Pathophysiological features in the brains of female Spontaneously Diabetic Torii (SDT) fatty rats

Diabetes mellitus (DM) and obesity are associated with neurodegenerative diseases such as Alzheimer’s disease and psychiatric disorders such as major depression. In this study, we investigated pathophysiological changes in the brains of female Spontaneously Diabetic Torii (SDT) fatty rats with diabetes and obesity. Brains of Sprague-Dawley (SD), SDT and SDT fatty rats were collected at 58 weeks of age. The parietal cortical thickness was measured and the number of pyramidal cells in the hippocampal cornu ammonis 1 and 3 (CA1 and CA3) and the number of granule cells in the dentate gyrus (DG) regions were counted. The area of glial fibrillary acidic protein (GFAP) positivity in CA1, CA3 and DG regions were measured. The parietal cortical thickness and the number of cells in CA3 and DG regions of SDT and SDT fatty rats did not show obvious changes. On the other hand, in the CA1 region, the number of cells in SDT rats and SDT fatty rats was significantly lower than that in SD rats, and that in SDT fatty rats was significantly lower than that in SDT rats. The GFAP-positive area in SDT fatty rats was significantly reduced compared to that in SD rats only in the DG region. Preliminarily result showed that the expression of S100a9, an inflammation-related gene, was increased in the brains of SDT fatty rats. These results suggest that female SDT fatty rat may exhibit central nervous system diseases due to obesity and DM

Role of methionine adenosyltransferase 2A in bovine preimplantation development and its associated genomic regions

Methionine adenosyltransferase (MAT) is involved in folate-mediated one-carbon metabolism, which is essential for preimplantation embryos in terms of both short-term periconceptional development and long-term phenotypic programming beyond the periconceptional period. Here, our immunofluorescence analysis of bovine oocytes and preimplantation embryos revealed the consistent expression of MAT2A (the catalytic subunit of the ubiquitously expressed-type of MAT isozyme) during this period. Addition of the MAT2A inhibitor FIDAS to the culture media of bovine preimplantation embryos reduced their blastocyst development, revealing the particular importance of MAT2A in successful blastocyst development. Exploration of MAT2A-associated genomic regions in bovine blastocysts using chromatin immunoprecipitation and sequencing (ChIP-seq) identified candidate MAT2A-associated genes implicated not only in short-term periconceptional embryo development, but also in long-term phenotypic programming during this period in terms of growth, metabolism, and immune functions. These results suggest the critical involvement of MAT2A in the periconceptional period in life-long programming of health and disease as well as successful preimplantation development

Effective RNA Regulation by Combination of Multiple Programmable RNA-Binding Proteins

RNAs play important roles in gene expression through translation and RNA splicing. Regulation of specific RNAs is useful to understand and manipulate specific transcripts. Pumilio and fem-3 mRNA-binding factor (PUF) proteins, programmable RNA-binding proteins, are promising tools for regulating specific RNAs by fusing them with various functional domains. The key question is: How can PUF-based molecular tools efficiently regulate RNA functions? Here, we show that the combination of multiple PUF proteins, compared to using a single PUF protein, targeting independent RNA sequences at the 3′ untranslated region (UTR) of a target transcript caused cooperative effects to regulate the function of the target RNA by luciferase reporter assays. It is worth noting that a higher efficacy was achieved with smaller amounts of each PUF expression vector introduced into the cells compared to using a single PUF protein. This strategy not only efficiently regulates target RNA functions but would also be effective in reducing off-target effects due to the low doses of each expression vector

Comparative proteomic analysis of Salmonella enterica serovar Typhimurium ppGpp-deficient mutant to identify a novel virulence protein required for intracellular survival in macrophages

