Faculty Bibliography

Background: Colorectal carcinoma (CRC) in patients with longstanding inflammatory bowel disease (IBD) is the culmination of an inflammation-dysplasia-cancer sequence. Its molecular pathogenesis is not fully understood, because in addition to adenomatous dysplasia analogous to conventional sporadic adenomatous lesions, dysplasia in IBD can assume other distinctive phenotypically heterogeneous changes, the clinical characteristics and significance of which are largely unknown. We characterized 2 types of unconventional dysplasias; Goblet Cell Deficient (GCD) and Crypt Cell/Terminally Differentiated (CC/TD) by sequencing DNA for cancerrelated hotspots.
Design(s): Nineteen mucosal samples of dysplasia from biopsies or resection specimens of patients with IBD, eleven ulcerative colitis, six Crohn's colitis, two indeterminate colitis) were evaluated at Mount Sinai Hospital. Fourteen as GCD featured tubular crypts lined by monotonous columnar cells with eosinophilic cytoplasm and mildly atypical nuclei and 5 as CC/TD featured tubular crypts lined by differentiated enterocytes, goblet cells, Paneth cells and/or endocrine cells with cytologically atypical nuclei as previously described (1). The slides were de-identified and the presence of dysplasia confirmed by two independent pathologist. DNA extracted from formalin-fixed, paraffin-embedded dysplastic samples containing at least 10% dysplastic cells and from adjacent non-dysplastic mucosa were analyzed for a 50-gene panel of hotspot mutation sequences on an Ion Torrent PGM sequencer with strict quality assurance (50,000 reads, 200X coverage, AQ20). Data was analyzed on Torrent Suite Analysis Software and annotated with Ion Reporter Software.
Result(s): Seven lesions (37%) harbored mutations that have been implicated in the development of carcinoma (GCD, n=4, CC/TD n=3) per Catalogue of Somatic Mutations in Cancer (COSMIC) database, including known mutations of KRAS, p53, IDH1 and FBXW7, however no correlations were observed between the mutations and dysplasia phenotypes or type of IBD. Two lesions contained variants of unknown significance (VUS) in addition to pathogenic mutations (GCD with IDH1, CC/TD with TP53) and two lesions harbored only VUS. (Table presented)
Conclusion(s): Unconventional dysplastic lesions corresponding to GCD and CC/TD harbor pathogenic mutations that are implicated in the development and progression of sporadic CRC

Introduction: Gut fermentation or auto-brewery syndrome is a relatively uncommon medical condition that presents a diagnostic and therapeutic challenge. Case Report: Here we present a unique case of a 45-yearold obese, male, diabetic patient treated with two courses of antibiotics for deviated nasal septum and dental procedure who reported episodes of diarrhea, vomiting, edema, seizures, hallucinations, intermittent fevers, chills, slurred speech, and loss of consciousness precipitated after meals for a duration of 14 months. The patient denied alcohol consumption at any time. His blood ethanol levels, measured on multiple occasions, were elevated and corroborated by a 24-hour meal test administered in the hospital. Small bowel and fecal contents collected during the endoscopic procedures were remarkable for the growth of Saccharomyces cerevisiae from both samples. Gram stain was negative for Salmonella, Shigella, or Campylobacter. A rapid membrane enzyme immunoassay for C difficile antigen and toxins A and B was also negative. The patient responded well to oral fluconazole but relapsed after a month and needed further assessment. Microbiological studies undertaken on gastric and small bowel contents collected during the follow-up upper and lower endoscopic procedures were positive for Candida intermedia, sensitive to fluconazole (0.250 mcg/ mL) and amphotericin B (0.250 mcg/mL). Bacterial cultures showed rare, positive growth for Klebsiella pneumoniae and Enterococcus faecium. The patient responded dramatically to a no-carbohydrate diet and intravenous administration of an antifungal agent through a peripherally inserted central catheter line. The patient has been asymptomatic ever since.
Summary: Auto-brewery syndrome in a setting of diabetes, obesity, and high carbohydrate intake presented with signs and symptoms of elevated serum ethanol levels. The importance of microbiological studies on carefully collected intestinal secretions is emphasized in this report, in the absence of validated diagnostic and treatment protocols in the literature

