Temperature dependent Transcriptome of L. monocytogenes EGDe in minimal medium
Ontology highlight
ABSTRACT:
ORGANISM(S): Listeria monocytogenes EGD-e
SUBMITTER: Stefanie MM-CM-<ller-Herbst
PROVIDER: E-GEOD-51193 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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Project description:Profound understanding of the mechanisms foodborne pathogenic bacteria utilize in adaptation to the environmental stress they encounter during food processing and storage is of paramount importance in design of control measures. Chill temperature is a central control measure applied in minimally processed foods; however, data on the mechanisms the foodborne pathogen Clostridium botulinum activates upon cold stress are scarce. Global gene expression analysis on the C.M-BM- botulinum ATCC 3502 strain upon temperature downshift from 37 M-BM-0C to 15M-BM- M-BM-0C was performed to identify the cold-responsive gene set of this organism. Significant up- or down-regulation of 16 and 11 genes, respectively, was observed already 1 h after the cold shock. At 5 h after the temperature downshift, 199 and 210 genes were up- and down-regulated, respectively. Thus, the cold shock rapidly affected the expression of a gene set of a relatively small size, indicating a targeted acute response to cold shock, whereas extensive metabolic remodeling took place after prolonged exposure to cold. Induction of genes related to fatty acid biosynthesis, oxidative stress response, and iron uptake and storage was observed, in addition to mechanisms previously characterized as cold-tolerance related in bacteria. Furthermore, induction of several uncharacterized DNA-binding transcriptional regulator-encoding genes was observed, suggesting involvement of novel regulatory mechanisms in the cold shock response of C.M-BM- botulinum. The role of such regulatory proteins, CBO0477 and CBO0558A, in cold tolerance of C.M-BM- botulinum ATCC 3502 was demonstrated by the deteriorated growth of mutants of the respective genes at 17M-BM- M-BM-0C. C. botulinum ATCC 3502 wild type cold-shocked vs. pre-cold-shock; 3 replicates; growth at 37C in TPGY broth batch culture and subjected to cold shock to 15C; sampling at mid-log growth phase before cold shock, and 1 h and 5 h after temperature downshift to 15C (= 3 time points). Dye-swapped hybridization.
2014-02-24 | E-GEOD-51465 | biostudies-arrayexpress
Project description:Escherichia coli O157 presents a number of specific problems in terms of food safety and public health. It has been found that E. coli O157 is more resistant to a number of the stresses encountered during food production such as heat, pH and osmotic shock. This greater resistance is thought to contribute to the low infectious dose of E. coli O157 (<100 organisms). Moreover, E. coli O157 is associated with debilitating conditions such as haemorrhagic colitis and haemoytic uraemic syndrome, particularly in children and the elderly. We have been studying the stress responses of E. coli O157:H7 (Sakai) and comparing with a commensal strain of E. coli K-12, MG1655. We found that E. coli O157 (Sakai) is more resistant to heat stress than MG1655. A microarray study of these strains subjected to sub-lethal heat-stress at 45M-BM-0C was carried out. In E. coli O157 (Sakai), 380 genes responded significantly to the treatment compared to 410 genes in MG1655. Overnight cultures of E. coli O157 (Sakai) and E. coli K-12 MG1655 were grown in Neidhardt's EZ Rich Defined Medium and diluted 1:100 in 50 ml fresh medium in 125 ml Ehrlenmeyer flasks. The cultures were shaken at 37M-BM-0C until the optical density (OD600) reached 0.4. Each culture was divided into 2 equal parts in identical flasks. One flask flask was transferred to a shaking water bath and incubated at 45M-BM-0C for 10 min; the other flask was incubated at 37M-BM-0C for 10 min. After incubation, the cultures were transferred to 50 mL centrifuge tubes and treated with RNAprotectM-bM-^DM-" to stabilise the mRNA. The experiment was performed 3 times on different days. Six custom-made microarray slides were used in this study; each slide was hybridised with labelled cDNA made from untreated and heated E. coli O157 (Sakai) or MG1655.
