Project description:The transcriptome of Blochmannia floridanus, the endosymbiont of the carpenter ant Camponotus floridanus, is presented during eight developmental stages of its holometabolous host by use of a whole genome DNA-macroarray. The detected transcription patterns indicate the presence of local transcription units as well as global regulatory mechanisms. Yet, the overall regulation scale is very modest, rarely exceeding a factor of three which is in line with the low number of transcriptional regulators present in this reduced genome. A large number of genes show differential expression in different life stages and a distinct expression pattern of genes possibly involved in symbiotic function as compared to housekeeping genes is apparent. However, these transcriptional changes are small as compared to the changes in the number of bacteria during host development which is highest in pupae and in young imagines. Control of replication of the bacteria in certain life stages may therefore be the decisive parameter influencing overall gene expression of Blochmannia in the animal. The few highly expressed genes like those encoding molecular chaperones exhibit a significantly higher G+C-content than moderately expressed genes. Keywords: Host stage-dependent gene expression C. floridanus laboratory colonies were maintained under constant conditions at 25°C with a 12 h light cycle in artificial nests and fed twice a week with cockroaches (Nauphoeta cinerea), Bhatkar agar (Bhatkar and Whitcomb, 1970), and honey water (50% wt/wt) ad libitum. Developmental stages were defined as L1 (very young larvae, still clustered), L2 (older larvae, approx. 2-4 mm), P1 (pupae before metamorphosis), P2 (pupae after metamorphosis, still uncolored), P3 (pupae shortly before eclosion with dark abdomens), W1 (workers shortly after eclosion, not completely melanized, no aggressive behavior), W2 (fully melanized adult workers from the nest, age not distinguishable), and W3 (adult workers from isolated subcolonies, at least 4 months old). For RNA preparations, midguts of the respective life stage (L2 - W3) were dissected and bacteria were isolated by homogenization and centrifugation. Endosymbionts from small larvae (L1) were isolated from whole animals. RNA for each cDNA array experiment was isolated from individual bacterial preparations; from L1 to W2, each developmental stage was represented by at least three independent experiments and samples from at least three different ant colonies. Two ant colonies (C79 and C118) were represented in every stage from L1 to W2 with additional samples from six other colonies. Old W3 workers were collected from two isolated subcolonies derived from the same mother colony, each sampled in duplicate.
Project description:The study comprises various components: Samples TD: We aims to screen out different gene expression profile in donor biopsies after revascularization , We aims to predict renal allograft dysfunction early after transplantation. Samples AR, ATN, Tx: We aim to screen out different gene expression profile in acute rejection on the kidney. We aim to screen out different gene expression profile in acute tubular necrosis on the kidney. Results from the various study components can help to diagnose renal allograft dysfunction with different causes by distinct gene expression profile. Keywords: acute rejection, acute tubular necrosis, donor biopsies, renal allograft dysfunction Samples AR1-AR17: This study has been accomplished with 17 patients of acute rejection on the kidney.Technical replicates: 2 replicates Samples ATN1-ATN5: This study has been accomplished with 5 patients of acute tubular necrosis on the kidney. Technical replicates: 2 replicates Samples Tx1-Tx14: This study has been accomplished with 14 patients of stable renal function on the kidney.Tecnical replicates:2 replicates(except Tx12) Samples TD1-TD12: This study has been accomplished with 12 patients of donor tissue with stable function early after transplantation on the kidney.Technical replicates: 2 replicates Samples TD13-TD21: This study has been accomplished with 9 patients of donor tissue with renal dysfunction early after transplantation on the kidney.Technical replicates: 2 replicates
Project description:The study comprises various components: We aim to screen out different gene expression profile in acute rejection on the kidney. We aim to screen out different gene expression profile in acute tubular necrosis on the kidney. We aim to screen out different gene expression profile in borderline change on the kidney. We aim to screen out different gene expression profile in non-rejection on the kidney. We aim to screen out different gene expression profile in presumed rejection on the kidney. We aim to screen out different gene expression profile in renal recipients with stable function. Results from the various study components can help to diagnose renal allograft dysfunction with different causes by distinct gene expression profile. Keywords: acute rejection, acute tubular necrosis, borderline change, non-rejection, presumed rejection, renal recipients with stable function, Samples GSM456771-GSM456800: This study has been accomplished with 15 patients of acute rejection on the kidney.Technical replicates: 2 replicates(except AR10 and AR13) Samples GSM456801-GSM456810: This study has been accomplished with 5 patients of acute tubular necrosis on the kidney. Technical replicates: 2 replicates(except ATN4) Samples GSM456811-GSM456831: This study has been accomplished with 11 patients of borderline change on the kidney.Tecnical replicates:2 replicates(except BL8 and BL11) Samples GSM456833-GSM456850: This study has been accomplished with 9 patients of non-rejection on the kidney.Technical replicates: 2 replicates Samples GSM456851-GSM456864: This study has been accomplished with 7 patients of presumed rejection on the kidney.Technical replicates: 2 replicates Samples GSM456865-GSM456894: This study has been accomplished with 15 patients of renal recipients with stable function.Technical replicates: 2 replicates
Project description:Several members of the Bacillus cereus group of bacteria carry lysogenic phages of the family Tectiviridae. The Bacilus cereus reference strain (ATCC 14579) harbor a linear plasmid (pBClin15) that is highly similar to the genomes of the Tectiviruses. Production of active phages has however never been reported for pBClin15 and the role of pBClin15 in B. cereus physiology has been enigmatic. We have for the first time demonstrated an effect of pBClin15 on the physiology of B. cereus ATCC 14579 whereby the wild type is more sensitive to DNA damaging antibiotics compared to a pBClin15 cured strain. The microarray experiments in this study were designed to highlight transcriptional differences between the B. cereus ATCC 14579 wild type and a plasmid cured variant that potentially could explain the impact of pBClin15 on the B. cereus physiology. Microarray experiments were carried out under non-stressful conditions (growth in logarithmic phase in LB medium) under which there were no phenotypic difference between the wild type and pBClin15 cured strain as well as under stressful conditions under which there were phenotypic differences between the two strain (growth in LB medium supplemented with 0.5M-5g norfloxacin per ml).
