7,284 research outputs found
Microcystin-LR stabilizes c-myc protein by inhibiting protein phosphatase 2A in HEK293 cells
No full textMicrocystin-LR is the most toxic and the most frequently encountered toxin produced by the cyanobacteria in the contaminated aquatic environment. Previous studies have demonstrated that Microcystin-LR is a potential carcinogen for animals and humans, and the International Agency for Research on Cancer has classified Microcystin-LR as a possible human carcinogen. However, the precise molecular mechanisms of Microcystin-LR-induced carcinogenesis remain a mystery. C-myc is a proto-oncogene, abnormal expression of which contributes to the tumor development. Although several studies have demonstrated that Microcystin-LR could induce c-myc expression at the transcriptional level, the exact connection between Microcystin-LR toxicity and c-myc response remains unclear. In this study, we showed that the c-myc protein increased in HEK293 cells after exposure to Microcystin-LR. Coexpression of protein phosphatase 2A and two stable c-myc protein point mutants (either c-myc(T58A) or c-myc(562A)) showed that Microcystin-LR increased c-myc protein level mainly through inhibiting protein phosphatase 2A activity which altered the phosphorylation status of serine 62 on c-myc. In addition, we also showed that Microcystin-LR could increase c-myc promoter activity as revealed by luciferase reporter assay. And the TATA box for P1 promoter of c-myc might be involved. Our results suggested that Microcystin-LR can stimulate c-myc transcription and stabilize c-myc protein, which might contribute to hepatic tumorigenesis in animals and humans. (C) 2014 Elsevier Ireland Ltd. All rights reserved.Microcystin-LR is the most toxic and the most frequently encountered toxin produced by the cyanobacteria in the contaminated aquatic environment. Previous studies have demonstrated that Microcystin-LR is a potential carcinogen for animals and humans, and the International Agency for Research on Cancer has classified Microcystin-LR as a possible human carcinogen. However, the precise molecular mechanisms of Microcystin-LR-induced carcinogenesis remain a mystery. C-myc is a proto-oncogene, abnormal expression of which contributes to the tumor development. Although several studies have demonstrated that Microcystin-LR could induce c-myc expression at the transcriptional level, the exact connection between Microcystin-LR toxicity and c-myc response remains unclear. In this study, we showed that the c-myc protein increased in HEK293 cells after exposure to Microcystin-LR. Coexpression of protein phosphatase 2A and two stable c-myc protein point mutants (either c-myc(T58A) or c-myc(562A)) showed that Microcystin-LR increased c-myc protein level mainly through inhibiting protein phosphatase 2A activity which altered the phosphorylation status of serine 62 on c-myc. In addition, we also showed that Microcystin-LR could increase c-myc promoter activity as revealed by luciferase reporter assay. And the TATA box for P1 promoter of c-myc might be involved. Our results suggested that Microcystin-LR can stimulate c-myc transcription and stabilize c-myc protein, which might contribute to hepatic tumorigenesis in animals and humans. (C) 2014 Elsevier Ireland Ltd. All rights reserved
The role of Ruditapes philippinarum glutathione transferases in the metabolism of microcystin-LR
No full textNo abstracts are to be cited without prior reference to the author. Glutathione transferases (GSTs) are phase II enzymes involved in the microcystin (MC) induced detoxication processes. In this study we analyze and compare the metabolism of MC-LR by the cytosolic GSTs from gills and hepatopancreas of Ruditapes philippinarum. Cytosolic GSTs were purified by glutathione (GSH)–agarose affinity chromatography from exposed and non-exposed bivalves to MC-LR (100 µg/L) representing the inducible and constitutive (Basal) GST fractions, respectively. For each mixture, we examined the in vitro cytosolic GST inhibition efficiency of the conjugation of CDNB to GSH by MC-LR and characterize the inhibition mechanism. Results support the important role of GST enzymes in detoxification of MCs in bivalve mollusk
Responses of the Proteome and Metabolome in Livers of Zebrafish Exposed Chronically to Environmentally Relevant Concentrations of Microcystin-LR
No full textIn
this study, for the first time, changes in expressions of proteins
and profiles of metabolites in liver of the small, freshwater fish Danio rerio (zebrafish) were investigated after
long-term exposure to environmentally relevant concentrations of microcystin-LR
(MC-LR). Male zebrafish were exposed via water to 1 or 10 μg
MC-LR/L for 90 days, and iTRAQ-based proteomics and 1H
NMR-based metabolomics were employed. Histopathological observations
showed that MC-LR caused damage to liver, and the effects were more
pronounced in fish exposed to 10 μg MC-LR/L. Metabolomic analysis
also showed alterations of hepatic function, which included changes
in a number of metabolic pathways, including small molecules involved
in energy, glucose, lipids, and amino acids metabolism. Concentrations
of lactate were significantly greater in individuals exposed to MC-LR
than in unexposed controls. This indicated a shift toward anaerobic
metabolism, which was confirmed by impaired respiration in mitochondria.