2018-03-20 | E-MTAB-5912 | biostudies-arrayexpress
Project description:The Thioacetamide-treated rat was first identified as a model of hepatotoxicity by Gupta in 1956 and is now well-established, not least because the histopathogical output closely mimics that seen in humans with chronic liver disease. Acute treatment of rats with Thioacetamide causes pronounced necrosis and inflammation. Animals received intraperitoneal (ip) doses of vehicle-only (0.9% (v/v) saline) (n=3), or 100 mg/kg Thioacetamide (n=3) and were sacrificed after 24 hours. Blood was withdrawn via the descending vena cava and immediately transferred into potassium/EDTA tubes. Following centrifugation (16,100g, 4M-BM-0C, 5 min) the plasma was collected and stored at -80M-BM-0C. miRNA microarray profiling of RNA extracted from the plasma of rats treated with Thioacetamide revealed that a subset of miRNAs were differentially expressed following treatment. These miRNAs appeared to mediate pathways involved in hepatic fibrosis and stellate cell activation, suggesting that they might function as predictive biomarkers following compound-induced hepatotoxicity. The changes correlated well with increases in ALT levels, which are the current gold standard method for determining the extent of liver injury. Furthermore, it is hypothesised that particular aetiologies of liver damage might cause differing expression profiles of miRNAs, thus certain miRNAs could be implemented in a panel-type expression study to distinguish between different types of hepatic injury. Single channel miRNA microarrays were performed on n= 3 samples, 2 treatment groups; control and test. Control animals received vehicle-only (0.9% (v/v) saline) via the ip route. Test animals received 100 mg/kg Thioacetamide dissolved in 0.9% (v/v) saline, via the ip route. 24 h after dosing animals were sacrificed using decapitation under terminal anaesthesia.
2014-03-14 | E-GEOD-55885 | biostudies-arrayexpress
Project description:Transcriptional profiling of the digestive gland tissue of female mussel Mytilus galloprovincialis exposed to copper along with a temperature gradient Background: Global warming is a major factor that may affect biological organization, especially in marine ecosystems and in coastal areas that are particularly subject to anthropogenic pollution. We evaluated the effects of simultaneous changes in temperature and copper concentrations on lysosomal membrane stability of the blue mussel Mytilus galloprovincialis (Lam.). Temperature and copper exerted additive effects on lysosomal membrane stability, exacerbating the toxic effects of metal cations present in non-physiological concentrations. Mussel lysosomal membrane stability was positively related toscope for growth, indicating possible effects of increasing temperature on mussel populations in metal-polluted areas. To clarify the molecular response to environmental stressors, we used a cDNA microarray with 1,700 sequences to measure the relative transcript abundances in the gills of mussels exposed to copper (40M-BM-5g/L) and a temperature gradient (16M-BM-0C, 20M-BM-0C, and 24M-BM-0C). In animals exposed only to heat stress, hierarchical clustering of the microarray data revealed three main clusters, which were largely dominated by down-regulation of translation-related differentially expressed genes, drastic up-regulation of folding protein-related genes, and genes involved in chitin metabolism. The response of mussels exposed to copper at 24M-BM-0 C was characterized by an opposite pattern of the genes involved in translation, most of which were up-regulated, as well asthe down-regulation of genes encoding heat shock proteins and microtubule-based movement proteins. Our data provide novel information on the transcriptomic modulations in mussels facing temperature increases and high copper concentrations; these data highlight the risk of marine life exposed to toxic chemicals in the presence of temperature increases due to climate change. Digestive gland tissue from individual animals in different experimental conditions were analyzed in a complete loop design. Dual color competitive hybridizations (Control M-bM-^@M-^\16M-BM-0CM-bM-^@M-^] vs M-bM-^@M-^\20M-BM-0CM-bM-^@M-^] vs M-bM-^@M-^\24M-BM-0CM-bM-^@M-^] vs M-bM-^@M-^\16M-BM-0C+CuM-bM-^@M-^], M-bM-^@M-^\20M-BM-0CM-bM-^@M-^] vs M-bM-^@M-^\24M-BM-0CM-bM-^@M-^] vs M-bM-^@M-^\20M-BM-0C+CuM-bM-^@M-^], M-bM-^@M-^\24M-BM-0CM-bM-^@M-^] vs M-bM-^@M-^\24M-BM-0C+CuM-bM-^@M-^], M-bM-^@M-^\16M-BM-0C+CuM-bM-^@M-^] vsM-bM-^@M-^]20M-BM-0C+CuM-bM-^@M-^] vs M-bM-^@M-^\24M-BM-0C+CuM-bM-^@M-^]; M-bM-^@M-^\20M-BM-0C+CuM-bM-^@M-^] vs M-bM-^@M-^\24M-BM-0C+CuM-bM-^@M-^]) including label swap. Single individuals. Four biological replicates. One replicate per array.
