Clostridium difficile strain 630 heat stress experiment
Ontology highlight
ABSTRACT:
ORGANISM(S): Clostridium difficile 630
SUBMITTER: Nigel Ternan
PROVIDER: E-GEOD-37442 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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Project description:By using a 44k chicken Agilent microarray, we systematically analyzed the chicken hypothalamus transcriptome response to thermal stress. Twelve hypothalamus samples were chosen from three groups (four samples per group) to be used in chicken genome microarray to examine differential gene expression.We compared the expression profiles between each pairs of the three groups using the microarray data. Totally, 2474 genes were found to be differentially expressed in the three comparisons with pM-oM-<M-^\0.05 and fold change (FC) higher than 1.5 and the genes were mainly involved in self-regulation and compensation required to maintain homeostasis, including heat shock protein family, enzyme and the hormone, neurotransmitter, cell-cell signaling, metabolism and cytokines. The transcripts of heat shock protein including Hsp 40 and Hsp 90 were significantly changed respond to thermal stress and genes involved in regulation of cell morphogenesis were significantly upregulated in heat stressed group with comparison to control and temperature recovery group. Additionally, the down-regulated genes in both heat stress and temperature recovery groups compared to control group were enriched in muscle organ development, striated muscle tissue development, cardiac muscle tissue development and muscle tissue development, which indicates that muscle development was inhibited during and in short-term after heat treatment. Most of genes dysregulated in heat stress group were found to be recovered in temperature recovery group, which confirmed their roles they could play in coping with heat stress. The present study provides a broader understanding of molecular mechanisms underlying the stress response in chicken and discovery of novel genes that are regulated in a thermal stress specific manner. Hypothalamus samples were collected from non-heat treated group (reared at 25C, used as control), 24h 34C treated group (heat stress treated group) and temperature recovery group (25C for 24h followed heat stress).
2012-10-01 | E-GEOD-37400 | biostudies-arrayexpress
Project description:Background: The in vivo gene response associated with hyperthermia and subsequent return to homeostasis or development of heat illness is poorly understood. Early activation of gene networks in the heat stress response is likely to lead to the systemic inflammation, multi-organ functional impairment, and other pathophysiological states characteristic of heat illness. Here, we perform an unbiased global characterization of the multi-organ gene response using an in vivo model of heat stress in the conscious rat. Results: Rats were subjected to elevated temperatures until implanted thermal probes indicated a maximal core temperature of 41.8 M-BM-0C (Tc,Max). Liver, lung, kidney, and heart were harvested at Tc,Max, 24 hours, and 48 hours after heat stress in groups of experimental animals and time-matched controls kept at ambient temperature. Clinical chemistries suggested abnormal function in liver, kidney, and lung at Tc,Max, and cardiac histopathology at 48 hours supported persistent cardiac damage in 3 out of 6 animals. Microarray analysis identified 78 differentially expressed genes common to all 4 organs at Tc,Max (i.e., the consensus heat stress response). Gene set enrichment analysis of gene ontology terms identified 25 biological processes in 4 general gene ontology categories: protein folding, regulation of apoptosis, response to cytokines, and transcriptional responses. Functional analysis clustering of the 78 differentially expressed genes in the consensus heat stress response also identified functional categories of protein folding and regulation of apoptosis. Self-organizing maps identified gene-specific signatures corresponding to protein folding disorders specific to only heat-stressed rats with histopathologic evidence of cardiac injury at 48 hours. Enrichment analysis of differentially expressed proteins in heat-injured hearts at 48 hours corroborated gene enrichment analysis results. Quantitative proteomics analysis by iTRAQ demonstrated that differential protein expression was not comparable to transcript expression at Tc,Max and 24 hours. However, the profile of differentially expressed proteins closely matched the transcriptomic profile in heat-injured animals at 48 hours. Pathway analysis at both the transcript and protein levels supported catastrophic deficits in energetics and cellular metabolism, chronic proteotoxic response, and activation of the unfolded protein response. Calculation of protein super-saturation scores demonstrated an increased propensity of proteins to aggregate in the hearts of heat-injured animals at 48 hours, consistent with accumulation of misfolded proteins. Conclusions: Global transcriptomic and proteomic analysis identified networks of genes and proteins initiating an unfolded protein response, metabolic dysfunction, and mitochondrial energy crisis in animals with histopathologic evidence of persistent heat injury, providing the basis for a systems-level physiological model of heat illness and recovery. To induce the pathophysiological effects of heat stress, conscious rats were placed in an incubator at 37M-BM-:C and their core temperature was monitored. Animals were sacrificed when their core temperature reached 41.8M-BM-:C (Tc,Max), or they were returned to their normal housing environment and were allowed to recover for up to 24 or 48 hours prior to sacrifice. Time-matched controls were euthanized at times corresponding to Tc,Max, 24 hours, and 48 hours. For each condition n=6 for a total of 36 animals. Four tissues (liver, lung, kidney and heart) were analyzed from each animal for a total of 144 arrays; however a few arrays did not pass QC therefore only 140 are reported here: 34 liver, 35 lung, 36 kidney, or 35 heart.
