AMATO, Antonella

BALDASSANO, Sara

CAPPELLO, Francesco

Lo Bello, M

MULE', Flavia

RAPPA, Francesca

Rotondo, A

Archivio istituzionale della ricerca - Università di Palermo

AGAAbstractstract is not known. In this study, we tested the hypothesis that sucrose may increase theexpression of cryptdin-5 and cryptdin-7. Methods: To test this hypothesis, we comparedcryptdin-5 and cryptdin-7 expression in the small intestine (proximal, mid and distal 1/3),colon and liver by real-time PCR. Twenty-four rats were randomized into 1 of 3 treatmentsconsisting of 24h access to regular rat chow (Control), uncooked red kidney bean-supple-mented chow (25%) (RKB) or uncooked red kidney bean-supplemented chow (25%) +sucrose in the drinking water (Sucrose) followed by tissue collection. Data generated byreal-time PCR are represented by 2-ΔΔCt values where Ct represents PCR cycle thresholds(Methods 2001;25:402-408). Lower 2-ΔΔCt values represent greater gene expression. Dataare mean + SD. Results: 1. In the small intestine, compared with Controls (proximal: 6.72+ 2.4, mid: 2.5 + 1.2, distal: 2.6 + 1.3), cryptdin-5 expression was increased in the Sucrosegroup across the entire length of the organ (proximal: 3.7 + 0.8, mid: 1.2 + 0.6, distal: 0.7+ 0.5)(p,0.05); 2. In the liver, compared to Controls (12.6 + 2.0), cryptdin-5 expressionwas increased in Sucrose (10.8 + 0.9)(p,0.05); 3. Cryptdin-5 expression was also greaterin Sucrose than RKB (p,0.05); 4. Similarly, cryptdin-7 expression in the small intestinewas greater in Sucrose when compared with Controls (p ,0.05) but there was only a trendfor this effect in the liver (p= 0.7); 5. In addition, cryptdin-7 expression was greater inSucrose than RKB in the liver (p ,0.05) but not in the small intestine; 6. There was noeffect of sucrose in the colon for either cryptdin-5 or cryptdin-7 expression. Conclusion:Sucrose increased expression of antimicrobial cryptdin-5 and cryptdin-7 in the small intestineand liver. Dr. Lin's research is supported by NIH, VA Research and the Department of Defense.Su1783Role of Endogenous GLP-2 in the Intestinal Adaptation to a Chronic High FatDietSara Baldassano, Francesca Rappa, Alessandra Rotondo, Margherita Lo Bello, FrancescoCappello, Antonella Amato, Flavia Mule'BACKGROUND AND AIM: Glucagon-like peptide-2 (GLP-2) is a nutrient-dependent peptidehormone with multiple effects on the intestine, including expansion of the mucosal surfacearea through stimulation of crypt cell proliferation and enhancement of nutrient digestionand absorption. Effects of GLP-2 occur via the GLP-2 receptor (GLP-2R), a G-protein-coupled receptor, mainly present in the gastrointestinal tract. We sought to determine ifGLP-2 is involved in the regulation of the gut morphological changes following chronichigh fat (HF) diet METHODS: C57BL6/J mice, fed a high fat diet for six weeks, and theage-matched control animals were injected once a day intraperitoneally (i.p.) with 100 μlof GLP-2 (3-33), a GLP-2 receptor antagonist (30 or 60 ng) or PBS (vehicle control) forfour weeks. Plasma GLP-2 concentrations, by ELISA, small intestinal GLP-2R expressionlevels by real-time RT-PCR and western blot analysis, and intestinal morphometric parameterswere determined. Immunohistochemistry for the proliferative and apoptotic markers, respec-tively Ki67 and caspase-3 was performed. RESULTS: Compare with age-matched controlanimals, HF fed mice exhibited increase of crypt-villus height, cell number per villus andKi-67 positive cell number of the crypt region. None difference was seen in the number ofgoblet cells per villus and in the thickness of the external muscular layer. The chronicexposure to HF diet also caused a significant increase in GLP-2 plasma levels