Achyranthes aspera leaf extract modulates the expression of genes involved in immune response and embryonic development etc in vitro in cultured human pancreatic cancer cell line MIA PaCa-2 (CRL1420).
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ABSTRACT: Examining of the global gene expression changes in cultured human pancreatic cancer cells effected by the extract of A. aspera leaves. Comparison of cells treated for 24, 48 and 72 hours with the respective controls (untreated) allowed to select key genes that may be of interest as a response markers. Two-condition experiment, treated vs. untreated cells at 3 time points. Biological replicates: 6 control and 6 treated replicates for 24 hours time point; 4 control and 4 treated replicated for 48 hours time point; 2 control and 2 treated replicates for 72 hours time point.
Project description:The aim of this study was to analyze the host responses to ionizing radiation by nuclear factor-κB (NF-κB) bioluminescence imaging-guided transcriptomic tool. Transgenic mice, carrying the NF-κB-driven luciferase gene, were exposed to a single dose of 8.5 Gy total-body irradiation. In vivo imaging showed that a maximal NF-κB-dependent bioluminescent intensity was observed at 3 h after irradiation and ex vivo imaging showed that liver, intestine, and brain displayed strong NF-κB activations. Microarray analysis of these organs showed that irradiation altered gene expression signatures in an organ-specific manner. Pathway analysis showed that pathways associated with metabolism and immune system were altered primarily in liver and intestine. the upregulation of fatty acid binding protein 4, serum amyloid A2, and serum amyloid A3, which are participated in both inflammation and lipid metabolism, suggesting that irradiation might affect the cross pathways of metabolism and inflammation. Moreover, The upregulation of chemokine (CC-motif) ligand 5, chemokine (CC-motif) ligand 20, and Jagged 1 genes suggested that these genes might contribute to the radiation enteropathy. Male transgenic mice (6 to 8 weeks old) were exposed to a single dose of whole-body X-rays generated at 6 MV (Clinac® 21EX medical linear accelerator, Varian, Palo Alto, CA, USA) and at a dose rate of 4 Gy/min. Mice were imaged at 0 h, 1 h, 3 h, 9 h, 24 h, 48 h, or 72 h, or on 7 d or 14 d after irradiation with 8.5 Gy. RNAs were extracted at 3 h after irradiation.
Project description:Genipin is a natural blue colorant in food industry. Inflammation is correlated with human disorders, and nuclear factor-κB (NF-κB) is the critical molecule involved in inflammation. In this study, the anti-inflammatory effect of genipin on the lipopolysaccharide (LPS)-induced acute systemic inflammation in mice was evaluated by NF-κB bioluminescence-guided transcriptomic analysis. Transgenic mice carrying the NF-κB-driven luciferase genes were administered intraperitoneally with LPS and various amounts of genipin. Bioluminescent imaging showed that genipin significantly suppressed LPS-induced NF-κB-dependent luminescence in vivo. The suppression of LPS-induced acute inflammation by genipin was further evidenced by the reductions of cytokine levels in sera and organs. Microarray analysis of these organs showed that the transcripts of 79 genes were differentially expressed in both LPS and LPS/genipin groups, and one third of these genes belonged to chemokine ligand, chemokine receptor, and interferon (IFN)-induced protein genes. Moreover, network analysis showed that NF-κB played a critical role in the regulation of genipin-affected gene expression. In conclusion, we newly identified that genipin exhibited anti-inflammatory effects in a model of LPSinduced acute systemic inflammation via downregulation of chemokine ligand, chemokine receptor, and IFN-induced protein productions. A total of 25 transgenic mice (female, 6 to 8 weeks old) were randomly divided into five groups of five mice: (1) mock, no treatment; (2) LPS (4 mg/kg), (3) LPS plus genipin (1 mg/kg), (4) LPS plus genipin (10 mg/kg), and (5) LPS plus genipin (100 mg/kg). Mice were challenged intraperitoneally with LPS and then with genipin 10 min later. Four hours later, mice were imaged for the luciferase activity, and subsequently sacrificed for ex vivo imaging, RNA extraction, and immunohistochemical staining.
Project description:In this study, a novel IR-binding protein (IRBP) from Momordica charantia, named as mcIRBP, was identified by analyzing the physical and functional interactions between mcIRBP and IR. The hypoglycemic effect and mechanism of mcIRBP were further evaluated in normal and streptozotocin-induced diabetic mice. Normal and diabetic mice were treated without or with mcIRBP and RNAs from muscle tissues were extracted for microarray analysis. Number of replicate was three.