<p>Abstract</p> <p>Background</p> <p>The global ppGpp-mediated stringent response in pathogenic bacteria plays an important role in the pathogenesis of bacterial infections. In <it>Salmonella enterica </it>serovar Typhimurium (<it>S</it>. Typhimurium), several genes, including virulence genes, are regulated by ppGpp when bacteria are under the stringent response. To understand the control of virulence genes by ppGpp in <it>S</it>. Typhimurium, agarose 2-dimensional electrophoresis (2-DE) combined with mass spectrometry was used and a comprehensive 2-DE reference map of amino acid-starved <it>S</it>. Typhimurium strain SH100, a derivative of ATCC 14028, was established.</p> <p>Results</p> <p>Of the 366 examined spots, 269 proteins were successfully identified. The comparative analysis of the wild-type and ppGpp⁰mutant strains revealed 55 proteins, the expression patterns of which were affected by ppGpp. Using a mouse infection model, we further identified a novel virulence-associated factor, STM3169, from the ppGpp-regulated and <it>Salmonella</it>-specific proteins. In addition, <it>Salmonella </it>strains carrying mutations in the gene encoding STM3169 showed growth defects and impaired growth within macrophage-like RAW264.7 cells. Furthermore, we found that expression of <it>stm3169 </it>was controlled by ppGpp and SsrB, a response regulator of the two-component system located on <it>Salmonella </it>pathogenicity island 2.</p> <p>Conclusions</p> <p>A proteomic approach using a 2-DE reference map can prove a powerful tool for analyzing virulence factors and the regulatory network involved in <it>Salmonella </it>pathogenesis. Our results also provide evidence of a global response mediated by ppGpp in <it>S. enterica</it>.</p

地域で生活している５５歳以上の方の慢性疾患，年齢の捉え方，睡眠の質，健康関連QOL，日常生活行動との関係性

Japan’s aging population rate is increasing and healthy life expectancy has decreases by 10 years shorter than average life expectancy. The aim of this study is to determine the relationship among chronic disease, sleep quality, health-related quality of life (HRQOL), and activities of daily living in people over 55 years old who live in the community. Subjects were 161 persons aged 57 to 90 years who were treated with chronic disease in the outpatient department of the A hospital. Exclusion criteria included patients with dementia, cancer and severe heart disease. The survey evaluation questionnaires included the Pittsburgh Sleep Quality Index (PSQI), HRQOL by Short-Form 8 Health Survey (SF-8), and activities of daily living. Variables associated with quality of sleep, HRQOL in univariate analysis with p < 0.05 were entered into multivariate analysis using logistic regression with a stepwise forward selection procedure to determine independent variables and their association with major causes. The logistic regression analysis was done using SPSS software and the post-hoc power of the study was estimated using G*power. The level of significance was set at p < 0.05. The risk factor of poor sleep quality was because of history of cancer [odds ratio (OR): 3.53, 95% confidence interval (CI): 1.06 - 11.77], and insomnia (OR: 3.25, 95% CI: 1.55 - 6.79). The risk factors of poor physical HRQOL were motor disease (OR: 2.62, 95% CI: 1.36 - 5.07), respiratory disease (OR: 3.24, 95% CI: 1.27 - 8.26) and having pain (OR: 11.71, 95% CI: 5.35 - 25.66). In addition, anemia was found to be a risk factor of poor mental HRQOL (OR: 4.87, 95% CI: 1.11 - 21.33). The feeling-for-their-body-age (OR: 0.30, 95% CI: 0.15-0.59) was as “younger than actual age” and advanced the risk factor of poor sleep quality. In addition, feeling-for-their-age (OR: 0.44, 95% CI: 0.21 - 0.92) resulted in reduced risk factor of poor physical HRQOL. The risk factor of poor sleep quality was due to a patient with history of cancer. The factor for good sleep quality and the good factor for physical HRQOL were indications of feeling younger than the actual age

