Sirtuin 6 regulates glucose-stimulated insulin secretion in mouse pancreatic beta cells

AIMS/HYPOTHESIS: Sirtuin 6 (SIRT6) has been implicated in ageing, DNA repair and metabolism; however, its function in pancreatic beta cells is unclear. The aim of this study is to elucidate the role of SIRT6 in pancreatic beta cells. METHODS: To investigate the function of SIRT6 in pancreatic beta cells, we performed Sirt6 gene knockdown in MIN6 cells and generated pancreatic- and beta cell-specific Sirt6 knockout mice. Islet morphology and glucose-stimulated insulin secretion (GSIS) were analysed. Glycolysis and oxygen consumption rates in SIRT6-deficient beta cells were measured. Cytosolic calcium was monitored using the Fura-2-AM fluorescent probe (Invitrogen, Grand Island, NY, USA). Mitochondria were analysed by immunoblots and electron microscopy. RESULTS: Sirt6 knockdown in MIN6 beta cells led to a significant decrease in GSIS. Pancreatic beta cell Sirt6 knockout mice showed a ~50% decrease in GSIS. The knockout mouse islets had lower ATP levels compared with the wild-type controls. Mitochondrial oxygen consumption rates were significantly decreased in the SIRT6-deficient beta cells. Cytosolic calcium dynamics in response to glucose or potassium chloride were attenuated in the Sirt6 knockout islets. Numbers of damaged mitochondria were increased and mitochondrial complex levels were decreased in the SIRT6-deficient islets. CONCLUSIONS/INTERPRETATION: These data suggest that SIRT6 is important for GSIS from pancreatic beta cells and activation of SIRT6 may be useful to improve insulin secretion in diabetes

Sirt6 Regulates Insulin Secretion from the Pancreatic Beta Cells

poster abstractSirt6 is an NAD-dependent histone deacetylase, which is involved in multiple biological processes, including aging, DNA repair, and metabolism; however, it is unclear what its functions in pancreatic beta-cells are. The beta cells play an essential role in metabolic regulation by secreting insulin in response to an elevated glucose concentration in the circulation. To examine the role of Sirt6 in beta cells, we initially used adenovirus-mediated shRNA to knock down the Sirt6 gene expression in a mouse pancreatic beta cell line - MIN6. Knockdown of the Sirt6 gene significantly reduced glucose-stimulated insulin secretion. To further validate this phenotype in vivo, we generated pancreatic beta-cell-specific Sirt6 knockout mice (bKO) using mouse genetic approach. Indeed, the bKO mice showed remarkable impairment in both first and second phases of insulin secretion in response to a glucose load. While morphometric analyses did not reveal significant difference in islet area between wild-type and bKO mice, biochemical analysis of ATP concentrations showed a 22% decrease in bKO mouse islets relative to control wild-type islets after glucose stimulation. To assess mitochondrial function in Sirt6-deficient beta cells, we also performed Seahorse bioenergetics assays in MIN6 cells after the Sirt6 gene was knocked down. Glucose oxidation in mitochondria was decreased 20-30% in Sirt6- knockdown MIN6 cells as compared to the control cells. Since calcium signaling is critical to insulin secretion, we also measured intracellular calcium concentrations using a fluorescent imaging approach. The results showed a significant decrease in cytoplasmic calcium in the bKO islets as compared to the wild-type controls. Overall, our data demonstrate that Sirt6 plays a critical role in the regulation of pancreatic insulin secretion. This work was supported in part by the NIDDK grant R01DK091592

Neurospora COP9 Signalosome Integrity Plays Major Roles for Hyphal Growth, Conidial Development, and Circadian Function

The COP9 signalosome (CSN) is a highly conserved multifunctional complex that has two major biochemical roles: cleaving NEDD8 from cullin proteins and maintaining the stability of CRL components. We used mutation analysis to confirm that the JAMM domain of the CSN-5 subunit is responsible for NEDD8 cleavage from cullin proteins in Neurospora crassa. Point mutations of key residues in the metal-binding motif (EXnHXHX10D) of the CSN-5 JAMM domain disrupted CSN deneddylation activity without interfering with assembly of the CSN complex or interactions between CSN and cullin proteins. Surprisingly, CSN-5 with a mutated JAMM domain partially rescued the phenotypic defects observed in a csn-5 mutant. We found that, even without its deneddylation activity, the CSN can partially maintain the stability of the SCFFWD-1 complex and partially restore the degradation of the circadian clock protein FREQUENCY (FRQ) in vivo. Furthermore, we showed that CSN containing mutant CSN-5 efficiently prevents degradation of the substrate receptors of CRLs. Finally, we found that deletion of the CAND1 ortholog in N. crassa had little effect on the conidiation circadian rhythm. Our results suggest that CSN integrity plays major roles in hyphal growth, conidial development, and circadian function in N. crassa

Moves recognition in abstract of research paper based on deep learning

The purpose of this work is to explore the applicability and effectiveness of deep learning methods for the task-moves recognition in abstract of research paper. We firstly build a large corpus for moves recognition. Then we choose the traditional machine learning method SVM as a benchmark, and develop four moves recognition methods based on DNN, LSTM, Attention-BiLSTM and BERT. Finally, we design two groups of experiments with sample size 10,000 and 50,000 and then compare experimental results. The results show that most of the deep learning methods outperform the traditional machine learning method SVM especially in large-scale sample experiments, in which the BERT with a re-pre-trained model achieves the best results in both groups of experiments. Deep learning methods are proved applicable and effective for moves recognition in research paper abstracts.</p

Purification and characterization of a new β-glucosidase from Penicillium piceum and its application in enzymatic degradation of delignified corn stover

A new β-glucosidase (Cel3B) was first isolated from cellulytic fungi, designated as PpCel3B. Although PpCel3B was classified to GH family 3 based on the homology sequence, PpCel3B had different biological functions in cellulose degradation and signaling molecules production. PpCel3B was constitutive and could form multiple soluble lignocellulose inducers for cellulase and hemicellulase synthesis via high tranglycosylation activity and new enzymatic activity. Moreover, PpCel3B showed apparent synergism with cellulases by removing several inhibitors. Supplementing low doses of PpCel3B (52 μg/g substrate) increased saccharification efficiency of cellulase produced by Trichoderma reesei and Penicillium piceum by 15% and 35%, respectively on delignified corn stover. PpCel3B had important application in boosting cellulase yield and efficiency

Degradation Behavior in vitro of Carbon Nanotubes (CNTs)/Poly(lactic acid) (PLA) Composite Suture

Poly(lactic acid) (PLA) suture can be absorbed by the human body, and so have wide applications in modern surgery operations. The degradation period of PLA suture is expected to meet with the healing time of different types of wounds. In order to control the degradation period of the PLA suture, the carbon nanotubes (CNTs) were composited with PLA suture, and the degradation experiment in vitro was performed on sutures. The structure and properties of sutures during degradation, such as surface morphology, breaking strength, elongation, mass and chemical structure, were tracked and analyzed. The results indicated that the degradation brought about surface defects and resulted in 13.5 weeks for the strength valid time of the original PLA suture. By contrast, the strength valid time of the CNTs/PLA suture was increased to 26.6 weeks. Whilst the toughness of both the pure PLA and CNTs/PLA sutures decreased rapidly and almost disappeared after 3 to 4 weeks of degradation. The mass loss demonstrated that the time required for complete degradation of the two sutures was obviously different, the pure PLA suture 49 weeks, while CNTs/PLA sutures 63 to 73 weeks. The research proved that CNTs delayed PLA degradation and prolonged its strength valid time in degradation