &#xC800; &#xC2B9; &#xB8E8;&#xBA58;&#xC5D0;&#xC11C; &#xC608;&#xC0C1; &#xB300;&#xB85C; &#xAC10;&#xC18C; &#xC0B0;&#xC18C; &#xD574;&#xB3C5; &#xACBD;&#xB85C;&#xC5D0;&#xC11C; &#xAD00;&#xCC30; &#xD588;&#xB2E4;. &#xC54C;&#xB824;&#xC9C4;&#xB41C; &#xB3C5;&#xC131; &#xC694;&#xC778;&#xC758; &#xAC00;&#xC7A5; &#xB180;&#xB77C;&#xC6B4; &#xBCC0;&#xD654;&#xB294; cysteine &#xAC00;&#xC218;&#xBD84;&#xD574; 4 &#xAC19;&#xC740; &#xC720;&#xC804;&#xC790; 20 35 &#xBC30; &#xC99D;&#xAC00; Gal/GalNAc lectin &#xBE5B; &#xC18C; &#xB2E8;&#xC704; &#xAC00;&#xC871;&#xC758; &#xB450; &#xBA64;&#xBC84;&#xC5D0; 2-3 &#xBC30; &#xAC10;&#xC18C; &#xD588;&#xB2E4;. MRNA &#xD48D;&#xBD80; &#xD558; &#xAC8C; &#xC18C;&#xC7A5;&#xC5D0;&#xC11C;&#xC5D0;&#xC11C; &#xAD00;&#xCC30; &#xB41C; &#xBCC0;&#xD654;&#xB97C; &#xC81C;&#xC5B4; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4; &#xC7A0;&#xC7AC;&#xC801;&#xC73C;&#xB85C; DNA &#xBC14;&#xC778;&#xB529; &#xB3C4;&#xBA54;&#xC778; &#xC544;&#xB798;&#xB85C; 6-16-&#xBC30; &#xC7A5; &#xD658;&#xACBD;&#xC5D0; &#xC788;&#xB358; 4 &#xAC1C;&#xC758; &#xAD00;&#xB828;&#xB41C; &#xB2E8;&#xBC31;&#xC9C8;&#xC744; &#xD568;&#xAED8; &#xD734;&#xC2DD;&#xC744;. &#xB05D;&#xC73C;&#xB85C;, E. histolytica transcriptome &#xCD5C;&#xCD08;&#xC758; &#xAC8C;&#xB188; &#xB113;&#xC740; &#xBD84;&#xC11D; &#xC2DC;&#xC5F0; &#xC720;&#xC804;&#xC790;&#xC758; &#xB300;&#xB2E4;&#xC218;&#xB294; trophozoites&#xC5D0;&#xC11C; &#xBCF5;&#xC0AC;&#xD560; &#xC218; &#xBC0F; &#xD638;&#xC2A4;&#xD2B8; &#xC18C;&#xC7A5; trophozoites &#xC2E0;&#xC9C4; &#xB300;&#xC0AC;, &#xC0B0;&#xC18C; &#xBC29;&#xC5B4;, &#xC138;&#xD3EC; &#xC2E0;&#xD638;, &#xB3C5;&#xC131;, &#xD56D;&#xADE0; &#xD65C;&#xB3D9; &#xBC0F; DNA &#xBC14;&#xC778;&#xB529;&#xC5D0;&#xC11C; &#xB0B4;&#xD3EC; &#xB41C; &#xC720;&#xC804;&#xC790;&#xC5D0; &#xB300; &#xD55C; Mrnas&#xC758; &#xC2DD;&#xC744; &#xBCC0;&#xACBD; &#xD569;&#xB2C8;&#xB2E4;.

&#xC7A5; &#xC2DD;&#xBBFC;&#xC9C0;&#xC640; Entamoeba histolytica transcriptome&#xC5D0; &#xCE68;&#xB7B5;&#xC758; &#xC601;&#xD5A5;.