and in GLP-2R intestinal expression. Treatment with GLP-2 (3-33) did not significantly modify thecrypt-villus height in control animals. On the contrary, in HF fed mice administration ofGLP-2 (3-33) significantly reduced the crypt-villus height, the number of cells per villus aswell as the number of Ki-67 positive cells, and it increased the percent of caspase-3 positivecells in the small intestine. CONCLUSIONS: This study suggests that GLP-2 may play arole in the murine intestinal adaptation to HF diet, leading to enhancing energy intake.Prolonged HF diet could induce dysregulation of GLP-2/GLP-2R system. Supported byMIUR, ItalySu1784Enteral Deprivation Leads to a Loss of Intestinal Epithelial Barrier FunctionMatthew W. Ralls, Daniel H. Teitelbaum, Eiichi A. Miyasaka, Kathy M. IgnatoskiObjective: Total parenteral nutrition (TPN) increases risk of infections versus enteral feeding.As a major source of these infections is the gastrointestinal tract, we hypothesized thatunderfed mucosa has decreased epithelial barrier function (EBF), and tested this in a seriesof human resection specimens.. Methods: Small bowel, excluding Crohn's disease, wasobtained from pediatric patients (aged 2 days to 19 years) undergoing intestinal resection.EBF was assessed in Ussing chambers for trans-epithelial resistance(TER) and passage ofFITC-Dextran(4kD). Immunofluorescence staining was used to evaluate intensity and distri-bution of tight junctional (TJ) and adherence junctional (AJ) proteins. Abundance of junc-tional proteins was also measured with Western Immunoblot. Statistical significance for TERused Student T-test. Results: Interestingly, initial examination of specimen(N=22) failed tocorrelate lack of feeding and TER(p=0.09). However, in fed samples, TER increased withpatient age (p,0.01). Therefore results were then stratified by patient's age. Fully fed bowelhad significantly higher TER versus unfed bowel in teenagers(p ,0.05). This trend was alsoobserved in infants(p=0.053). These changes in TER correlated with permeation of FITC-Dextran(p=0.03). We next correlated these findings with Western immunoblot. There wasan increase in protein abundance in unfed patients. To localize these protein, we next lookedat immunofluorescence staining. Dramatic declines in staining of ZO-1, occludin, E-cadherinand Claudin-1 were observed in unfed segments. Conclusion: EBF declined in unfed segmentsof human bowel. This finding may explain increased infectious complications seen in patientsnot receiving enteral feeds.S-476AGA AbstractsSu1785Differences in the Degree of Cerulein Induced Chronic Pancreatitis in C57BL/6 Mouse Substrains Lead to New Insights in Identification of Potential RiskFactors in the Development of Chronic PancreatitisBarbara Ulmasov, Kiyoko Oshima, Michael G. Rodriguez, Roger D. Cox, Brent A.Neuschwander-TetriA frequently used experimental model of chronic pancreatitis (CP) recapitulating humandisease is repeated injection of cerulein to mice. C57BL/6 is the most commonly used inbredstrain for biomedical research, but widespread demand led to the creation of several substrainswith subtly different phenotypes. In this study we found that two common substrains ofC57BL/6, C57BL/6J and C56BL/6NHsd , exhibit different degrees of CP with C57BL/6Jbeing more susceptible to repetitive cerulein-induced CP as assessed by pancreatic atrophy,pancreatic morphological changes and fibrosis. We hypothesized that the deficiency ofnicotinamide nucleotide transhydrogenase (NNT) protein in C57BL/6J was responsible forthe more severe C57BL/6J phenotype but the parameters of CP in NNT expressing transgenicmice created on C57BL6/J background were undistinguished from the wild type C57BL/6J.The highly