Project description:Transcriptome analysis in nodules of Medicago truncatula with customn 2.3K cDNA arrays.<br><br>Study of different nodule developmental stages in wild type.<br><br>Study of various mutants forming non-functional nodules.
Project description:Umbilical cord mesenchymal stem cells were stimulated to osteogenesis by 24h, 48h and 7 days. Undifferentiated and treated cells have their total RNA extracted and 25 ug were used to extract micro RNAs by Flash PAGE Fractionator (Ambion). These miRNA were labeled and used to hybridize to 662 oligo arrays corresponding to miRNA sequences from human, rat and mouse. The hybridized slides were scanned in the Amersham Automatic Slide Processor, quantified by TIGR Spotfinder software and submitted to stathiscal treatment in TIGR MEV software.
Project description:The endometrium provides optimal conditions for the transport of sperm to the oviduct, to the site of fertilization, and later on for the reception of the embryo. To study these changes on the level of gene expression, a messenger RNA expression profiling of endometrium tissue samples collected from 19 cyclic heifers at five stages of the estrous cycle (days 0, 3.5, 12, 18, 20) was performed. RNA was extracted from these tissue samples and analyzed with a custom-made bovine oviduct and endometrium (BOE) cDNA array. The cDNAs present on the array were derived from several previously conducted differential gene expression studies of bovine endometrium between different stages of the estrous cycle, during early pregnancy, and from studies of bovine oviduct epithelial cells. In all of these studies cDNAs of differentially expressed genes were identified using a combination of subtracted cDNA libraries and cDNA array hybridization. 1,440 cDNA fragments are located on the array. Twenty radioactively labeled cDNA samples (n=4 for each cycle stage) were hybridized with BOE arrays. Raw data were normalized using the BioConductor package vsn. Keywords: time course of gene expression in bovine endometrium during estrous cycle Nineteen German Fleckvieh (Simmental) heifers were slaughtered at four different days of the estrous cycle, four animals at day 0, four at day 3.5, three at day 12 and eight animals at day 18. Four of the day 18 animals showed high and the other four low serum progesterone (P4) levels, respectively. This resulted in five groups of endometrial tissue samples corresponding to five stages of the estrous cycle.
Project description:Although somatic cell nuclear transfer (SCNT) cloning is more efficient in bovine than in all other species tested so far, there is a high rate of pregnancy failure that has been linked to structural and functional abnormalities of the placenta. We tested the hypothesis that these changes may originate from disturbed embryo-maternal interactions in the pre-implantation period. Therefore, we evaluated the transcriptome response of the endometrium to SCNT embryos (produced from five different donor cell cultures) as compared to embryos derived from in vitro fertilization (IVF). SCNT embryos and IVF embryos were cultured under identical conditions to the blastocyst stage (Day 8) and transferred to recipients. The recipients were slaughtered at day 18 of pregnancy and the uterus was recovered. Pregnancy was verified by the presence of at least one normally developed embryo. Transcriptome profiling of endometrium samples using a custom cDNA microarray covering transcripts expressed in the endometrium and/or oviduct epithelium revealed 58 transcripts that were differently abundant between endometrium samples from SCNT vs. IVF pregnancies. Prominent examples are NR2F2 (encoding the orphan nuclear receptor COUP-TFII) and GJA1 (encoding connexin 43). Both transcripts are known to play important roles in placentation and were significantly less abundant in endometrium from SCNT vs. IVF pregnancies. These findings suggest that placental failure in bovine clone pregnancies may originate from abnormal embryo-maternal communication already in the pre- or peri-implantation period. Endometrium transcriptome profiles may serve as a novel readout to evaluate SCNT embryos for their ability to induce pregnancy with a functional placenta. Keywords: response to different embryos Nineteen German Fleckvieh (Simmental) heifers were slaughtered at day 18 of pregnancy. Cycle-synchronized recipient heifers received either IVP or SCNT embryos at day 7 of the estrous cycle. Animals were slaughtered at day 18. Endometrial (intercaruncular) tissue samples were obtained from 10 pregnant animals after transfer of IVP embryos and from 9 pregnant animals after transfer of SCNT embryos.
Project description:Small interfering RNA ( siRNA) was used to knockdown Autoimmune regulator (Aire) gene by in vivo electroporation of the thymus of BALB-c mice. In this set of data we include control and Aire-knockdown mTEC cells isolated from thymus of BALB-c mice.