Testicular toxicity induced by carbendazim, iprodione, alone or in combination
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ABSTRACT: Using an ex-vivo testicular culture from rats, we carried out transcriptomic experiments to identify the pathway of toxicity ellicited by the fungicides carbendazim, iprodione alone or in combination. we used commercial Agilent Microarray GE 4x44K Rat (V3) Gene Expression Microarray (G2514F) AMADID : 028282 Cultures were performed with and without fungicides. When required, CBZ, IPR or mixture was added beginning from day 2, at 50 nM or 500 nM for CBZ and at 50 nM or 500 nM for IPR; 50 nM each or 500 nM each for the mixture. For microarray experiments, three different pools of seminiferous tubules were exposed to two concentrations of CBZ, IPR or their mixture (50 nM and 500 nM) or to complete medium with vehicle (control cells) for 7, 14, and 21 days. Each condition of exposure to fungicides was compared with control cells at the same time point.
Project description:Carbamazepine (CBZ) is widely used in the treatment of neuropsychiatric disorders as an anticonvulsant. A recent report revealed that CBZ has an anti-fibrotic effect on the livers of M-NM-11-antitrypsin-deficient mice, raising the possibility of a hepatology-related therapeutic use for the drug.Given the fact that CBZ has complicated pharmacokinetic properties, to comprehensively understand the its mechanism of action, we performed microarray analysis of the mouse liver tissues 2 hours after CBZ administration. The mice were orally administered 250 mg/kg of CBZ or DMSO and examined for the gene expression 2 hours later; 3 mice per group.
Project description:Changes in the transcriptional profiles of Xiphophorus maculatus skin was assessed by RNA-Seq after exposure to fluorescent (i.e., FL, 4,100K or “cool white”), or narrow wavelength regions of light between 300-600 nm (i.e., 50 nm or 10 nm regions, herein termed ”wavebands”). Exposure to varied 50 nm wavebands allowed identification of waveband specific transcriptional modulation within gene sets representing discrete functional pathways within Xiphophorus skin. For example, exposure to either 350-400 or 450-500 nm wavebands resulted in opposite transcriptional effects in necrosis and apoptosis gene sets that predict selective differential suppression or activation of these pathways (i.e., 350-400 nm; necrosis suppression, apoptosis activation, and 450-500 nm; apoptosis suppression, necrosis activation). Further investigation of waveband specific transcription employing successive 10 nm waveband exposures within the genetically responsive 500-550 nm waveband show; (a) greater numbers of genes may be transcriptionally modulated after 10 nm exposures, than observed for either 50 nm or FL exposures. (b) the 10 nm wavebands incite greater functional specificity than either 50 nm or FL exposures. (c) The principal genetic effects of FL are primarily due to the 30 nm between 500 to 530 nm.
Project description:we assessed characteristic molecular and proteomic signatures in rat liver treated with drugs (pyrazinamide, ranitidine, enalapril, carbamazepine, and chlorpromazine) that are known to cause DILI in humans. In the present study, we assessed the characteristic gene expression signature for DILI in a rat model. Rats were administered representative drugs that are already known to induce DILI in humans and transcriptomic changes in rat liver were analyzed. The representative drugs, which induce three types (hepatocellular, mixed, and cholestatic) of DILI, that were used in this study were pyrazinamide (PZA, 150~1500 mg/kg), ranitidine (RAN, 209.5~2095 mg/kg), enalapril (ENA, 148.65~1486.5 mg/kg), carbamazepine (CBZ, 97.85~978.5 mg/kg), and chlorpromazine (CPZ, 7.1~71 mg/kg).
Project description:Gene expression profiles of Bacillus subtilis strain AG174 were compared as a function of steady-state external pH during growth with aeration. Aerated overnight cultures were diluted 1:500 and incubated in 250 mL baffled flasks containing potassium-modified Luria-Bertani medium (LBK) buffered with 50 mM HOMOPIPES at pH 6.0, pH 7.0, and pH 9.0. Tubes were incubated at 37°C with aeration (260 rpm) to an optical density at 600 nm of 0.2. For each of the three pH conditions, RNA was isolated from five independent cultures. Overnight cultures were diluted 1:500 and incubated in 250 mL baffled flasks containing potassium-modified Luria-Bertani medium (LBK) buffered with 50 mM HOMOPIPES at pH 6.0, pH 7.0, and pH 9.0. Cultures were incubated at 37°C with aeration (260 rpm) to an optical density at 600 nm of 0.2. Five replicate cultures were grown at each pH.
