The aim of the current study was to evaluate the transport system in EMT-6 cell for the uptake of the methylmercury(MeHg). Several inhibitors ere used to test used to test which potential transport system might be involoved in MeHg uptake. Probenecid was used to test the organic transport system, valinomycin for testing the effect of the membrane potential, cytochalasin B for testing the facilitated diffusive D-glucose transport system and colchicine for testing the microtubule system. Ouabain for evaluating active transport system, 4',4-diisothiocyano-2',2-stilbenedisulfonic acid(DIDS) the Cl- ion transport system and verapamil for the $Ca^{2+}$ transprot system. Significantly, MeHg decreased the synthesis of nitric oxcide(NO) and intracellular ATP in ENT-6 cells. In the condition of ouabain containing with MeHg decreased the production of NO and intracelluar ATP. In the treatment of inhibitors, ouabain showed protective effect against cytotoxicity of MeHg but ather inhibitors not showed protective effects. The protective effects of ouabain against the cytotoxicity of MeHg deoended on the concentration of added ouabain to the culture medium for MET-6 cells. These result showed that the uptake of MeHg might be involved in the active transport system. Active transports system seems to share similarities with the transport systems for the uptake of MeHg when using MeHg and MeHg-glutathione complex.x.
Over the last century the mercury (Hg) concentration in the environment has been increased by human activities with inputs from sources such as atmospheric deposition, urban runoff, and industrial effluents. Mercury can be transformed to methylmercury (MeHg) in anaerobic conditions by sulfate reducing bacteria (SRB) and sediments are the principal location for MeHg production in aquatic environments. Interest in bioaccumulation of Hg and MeHg into lower trophic levels of benthic and pelagic organisms stems from public health concerns as these organisms provide essential links for higher trophic levels of food chains such as fish and larger invertebrates. Fish consumption is the major exposure route of MeHg to humans. Recently, it was reported that blood samples in Korea showed much higher Hg levels (5-8 times) than those in USA and Germany. Although this brings much attention to Hg research in Korea, there are very few studies on Hg biogeochemical cycling and bioaccumulation in aquatic environments. Given the importance of Hg methylation and MeHg transfer through food chains in aquatic environments, it is imperative that studies should be done in much detail looking at the fate, transport, and bioaccumulation of Hg and MeHg in the environment. Moreover, there should be long-term monitoring plans in Korea to evaluate the environmental and health effects of Hg and MeHg.
The methylmercury (MeHg) is a toxic environmental pollutant, causing serious neurological and developmental effects in humans. Recent epidemiological studies have indicated that ingestion of MeHg in fish during pregnancy can result in neuroethological effects in the offspring. However, the mechanism underlying the MeHg-toxicity is not fully understood. To elucidate the mechanisms of toxicity of MeHg and of defense against MeHg, we searched for factors that determine the sensitivity of yeast cells to MeHg, and found that overexpression of Cdc34, a ubiquitin-conjugating enzyme (E2) that is a component of the ubiquitin-proteasome (UP) system, induces a resistance to MeHg toxicity in both yeast and human cells. The UP system is involved in the intracellular degradation of proteins. When Cdc34 is overexpressed in cells, ubiquitination reactions are activated and the degradation of certain proteins by the UP system is enhanced. Therefore, it seems likely that certain as-yet-unidentified proteins that increase MeHg toxicity might exist in cons and that toxicity might be reduced by the enhanced degradation of such proteins, mediated by the UP system, when Cdc34 is overexpressed. SCF ubiquitin-ligase is a component of UP system and consists of Skpl, the scaffold protein Cdc53, the RING-finger protein Hrt1, and one member of the family of F-box proteins. The F-box proteins directly bind to the substrates and are the determinants of substrate specificity of SCF. Therefore, we searched for the f-box protein that cofers resistance to MeHg, and found that overexpression of Hrt3 or Yi1224w induced resistance to MeHg toxicity in yeast cells. Since the protein(5) that enhance toxicity of MeHg might plausibly be induced in substrates of both f-box proteins, we next searched for substrate proteins that are recognized by Hrt3 or Y1r224w using two-hybrid screen. We found that Did3 or Crsl interacts with Hrt3; and Eno2 interacts with Yir224w. The yeast cells that overexpressed each those proteins showed hypersensitivity to MeHg, respectively, indicating that those proteins enhance the MeHg toxicity. Both Dld3 and Eno2 are proteins involved in the synthesis of pyruvate, and overexpression of both proteins might induce increase in interacellular levels of pyruvate. Deletion of Yi1006w that transports pyruvate into the mitochondria induced aresistance to MeHg. These results suggest that the promotion of the pyruvate irdlowinto the mitochondria might enhance MeHg toxicity. This study providesimportant keyfor the elucidauon of the molecular mechanism of MeHg toxicity.