Proteomics revealed that MC-LR significantly influenced multiple proteins,
including those involved in folding of proteins and metabolism. Endoplasmic
reticulum stress contributed to disturbance of metabolism of lipids
in liver of zebrafish exposed to MC-LR. Identification of proteins
and metabolites in liver of zebrafish responsive to MC-LR provides
insights into mechanisms of chronic toxicity of MCs
Transcriptional responses of glutathione transferase genes in Ruditapes philippinarum exposed to microcystin-LR
No full textNo abstracts are to be cited without prior reference to the author. Microcystins (MCs) are potent hepatotoxins produced by bloom-forming species of toxic cyanobacteria. Among these, MC-LR is the most commonly found and toxic variant. Bivalves, due to their benthic and sedentary mode of life, are one of the most threatened organisms by these environmental stressors. Glutathione transferases (GSTs) play a major role in cellular defense against MCs toxicity. The aim of this study was to compare the relative changes of gene expression of the different GSTs isoforms in mollusc bivalves exposed to MCs. The time-dependent changes of relative transcription abundance of several GST isoforms in parallel with enzymatic activity of total GST were investigated in gills and hepatopancreas of Ruditapes philippinarum exposed to dissolved MC-LR. The relative changes of gene expression and enzyme activity were analyzed by quantitative real-time PCR and colorimetric assays respectively. We found that MC-LR could affect the transcriptional activities of these detoxification enzymes in gills and hepatopancreas of the tested bivalves. Most GST isoforms showed differential response profiles depending on the concentrations of MC-LR and exposure times for clams. These results highlight the important role of GSTs in counteracting the potential deleterious effects induced by MCs in bivalve
The role of cysteine conjugation in the detoxification of microcystin-LR in liver of bighead carp (Aristichthys nobilis): a field and laboratory study
No full textThe role of glutathione (GSH) and cysteine (Cys) conjugates in the detoxification of microcystin-LR (MC-LR) in bighead carp (Aristichthys nobilis) was examined under laboratory and field conditions. Wild individuals of bighead carp were collected from 5 eutrophic lakes along the Yangtze River, while in laboratory experiment, bighead carp were injected intraperitoneally with 500 mu g purified MC-LR/kg body weight (bw). Contents of MC-LR and its glutathione (MC-LR-GSH) and cysteine conjugates (MC-LR-Cys) in the liver of bighead carp were determined by liquid chromatography electrospray ionization mass spectrum (LC-ESI-MS). In laboratory experiment, low concentrations of MC-LR-GSH (mean: 0.042 mu g/g dry weight (DW)) were always detectable, and the mean ratio of MC-LR-Cys to MC-LR-GSH was 6.55. While, in field study, relatively high MC-LR-Cys concentration (mean: 0.22 mu g/g DW) was detected, whereas MC-LR-GSH was occasionally detectable, and the average ratio of MC-LR-Cys to MC-LR-GSH was as high as 71.49. A positive correlation was found between MC-LR-Cys concentration in the liver of bighead carp and MC-LR content in seston from the five lakes (r = 0.85). These results suggest that MC-LR-Cys might be much more important than MC-LR-GSH in the detoxification of MC-LR in fish liver, and that cysteine conjugation of MC-LR might be a physiological mechanism for the phytoplanktivorous bighead carp to counteract toxic cyanobacteria.The role of glutathione (GSH) and cysteine (Cys) conjugates in the detoxification of microcystin-LR (MC-LR) in bighead carp (Aristichthys nobilis) was examined under laboratory and field conditions. Wild individuals of bighead carp were collected from 5 eutrophic lakes along the Yangtze River, while in laboratory experiment, bighead carp were injected intraperitoneally with 500 mu g purified MC-LR/kg body weight (bw). Contents of MC-LR and its glutathione (MC-LR-GSH) and cysteine conjugates (MC-LR-Cys) in the liver of bighead carp were determined by liquid chromatography electrospray ionization mass spectrum (LC-ESI-MS). In laboratory experiment, low concentrations of MC-LR-GSH (mean: 0.042 mu g/g dry weight (DW)) were always detectable, and the mean ratio of MC-LR-Cys to MC-LR-GSH was 6.55. While, in field study, relatively high MC-LR-Cys concentration (mean: 0.22 mu g/g DW) was detected, whereas MC-LR-GSH was occasionally detectable, and the average ratio of MC-LR-Cys to MC-LR-GSH was as high as 71.49. A positive correlation was found between MC-LR-Cys concentration in the liver of bighead carp and MC-LR content in seston from the five lakes (r = 0.85). These results suggest that MC-LR-Cys might be much more important than MC-LR-GSH in the detoxification of MC-LR in fish liver, and that cysteine conjugation of MC-LR might be a physiological mechanism for the phytoplanktivorous bighead carp to counteract toxic cyanobacteria