2015-01-14 | E-GEOD-56740 | biostudies-arrayexpress
Project description:Exiqon miRNA array contains LNA-modified oligos which minimizes the Tm-range within which probe hybridizes with the transcript in query, thus minimizing the mis-match. Taking advantage of this, we used the miRCURY LNA array to investigate genome wide signatures for miRNA expression in response to heat shock stress. We also did mRNA expression profiling using the same RNA to look for possible miRNA-mRNA interaction in response to heat shock stress. For the differentially expressed miRNAs, target predictions were done using both miRanda and TargetScan. The consensus set was used for subsequent experimental validation. In total, 6 comparative hybridizations of heat-shock treated versus control were profiled using the Exiqon arrays. These consisted of 3 biological replicates, each with its dye-swap pair.
2015-01-01 | E-GEOD-60472 | biostudies-arrayexpress
Project description:Genome wide expression profiles of 10-day-old seedlings in response to prior exposure with OS followed by heat stress or cold stress as well simultaneous exposure to OS along with HS/CS was done to study the transcriptional changes in response to combination of stresses. 10-day old rice seedlings were used for experiments. Transcript profiling was done in 10-day old rice seedling following oxidative Stress prior to High Temperature Stress (PO_HS) or Low Temperature Stress (PO_CS) and Oxidative Stress during High Temperature Stress (DO_HS)/ during Low temperature stress (DO_CS). Unstressed seedlings were used as control. For each condition, three biological replicates were processed in a similar manner for the microarray analysis.
2012-12-26 | E-GEOD-32704 | biostudies-arrayexpress
Project description:Purpose: To determine how divergent strains of C. difficile respond to environmental changes, the transcriptomes of two historic and two recently isolated hypervirulent strains were analyzed following nutrient shift and osmotic shock. Methods: Following nutrient shift and osmotic shock, mRNA profile were determined using sequing of transcriptome in biological duplicates, using Illumina Hi-seq 2000. Results: After applying the quality control steps, for each sample, sequence reads were 90 nucleotides in length and the total number of reads per sample was ~ 26.6 million on average. Expression of more than 90% CDS was detected under the three experimental conditions combined. About 20% of these genes were indentified to be differentially expressed at 1.5 fold or more. Conclusions: Our results reveal that although C. difficile strains contain a large number of shared and strain specific genes, the majority of the differentially expressed genes were core genes. We also detected a number of transcriptionally active regions that were not part of the primary genome annotation. Some of these are likely to be small regulatory RNAs. Differential stress transcriptomes of two historic (strain 630 and 196) and two recently emerged hypervirulent strains (stain R20291 and 32g58) of C. difficile was determined by deep sequencing, in duplicate, using Illumina Hi-Seq 2000.