Project description:Inflammatory bowel disease is a chronic colonic inflammation that displays symptoms like diarrhea and weight loss. Acupuncture has been widely accepted by Western countries for the treatment of pain. In this study, we analyzed the efficacy and mechanism of electroacupuncture (EA) on trinitrobenzene sulphonic acid (TNBS)-induced colitis in mice. Mice were intrarectally administered 250 mg/kg TNBS and electroacupunctured at Quze (PC3) and Neiguan (PC6) acupoints, which have been applied for gastrointestinal disorders. Gene expression profiles in colons and spleens were analyzed by microarray for the elucidation of mechanism of EA. Our data showed that EA at PC3 and PC6 improved macroscopic and microscopic features of colitis, and the improvement displayed a frequency-dependent manner. Administration of TNBS upregulated the expression of most cytokine genes in colons, while EA downregulated the expression of TNBS-induced cytokine genes. Pathway analysis showed that EA significantly affected inflammatory pathways in colons and immunity-associated pathway in spleens. Immunohistochemical staining further showed that EA decreased the expression of interleukin-1? and nuclear factor-?B. In conclusion, this is the first study reporting the global gene expression profiles of EA on TNBS-induced colitis. Our findings suggested that inflammatory and immunity pathways were involved in the anti-inflammatory mechanism of EA on colitis induced by TNBS. In this study, we analyzed the efficacy and mechanism of electroacupuncture (EA) on trinitrobenzene sulphonic acid (TNBS)-induced colitis in mice. Mice were intrarectally administered 250 mg/kg TNBS and electroacupunctured at Quze (PC3) and Neiguan (PC6) acupoints, which have been applied for gastrointestinal disorders. Gene expression profiles in colons and spleens were analyzed by microarray for the elucidation of mechanism of EA.
Project description:An important, but rarely performed, test of Koch’s molecular postulates involves evaluating the capacity of candidate virulence genes to confer pathogenicity in otherwise non-virulent species. Unbiased genomic surveys of avirulent natural isolates might reveal rare variants possessing specific virulence features, which might prove useful in testing their functional sufficiency. Using a custom pan-genome array, we analyzed a panel of avirulent Burkholderia thailandensis (Bt) isolates related to Burkholderia pseudomallei (Bp), the causative agent of the often fatal human and animal disease melioidosis. We report the discovery of variant Bt isolates exhibiting isolated acquisition of a capsular polysaccharide biosynthesis gene cluster (BpCPS), long regarded as an critical species-specific virulence factor essential for Bp mammalian virulence. BpCPS-expressing Bt strains exhibited certain pathogen-related phenotypes including resistance to human complement binding, but did not exhibit enhanced virulence when assessed in two different in vivo animal infection models. Phylogenetic analysis revealed that the BpCPS-expressing Bt strains likely reside within an evolutionary subgroup distinct from the majority of previously-described Bt strains. Our findings suggest that BpCPS acquisition alone is unlikely to fully explain the ability of Bp to colonize humans and animals, highlighting the importance of other collaborating factors in the pathogenesis of mammalian melioidosis. Genomic DNA of several Bt strains were hybridized against a common reference strain (Bt E264), to see gain/loss
Project description:Aiming at the development of a micropollutant biosensor in the frame of the Micro-Ecological Life Support System Alternative (MELiSSA), a pilot study was initiated to identify triclosan (TCS)-responsive biomarker genes in the MELiSSA carbon-mineralizing microorganism, Rhodospirillum rubrum S1H. TCS is a biocide, commonly found in human excrements and is considered an emerging pollutant in wastewater and the environment. Chronic exposure to MELiSSA-relevant concentrations (≥25 µg L-1) TCS caused a significant extension of the lag phase without affecting the growth rate. Analytical determination gave no indication of TCS biodegradation during the growth experiment and flow cytometric viability analyses revealed that TCS is only slightly toxic to R. rubrum. Through microarray analyses, the genetic mechanisms supporting the reversibility of TCS-induced inhibition were scrutinized. Hence, an extremely TCS-responsive cluster of four small adjacent genes was revealed, with up to 34-fold induction at 25 µg L-1. These four genes, for which the name micropollutant-upregulated factor (muf) was proposed; appear to be unique to R. rubrum and are shown for the first time to be involved in the response to stress. Moreover, numerous other systems that are associated with the proton-motive force were shown to be responsive to TCS, but never as highly upregulated as the muf genes. Hence, R. rubrum induced the phage shock protein operon (pspABC), numerous major facilitator efflux systems, cell envelope consolidation mechanisms, oxidative stress response, beta-oxidation, and carbonic anhydrase; while downregulating bacterial conjugation- and carboxysome synthesis genes. The muf genes and three efflux-related genes showed most potential as low-dose biomarkers. The two microarray experiments (10 and 25 µg l-1 Triclosan) were all performed in biological triplicate and containing three (in-slide) technical repeats. For all conditions, the Triclosan exposed sample (Cy5) was compared with the non-exposed solvent control (Cy3) to identify those genes that were differentially expressed upon Triclosan exposure.