Higher methylmercury (MeHg) accumulation and susceptibility to toxicity in the fetus than in the mother at parturition is well known. However, the difference in MeHg exposure to fetus and offspring throughout gestation and suckling is not well established. In the human, the effects of MeHg exposure on pregnant and breast-feeding women remain an important issue for elucidation, especially those of continuous uptake in high-fish-consumption populations. The purpose of this paper was to evaluate the difference in MeHg exposure to fetus and offspring throughout gestation and lactation using our recent animal and human studies data. In the animal study, adult female rats were given a diet containing 5 ${\mu}$g/g Hg (as MeHg) for 8 weeks. Then they were mated and subsequently given the same diet throughout gestation and suckling. On embryonic days 18, 20, 22 and at parturition, the concentrations of Hg in the brains of fetus were approximately 1.5-2.0 times higher than those in the mothers. However, during the suckling period Hg concentrations in the brain rapidly declined to about 1/10 of that during late pregnancy. Hg concentrations in blood also decreased rapidly after birth. In human study, Hg concentrations in red blood cells (RBCs-Hg) in 16 pairs of maternal and umbilical cord blood samples were compared at birth and 3 months of age after parturition. RBCs-Hg concentration in the umbilical cords was about 1.6 times higher than those in the mothers at parturition. However, all the infants showed declines in Hg concentrations throughout the breast-feeding period. The Hg concentration in RBCs-Hg at 3 months of age was about half that at birth. Both the animal and human studies indicated that MeHg exposure to the fetus might be especially high but it dramatically decreases during the suckling period. Therefore, close attention should be paid to the gestation rather than the breast-feeding period to avoid the risk of MeHg to human infants.
Background: Studies on the risk of mercury (Hg) in Korean fishery products focus primarily on total Hg levels as opposed to methylmercury (MeHg) levels. None of the few studies on MeHg in tuna investigated tuna from Japanese restaurants. Few have evaluated lead (Pb), cadmium (Cd) and arsenic (As) in tuna. Thus, this study aimed to conduct a risk assessment by evaluating heavy metal concentrations in tuna from Japanese restaurants. Methods: Thirty-one tuna samples were collected from Japanese restaurants in the Republic of Korea. They were classified according to region and species. The concentration of heavy metals in the samples was analyzed using the Ministry of Food and Drug Safety Food Code method. The rate of exceedance of maximum residue levels (MRLs) and the risk compared to the provisional tolerable weekly intake (PTWI) set by the Joint Food and Agriculture Organization/World Health Organization Expert Committee on Food Additives (%PTWI) were evaluated for risk assessment. Results: The mean of MeHg, Pb, Cd and As concentrations were 0.56 ± 1.47 mg/kg, 33.95 ± 3.74 ㎍/kg, 14.25 ± 2.19 ㎍/kg and 1.46 ± 1.89 mg/kg, respectively. No sample exceeded the MRLs of Pb and Cd, but 9.7% of the samples exceeded the MRL of MeHg. The %PTWIs of MeHg, Pb, Cd and As were 4.2037, 0.0162, 0.0244 and 1.1627, respectively. The %PTWI of MeHg by age group and sex was highest among men aged 19-29 years (10.6494), followed by men aged 30-49 years (7.2458) and women aged 19-29 years (4.8307). Conclusions: We found that 3 out of 31 samples exceeded the MRL of MeHg. The %PTWI of MeHg showed significant differences based on age and sex, and the value was likely to exceed a safe level depending on individuals' eating behaviors. Therefore, improved risk management for MeHg is required.
Total mercury (T-Hg) and methyl mercury (MeHg) were determined in marine fisheries (41 species, n=87) commonly consumed in Korea, using a gold amalgamation method and gas chromatography-cold vapor atomic fluorescent spectroscopy, respectively. Concentrations of T-Hg and MeHg in all samples (31 fish, 4 crustaceans, 4 cephalopods, and 2 gastropod species) were in the range of 0.016-0.495 (mean, 0.093) mg/kg-wet and not detected-0.338 (mean, 0.067) mg/kg-wet, respectively. The concentrations of MeHg in marine fisheries were significantly correlated with T-Hg concentrations (P<0.001). The highest mean concentrations of T-Hg and MeHg were found in fish species, followed by crustaceans. The contribution of MeHg to T-Hg was in the range of 64-95% (mean, 83%) in cephalopods, 28-98% (mean, 69%) in fish, and 26-88% (mean, 57%) in crustaceans. The weekly intakes of T-Hg and MeHg by fisheries consumption for the Korean general population were estimated to be 0.463 and $0.338{\mu}g/kg$ body weight/week, respectively. The concentrations and intakes of T-Hg and MeHg were less than the allowable residue levels and in the range of 12 to 17% of the provisional tolerable weekly intake (PTWI) applied in Korea.