Se realiz&oacute; un an&aacute;lisis transcripcional de genoma de Entamoeba histolytica en trofozoitos aislados de los dos puntos de seis ratones infectados y de cultivo in vitro. Una matriz de expresi&oacute;n de genes de plataforma fue dise&ntilde;ada para este an&aacute;lisis que incluy&oacute; sonda de Affymetrix establece 9435 abierto leyendo Marcos (ORFs) y 9066 5&#39; y 3&#39; que flanquean las regiones. Las transcripciones fueron detectadas &gt; 80% de las ORFs todos. Un total de 523 transcripciones (5,2% de todos los genes de E. histolytica) cambiaron significativamente en amebas aislado del intestino en los d&iacute;as 1 y 29 despu&eacute;s de la infecci&oacute;n: 326 y 109 &uacute;nicamente en los d&iacute;as 1 y 29 y 88 en ambos d&iacute;as. La transcriptasa inversa en tiempo real cuantitativa PCR confirm&oacute; estos cambios en 11/12 genes analizados utilizando mRNA aislado de un adicional seis ratones. Adaptaci&oacute;n al ambiente intestinal fue acompa&ntilde;ada por aumentos en un subconjunto de genes incluyendo transmembrana quinasas, ras y familia rho GTPasas y prote&iacute;nas de uni&oacute;n de calcio de la se&ntilde;alizaci&oacute;n de la c&eacute;lula. Disminuciones significativas en la abundancia de mRNA de genes implicados en la glicolisis y aumento concomitante de lipasas era constante con un cambio en el metabolismo energ&eacute;tico. Defensa contra las bacterias presentes en el intestino (pero que carecen de cultivo in vitro) fue sugerido por alteraciones en los niveles de mRNA de genes similares a las prote&iacute;nas antibacterianas de la planta AIG1. Se observaron disminuciones en las v&iacute;as de desintoxicaci&oacute;n de ox&iacute;geno como se esperaba en el lumen col&oacute;nico anaerobio. De los factores de virulencia conocidos los cambios m&aacute;s notables fueron un aumento de 20-35 veces en un gen de cuatro-como ciste&iacute;na proteinasa y una disminuci&oacute;n de 2-3 veces en dos miembros de la familia de luz de la subunidad de lectinas Gal/GalNAc. Control de los cambios observados en la abundancia de mRNA en el intestino puede estar potencialmente con cuatro prote&iacute;nas relacionadas con dominios de uni&oacute;n a ADN que regula 6-16-doblez en el ambiente intestinal. En conclusi&oacute;n, el primer an&aacute;lisis del genoma del transcriptoma de E. histolytica demostr&oacute; que la gran mayor&iacute;a de los genes se transcribe en trofozoitos, y que en la hostia trofozoitos del intestino alteraci&oacute;n la expresi&oacute;n de los mRNAs de genes implicados en el metabolismo, defensa de ox&iacute;geno, se&ntilde;alizaci&oacute;n celular, virulencia, actividad antibacteriana y enlace de ADN.

Impacto de la colonizaci&oacute;n intestinal e invasi&oacute;n en el transcriptoma de Entamoeba histolytica.

Resistance to intestinal amoebiasis is mouse strain dependent. C57BL/6 (B6) mice clear Entamoeba histolytica within hours of challenge, whereas C3H and CBA strains are susceptible to infection and disease. In this study, we show using bone marrow (BM) chimeric mice that mouse strain-dependent resistance is mediated by nonhemopoietic cells; specifically, B6 BM --> CBA recipients remained susceptible as measured by amoeba score and culture, whereas CBA BM --> B6 recipients remained resistant. Interestingly, hemopoietic IL-10 was required for maintaining the resistance of B6 mice, in that B6 IL-10-deficient mice and IL-10(-/-) BM --> wild-type recipients, but not IL-10(+/+) BM --> IL-10(-/-) recipients, exhibited higher amoeba scores than their wild-type controls. Additionally, C57BL/10 IL-10(-/-)Rag2(-/-) mice exhibited diminished amoeba scores and culture rates vs IL-10(-/-) mice, indicating that lymphocytes potentiated the susceptibility of IL-10-deficient mice. We conclude that nonhemopoietic cells mediate the natural resistance to intestinal amoebiasis of B6 mice, yet this resistance depends on hemopoietic IL-10 activity.

Resistance of C57BL/6 mice to amoebiasis is mediated by nonhemopoietic cells but requires hemopoietic IL-10 production.