2013-11-06 | E-GEOD-50497 | biostudies-arrayexpress
Project description:Corynebacterium pseudotuberculosis strain 258, isolated from a caprine host in Brazil, was maintained in brain-heart-infusion broth (BHI HiMedia Laboratories Pvt. Ltda, India) at 37C, under rotation. Then, the bacteria was added to 20mL of BHI-Broth with 0.05% of 80% Tween and was maintained at 37C overnight, under rotation. The sample was then diluted 1/100 in 100mL of BHI broth, maintained at 37C, under rotation, and it was monitored until it reached the beginning of the exponential fase (A600 = 0.2). Afterwards, the sample was equally splitted in 4 aliquots of 20mL, centrifuged for 3 minutes under 8000rpm at 37C, and the pellets were re-suspendend in different medias according to the following conditions: acid stress, with HCl (fitting the pH to 5); osmotic stress, with 2M of NaCl; heat shock, where the pellet was re-suspended in pre-heated BHI (50C); and control, where the pellet was re-suspended in BHI under physiologic condition. Then, the samples for the acid stress, osmotic stress and control condition were maintained at 37C for 15 minutes, under rotation, whereas the sample under heat shock condition was maintained at 50C. Next, the samples were serially diluted from 10e-1 to 10e-6, where the samples from 10e-4 to 10e-6 were spread in BHI with agar (1.5%), which was maintained at 37C for 48hs for colony counting (duplicated samples); and the broth was centrifuged at 37C for 3 minutes under 8000rpm and the final pellet was re-suspended in 2mL of RNAlater, following the protocol of the manufacturer. Briefly, the bacteria suspended in RNAlater buffer were subjected to total RNA extraction using the ChargeSwitch total RNA cell kit (Invitrogen, USA) in accordance with the manufacturers recommendations, including the following adaptations: after the addition of the lysis buffer (Invitrogen), the material was transferred to 2mL tubes partially filled with 1mm diameter glass microbeads (Bertin Technologies). The cells were lysed mechanically using a Prescellys 24 homogenizer, set at 6,500 rpm, for 2 cycles (15 seconds per cycle) with an interval of 30 seconds between the cycles. The samples were centrifuged for 1 minute, and the supernatant transferred to fresh 2ml tubes and incubated in a dry bath at 60C for 15 minutes (representing the complete original protocol). DNAse was added to eliminate the residual genomic DNA. The elution of the total RNA from the magnetic beads was performed using 100 uL of milli-Q RNase-free water. The amount of total RNA was assessed using a Qubit 2.0 fluorometer (Invitrogen). In order to construct the library, the SOLiD Total RNA-Seq Kit was used. Briefly, after removing rRNAs, approximately 245ng of RNA was fragmented with RNAIII through incubation in a MJ Research thermocicler at 37C for 10 min and the reaction was interrupted with the addition of 90uL of nuclease-free water. The RNA was, then, concentrated through precipitation with ethanol according to the RiboMinus Concentration Module (Invitrogen - EUA), for subsequent hybridization with adapters and amplification of the cDNA library, which was obtained through reverse transcription using P1 adapters linked to the RNA extremities, according to the protocol SOLiD Total RNA-Seq Kit (Life Technologie, EUA). Next, the cDNA was denatured in polyacrylamide gel electrophoresis (6%) for obtaining fragments in the range from 150 to 250 base pairs for PCR amplification. The PCR amplifications were carried out in a MJ Research thermocycler as follow: the mixture was denatured at 95C for 30 sec, followed by 15 cycles of amplification at 95C for 30 sec, 62C for 30 sec and 72C for 30 sec, with a final extension of 72C for 7 min. Finally, the amplicons were purified using the kit PureLinkTM PCR Micro (Invitrogen - EUA), quantified in Qubit 2.0 Fluorometer-Invitrogen (EUA) and assessed through agarose electrophoresis 2%. The samples were fitted to the concentration of 60pg/mL, which is the suggested concentration for emulsion PCR. After the emulsion PCR, an enrichment reaction was performed for capturing only microbeads with fragments inserted. Next, a workflow analysis (WFA) was performed, where 15,000,000 microbeads were deposited to the slide and have undergone a binding cycle for quantitative and qualitative evaluation. According to this evaluation, 44,000,000 microbeads were deposited for sequencing, based on the manufacturer's description, generating reads of 50 nucleotides in range.
2020-06-20 | E-MTAB-9217 | biostudies-arrayexpress
Project description:This project investigates the changes in proteome profiles that occurred in the same set of S. mutans wild-type and lrgAB mutant samples used in our recently published RNA-seq study. To this end, mass spectrometry-based label-free quantitative proteomics, a technology known to enable comprehensive identification and quantification of complete bacterial proteomes, was adapted to study changes in S. mutans intracellular protein levels in response to aeration, heat, and vancomycin stress. Furthermore, the degree of correlation between S. mutans protein abundance profiles and gene expression changes was determined. Further understanding of these combined “-omics” data at the cellular or molecular level will enhance our knowledge of Cid/Lrg-mediated cellular responses of S. mutans to adverse environments. Furthermore, these “-omics” data will serve as a valuable resource that can be mined to help clarify the role of the S. mutans stress response and physiological activity to its dynamic survival in the oral cavity.
2018-10-24 | PXD006735 | Pride
Project description:Heat stress is the primary obstacle for landscape application of azalea in hot-summer areas. Heat acclimation can confer acquired thermotolerance to R. hainanense even after a long recovery. The mechanisms and key factors involved in the heat acclimation memory were investigated.