Project description:The genomic distribution of post-translationally modified histones in the zebrafish (Danio rerio) genome remain largely unraveled. The objective of this study was to (1) design a microarray covering all zebrafish gene promoters and 5' end of coding regions, and (ii) using these arrrays, map the genome-wide distribution of trimethylated H3K4 and H3K27 in the embryo derived ZF4 cell line. Specifically, we determined the enrichment profile of H3K4me3, H3K27me3 and of both (co-enriched) modifications in ZF4 cells. We determined average enrichment profiles through metagene analysis, functional terms enriched among genes marked by either modification, pathway analysis, and in relation to gene expression. Gene expression experiments were performed on three biological replicates.
Project description:Transcriptional profiling of salivary gland, midgut and ovary tissues isolated from Rhipicephalus microplus females at day 20 post infestation. This enabled the identification of transcripts that are tissue-specific or shared among the tissues tested. Reference pool design: Each tissue tested was compared to a reference pool comprising ticks (immature to adult stages) sampled on day 4, 5, 7, 13, 15 and tissues collected on day 20 post infestation. Biological replicates: 2; Technical replicates: 2.
Project description:Background: The high number of heavy metal resistance genes in the soil bacterium Cupriavidus metallidurans CH34 makes it an interesting model organism to study microbial responses to heavy metals. Results: In this study the transcriptional response of this bacterium was measured after challenging it to a wide range of sub-lethal concentrations of various essential or toxic metals. Considering the global transcriptional responses for each challenge as well as by identifying the overlap in upregulated genes between different metal responses, the sixteen metals could be clustered in three different groups. Additionally, next to the assessment of the transcriptional response of already known metal resistance genes, new metal response gene clusters were identified. The majority of the metal response loci showed similar expression profiles when cells were exposed to different metals, suggesting complex cross-talk at transcriptional level between the different metal responses. The highly redundant nature of these metal resistant regions – illustrated by the large number of paralogous genes – combined with the phylogenetic distribution of these metal response regions within evolutionary related and other metal resistant bacteria, provides important insights on the recent evolution of this naturally soil dwelling bacterium towards a highly metal-resistant strain found in harsh and anthropogenic environments. Conclusions: The metal-resistant soil bacterium Cupriavidus metallidurans CH34 displays myriads of gene expression patterns when exposed to a wide range of heavy metals at non-lethal concentrations. The interplay between the different gene expression clusters points towards a complex cross-regulated regulatory network governing heavy metal resistance in C. metallidurans CH34. Keywords: Cupriavidus metallidurans CH34, transcriptional regulation, heavy metal resistance Two-condition experiments. Comparing samples after induction with heavy metals versus non-induced samples. Biological duplicate or triplicate. Each array contains 3 or 4 technical replicates.
Project description:A total of 30 male IL10-/- (B6.129P2-Il10<tm1Cgn>/J) and 30 male C57 control (C57BL/6J) mice were obtained from The Jackson Laboratory (Bar Harbor, Maine, USA). All mice were between 28 and 35 days of age at the start of the study. For convenience and consistency in reporting, their age was defined as 35 days or 5 weeks of age. The mice were housed individually in standard shoebox size cages containing untreated wood shavings and maintained in an air-conditioned animal room with a 12-h light-dark cycle. Animals had free access to water and were fed an AIN-76A standard powder diet prepared in-house. This study was approved by the AgResearch Ruakura Animal Ethics Committee in Hamilton, New Zealand according to the Animal Protection Act (1960) and Animal Protection Regulations (1987) and amendments. Four days after arrival, all mice were inoculated orally with a mixture of Enterococcus faecalis strains and bacteria commonly found in the intestinal lumen to obtain a more consistent and reproducible intestinal inflammation. Mice were randomly assigned to five sampling groups (7, 8.5, 10, 12 and 14 weeks of age). Experiment Overall Design: For the comparison of gene expression levels, colonic RNA of IL10-/- and C57 at 7 and 12 weeks of age were compared because of a low HIS variability in the colon within those time points. A reference design with 15 arrays was used, where each individual RNA sample was hybridized with a reference sample onto the array, in a total of 3 or 4 biological replicates per treatment. Reference RNA was prepared by pooling equal amounts of purified total RNA extracted from small intestine, colon, kidney, liver and fetuses from normal, healthy Swiss mice.