Microarray analysis of pancreatic gene expression during biotin repletion in biotin-deficient rats

Biotin is a B vitamin involved in multiple metabolic pathways. In humans, biotin deficiency is relatively rare but can cause dermatitis, alopecia and perosis. Low biotin levels occur in type II diabetes, and biotin supplementation with chromium may improve blood sugar control. The acute effect on pancreatic gene expression of biotin repletion following chronic deficiency is unclear, therefore we induced biotin deficiency in adult male rats via a 20% raw egg white diet for 6 weeks. Animals were then randomized: one group received a single biotin supplement and returned to normal chow lacking egg white, while the second group remained on the depletion diet. After one week, pancreata were removed from biotin-deficient (BD) and biotin-repleted (BR) animals and RNA isolated for microarray analysis. Biotin depletion altered gene expression indicative of inflammation, fibrosis and defective pancreatic function. Conversely, biotin repletion activated numerous repair and anti-inflammatory pathways reduced fibrotic gene expression and induced multiple genes involved in pancreatic endocrine and exocrine function. A subset of results was confirmed by qPCR analysis, as well as by biotin treatment of pancreatic AR42J cells. The results indicate that biotin repletion, even after lengthy deficiency, results in the rapid induction of repair processes in the pancreas.The accepted manuscript in pdf format is listed with the files at the bottom of this page. The presentation of the authors' names and (or) special characters in the title of the manuscript may differ slightly between what is listed on this page and what is listed in the pdf file of the accepted manuscript; that in the pdf file of the accepted manuscript is what was submitted by the author

Characterizing changes in transcriptome and kinome responses in testicular cells during infection by Ebola virus

Abstract Ebola virus (EBOV) is able to persist and actively replicate in the reproductive tract of male disease survivors months or years after recovery from Ebola virus disease (EVD)1. Persistent EBOV infections are usually asymptomatic and can be transmitted sexually, but the host and viral factors that mediate these infections have not been characterized2,3. We investigated the interaction between host and viral factors during EBOV infection of the blood testis barrier (BTB), with a focus on Sertoli cells as a potential reservoir for viral persistence. We assessed viral replication kinetics and host responses of mouse testicular Leydig cells and Sertoli cells infected with EBOV Makona (i.e. infectious EBOV) and collected samples up to 28 days post-infection. Viral replication was apparent in both cell lines, but intracellular early viral loads were much higher in Leydig cells compared to Sertoli cells. We used RNAseq analysis to characterize transcriptomic responses of Leydig cells and Sertoli cells to EBOV infection over time. Further investigation of early interactions between host cells and EBOV was performed using virus-like particles (EBOV trVLP) and assays of phosphorylation-based cell signaling. Our findings indicate that virus-treated Sertoli cells responded more rapidly and robustly than Leydig cells, and with a particular emphasis on detection of, and response to, external stimuli. We discuss how the roles played by Sertoli cells in immune privilege and spermatogenesis may affect their initial and continued response to EBOV infection in a manner that could facilitate asymptomatic persistence

Additional file 4: Figure S3. of The transcription factor scleraxis is a critical regulator of cardiac fibroblast phenotype

Capillary density and cardiomyocyte cross-sectional area analysis of cardiac sections from WT and scleraxis KO mice. (A) Cardiac tissue sections (6 μm) from WT and scleraxis KO mice were stained with Rhodamine-labeled GSL I and capillaries counted to assess density. (B) Results for n = 3 independent samples per genotype (two fields/sample); mean ± SEM; *P < 0.05 vs WT. (C) Cardiac tissue sections (6 μm) from WT and scleraxis KO mice were stained with fluorescein-labeled peanut agglutinin for measurement of cardiomyocyte cross-sectional area. (D) Results for n = 3 independent samples per genotype (50 cells/sample); mean ± SEM; *P < 0.05 vs WT; 40× objective, scale bar = 25 μm. (EPS 19611 kb

Additional file 7: of The transcription factor scleraxis is a critical regulator of cardiac fibroblast phenotype

Supporting data. (XLSX 284 kb

Additional file 2: Table S1. of The transcription factor scleraxis is a critical regulator of cardiac fibroblast phenotype

Echocardiographic parameters. Table S2. Antibodies used in the study. Table S3. Primers used for qPCR analysis. Table S4. Primers used for site-directed mutagenesis of αSMA proximal promoter. Table S5. Primers used for nested PCR generation of ScxΔΔ. Table S6. Probes used for electrophoretic mobility shift assay. Table S7. Primers used for chromatin immunoprecipitation assay. (DOCX 34 kb

Additional file 6: Figure S5. of The transcription factor scleraxis is a critical regulator of cardiac fibroblast phenotype

Induction of fibroblast, mesenchyme and tendon markers in C3H10T1/2 cells by scleraxis. C3H10T1/2 cells were treated with AdGFP or AdScx for 48 h, and mRNA assayed by qPCR for representative fibroblast, mesenchyme and tendon marker gene expression. Results were normalized to Gapdh and AdGFP-infected samples; mean ± SEM; n = 3; *P < 0.05 vs AdGFP. (EPS 978 kb

Baseline characteristics of people deprived of liberty with active TB, latent tuberculosis infection and non-infected according to the time of incarceration.

Baseline characteristics of people deprived of liberty with active TB, latent tuberculosis infection and non-infected according to the time of incarceration.</p

Specific differential expression in each group.

LTBI-LI: Latent tuberculosis infection with long incarceration (people already had ≥ 1 year in prison when entering the study). LTBI-SI: Latent tuberculosis infection with short incarceration (people started the follow-up with less than three months of incarceration). ATB: active tuberculosis. NI-SI: non-infected with short incarceration. NI-LI: non-infected with long incarceration. (PDF)</p

Database including the participants’ sociodemographic information.

(Excel, sheet 1); Excel, sheet 2 shows the definition of variables. (XLSX)</p

Principal component analysis among the five groups, to evaluate the heterogeneity.

LTBI-LI: Latent tuberculosis infection with long incarceration (people already had ≥ 1 year in prison when entering the study). LTBI-SI: Latent tuberculosis infection with short incarceration (people started the follow-up with less than three months of incarceration). ATB: active tuberculosis. NI-SI: non-infected with short incarceration. NI-LI: non-infected with long incarceration. (TIF)</p