地域で生活している５５歳以上の方の慢性疾患，年齢の捉え方，睡眠の質，健康関連QOL，日常生活行動との関係性

Japan’s aging population rate is increasing and healthy life expectancy has decreases by 10 years shorter than average life expectancy. The aim of this study is to determine the relationship among chronic disease, sleep quality, health-related quality of life (HRQOL), and activities of daily living in people over 55 years old who live in the community. Subjects were 161 persons aged 57 to 90 years who were treated with chronic disease in the outpatient department of the A hospital. Exclusion criteria included patients with dementia, cancer and severe heart disease. The survey evaluation questionnaires included the Pittsburgh Sleep Quality Index (PSQI), HRQOL by Short-Form 8 Health Survey (SF-8), and activities of daily living. Variables associated with quality of sleep, HRQOL in univariate analysis with p < 0.05 were entered into multivariate analysis using logistic regression with a stepwise forward selection procedure to determine independent variables and their association with major causes. The logistic regression analysis was done using SPSS software and the post-hoc power of the study was estimated using G*power. The level of significance was set at p < 0.05. The risk factor of poor sleep quality was because of history of cancer [odds ratio (OR): 3.53, 95% confidence interval (CI): 1.06 - 11.77], and insomnia (OR: 3.25, 95% CI: 1.55 - 6.79). The risk factors of poor physical HRQOL were motor disease (OR: 2.62, 95% CI: 1.36 - 5.07), respiratory disease (OR: 3.24, 95% CI: 1.27 - 8.26) and having pain (OR: 11.71, 95% CI: 5.35 - 25.66). In addition, anemia was found to be a risk factor of poor mental HRQOL (OR: 4.87, 95% CI: 1.11 - 21.33). The feeling-for-their-body-age (OR: 0.30, 95% CI: 0.15-0.59) was as “younger than actual age” and advanced the risk factor of poor sleep quality. In addition, feeling-for-their-age (OR: 0.44, 95% CI: 0.21 - 0.92) resulted in reduced risk factor of poor physical HRQOL. The risk factor of poor sleep quality was due to a patient with history of cancer. The factor for good sleep quality and the good factor for physical HRQOL were indications of feeling younger than the actual age

Functionally confirmed compound heterozygous ADAM17 missense loss-of-function variants cause neonatal inflammatory skin and bowel disease 1

A disintegrin and metalloprotease 17 (ADAM17) is the major sheddase that processes more than 80 substrates, including tumour necrosis factor-α (TNFα). The homozygous genetic deficiency of ADAM17 causing a complete loss of ADAM17 expression was reported to be linked to neonatal inflammatory skin and bowel disease 1 (NISBD1). Here we report for the first time, a family with NISBD1 caused by functionally confirmed compound heterozygous missense variants of ADAM17, namely c.1699T>C (p.Cys567Arg) and c.1799G>A (p.Cys600Tyr). Both variants were detected in two siblings with clinical features of NISBD1, such as erythroderma with exudate in whole body, recurrent skin infection and sepsis and prolonged diarrhoea. In a cell-based assay using Adam10/17 double-knockout mouse embryonic fibroblasts (Adam10/17−/− mEFs) exogenously expressing each of these mutants, phorbol 12-myristate 13-acetate-stimulated shedding was strongly reduced compared with wild-type ADAM17. Thus, in vitro functional assays demonstrated that both missense variants cause the loss-of-function of ADAM17, resulting in the development of NISBD1. Our study further expands the spectrum of genetic pathology underlying ADAM17 in NISBD1 and establishes functional assay systems for its missense variants

Extensive Atrophic Gastritis Increases Intraduodenal Hydrogen Gas

Objective. Gastric acid plays an important part in the prevention of bacterial colonization of the gastrointestinal tract. If these bacteria have an ability of hydrogen (H2) fermentation, intraluminal H2 gas might be detected. We attempted to measure the intraluminal H2 concentrations to determine the bacterial overgrowth in the gastrointestinal tract. Patients and methods. Studies were performed in 647 consecutive patients undergoing upper endoscopy. At the time of endoscopic examination, we intubated the stomach and the descending part of the duodenum without inflation by air, and 20 mL of intraluminal gas samples of both sites was collected through the biopsy channel. Intraluminal H2 concentrations were measured by gas chromatography. Results. Intragastric and intraduodenal H2 gas was detected in 566 (87.5%) and 524 (81.0%) patients, respectively. The mean values of intragastric and intraduodenal H2 gas were 8.5 ± 15.9 and 13.2 ± 58.0 ppm, respectively. The intraduodenal H2 level was increased with the progression of atrophic gastritis, whereas the intragastric H2 level was the highest in patients without atrophic gastritis. Conclusions. The intraduodenal hydrogen levels were increased with the progression of atrophic gastritis. It is likely that the influence of hypochlorhydria on bacterial overgrowth in the proximal small intestine is more pronounced, compared to that in the stomach