2020-04-02 | PXD017005 | Pride
Project description:ABSTRACT: Exposure to abiotic stresses triggers global changes in the expression of thousands of eukaryotic genes at the transcriptional 70 and post-transcriptional levels. Small RNA (smRNA) pathways and splicing both function as crucial mechanisms regulating stress-responsive gene expression. However, examples of smRNAs regulating gene expression remain largely limited to effects on mRNA stability, translation, and epigenetic regulation. Also, our understanding of the networks controlling plant gene expression in response to environmental changes, and examples of these regulatory pathways intersecting, remains limited. Here, to investigate the role of smRNAs in stress responses we examined smRNA transcriptomes of Brachypodium distachyon plants subjected to various abiotic stresses. We found that exposure to different abiotic stresses specifically induced a group 75 of novel, endogenous small interfering RNAs (stress-induced, UTR-derived siRNAs, or sutr-siRNAs) that originate from the 3M-bM-^@M-2 UTRs of a subset of coding genes. Our bioinformatics analyses predicted that sutr-siRNAs have potential regulatory functions and that over 90% of sutr-siRNAs target intronic regions of many mRNAs in trans. Importantly, a subgroup of these sutr- siRNAs target the important intron regulatory regions, such as branch point sequences, that could affect splicing. Our study indicates that in Brachypodium, sutr-siRNAs may affect splicing by masking or changing accessibility of specific cis-elements 80 through base-pairing interactions to mediate gene expression in response to stresses. We hypothesize that this mode of regulation of gene expression may also serve as a general mechanism for regulation of gene expression in plants and potentially in other eukaryotes. Analysis of smRNA populations in Brachypodium plants challenged by abiotic stresses: To profile the populations of smRNAs in the model monocot Brachypodium distachyon and examine their regulation in response to abiotic stresses, we conducted high-throughput sequencing of small RNAs from plants exposed to four different abiotic stress conditions, cold, heat (air), heat (water immersion), and salt, in the wild type Brachypodium cultivar Bd21. For our experiments we used information from the literature to select two time-points for stress durations, short and long, which differed for each stress: cold (6 and 24 hours), heat-air (1 and 3 hours), heat-water (1 and 3 hours), and salt (48 hours). We generated small RNA libraries for Illumina sequencing (GAII) from the leaves of Brachypodium plants subjected to stresses and selected smRNAs between 15 and 40 nt in length, which we mapped to the Brachypodium genome.
2015-01-20 | E-GEOD-55217 | biostudies-arrayexpress
Project description:This study was conducted to determine the impact of extreme temperatures, aiming at controlling microorganisms by cooking, on the physiology and the transcriptome of Escherichia coli. ABSTRACT: Because the genome of Escherichia coli K12 is well understood, this organism was used as a model to investigate physiological and molecular changes during cell adaptation and survival to cooking temperatures used in the food industry. Bacteria grown at 37°C to stationary phase in Brain Heart Infusion (BHI) broth, were heated at 58°C or 60°C to reach a process lethality value (F) of 2 and 3, or until an internal core temperature of 71°C was reached (control cooking temperature). Both growth recovery and cell membrane integrity were evaluated immediately after heating and a global transcription analysis was performed by using gene expression microarrays. Only cells heated at 58°C F = 2 were still able to grow on liquid or solid BHI after heat treatment. However, their transcriptome did not differ significantly compared to those of bacteria heated at 58°C F = 3 (P-FDR > 0.01) where no growth recovery was observed post treatment. Genome-wide transcriptomic data obtained at 71°C were distinct from the other treatments. Quantification of heat-shock genes expression by real-time PCR revealed that dnaK and groEL mRNA levels were significantly lower at 71°C than at 58°C and 60°C (P < 0.0001). Furthermore, despite of similar cell inactivation but growth on BHI media after heating, 132 and 8 genes were differentially expressed at 71°C when compared to 58°C and 60°C at F = 3 respectively (P-FDR < 0.01). Among them, genes like aroA, citE, glyS, oppB and asd, whose expression was up-regulated at 71°C, may be considered as good biomarkers to determine more accurately the efficiency of heat treatments. Bacteria were submitted to four different heat-treatments. Biological replicates: 5 Controls grown to an optimal temperature of 37°C (and used to prepare a common reference for each array hybridization), 5 Treatment 1 (58°C F = 2), 4 Treatment 2 (58°C F = 3), 4 Treatment 3 (60°C F = 3), 5 Treatment 4 (core temperature of 71°C). Treatment replicates were randomly hybridized on 3 microarray slides Agilent 8x15K (total of 24 array areas). Equal amounts of the labeled control cDNA pool and labeled cDNA from one of the heated groups were mixed and hybridized on each array.
2012-09-15 | E-GEOD-40557 | biostudies-arrayexpress