Human cell toxicogenomic analysis links reactive oxygen species to the toxicity of monohaloacetic acid drinking water disinfection byproducts
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ABSTRACT: The monohaloacetic acids (monoHAAs) are generated as byproducts during the disinfection of drinking water and are cytotoxic, genotoxic, mutagenic, and teratogenic. Iodoacetic acid (IAA) toxicity was mitigated by antioxidants, suggesting the involvement of oxidative stress. Other monoHAAs may share a similar mode of action. Human oxidative stress and antioxidant defense gene arrays (SA biosciences) were used to evaluate changes in transcriptome profiles in the human intestinal epithelial cell line FHS 74 INT generated by three compounds, chloroacetic acid (CAA), bromoacetic acid (BAA) and IAA at two time points (30 min and 4 h). Twelve samples were evaluated. Each treated sample was paired with a concurrent negative control (cells treated in medium only). Three technical repeats were included for each sample and Ct values were calculated from the average of the three repeats. Samples 1,2,and 3 were isolated from 30 min negative controls for CAA, BAA, and IAA respectively. Samples 4, 5, and 6 were isolated from cells treated for 30 min with CAA, BAA, and IAA respectivley. Samples 7, 8, and 9 were isolated from 4h negative controls for CAA, BAA, and IAA respectively. Samples 10, 11, and 12 were isolated from cells treated for 4 h with CAA, BAA and IAA respectively. Ct values were normalized against the average of the 5 housekeeping genes included in the array to generate M-NM-^TCt values. Fold changes for each gene were calculated as a ratio of 2^-M-NM-^TCttest / 2^-M-NM-^TCtcontrol.
Project description:Wild type (WT) and Pglyrp1-/- mice were treated with PBS or sensitized 5 days/week for 3 or 5 weeks with 10 M-BM-5l per application of 2.5 mg/ml of purified house dust mite allergen. 3 days after the last sensitization the lungs were removed and homogenized, and RNA was isolated from the right lobes using the TRIZOL method. Quantitative reverse transcription real-time PCR (qRT-PCR) was used to quantify the amounts of mRNA in the lungs using custom RT2 Profiler PCR Arrays designed by us and manufactured by Qiagen/SA Biosciences. qRT-PCR gene expression profiling
Project description:Wild type (WT), Nod2-/-, Pglyrp3-/-, and Pglyrp3-/-Nod2-/- mice (BALB/c) were treated with oral 5% dextran sodium sulfate (DSS) for 48, 72, or 96 hrs, their colons were removed and homogenized, and RNA was isolated using the TRIZOL method. Quantitative reverse transcription real-time PCR (qRT-PCR) was used to quantify the amounts of mRNA in the colon using the inflammatory gene expression RT2 Profiler PCR Array from Qiagen/SA Biosciences. qRT-PCR gene expression profiling
Project description:Background: Identifying the immune components that are regulated by a2NTD, a peptide cleaved from the N-terminus of the a2 vacuolar ATPase. Methods: In this study, we used pathway-focused PCR arrays to determine the genes that are regulated in human monocytic cell line, THP-1 after a2NTD stimulation over time. Results: a2NTD up-regulated several cytokines including IL-1 alpha, IL-1 beta, and IL-10. Several chemokines were also upregulated including the MCP and MIP families. Conclusion: We believe a2NTD to be an immune modulator made by tumor cells that aid in preventing immune surveillance by up-regulating proteins in monocytes that are both pro-and anti-inflammatory in nature. The Human Inflammatory Response and Autoimmunity RTM-BM-2 ProfilerM-bM-^DM-" PCR Array profiles the expression of 84 key genes involved in autoimmune and inflammatory immune responses. It represents the expression of inflammatory cytokines and chemokines as well as their receptors. It also contains genes related to the metabolism of cytokines and involved in cytokine-cytokine receptor interactions. Thoroughly researched panels of genes involved in the acute-phase response, inflammatory response, and humoral immune responses are represented as well. Using real-time PCR, you can easily and reliably analyze expression of a focused panel of genes related to inflammatory and autoimmune responses with this array.