2012-10-31 | E-GEOD-41899 | biostudies-arrayexpress
Project description:To investigate the regulatory mechanisms governing the malignant signature of different gliomas we analyzed microRNA expression profiles in human tumor samples of world health organization (WHO) grade I (benign tumors), II (low grade tumors) and IV (high grade tumors) and from primary cultures obtained from tumor samples of grade II and IV. Patients This study included tumor samples histologically verified as astrocytic gliomas obtained from patients who had undergone craniotomy for microsurgical tumor removal. According to the revised WHO classification, tumors were diagnosed as: grade I or pilocytic astrocytomas; grade II or diffuse fibrillary astrocytomas; grade IV or glioblastoma multiforme. Primary cell cultures from grade II and grade IV gliomas were also obtained and miRNA expression in these cultures were analyzed RNA extraction Total RNA, including small RNA, was isolated from tissue samples using the mirVanaTM miRNA Isolation Kit (Ambion) following the standard protocol. The quantity and quality of the purified RNA was evaluated by spectrophotometric analysis and electrophoresis on denaturing gel of acrylamide. Multiplex Real-Time Quantitative Reverse-Transcriptase Polymerase Chain Reaction (RT-PCR) The miRNAs were first converted to cDNA using Multiplex RT for TaqMan Array Human MicroRNA Panel. The RT Master mix included 100 mM each of dNTPs , 50 U/ml MultiScrabe reverse transcriptase (Applied Biosystems), 20 U/M-BM-5l RNase inhibitor (Applied Biosystems) and 10X RT Buffer. The 10 M-BM-5l reactions, including 7 M-BM-5l of RT master mix, 2 M-BM-5l of purified microRNA and 1 M-BM-5l of Multiplex RT Human primer pool (Applied Biosystem), were incubated in ice for 5 min and then in a thermal cycler for 30 min at 16M-BM-0C, 30 min at 42M-BM-0C, 5 min at 85M-BM-0C, and then hold at 4M-BM-0C. miRNA levels were normalized to the expression of small nucleolar RNAs, RNU44, RNU48 and RNU6B. All reverse transcriptase reactions, including no-template controls and RT controls, were run in duplicate. Real-time PCR was performed using a standard TaqMan PCR kit procedure on an M-bM-^@M-^\Real Time Fast 7900 HTM-bM-^@M-^] PCR System (Applied Biosystems). The 100 M-BM-5l PCR included 50 M-BM-5l RT product (before diluited 1:60) and 50 M-BM-5l TaqMan Universal PCR Master Mix (2X) (Applied Biosystems). The total volume were loaded into Card TaqMan Low Density Array Human MicroRNA Panel (Applied Biosystem) including a total of 384 human microRNAs publicated on databases www.sanger.ac.uk. The reaction cards was runned at 50M-BM-0C for 2 min and 95M-BM-0C for 10 min, followed by 40 cycles of 97M-BM-0C for 30s and 59,7M-BM-0C for 1 min. All reactions were run in triplicate. Analysis of data was performed using the SDS 2.3 software using the 2-M-bM-^HM-^FM-bM-^HM-^FCt (relative quantitative) method . The M-bM-^HM-^FCt of every miRNA was determined in relation to the endogenous control RNA U6 that was invariably expressed in all samples. The M-bM-^HM-^FM-bM-^HM-^FCt value was determined in relation to the calibrator, namely the normal brain tissue. Resulting data were grouped according to the tumor grading e selectioned using a cut-off value of 3. Results were expressed as M-bM-^@M-^\fold changeM-bM-^@M-^] over normal brain tissue. We analyzed two samples of grade I, two samples of grade II, two samples of grade IV gliomas. Four samples form norma brain were used as norma control. Primary cell cultures form grade II and grade IV samples were used for the analysis. All reverse transcriptase reactions, including no-template controls and RT controls, were run in triplicate.
