Type 2 diabetes (T2D) is a complex metabolic syndrome characterized by insulin dysfunction and abnormalities in glucose and lipid metabolism. The gut microbiome has been recently identified as an important factor for development of T2D. In this study, a total of 102 subjects were recruited, and we have looked at the gut microbiota of prediabetics (PreDMs) (n = 17), newly diagnosed diabetics (NewDMs) (n = 11), and diabetics on antidiabetic treatment (KnownDMs) (n = 39) and compared them with healthy nondiabetics (ND) (n = 35). Twenty-five different serum biomarkers were measured to assess the status of diabetes and their association with gut microbiota. Our analysis revealed nine different genera as differentially abundant in four study groups. Among them, Akkermansia, Blautia, and Ruminococcus were found to be significantly (P < 0.05) decreased, while Lactobacillus was increased in NewDMs compared to ND and recovered in KnownDMs. Akkermansia was inversely correlated with HbA1c and positively correlated with total antioxidants. Compared to ND, there was increased abundance of Megasphaera, Escherichia, and Acidaminococcus and decreased abundance of Sutterella in KnownDMs. Among many taxa known to act as community drivers during disease progression, we observed genus Sutterella as a common driver taxon among all diabetic groups. On the basis of the results of random forest analysis, we found that the genera Akkermansia and Sutterella and that the serum metabolites fasting glucose, HbA1c, methionine, and total antioxidants were highly discriminative factors among studied groups. Taken together, our data revealed that gut microbial diversity of NewDMs but not of PreDMs is significantly different from that of ND. Interestingly, after antidiabetic treatment, the microbial diversity of KnownDMs tends to recover toward that of ND. IMPORTANCE Gut microbiota is considered to play a role in disease progression, and previous studies have reported an association of microbiome dysbiosis with T2D. In this study, we have attempted to investigate gut microbiota of ND, PreDMs, NewDMs, and KnownDMs. We found that the genera Akkermansia and Blautia decreased significantly (P < 0.05) in treatment-naive diabetics and were restored in KnownDMs on antidiabetic treatment. To the best of our knowledge, comparative studies on shifts in the microbial community in individuals of different diabetic states are lacking. Understanding the transition of microbiota and its association with serum biomarkers in diabetics with different disease states may pave the way for new therapeutic approaches for T2D.

No abstract

Oct-4 is a POU family transcription factor associated with potentially totipotent cells. Genes expressed in the trophectoderm but not in embryos prior to blastocyst formation may be targets for silencing by Oct-4. Here, we have tested this hypothesis with the tau interferon genes (IFNT genes), which are expressed exclusively in the trophectoderm of bovine embryos. IFNT promoters contain an Ets-2 enhancer, located at ؊79 to ؊70, and are up-regulated about 20-fold by the overexpression of Ets-2 in human JAr choriocarcinoma cells, which are permissive for IFNT expression. This enhancement was reversed in a dose-dependent manner by coexpression of Oct-4 but not either Oct-1 or Oct-2. When cells were transfected with truncated bovine IFNT promoters designed to eliminate potential octamer sites sequentially, luciferase reporter expression from each construct was still silenced by Oct-4. Full repression required both the N-terminal and POU domains of Oct-4, but neither domain used alone was an effective silencer. Oct-4 and Ets-2 formed a complex in vitro in the absence of DNA through binding of the POU domain of Oct-4 to a site located between the "pointed" and DNA binding domains of Ets-2. The two transcription factors were also coimmunoprecipitated after being expressed together in JAr cells. Oct-4, therefore, silences IFNT promoters by quenching Ets-2 transactivation. The POU domain most probably binds to Ets-2 directly, while the N-terminal domain inhibits transcription. These findings provide further evidence that the developmental switch to the trophectoderm is accompanied by the loss of Oct-4 silencing of key genes.

We previously reported that either oxidation or alkylation of NF-cB in vitro abrogates DNA binding. We used this phenomenon to help elucidate structural determinants of NF-KB binding. We now demonstrate that Cys-62 of NF-KB p50 mediates the redox effect and lies within an N-terminal region required for DNA binding but not for dimerization. Several (16,23,33,40). All of these proteins are members of the Rel family of proteins, also denoted the NRD family, for NF-KB/Rel/dorsal. More recently recognized members of this family are RelB/I-Rel (41, 42) and p49 (also denoted p5OB), which is similar to p50 in that both are derived from larger precursor proteins and can interact functionally with p65 (10, 43).In unstimulated cells, NF-KB is present in the cytoplasm in a latent form consisting of p50 (or p49) and p65 and bound via p65 to an inhibitory molecule, IKB (4-6, 18, 22). Cellular activation results in dissociation of NF-KB from IKB and its translocation to the nucleus as an active DNA-binding complex (4, 5). In vitro, p50, p65, and c-Rel can bind DNA as homodimers or as pSO-p65 or p50-c-Rel heterodimers (8,23,33,49 Fig. 1, lane 1), a p50 construct corresponding to amino acids 1 to 399 (HindIII-RsaI fragment) (P501_399) was used. The binding properties of P501-399 and P501-502 are indistinguishable (23; data not shown). p65 constructs were derived from a BamHI fragment spanning the entire human cDNA coding sequence (40). Single point mutation, N-terminal deletion, and pSONTmyc constructs were prepared by site-directed mutagenesis as previously described (47) with a Bio-Rad kit. Following subcloning into pBluescript SK(-) (Stratagene), mutant constructs were resequenced. Chimeras between p50 and p65 were prepared by a two-step polymerase chain reaction method (19), and correctness was confirmed by sequencing the N-terminal regions extending through the point of "fusion" between p50 and p65.In vitro transcription and translation. In vitro transcription was carried out with constructs in which cDNAs were subcloned in pBluescript SK(-) and the mRNA cap kit (Stratagene). p50 and chimeric constructs were linearized with XbaI and transcribed with T7 RNA polymerase. p65 constructs were linearized with HindIII and transcribed with T3 RNA polymerase. Wheat germ extracts were used for the experiments shown in Fig. 1. Reticulocyte lysates were used for the experiments shown in Fig. 3 and 4. The data summarized in Fig. 2

Recent advances in the understanding of eukaryotic gene regulation have produced an extensive body of transcriptionally-related sequence information in the biological literature, and have created a need for computing structures that organize and manage this information. The 'relational model' represents an approach that is finding increasing application in the design of biological databases. This report describes the compilation of information regarding eukaryotic transcription factors, the organization of this information into five tables, the computational applications of the resultant relational database that are of theoretical as well as experimental interest, and possible avenues of further development.