The advent of anti-tumor necrosis factor (TNF) therapies revolutionized the treatment of inflammatory bowel disease. Adalimumab is a subcutaneous anti-TNF agent indicated for use in patients with moderate-to-severe Crohn's disease and those with moderate-to-severe ulcerative colitis. In both diseases, it can be used for both induction of remission and for maintenance of remission. This review focuses on its use in Crohn's disease as described in the EXTEND (Extend the Safety and Efficacy of Adalimumab through Endoscopic Healing) trial. Several clinical trials using traditional instruments to measure clinical response have had endoscopic substudies looking for endoscopic healing. The EXTEND trial is the first to use mucosal healing on endoscopy as a primary endpoint for patients with moderate-to-severe Crohn's disease and baseline ulcerative disease treated with continuous adalimumab. In this well designed trial, the primary endpoint was narrowly missed, but the secondary endpoints further the notion that mucosal healing should be a more mainstream measure of drug efficacy. How this will translate from clinical trials to the clinic is not yet clear, but identifying noninvasive markers for mucosal healing, and understanding the implications of mucosal healing for safety, resource utilization, and quality of life are all worthy targets for further study. The aim of this review is to understand the role of mucosal healing, safety profile, and efficacy in patients treated with anti-TNF therapy, with particular attention to adalimumab and the EXTEND trial.

Adalimumab treatment in Crohn's disease: an overview of long-term efficacy and safety in light of the EXTEND trial.

The physiological role of the TGR5 receptor in the pancreas is not fully understood. We previously showed that activation of TGR5 in pancreatic β cells by bile acids induces insulin secretion. Glucagon released from pancreatic α cells and glucagon-like peptide 1 (GLP-1) released from intestinal L cells regulate insulin secretion. Both glucagon and GLP-1 are derived from alternate splicing of a common precursor, proglucagon by PC2 and PC1 respectively. We investigated whether TGR5 activation in pancreatic α cells enhances hyperglycemia- induced PC1 expression thereby releasing GLP-1 which, in turn increases β cell mass and function in a paracrine manner. TGR5 activation augmented hyperglycemia-induced switch from glucagon to GLP-1 synthesis in human and mouse islet α cells by Gs/cAMP/PKA/CREB-dependent activation of PC1. Furthermore, TGR5-induced GLP-1 release from α cells was via an Epac- mediated PKA-independent mechanism. Administration of TGR5 agonist, INT-777 to db/db mice attenuated the increase in body weight and improved glucose tolerance and insulin sensitivity. INT-777 augmented PC1 expression in α cells and stimulated GLP-1 release from islets of db/db mice compared to control. INT-777 also increased pancreatic β cell proliferation and insulin synthesis. The effect of TGR5-mediated GLP-1 from α cells on insulin release from islets could be blocked by GLP-1 receptor antagonist. These results suggest that TGR5 activation mediates cross-talk between α and β cells by switching from glucagon to GLP-1 to restore β cell mass and function under hyperglycemic conditions. Thus INT-777 mediated TGR5 activation could be leveraged as a novel way to treat type 2 diabetes mellitus.

Activation of transmembrane bile acid receptor TGR5 modulates pancreatic islet α cells to promote glucose homeostasis.

The lack of a preclinical model of progressive non-alcoholic steatohepatitis (NASH) that recapitulates human disease is a barrier to therapeutic development.

Article	Year
Bile acids: emerging role in management of liver diseases. Hepatology international\| PubMed ID: 26320013	2015
Resistance to intestinal Entamoeba histolytica infection is conferred by innate immunity and Gr-1+ cells. Infection and immunity\| PubMed ID: 16040963	2005
Short report: asymptomatic Cryptosporidium hominis infection among human immunodeficiency virus-infected patients in Tanzania. The American journal of tropical medicine and hygiene\| PubMed ID: 16172475	2005
Impact of intestinal colonization and invasion on the Entamoeba histolytica transcriptome. Molecular and biochemical parasitology\| PubMed ID: 16569449	2006
Resistance of C57BL/6 mice to amoebiasis is mediated by nonhemopoietic cells but requires hemopoietic IL-10 production. Journal of immunology (Baltimore, Md. : 1950)\| PubMed ID: 16818779	2006
Adalimumab treatment in Crohn's disease: an overview of long-term efficacy and safety in light of the EXTEND trial. Clinical and experimental gastroenterology\| PubMed ID: 24039442	2013
Activation of transmembrane bile acid receptor TGR5 modulates pancreatic islet α cells to promote glucose homeostasis. The Journal of biological chemistry\| PubMed ID: 26757816	2016
A diet-induced animal model of non-alcoholic fatty liver disease and hepatocellular cancer. Journal of hepatology\| PubMed ID: 27261415	2016

Amon Asgharpour

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