2013-09-18 | E-GEOD-50924 | biostudies-arrayexpress
Project description:Penicillium marneffei (Talaromyces marneffei) is an opportunistic human pathogen that can grow in a multicellular hyphal form at 25M-BM-0C or a unicellular fission yeast form at 37M-BM-0C, and can switch between these two forms in response to temperature. The yeast form is found in infected individuals and represents the pathogenic phase. In response to specific environmental cues, the hyphal form can undergo asexual development (conidiation) the produce a differentiated multicellular structure called a conidiophore which produces dormant spores (conidia). Inhaled conidia initiate infection. To identify genes that are important during the early stages in the transition from hyphal to yeast and yeast to hyphal cells transcriptional profiling was performed with a custom microarray consisting of ~42% of the predicted P. marneffei genes and RNA from P. marneffei hyphal cells switched to growth at 37M-BM-0C for 6 hours (hyphal-yeast switch) and yeast cells switched to growth at 25M-BM-0C for 6 hours (yeast-hyphal switch). A custom microarray consisting of short, random genomic fragments from P. marneffei (~42% of the predicted genes) was generated using PCR products of the inserts from two independent DNA libraries constructed from genomic DNA of the P. marneffei type strain FRR2161. PCR products from previously cloned P. marneffei genes with known expression profiles were included as controls on the microarray. Total RNA from hyphal cells before and after switching to growth at 37M-BM-0C for 6 hours (hyphal-yeast switch) and yeast cells before and after switching to growth at 25M-BM-0C for 6 hours (yeast-hyphal switch) were used in pairwise combinations on the microarrays. Three biological replicate cultures were prepared for each experiment.
2013-11-21 | E-GEOD-51110 | biostudies-arrayexpress
Project description:Propionibacterium freudenreichii is used as a ripening culture in Swiss cheese manufacture. It produces flavor compounds over the whole ripening period. During cheese ripening, P. freudenreichii is exposed to a temperature downshift, especially when cheeses are transferred from warm temperature (about 24M-BM-0C) to cold temperature (about 4M-BM-0C). The cold adaptation of the type strain was studied previously. The aim of this study was to investigate the adaptation of 6 other P. freudenreichii strains at cold temperature by means of a global gene expression profile. The temporal transcriptomic response of 6 P. freudenreichii strains was analyzed at 2 times of growth, during growth at 30M-BM-0C then after 3 days 4M-BM-0C, in the constant presence of lactate as the main carbon source. Six strains were used: CIRM-BIA9, CIRM-BIA118, CIRM-BIA122, CIRM-BIA123, CIRM-BIA472, CIRM-BIA482. Gene expression was measured in the middle of exponential growth phase at 30M-BM-0C (20h, OD650 M-bM-^IM-^H 0.5), and after 3days of incubation at 4M-BM-0C. Three independent biological experiments were performed for each strain and at each time, and were labelled A, B, C. Five technical repetitions were performed using the RNA of 3J_122B, 3J_123A, 3J_472C, 20H_9A and 20H_482C samples. These technical repetitions were labelled 3J_122Bii, 3J_123Aii, 3J_472Cii, 20H_9Aii and 20H_482Cii respectively. The transcriptomic data for 20H_122A and 3J_123C samples were abberant and were thus not considered for further analysis.
2012-07-05 | E-GEOD-34227 | biostudies-arrayexpress
Project description:Transcriptional profiling of the digestive gland tissue of female mussel Mytilus galloprovincialis exposed to nickel along with a temperature gradient Background: The exposure of marine organisms to stressing agents may affect the level and pattern of gene expression. Although many studies have examined the ecological effects of heat stress on mussels, little is known about the physiological mechanisms that might be affected by co-exposure to heat stress and environmental contaminants such as nickel (Ni). In the present work we investigated the effects of simultaneous changes in temperature and Ni supply on lysosomal membrane stability (LMS) and malondialdehyde accumulation (MDA) in the digestive gland (DG) of the blue mussel Mytilus galloprovincialis (Lam.). To shed some light into how the molecular response to environmental stressors is modulated, we employed a cDNA microarray with 1,673 sequences to measure the relative transcript abundances in the DG of mussels exposed to Ni along with the temperature increase. Temperature and Ni rendered additive effects on LMS and MDA accumulation, increasing the toxic effects of metal cations. Ni loads