Project description:The combination of increasing consumption rates and limited elimination under conventional waste water treatment practices of many pharmaceutical compounds has now led to their detection in aquatic environments. Three of the most frequently detected pharmaceuticals in the environment are Acetaminophen (APAP), Atenolol (AT) and Carbamazepine (CBZ). Atlantic salmon (parr) was exposed to environmentally relevant levels of Acetaminophen (APAP) (54.77 ± 34.67 µg·L-1), Atenolol (AT) (11.08 ± 7.98 µg·L-1) and Carbamzepine (CBZ) (7.85 ± 0.13 µg·L-1). Gene expression was analyzed in liver tissues using a 16K GRASP (University of Victoria, Canada) cDNA microarray. GRASP 16K v.2 cDNA microarrays were used for this study (Accession # A-GEOD-2716). A dual-labelled experimental design was employed for the microarray hybridisations. Each experimental cDNA sample (Cy3 labeled) was competitively hybridised against a common pooled-reference sample (Cy5 labeled). The entire experiment comprised 20 hybridisations - 4 states (APAP, AT, CBZ, control) × 1 time-point ( at 5 days) × 5 biological replicates (males only). Hybridisations were undertaken concurrently.
Project description:We profiles the effects of six protein phosphatase inhibitors on EGF-dependent phosphoproteome dynamics. The cells were treated with 20 nM EGF for 8 or 20 min with or without 15 min pre-treatment of one of the phosphatase inhibitors (100 nM or 1 M; Raphin1 acetate, sanguinarine chloride, KY-226, SHP099, NSC95397, or BCI). We further focused on sanguinarine chloride, and Hela cells were treated with 20 nM EGF for 3, 8, or 20min with or without 15 min pre-treatment of sanguinarine (5, 50, or 500 nM). We also examined combination of sanguinarine chloride (500 nM) and BIRB796 (500 nM), a p38-kinase inhibitor.
Project description:We performed RNA-seq of MDA-MB-231 cells that were treated with MS645 or JQ1 at 50 nM and 500 nM in an effort to understand how MS645 exerts such a profound cell growth inhibition on cancer cells.
Project description:Genotoxic agents cause cellular DNA damage and stress responses, including transcriptional changes. Here we focused on the early transcriptional responses of human cells to benzo(a)pyrene diol epoxide (BPDE), which causes bulky DNA adduct damage. Human amnion epithelial FL cells were exposed to three doses of BPDE (5, 50, and 500 nM) and the vehicle control DMSO, and differential gene expression profiles were obtained 4 h after exposure using oligonucleotide microarrays followed by validation with quantitative real-time RT-PCR. Compared with a few genes affected by the low and medium-dose exposure, extensive and robust changes in gene expression were induced by the high-dose BPDE. We found that the expression of cell cycle-regulators, signaling molecules and transcription factors were significantly altered and important signaling pathways related to cell survival or apoptosis were affected by BPDE. Several genes and related regulatory pathways that were previously not known to be responsive to this genotoxic agent have now been implicated, which helps to draw the whole picture of how cells respond to environmental chemical exposure via transcriptional regulation. Experiment Overall Design: Human amnion epithelial FL cells were exposed to vehicle control (dimethyl sulfoxide) and increasing doses (5, 50, 500 nM) of anti-benzo(a)pyrene diol epoxide (anti-BPDE), respectively. The transcriptomes of the three treatments were compared to that of the control, respectively, to test the hypothesis that a characterized differential expression profile would be generated by exposure to various doses of this genotoxic agent.
Project description:Primary HUVEC were treated with 500 nM MLN7243, 500 nM MLN4924 or DMSO (control) for 6 h. The experiment aims to identify up- or downregulated proteins after treatment with the E1 ligase inhibitor MLN7243 and the cullin inhibitor MLN4924 in HUVEC. Proteins that are regulated by ubiquitination or more specifically by cullin-based E3 ligases in the quiescent endothelium, will be identified.
Project description:Neurosphere cultures were established from individually isolated E 14.5 forebrains of mouse embryos that carry a bcl2 transgene. Single-cell suspensions were prepared, seeded at 1x105 cells/ml and treated for two days with 150 nM Trichostatin A (TSA; histone deacetylase inhibitor), 500 nM 5-Aza-2-deoxycytidine (AzaC; demethylating agent) or both compounds, or left untreated. Two independent experiments were performed.