Comparative Studies on the pH Dependence of DOW of Microcystin-RR and -LR using LC-MS
No full textMicrocystins (MCs) are well known worldwide as hepatotoxins produced by cyanobacteria, but little is known about the physicochemical properties of these compounds. The dependence of the n-octanol/water distribution ratio (DOW) of MC-RR and -LR to pH was measured by high-performance liquid chromatography combined with mass spectrometry (LC-MS). There was a remarkable difference in such relationships between MC-RR and -LR. The log DOW of MC-LR decreased from 1.63 at pH 1.0 to -1.26 at pH 6.5, and stabilized between -1.04 and -1.56 at a pH of 6.5 similar to 12.0; log DOW of MC-RR varied between -1.24 and -0.67 at a pH of 1.00 similar to 4.00, and stabilized between -1.20 and -1.54 at a pH of 4.00 similar to 12.00. The difference of hydrophobicity in acidic condition between MC-RR and -LR is important, not only for the analytical method of both toxins, but perhaps also for understanding the difference of toxicity to animals between the two toxins
Eteone delta Wu & Chen
No full textEteone delta Wu & Chen Eteone delta Wu & Chen, 1963 Ecology: FT Distribution: PA, OL Habitat: LR, ES Additional references: Shen & Qi (1982)Published as part of Glasby, Christopher J., Timm, Tarmo, Muir, Alexander I. & Gil, João, 2009, Catalogue of non-marine Polychaeta (Annelida) of the World, pp. 1-52 in Zootaxa 2070 on page 18, DOI: 10.5281/zenodo.18708
Fast removal of cyanobacterial toxin microcystin-LR by a low-cytotoxic microgel-Fe(III) complex
No full textEutrophication has become a serious environmental threat throughout the world. In particular, the presence of cyanobacteria toxins, especially microcystins (MCs), has become a severe problem. Inhibition of Microcystis growth in water resources is the most effective way to reduce MCs, but it is a long-term investment. In the present study, a microgel-Fe(III) complex was developed for the fast removal of MC-LR. The microgel-Fe(III) characteristics and the MC-LR removal dynamics in Milli-Q water and natural water were evaluated. The removal efficiency negatively correlated to the initial MC-LR concentration and pH value (2.0-11.5), but the kinetics was not significantly influenced. The presence of natural organic matter (NOM) in water slightly reduced MC-LR removal using microgel-Fe(III). In addition, microgel-Fe(III) removed 98.99% of MC-LR in 12 min, while for activated carbon, it took 15-24 h to reach equilibrium. Furthermore, methanol was found to regenerate the microgel-Fe(III) after MC-LR removal for at least five regeneration cycles. Finally, the microgel-Fe(III) material was made into a membrane so that MCs could be removed by filtration. Therefore, microgel-Fe(III) is an effective technology and has a great potential in removing MC-LR from drinking water resources. (C) 2011 Elsevier Ltd. All rights reserved.Eutrophication has become a serious environmental threat throughout the world. In particular, the presence of cyanobacteria toxins, especially microcystins (MCs), has become a severe problem. Inhibition of Microcystis growth in water resources is the most effective way to reduce MCs, but it is a long-term investment. In the present study, a microgel-Fe(III) complex was developed for the fast removal of MC-LR. The microgel-Fe(III) characteristics and the MC-LR removal dynamics in Milli-Q water and natural water were evaluated. The removal efficiency negatively correlated to the initial MC-LR concentration and pH value (2.0-11.5), but the kinetics was not significantly influenced. The presence of natural organic matter (NOM) in water slightly reduced MC-LR removal using microgel-Fe(III). In addition, microgel-Fe(III) removed 98.99% of MC-LR in 12 min, while for activated carbon, it took 15-24 h to reach equilibrium. Furthermore, methanol was found to regenerate the microgel-Fe(III) after MC-LR removal for at least five regeneration cycles. Finally, the microgel-Fe(III) material was made into a membrane so that MCs could be removed by filtration. Therefore, microgel-Fe(III) is an effective technology and has a great potential in removing MC-LR from drinking water resources. (C) 2011 Elsevier Ltd. All rights reserved
Effects of microcystins on and toxin degradation by Poterioochromonas sp.