Kim, Youn-Jung;Kim, Mi-Soon;Jeon, Hee-Kyung;Ryu, Jae-Chun
Molecular & Cellular Toxicology
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제4권2호
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pp.164-169
/
2008
Methylmercury (MeHg) is known to have devastating effects on the mammalian nervous system. In order to characterize the mechanism of MeHg-induced neurotoxicity, we investigated the analysis of transcriptional profiles on human 8k cDNA microarray by treatment of $1.4{\mu}M$ MeHg at 3, 12, 24 and 48h in human neuroblastoma SH-SY5Y cell line. Some of the identified genes by MeHg treatment were significant at early time points (3h), while that of others was at late time points (48h). The early response genes that may represent those involved directly in the MeHg response included pantothenate kinase 3, a kinase (PRKA) anchor protein (yotiao) 9, neurotrophic tyrosine kinase, receptor, type 2 gene, associated with NMDA receptor activity regulation or perturbations of central nervous system homeostasis. Also, when SH-SY5Y cells were subjected to a longer exposure (48h), a relative increase was noted in a gene, glutamine-fructose-6-phosphate transaminase 1, reported that overexpression of this gene may lead to the increased resistance to MeHg. To confirm the alteration of these genes in cultured neurons, we then applied real time-RT PCR with SYBR green. Thus, this result suggests that a neurotoxic effect of the MeHg might be ascribed that MeHg alters neuronal receptor regulation or homeostasis of neuronal cells in the early phase. However, in the late phase, it protects cells from neurotoxic effects of MeHg.
To elucidate contamination levels and distribution of methyl mercury (Me-Hg) in Korean coastal areas, 126 sediment samples were collected from Kyeonggi Bay, Namyang Bay, Chinhae Bay, and Lake Shihwa during 1995-1996, and the Me-Hg concentrations were determined by cold vapor atomic fluorescence spectrometry (CVAFS). Contamination levels of Me-Hg in sediments from Kyeonggi Bay, Namyang Bay, Chinhae Bay, and Lake Shihwa were 274 ${\pm}$ 990, 108 ${\pm}$ 24, 294 ${\pm}$ 342, and 1080 ${\pm}$ 760 pg/g, respectively. Concentrations of Me-Hg in sediments were significantly correlated with total organic carbon and sulfur contents, but were independent of mud contents and mean grain size. The highest concentration of Me-Hg (7100 pg/g) was observed at Incheon North Harbor (Site Kl9) in Kyeonggi Bay. This Me-Hg concentration was one or two orders of magnitude higher than those in other Kyeonggi Bay sediments were. The average concentration of Me-Hg in sediments from Lake Shihwa was higher than in those from other study areas. The three peaks of Me-Hg concentrations were observed on three sites (55, 56,and 510) in Lake Shihwa and gradually decreased in distance-dependent manner around these sites. High concentrations of Me-Hg at surface and 10-cm sediment depth in Chinhae Bay maybe due to higher rates of methylation process by active sulfate-reducing bacteria or higher concentrations of total mercury available to sulfate-reducing bacteria.
In this study, total mercury and methyl mercury in whole blood of Korean was analyzed so as to investigate the correlation between total mercury (T-Hg) and methyl mercury (Me-Hg). 4000 whole blood samples were divided in four groups, according to T-Hg concentration in percentile: group I (p25-p50), group II (p50-p75), group III (p75-p95) and group IV (p95-p100). 100 samples were randomly selected from the each group, and Me-Hg concentration was measured. T-Hg concentration in whole blood was analyzed using a Direct Mercury Analyzer-80 and obtained limit of detection (LOD) was $0.2{\mu}gL^{-1}$. Me-Hg concentration was analyzed with ethylate derivatization using headspace-gas chromatography-mass spectrometry, and obtained LOD of methyl mercury was $0.5{\mu}gL^{-1}$. The geometric means of T-Hg and Me-Hg were $6.35{\mu}gL^{-1}$ and $4.44{\mu}gL^{-1}$, respectively, and 71.91% of T-Hg was presented as Me-Hg.
Kim, Youn-Jung;Chang, Suk-Tai;Yun, Hye-Jung;Ryu, Jae-Chun
한국환경독성학회:학술대회논문집
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한국환경독성학회 2003년도 춘계학술대회
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pp.187-187
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2003
Methylmercury (MeHg), one of the heavy metal compounds, can cause severe damage to the central nervous system in humans. Many reports have shown that MeHg is poisonous to human body through contaminated foods and has released into the environment. Despite many studies on the pathogenesis of MeHg-induced central neuropathy, no useful mechanism of toxicity has been established so far. In this study, two methods, cDNA Microarray and SSH, were performed to assess the expression profile against MeHg and to identify differentially expressed genes by MeHg in neuroblastoma cell line. TwinChip Human-8K (Digital Genomics) was used with total RNA from SH-SY5Y (human neuroblastoma cell line) treated with solvent (DMSO) and 6.25 uM (IC50) MeHg. And we performed forward and reverse SSH method on mRNA derived from SH-SY5Y treated with DMSO and MeHg (6.25 uM). Differentially expressed cDNA clones were sequenced and were screened by dot blot and ribonuclease protection assay to confirm that individual clones indeed represent differentially expressed genes. These sequences were identified by BLAST homology search to known genes or expressed sequence tags (ESTs). Analysis of these sequences may provide an insight into the biological effects of MeHg in the pathogenesis of neurodegenerative disease and a possibility to develop more efficient and exact monitoring system of heavy metals as environmental pollutants.
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