scholarly journals Effect of Chloride Concentration on the Cytotoxicity, Bioavailability, and Bio-Reactivity of Copper and Silver in the Rainbow Trout Gut Cell Line (RTgutGC)

2021 ◽  
Author(s):  
Md Ibrahim ◽  
Matteo Minghetti
Keyword(s):  
Rainbow Trout ◽  
Cell Line ◽  
Metal Speciation ◽  
Membrane Integrity ◽  
Chloride Concentration ◽  
Mrna Levels ◽  
Chloride Complexes ◽  
Visual Minteq ◽  
Cu Uptake ◽  
High Chloride

Abstract Chloride (Cl-) influences the bioavailability and toxicity of metals in fish, but the mechanisms by which it influences these processes is poorly understood. Here, we investigated the effect of chloride on the cytotoxicity, bioavailability (i.e., accumulation) and bio-reactivity (i.e., induction of mRNA levels of metal responsive genes) of copper (Cu) and silver (Ag) in the rainbow trout gut cell line (RTgutGC). Cells were exposed to metals in media with varying Cl- concentrations (0, 1, 5 and 146 mM). Metal speciation in exposure medium was analyzed using Visual MINTEQ software. Cytotoxicity of AgNO3 and CuSO4 was measured based on two endpoints: metabolic activity and membrane integrity. Cells were exposed to 500 nM of AgNO3 and CuSO4 for 24 hours in respective media to determine metal bioavailability and bioreactivity. Ag speciation changes from free ionic (Ag+) to neutral (AgCl), to negatively charged chloride complexes (AgCl2-, AgCl3-) with increasing Cl- concentration in exposure media whereas Cu speciation remains in two forms (Cu2+ and CuHPO4) across all media. Chloride does not affect Ag bioavailability but decreases metal toxicity and bio-reactivity. Cells exposed to Ag expressed significantly higher metallothionein mRNA levels in low Cl- media (0, 1, and 5 mM) than in high Cl- medium (146 mM). This suggests that chloride complexation reduces silver bio-reactivity and toxicity. Conversely, Cu bioavailability and toxicity were higher in the high chloride medium (146 mM) than in the low Cl- (0, 1, and 5 mM) media, supporting the hypothesis that Cu uptake may occur via a chloride dependent mechanism.

HIF1A is Overexpressed in Medulloblastoma and its Inhibition Reduces Proliferation and Increases EPAS1 and ATG16L1 Methylation

2018 ◽  
Vol 18 (3) ◽  
pp. 287-294 ◽  
Author(s):  
Gustavo Alencastro Veiga Cruzeiro ◽  
Maristella Bergamo dos Reis ◽  
Vanessa Silva Silveira ◽  
Regia Caroline Peixoto Lira ◽  
Carlos Gilberto Carlotti Jr ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Cell Line ◽  
Cellular Proliferation ◽  
Mrna Level ◽  
Relevant Information ◽  
Protein Stabilization ◽  
Mrna Levels ◽  
Medulloblastoma Cell Line ◽  
Pediatric Medulloblastoma ◽  
Fresh Frozen

Background: Genetic and epigenetic modifications are closely related to tumor initiation and progression and can provide guidance for understanding tumor functioning, potentially leading to the discovery of new therapies. Studies have associated hypoxia-related genes to tumor progression and chemo/radioresistance in brain tumors. Information on the expression profile of hypoxiarelated genes in pediatric medulloblastoma, although scarce, may reveal relevant information that could support treatment decisions. Objective: Our study focused on evaluation the of CA9, CA12, HIF1A, EPAS1, SCL2A1 and VEGF genes in 41 pediatric fresh-frozen medulloblastoma sample. Additionally, we analyzed the effect of hypoxia and normoxia in the pediatric medulloblastoma cell-line UW402. Furthermore, we assessed the effects of HIF1A knockdown in cell-proliferation and methylation levels of genes related to hypoxia, apoptosis and autophagy. Method: qPCR was performed to evaluate mRNA levels, and Western blot to confirm HIF1A silencing in both patient samples and cell line. Pyrosequencing was performed to asses the methylation levels after HIF1A knockdown in the UW402 cell line. Results: A higher HIF1A mRNA level was observed in MB patients when compared to the cerebellum (non-tumor match). In UW402 MB cell-line, chemically induced hypoxic resulted in an increase of mRNA levels of HIF1A, VEGF, SCL2A1 and CA9 genes. Additionally, HIF1A knockdown induced a decrease in the expression of hypoxia related genes and a decrease of 30% in cell proliferation was also observed. Also, a significant increase in the methylation of ATG16L1 promoter and decrease in the methylation of EPAS1 promoter were observed after HIF1A knockdown. Conclusion: HIF1A knockdown in medulloblastoma cells lead to decreased cellular proliferation, suggesting that HIF1A can be a potential therapeutic target to be explored in the medulloblastoma. However, the mechanisms behind HIF1A protein stabilization and function are very complex and more data need to be generated to potentially use HIF1A as a therapeutical target.