similar genetic backgrounds but different CP phenotypes of these two substrainspresents a unique opportunity to discover genes important in pathogenesis of CP. To exploitthis opportunity, we conducted whole mouse genome Affymetrix microarray analysis ofpancreatic gene expression of C57BL/6J and C57BL/6NHsd before and after the inductionof CP. Differentially regulated gene expression between the two substrains that might becandidates in CP progression included Mmp7, Pcolce2, Itih4 and Vnt. Additionally, weidentified several genes associated with the development in CP in both substrains includingRIKEN 1810009J06 (trypsinogen5), Ccl8 and Ccl6.Su1786Increased Peripheral Cd19+CD24HiCd38Hi Regulatory B Cells May Be Involvedin the Pathophysiology of Type 1 Autoimmune PancreatitisKimi Sumimoto, Kazushige Uchida, Toshiyuki Mitsuyama, Yuri Fukui, Takeo Kusuda,Hideaki Miyoshi, Norimasa Fukata, Masanori Koyabu, Yutaku Sakaguchi, Tsukasa Ikeura,Masaaki Shimatani, Toshiro Fukui, Mitsunobu Matsushita, Makoto Takaoka, AkiyoshiNishio, Kazuichi OkazakiBackground: Autoimmune pancreatitis (AIP) is a newly recognized pancreatic disorder. In2011, International Consensus Diagnostic Criteria for AIP (ICDC) was published. In thisICDC, AIP was classified into type 1 and type 2. Patients with type 1 AIP have severalimmunologic and histologic abnormalities specific to the disease, including increased levelsof serum immunoglobulin G4 (IgG4) and storiform fibrosis with infiltration of lymphocytesand IgG4-positive plasmacytes in the involved organs. We previously reported that increasednumbers of inducible costimulatory molecule (ICOS) (+) regulatory T cells (Tregs) mightinfluence IgG4 production via interleukin-10 (IL-10) in type 1 AIP. In addition to Tregs,it has been reported that regulatory B cells (Bregs) may also produce IL-10, and be involvedin the development of various autoimmune diseases such as systemic lupus erythematosus(SLE), immune thrombocytopenia (ITP), and sarcoidosis. However, Bregs has not beeninvestigated in type 1 AIP. On the other hand, depletion of B cells by rituximab (RTX) iseffective for treatment of IgG4-related systemic disease such as type 1 AIP. However, inaddition to B cell function deleted by RTX, it is unclear whether Bregs are involved in thedevelopment of type 1 AIP or not. To clarify the role of Bregs in the pathophysiology oftype 1 AIP, we analyzed circulating Bregs in type 1 AIP. Methods: We recruited 21 patientswith type 1 AIP on the basis of ICDC for this study. All patients were untreated withcorticosteroids. For comparison, we also recruited 14 patients with chronic pancreatitis(CP), 18 patients with pancreatic cancer, and 25 healthy subjects as controls. We analyzedBregs as CD19+CD24hiCD38hi and IL10+ B cells from peripheral blood by flow cytometry.Results: In peripheral blood, CD19+CD24hiCD38hi Bregs were significantly increased in type1 AIP patients (6.44% ± 3.19%) compared with CP (4.58% ± 2.16%, P , 0.05), pancreaticcancer (3.79% ± 1.97%, P , 0.01), and healthy control (4.84% ± 2.20%, P , 0.05). IL10+B cells were not significantly different from type 1 AIP and healthy control. In untreatedtype 1 AIP patients, the number of CD19+CD24hiCD38hi Bregs and IgG4 were not correlated(R = 0.334, P 0.14). Conclusions: Our data suggested that CD19+CD24hiCD38hi Bregs maybe involved in the pathophysiology of type 1 AIP via another mechanism rather than IL-10 production.

Role of Endogenous GLP-2 in the Intestinal Adaptation to a Chronic High Fat Diet

https://iris.unipa.it/retrieve/handle/10447/75286/76389/abstract%20DDW%202013.pdf

Role of Endogenous GLP-2 in the Intestinal Adaptation to a Chronic High Fat Diet

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