OBJECTIVE: This study investigated the utility of advanced computational techniques to large-scale genome-based data to identify novel genes that govern murine pancreatic development. METHODS: An expression data set for mouse pancreatic development was complemented with high-throughput data analyzer to identify and prioritize novel genes. Quantitative real-time polymerase chain reaction, in situ hybridization, and immunohistochemistry were used to validate selected genes. RESULTS: Four new genes whose roles in the development of murine pancreas have not previously been established were identified: cystathionine beta-synthase (Cbs), Meis homeobox 1, growth factor independent 1, and aldehyde dehydrogenase 18 family, member A1. Their temporal expression during development was documented. Cbs was localized in the cytoplasm of the tip cells of the epithelial chords of the undifferentiated progenitor cells at E12.5 and was coexpressed with the pancreatic and duodenal homeobox 1 and pancreas-specific transcription factor, 1a-positive cells. In the adult pancreas, Cbs was localized primarily within the acinar compartment. CONCLUSIONS: In silico analysis of high-throughput microarray data in combination with background knowledge about genes provides an additional reliable method of identifying novel genes. To our knowledge, the expression and localization of Cbs have not been previously documented during mouse pancreatic development.

More than one-half of the ~50 human chemokines have been associated with or implicated in the pathogenesis of type 1 diabetes, yet their actual expression patterns in the islet environment of type 1 diabetic patients remain, at present, poorly defined. Here, we have integrated a human islet culture system, murine models of virus-induced and spontaneous type 1 diabetes, and the histopathological examination of pancreata from diabetic organ donors with the goal of providing a foundation for the informed selection of potential therapeutic targets within the chemokine/receptor family. Chemokine (C-C motif) ligand (CCL) 5 (CCL5), CCL8, CCL22, chemokine (C-X-C motif) ligand (CXCL) 9 (CXCL9), CXCL10, and chemokine (C-X3-C motif) ligand (CX3CL) 1 (CX3CL1) were the major chemokines transcribed (in an inducible nitric oxide synthase-dependent but not nuclear factor-kappaB-dependent fashion) and translated by human islet cells in response to in vitro inflammatory stimuli. CXCL10 was identified as the dominant chemokine expressed in vivo in the islet environment of prediabetic animals and type 1 diabetic patients, whereas CCL5, CCL8, CXCL9, and CX3CL1 proteins were present at lower levels in the islets of both species. Of importance, additional expression of the same chemokines in human acinar tissues emphasizes an underappreciated involvement of the exocrine pancreas in the natural course of type 1 diabetes that will require consideration for additional type 1 diabetes pathogenesis and immune intervention studies.

OBJECTIVE: The SLC30A8 gene encodes the islet-specific transporter ZnT-8, which is hypothesized to provide zinc for insulin-crystal formation. A polymorphic variant in SLC30A8 is associated with altered susceptibility to type 2 diabetes. Several groups have examined the effect of global Slc30a8 gene deletion but the results have been highly variable, perhaps due to the mixed 129SvEv/C57BL/6J genetic background of the mice studied. We therefore sought to remove the conflicting effect of 129SvEv-specific modifier genes. METHODS: The impact of Slc30a8 deletion was examined in the context of the pure C57BL/6J genetic background. RESULTS: Male C57BL/6J Slc30a8 knockout (KO) mice had normal fasting insulin levels and no change in glucose-stimulated insulin secretion (GSIS) from isolated islets in marked contrast to the approximately 50% and approximately 35% decrease, respectively, in both parameters observed in male mixed genetic background Slc30a8 KO mice. This observation suggests that 129SvEv-specific modifier genes modulate the impact of Slc30a8 deletion. In contrast, female C57BL/6J Slc30a8 KO mice had reduced ( approximately 20%) fasting insulin levels, though this was not associated with a change in fasting blood glucose (FBG), or GSIS from isolated islets. This observation indicates that gender also modulates the impact of Slc30a8 deletion, though the physiological explanation as to why impaired insulin secretion is not accompanied by elevated FBG is unclear. Neither male nor female C57BL/6J Slc30a8 KO mice showed impaired glucose tolerance. CONCLUSIONS: Our data suggest that, despite a marked reduction in islet zinc content, the absence of ZnT-8 does not have a substantial impact on mouse physiology.