Use of the Rainbow Trout Hepatoma Cell Line, RTH-149, in a Cytotoxicity Assay

1989 ◽  
Vol 17 (2) ◽  
pp. 67-71
Author(s):  
Harvey Babich ◽  
Nieves Martin-Alguacil ◽  
Ellen Borenfreund
Keyword(s):  
Polycyclic Aromatic Hydrocarbons ◽  
Rainbow Trout ◽  
Cell Line ◽  
Aromatic Hydrocarbons ◽  
Hepatoma Cell ◽  
Neutral Red ◽  
Cytotoxicity Assay ◽  
Hepatoma Cell Line ◽  
Polycyclic Aromatic ◽  
Direct Acting

The rainbow trout hepatoma cell line, RTH-149, was evaluated for use as a bioindicator cell type in the neutral red cytotoxicity assay. The cells were exposed for six days to various polycyclic aromatic hydrocarbons, including chemicals that are direct-acting toxicants and chemicals that require enzymatic biotransformation to cytotoxic metabolites. Whereas benzo[a]pyrene was only slightly cytotoxic, its metabolites — (±)trans-7,8-diol-benzo[a]pyrene and 3-hydroxy-benzo[a]pyrene — were highly cytotoxic. 7,12-Dimethylbenz[a]anthracene was cytotoxic, but cytotoxicity did not occur with benzo[a]anthracene, benzo[b]fluoranthene and benzo[k]fluoranthene. This cell line appears to lack sufficient xenobiotic metabolising capacity to biotransform many of these polycyclic aromatic hydrocarbons to activated cytotoxic metabolites.

Column Study on the Effect of Nitrate and Chloride on TCE Removal by Granular Iron

2011 ◽  
Vol 356-360 ◽  
pp. 1642-1646
Author(s):  
Xue Qiang Zhu ◽  
Bao Ping Han ◽  
Guo Jun Wu ◽  
Xiao Qing Zhang
Keyword(s):  
Pitting Corrosion ◽  
Nitrate Concentration ◽  
Chloride Concentration ◽  
Inorganic Anions ◽  
Dual Effect ◽  
Column Study ◽  
Granular Iron ◽  
High Chloride ◽  
Chloride Concentrations

The effects of individual inorganic anions (nitrate and chloride) on the reactivity of granular iron were investigated using plexiglass columns packed with granular iron. The results show that TCE removal decreases apparently with increasing nitrate concentration due to competition for reactive sites. Chloride exhibits dual-effect on the TCE removal by Fe0. In the studied condition, the TCE dechlorination is enhanced at the low chloride concentration due to pitting corrosion and is dampened at the high chloride concentrations such as 59.98 and 110.45 mg/L as Cl-.

scholarly journals The human hepatoma Hep3B cell line as an experimental model in the study of the long-term regulation of acute-phase proteins by cytokines

1992 ◽  
Vol 287 (1) ◽  
pp. 255-259 ◽  
Author(s):  
M Hiron ◽  
M Daveau ◽  
P Arnaud ◽  
J Bauer ◽  
J P Lebreton
Keyword(s):  
Cell Line ◽  
Acute Phase ◽  
Culture System ◽  
Acute Phase Protein ◽  
Hepatoma Cell Line ◽  
Mrna Levels ◽  
Human Hepatoma ◽  
Long Term Culture ◽  
Phase Protein

The regulation of the synthesis by the cytokines interleukin-1 (IL-1) and IL-6 of the positive acute-phase protein alpha 1-acid glycoprotein (AGP) and of the negative acute-phase protein alpha 2-HS glycoprotein (AHSG) has been studied in a long-term culture system of the human hepatoma cell line Hep3B. The culture system contained 30 nM-sodium selenite as the only supplement. This allowed maintenance of the synthesis of the proteins under study at a near steady state for over 3 months. An increase in AGP mRNA and a decrease in AHSG mRNA were observed when cells were treated for two successive 48 h-periods with monocyte-conditioned medium. A return to basal levels was obtained after cessation of the cytokine addition. Two further additions of cytokines led to alterations in mRNA levels similar to those observed following the first cytokine treatment. The amounts of AGP and AHSG secreted were altered in accordance with the mRNA modifications. These results suggest that new cytokine receptors were being constantly synthesized during cell culture. When cytokines were present in the culture medium for 10 days, maximum alterations in AGP and AHSG synthesis were obtained following 2 and 4 days of treatment respectively, but further alterations in protein levels could not be observed afterwards. Expression of IL-6 receptor mRNA was not up-regulated by cytokines, but only by 1 microM-dexamethasone. Our results show that, in this long-term culture system, cytokines induce a response in hepatoma cells similar to that observed in vivo during human inflammatory states. This model could be used to evaluate the effects of agonists or antagonists of cytokines responsible for the hepatic acute-phase protein response.

