Obesity-induced overexpression of miR-802 impairs insulin transcription and secretion.

B cell dysfunction due to obesity can be associated with alterations in the levels of micro-RNAs (miRNAs). However, the role of miRNAs in these processes remains elusive. Here, we show that miR-802 is increased in the pancreatic islets of obese mouse models and demonstrate that inducible transgenic overexpression of miR-802 in mice causes impaired insulin transcription and secretion. We identify Foxo1 as a transcription factor of miR-802 promoting its transcription, and NeuroD1 and Fzd5 as targets of miR-802-dependent silencing. Repression of NeuroD1 in ß cell and primary islets impairs insulin transcription and reduction of Fzd5 in ß cell, which, in turn, impairs Ca2+ signaling, thereby repressing calcium influx and decreasing insulin secretion. We functionally create a novel network between obesity and ß cell dysfunction via miR-802 regulation. Elucidation of the impact of obesity on microRNA expression can broaden our understanding of pathophysiological development of diabetes.

Achieving Stable Partial Nitritation in an Acidic Nitrifying Bioreactor.

Partial nitritation providing a suitable effluent for subsequent anammox is a critical step in a two-stage autotrophic nitrogen removal system. This study demonstrates an innovative approach for attaining partial nitritation in an acidic bioreactor operating at a slightly low pH (i.e., 5-6). This approach is based on our hypothesis in this study that acid-tolerant ammonia-oxidizing bacteria (AOB) can produce nitrite and protons to self-sustain free nitrous acid (FNA, NO2- + H+ ↔ HNO2) at a parts per million level, as an inhibitor of nitrite-oxidizing bacteria (NOB). With influent nitrogen of about 200 mg/L and operating conditions of high dissolved oxygen, long sludge retention time, and moderate temperature, a lab-scale acidic bioreactor with FNA up to 2 mg of HNO2-N/L successfully established stable nitrite accumulation in the effluent for 200 days, with an average ratio [NO2-/(NO2- + NO3-)] exceeding 95%. A 16S rRNA amplicon sequencing analysis showed that Nitrosospira was the dominant AOB in the biomass of the bioreactor, while Nitrosomonas and Nitrospira, two typical nitrifying genera in neutral wastewater treatment, both disappeared after the startup of partial nitritation. Kinetic characterization revealed that Nitrosospira had a substrate affinity of 11.4-16.5 mg of total ammonia (NH4+ + NH3)/L. It also revealed that less than 3.5 mg of HNO2-N/L FNA did not inhibit AOB activity significantly. Acidic operation is economically attractive because it can be achieved via acidophilic ammonia oxidation without adding chemical acid. However, hazardous gas, nitric oxide (NO), should be removed from gas produced by acidic nitrifying bioreactors.

Expansion and Maintenance of CD133-Expressing Pancreatic Ductal Epithelial Cells by Inhibition of TGF-ß Signaling.

Restoring ß-cell mass by the transplantation of pancreatic islets is an effective diabetes treatment, but it is limited by the shortage of donor organs. CD133-expressing pancreatic ductal epithelial cells (PDECs) have the ability to generate insulin-producing cells. The expansion of these cells is dependent on extrinsic niche factors, but few of those signals have been identified. In this study, CD133-expressing PDECs were purified by sorting from adult wild-type C57BL/6 mice and TGFßRIInull/null mice. Furthermore, using immunofluorescence and transplantation assays, we found that the inhibition of the transforming growth factor-ß (TGF-ß) pathway promoted the expansion of CD133-expressing PDECs for many generations and maintained the ability of CD133-expressing PDECs to generate insulin-producing cells. Moreover, western blot, qRT-PCR, and dual luciferase assays using TGF-ß inhibitors were performed to identify the mechanisms by which TGF-ß signaling regulates proliferation and differentiation. The results showed that the inhibition of TGF-ß signaling enhanced Id2 binding to the promoter region of the cell proliferation repressor p16 and promoted the expansion of CD133-expressing PDECs, and the increased Id2 binding to NeuroD1 decreased the transcription of Pax6 to maintain CD133-expressing PDECs in the Pdx1-expression stage. Taken together, the effect of TGF-ß antagonists on CD133-expressing PDECs reveals a novel paradigm of signaling that explains the balance between the expansion and differentiation of pancreatic duct epithelial progenitors.