Project description:ATP-binding cassette (ABC) transporters contribute to development of resistance to anti-cancer drugs via ATP-dependent drug efflux. A major goal of our study was to investigate associations between expression of ABC transporters and outcome of breast carcinoma patients. ABCA5/6/8/9/10, ABCB1/5/11, ABCC6/9, ABCD2/4 and ABCG5/G8 were significantly downregulated and ABCA2/3/7/12, ABCB2/3/8/9/10, ABCC1/4/5/10/11/12, ABCD1/3, ABCE1, ABCF1/2/3 and ABCG1 upregulated in post-treatment tumors compared with non-neoplastic tissues. Significant associations of ABCC1 and ABCC8 levels in tumors with grade and expression of hormonal receptors were found. ABCA13 and ABCD2 levels significantly associated with the response to neoadjuvant chemotherapy in post-treatment patients. Transcript levels of all forty nine human ABCs were determined by qPCR in post-treatment tumor and non-neoplastic tissue samples from 68 breast carcinoma patients treated by neoadjuvant chemotherapy. EIF2B1, MRPL19, IPO8, and UBB were used as reference genes for data normalization.
Project description:The monohaloacetic acids (monoHAAs) are generated as byproducts during the disinfection of drinking water and are cytotoxic, genotoxic, mutagenic, and teratogenic. Iodoacetic acid (IAA) toxicity was mitigated by antioxidants, suggesting the involvement of oxidative stress. Other monoHAAs may share a similar mode of action. Human oxidative stress and antioxidant defense gene arrays (SA biosciences) were used to evaluate changes in transcriptome profiles in the human intestinal epithelial cell line FHS 74 INT generated by three compounds, chloroacetic acid (CAA), bromoacetic acid (BAA) and IAA at two time points (30 min and 4 h).
Project description:The goal of this study was to identify cerebellar gene expression differences between mutant MeCP2 A140V mice and their wild type littermates.The PCR array used for these experiments was the Mouse Epigenetic Chromatin Modification Enzymes Array (Catalog# PAMM-085A) from SABiosciences. Mice used in this study were 4 week old males. Total RNA was extracted from whole cerebella of 4 week old male MeCP2 A140V hemizygous mutant mice and their wild type littermates. For this study, cerebella from 3 mice of each genotype were used and RNA from the cerebellum of each mouse was analyzed separately (ie., triplicate biological replicates). The RNA was purified to remove genomic DNA and was then reverse transcribed for use in quantitative RT-PCR analysis of gene expression using the SABiosciences RT2 profiler PCR array system. The PCR array used for these experiments was the Mouse Epigenetic Chromatin Modification Enzymes Array (Catalog# PAMM-085A).
Project description:The goal of this study was to identify cerebellar gene expression differences between mutant MeCP2 A140V mice and their wild type littermates. The PCR array used for these experiments was the Mouse Epigenetic Chromatin Modification Enzymes Array (Catalog# PAMM-085A) from SABiosciences. Mice used in this study were 2 week old males. Total RNA was extracted from whole cerebella of 2 week old male MeCP2 A140V hemizygous mutant mice and their wild type littermates. For this study, cerebella from 3 mice of each genotype were used and RNA from the cerebellum of each mouse was analyzed separately (ie., triplicate biological replicates). The RNA was purified to remove genomic DNA and was then reverse transcribed for use in quantitative RT-PCR analysis of gene expression using the SABiosciences RT2 profiler PCR array system. The PCR array used for these experiments was the Mouse Epigenetic Chromatin Modification Enzymes Array (Catalog# PAMM-085A).
Project description:The goal of this study was to identify brain cortex gene expression differences between mutant MeCP2 A140V mice and their wild type littermates.The PCR array used for these experiments was the Mouse Cytoskeleton Regulators Array (Catalog# PAMM-088A) from SABiosciences. Mice used in this study were 2 week old males. Total RNA was extracted from the somatosensory/motor cortex of 2 week old male MeCP2 A140V hemizygous mutant mice and their wild type littermates. For this study, cortical tissue from 3 mice of each genotype was used and RNA from the cortex of each mouse was analyzed separately (ie., triplicate biological replicates). The RNA was purified to remove genomic DNA and was then reverse transcribed for use in quantitative RT-PCR analysis of gene expression using the SABiosciences RT2 profiler PCR array system. The PCR array used for these experiments was the Mouse Cytoskeleton Regulators Array (Catalog# PAMM-088A).