DNA rearrangement causes a high rate of spontaneous mutation at the immunoglobulin heavy-chain locus of a mouse myeloma cell line

1986 ◽  
Vol 6 (12) ◽  
pp. 4228-4235
Author(s):  
H Yu ◽  
L A Eckhardt
Keyword(s):  
Cell Line ◽  
Heavy Chain ◽  
Spontaneous Mutation ◽  
Immunoglobulin Heavy Chain ◽  
Myeloma Cell ◽  
Chain Gene ◽  
Parent Cell ◽  
Mrna Levels ◽  
Dna Rearrangement ◽  
Myeloma Cell Line

The spontaneous mutation rate of immunoglobulin genes expressed in myeloma cells is well above that of other genes expressed in these or in other cell types. The nature of such mutations in one myeloma cell line, MPC11, was explored at the molecular level. Included in this study were MPC11 variants representing 24 independent and spontaneous mutations affecting immunoglobulin secretion. Of the mutants studied, 19 had ceased immunoglobulin heavy chain (IgH) production (nonproducers), and 5 produced from as little as 1/1,000 to as much as 1/10 the amount of immunoglobulin produced by MPC11 (low producers). Only one of the MPC11 mutants (a nonproducer) showed no evidence of DNA rearrangement in or near the expressed IgH gene. The formerly expressed gamma 2b gene had been deleted in 18 of the 19 nonproducers. All of the low producers had undergone DNA rearrangement in or near the expressed IgH gene, and three of them produced immunoglobulin of a new heavy chain class. The cause for reduced heavy-chain synthesis in the low producers is not yet known. However, in several of these mutants, the defect appeared to be posttranscriptional. In these cell lines, steady-state IgH mRNA levels were much lower than in the parent cell line, while the heavy-chain gene transcription rate remained unchanged.

scholarly journals Regulation of β-adrenoceptor density and mRNA levels in the rat heart cell-line H9c2

1996 ◽  
Vol 317 (3) ◽  
pp. 925-931 ◽  
Author(s):  
Volker DANGEL ◽  
Jeanette GIRAY ◽  
Dieter RATGE ◽  
Hermann WISSER
Keyword(s):  
Cell Line ◽  
Half Life ◽  
Rat Heart ◽  
Radioligand Binding ◽  
Mrna Level ◽  
Fold Increase ◽  
Heart Cell ◽  
H9c2 Cell ◽  
Heart Cells ◽  
Mrna Levels

The regulation of the expression of β-adrenoceptors (β-ARs) is not thoroughly understood. We demonstrate that the rat heart cell-line H9c2 expresses both β1- and β2-ARs. In radioligand-binding experiments, the maximal binding capacity of (-)-[125I]-iodocyanopindolol was determined as 18±0.6 fmol/mg of protein with a KD of 35.4±4.1 pM. Competitive radioligand-binding experiments with subtype-specific β-antagonists reveal a subtype ratio of β1- to β2-ARs of 29%:71%. With competitive reverse-transcriptase PCR we found β2-mRNA to be up to 1600 times more frequent than β1-mRNA. Treatment of the H9c2 cell-line with the β-adrenergic agonist (-)-isoproterenol (10-6 M), the antagonist (-)-propranolol (10-6 M) and the glucocorticoid dexamethasone (500 nM) induces regulatory effects on both the β-AR protein and mRNA level. Isoproterenol treatment leads to down-regulation of the total receptor number by 56±4%, due to a decrease in β2-ARs, while maintaining the β1-AR number constant. On the transcription level, both β1-and β2-mRNAs are decreased by 30% and 42% respectively. mRNA stability measurements reveal a reduced half-life of β2-mRNA from 9.3 h to 6.5 h after isoproterenol treatment. Incubation of cells with (-)-propranolol does not affect the amounts of β-ARs and their mRNAs. Dexamethasone induces a 1.8±0.2-fold increase in β-AR number over the basal level as well as a 1.9±0.2-fold increase in the amount of β2-mRNA. Because the half-life of β2-mRNA was unaffected by dexamethasone, the increased β2-mRNA level must be due to an enhanced transcription rate. The β1-mRNA levels are unchanged during dexamethasone-incubation of the cells. Our data clearly demonstrate that treatment of H9c2 rat heart cells with isoproterenol and dexamethasone induces alterations in the level of RNA stability as well as gene transcription, leading to altered receptor numbers.

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