in DG tissues was also affected by co-exposure to 26M-BM-0C. In animals exposed only to heat stress, functional genomics analysis of the microarray data (171 DEGs) revealed 7 biological processes, largely dominated by the up-regulation of folding protein-related genes, and the down regulation of genes involved in cell migration and cellular component assembly. Exposure to Ni at 18M-BM-0C and 26M-BM-0C rendered respectively 188 and 262 DEGs showing distinct pattern in term of biological processes. In particular, the response of mussels exposed to Ni at 26M-BM-0C was characterized by the up regulation of proteolysis, ribosome biogenesis, response to unfolded proteins and catabolic-related genes as well as the down-regulation of genes encoding cellular metabolic processes. Our data provide new insights on the transcriptomic response in mussels challenging temperature increases and Ni exposure and should be carefully considered in view of the biological effects of heat stress and particularly in polluted areas. Digestive gland tissue from individual animals in different experimental conditions were analyzed in a complete loop design. Dual color competitive hybridizations (Control M-bM-^@M-^\16M-BM-0CM-bM-^@M-^] vs M-bM-^@M-^\20M-BM-0CM-bM-^@M-^] vs M-bM-^@M-^\24M-BM-0CM-bM-^@M-^] vs M-bM-^@M-^\16M-BM-0C+CuM-bM-^@M-^], M-bM-^@M-^\20M-BM-0CM-bM-^@M-^] vs M-bM-^@M-^\24M-BM-0CM-bM-^@M-^] vs M-bM-^@M-^\20M-BM-0C+CuM-bM-^@M-^], M-bM-^@M-^\24M-BM-0CM-bM-^@M-^] vs M-bM-^@M-^\24M-BM-0C+CuM-bM-^@M-^], M-bM-^@M-^\16M-BM-0C+CuM-bM-^@M-^] vsM-bM-^@M-^]20M-BM-0C+CuM-bM-^@M-^] vs M-bM-^@M-^\24M-BM-0C+CuM-bM-^@M-^]; M-bM-^@M-^\20M-BM-0C+CuM-bM-^@M-^] vs M-bM-^@M-^\24M-BM-0C+CuM-bM-^@M-^]) including label swap. Single individuals. Four biological replicates. One replicate per array.
2013-06-19 | E-GEOD-48062 | biostudies-arrayexpress
Project description:Cinnamaldehyde is a natural antimicrobial and has been found to be effective against many foodborne pathogens including Escherichia coli O157:H7. Although its antimicrobial effects have been well investigated, limited information is available on its effects at the molecular level. Sublethal treatment at 200 mg/l cinnamaldehyde inhibited growth of E. coli O157:H7 at 37oC and for M-bM-^IM-$ 2 h caused cell elongation, but from 2 to 4 h growth resumed and cells reverted to normal length. To understand this transient behaviour, genome-wide transcriptional analysis of E. coli O157:H7 was performed at 2 and 4 h exposure to cinnamaldehyde. Drastically different gene expression profiles were obtained at 2 and 4 h. At 2 h exposure, cinnamaldehyde induced overexpression of many oxidative stress-related genes, reduced DNA replication, and synthesis of protein, O-antigen and fimbriae. At 4 h, many cinnamaldehyde-induced repressive effects on E. coli O157:H7 gene expressions were reversed and oxidatve stress genes were nolonger differentially expressed. Duplicate E. coli O157:H7 cultures with or without 200 mg/l cinnamaldehyde were incubated at 37M-BM-0C for M-bM-^IM-$ 4 h. Cinnamaldehyde-induced changes in gene expression profiles were compared at 2 and 4 h using Affymetrix Ginechip 2.0 microarrays.
2012-09-08 | E-GEOD-40693 | biostudies-arrayexpress
Project description:Several bacterial human pathogens regulate the production of virulence factors by temperature, expressing them only at 37 M-BM-0C. Accordingly we show that the production of all P. aeruginosa virulence factors that are dependent on the QS transcriptional regulator RhlR, but only a fraction that are activated by LasR, are induced at 37 M-BM-0C compared to 30 M-BM-0C or 25 M-BM-0C. The RhlR-dependent induction at 37 M-BM-0C is a posttranscriptional effect due to an RNA thermometer of the ROSE family that thermoregulates the expression of rhlAB operon involved in rhamnolipids production, a virulence associated trait. This RNA structure also affects the expression of the downstream rhlR gene. A second thermometer is present upstream lasI and causes a reduced expression of this gene at lower temperatures without causing a significant decrease of the autoinducer 3-oxo-dodecanoyl homoserine lactone. Using transcriptomic analysis and quantitative real time PCR we show that these RNA thermometers are the main mechanism of thermoregulation of P. aeruginosa virulence factor production For the transcriptome analysis, cells were growth in PPGAS media at 25 M-BM-0C and 37 M-BM-0C, respectively. Samples were harvested at O.D600 of 1.5. Three biological replicates of each growth condition was used for RNA extraction and hybridization on Affimetrix microarrays.
2013-04-02 | E-GEOD-45695 | biostudies-arrayexpress