Gene Expression analysis of znuA of Moraxella catarrhalis
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ABSTRACT: Analysis of the gene expression pattern of a znuA mutant constructed in the O35E background. It's pattern of expression was compared to that of O35E. 3 biological replicates were prepared. Each replicate was grown in BHI broth and RNA extracted with the Ambion Ribopure bacterial kit.
Project description:This series of microarrays explores the genes regulated by the Zur gene of Moraxella catarrhalis. The gene expression profile of O35E Wild-type cells was utilized as the baseline and compared with the gene expression profile of a non-polar mutant of the Zur gene constructed in the O35E background. There were two biologic replicates and two internal dye swaps performed.
Project description:M. catarrhalis strain O35E.rpsl was inoculated into the nasopharynx of healthy, male adult chinchillas. 24 hours later the nasopharyngeal tissues were extracted and homogenized. Total RNA was extracted from these tissue samples. Subsequently, M. catarrhalis genome directed primers were utilized to synthesize cDNA from the total RNA sample. As a control, M.catarrhalis strain O35E.rpsl was grown in BHI broth to a Klett density of 200 units and underwent RNA extraction per standard protocols. The genome directed primers mentioned above were utilized to synthesize cDNA. Both cDNA samples were subsequently labelled with either Cy3 or Cy5 and hybridized to a custom Microarrays, Inc. gene chip and scanned after 16 hours. Differential gene expression was measured utilizing the broth grown cells as the baseline and the chinchilla isolated cells as the experimental variable. There are 5 individual sample results included in this series. These represent the data from four individual biological replicates (i.e. 4 different sets of inoculated animals). For each replicate the control samples are simultaneously grown broth samples. Three dye swap experiments were performed.
Project description:Series of DNA microarrays (4) comparing the M. catarrhalis strain 035E and M. catarrhalis oxyR mutant response to 50 mM hydrogen peroxide. Wild-type cells and oxyR mutant cells were exposed to 50 mM hydrogen peroxide for 15 minutes. RNA was extracted and DNA microarray analysis performed. 4 biologic replicates were studied. One dye swap was included in this series analysis. Control oxyR mutant cells exposed to water
Project description:Series of DNA microarrays (4) comparing the M. catarrhalis strain 035E response to 50 mM hydrogen peroxide relative to a water-only control. Wild-type cells were exposed to either 50 mM hydrogen peroxide or a water-only control for 15 minutes. RNA was extracted and DNA microarray analysis performed. 4 biologic replicates were studied. One dye swap was included in this series analysis.
Project description:Comparison of the gene expression profile of Moraxella catarrhalis grown in the presence of 20% pooled human sputum in chemically-defined medium relative to Moraxella catarrhalis grown in chemically-defined medium alone. Moraxella catarrhalis ATCC43617 was grown to mid-logarithmic phase either in the presence of 20% pooled human sputum in chemically-defined medium or in chemically-defined medium alone. Total RNA was extracted from bacterial cells exposed to each of these conditions and cDNA was generated for CyDye labelling. 3 biologic replicates were generated and each replicate underwent a dye swap (total of 6 experimental data collections). The gene expression profile reported is that of Moraxella catarrhalis grown in the presence of pooled human sputum in a chemically-defined medium relative to Moraxella catarrhalis grown only in the presence of the chemically-defined medium.