No full textA Chrysophyceae species, Poterioochromonas sp., was isolated from Microcystis cultures. This species can efficiently prey on Microcystis and can grow faster phagotrophically than autotrophically. The growth of Poterioochromonas sp. was stimulated in the presence of microcystin-LR and microcystin-RR (in concentrations ranging from 0.1 to 4 mg/L). The growth rate of Poterioochromonas was 4-5 times higher than the control, indicating the toxins serve as growth stimuli for this organism. A subculture of toxin-treated cells, however, showed low cellular viability, suggesting that growth enhancement by microcystins was not a normal process. The antioxidant enzymatic activity of Poterioochromonas sp. was screened for toxicology analysis. Glutathione; malondialdehyde, and superoxide dismutase (SOD) content was up-regulated within,8 h of exposure to microcystin-LR (500 mu g/L). A high level of SOD activity during exposure to the toxin indicated that SOD was involved in decreasing oxidative stress caused by microcystin-LR. Simultaneously with growth, Poterioochromonas was able to degrade microcystin-LR even, at a toxin concentration of 4 mg/L. This putative degradation mechanism in Poterioochromonas is explored further and discussed in this article. Our findings may shed light on understanding the role of Poterioochromonas in the aquatic ecosystem, in particular, as a grazer of toxic cyanobacteria and a biodegrader for microcystins. (c) 2005 Wiley Periodicals, Inc
Identification of cda gene in bighead carp and its expression in response to microcystin-LR
No full textMicrocystin-LR (MCLR) is a widespread cyanotoxin, which can influence genes transcription and cause nucleic acid damage in different organisms. To identify MCLR induced transcriptionally changed hepatic genes in bighead carp by subtractive suppression hybridization, we obtained the cDNA fragment of cda. Then we cloned its full-length cDNA, which encodes a cytidine deaminase (CDA). 3D structure prediction showed that the 3D structure and amino acid residues related to function sites of bighead carp CDA were highly conserved. Bighead carp CDA shared high identities with other CDA sequences, and evolved closely to non-mammalian CDAs. Bighead carp expressed cda in all tested tissues under normal situation, and changed its expression profile in a time inversely dependent and dose dependent manner to MCLR, so as to protect itself from MCLR induced toxic damage. These indicated that cda might be involved in anti-MCLR response, especially in the regulation of cytidine and dexocytidine metabolism pathway. (C) 2012 Elsevier Inc. All rights reserved.Microcystin-LR (MCLR) is a widespread cyanotoxin, which can influence genes transcription and cause nucleic acid damage in different organisms. To identify MCLR induced transcriptionally changed hepatic genes in bighead carp by subtractive suppression hybridization, we obtained the cDNA fragment of cda. Then we cloned its full-length cDNA, which encodes a cytidine deaminase (CDA). 3D structure prediction showed that the 3D structure and amino acid residues related to function sites of bighead carp CDA were highly conserved. Bighead carp CDA shared high identities with other CDA sequences, and evolved closely to non-mammalian CDAs. Bighead carp expressed cda in all tested tissues under normal situation, and changed its expression profile in a time inversely dependent and dose dependent manner to MCLR, so as to protect itself from MCLR induced toxic damage. These indicated that cda might be involved in anti-MCLR response, especially in the regulation of cytidine and dexocytidine metabolism pathway. (C) 2012 Elsevier Inc. All rights reserved
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