Project description:Gene expression comparison of L. reuteri ATCC PTA 6475 and cyclopropane-fatty-acyl-phospholipid synthase mutant in MRS medium in anaerobic condition at 37C Include 3 biological replicate and dye-swap for each comparison. Reference time point = L. reuteri ATCC PTA 6475
Project description:Study Objectives: Sleep deprivation is highly prevalent and caused by conditions such as night shift work or illnesses like obstructive sleep apnea. Compromised sleep is proposed to play a role in several cardiovascular, immune related and neurodegenerative disorders. We recently published human serum proteome changes after a simulated night shift. This study aimed to further explore changes in the human blood serum after 6h of sleep deprivation at night by proteomics and systems biological databases. Methods: Human blood serum samples from 8 self-declared healthy females were analyzed using mass spectrometry and high-pressure liquid chromatography. Each subject was their own control, and two samples were taken from each subject, the first one after 6h of sleep at night and the second one after 6h of sleep deprivation the following night. Biological databases and bioinformatic software were used for systems biological analyzes and comparative analysis with other published sleep-related datasets. Results: Of 494 proteins, 66 were found to be differentially expressed after 6h of sleep deprivation at night. Functional enrichment analysis revealed associations of these proteins with several biological functions related to the regulation of cellular processes like protein- and ion-binding connected to platelet degranulation and blood coagulation, as well as associations with different curated gene sets. Conclusions: This study presents serum proteomic changes after 6h of sleep deprivation, supports previous findings that only 6h of sleep deprivation affects several biological processes and revealed a molecular signature of protein changes related to pathological conditions like altered coagulation and platelet function, impaired lipid and immune function and cancer. Keywords: Human blood serum, proteomics, sleep deprivation, cellular stress, functional enrichment analysis
Project description:Background In broilers, heat stress can result in reduced feed consumption, digestive inefficiency, impaired metabolism, and even death. The broiler sector of the U.S. poultry industry incurs approximately $52 million in heat stress-related losses annually. The objective of this study is to characterize the effects of chronic, cyclic heat stress on the transcriptome of a metabolically active organ, the liver. Characterizing the liver transcriptome of heat-stressed broilers will help clarify the effects of heat stress on metabolism. This information will provide a platform for future investigations that further elucidate physiologic responses to heat stress and seek methods to ameliorate the negative impacts of heat. Results Transcriptome sequencing of the livers of 8 broiler males using Illumina HiSeq 2000 technology, resulted in a total of 138 million, 100 base pair single end reads, yielding 13.8 gigabases of sequence. Forty genes were differentially expressed at a significance level of P-value < 0.05 and a fold change ≥ 2 in response to chronic, cyclic heat stress (mid-point of the last day of a 7-day cyclic heat stress of 7 hours per day), with 27 down-regulated and 13 up-regulated. Two gene networks were created from the function-based Ingenuity Pathway Analysis (IPA) of the differentially expressed genes; “Cell Signaling, Molecular Transport, Small Molecule Biochemistry” and “Endocrine System Development and Function, Small Molecule Biochemistry Cell Signaling”. Members of the MAPK signaling pathway and differentially expressed genes that are associated with MAPK-related functions were prominent in the networks. Cellular proliferation and differentiation, inflammationand stress-related signaling, and apoptosis-associated genes were down-regulated in response to heat stress. Genes responsible for inhibiting feed intake and sphingolipidrelated signaling were up-regulated. Genes involved with the regulation of inflammation, stress, thyroid hormone level, and body temperature were both up- and down-regulated. Conclusions Chronic, cyclic heat stress of broilers results in metabolic changes that can be characterized through RNA-seq analysis of the liver transcriptome. The primary affected pathways included cell signaling, molecular transport, endocrine system development and signaling, and small molecule biochemistry. Examination of 2 heat treatments. Four heat stressed liver samples and 4 control liver samples analyzed.
Project description:Astrocytes can support neuronal survival through a range of secreted signals that protect against neurotoxicity, oxidative stress, and apoptotic cascades. To identify proteins contributing to protective intracellular neuronal signalling originating from astrocytes, endogenous PI3K was immunoprecipitated from Ht22 cells exposed to primary astrocyte conditioned media (ACM) or cell free media (CFM), followed by iTRAQ-based quantitative proteomic analysis.
Project description:Gene expression comparison of L. reuteri ATCC PTA 6475 grown (stationary phase) in a defined media with either glucose or sucrose as the carbon source, in anaerobic conditions at 37C. Includes 2 biological replicates and dye-swaps for each comparison. Reference time point is is L. reuteri ATCC PTA 6475 grown in a glucose-based media.