metallothionein and Heavy-Metal-Poisoning

Metallothioneins are peculiar cysteine rich, heat resistant, small cellular plasma proteins expressed through almost all life forms. The currently established biological functions of metallothioneins are the homeostasis of essential metals and protection against toxic transitional metals (TM) alongside defence from oxidative stress by direct scavenging of reactive oxygen and nitrogen species (ROS and RNS). In mammals, among the four main evolutionary conserved forms, only the ubiquitously expressed metallothionein 1 and 2 (here abbreviated as MT) are inducible by TM, oxidative stress, glucocorticoids and starvation among various other stimuli. However, more than sixty years after being discovered, metallothioneins still bear unresolved issues about their possible physiological function and regulation. The biological function of MTs has still not been associated with the in vitro-demonstrated capacity of MT interaction with cellular molecules glutathione (GSH) or adenosine triphosphate (ATP), or with the possibility of direct iron-MT binding in the reducing intracellular environment of some organelles, e.g. lysosomes. Iron as the most abundant cellular TM is also one of the main physiological sources of ROS. Moreover, iron exhibits strain, sex and age differences that reflected ROS generation and MT induction in (patho)physiology and toxicology studies. A recent study showed that iron sex differences follows expression of both ferritin and MT leading to wide implications from essential TM interconnectivity to aging. This review places emphasis on biochemically proven but physiologically ignored interactions of MT with iron to stimulate advanced research for establishing a wide frame of the biological roles of MTs important for health and longevity.

Topics: Animals; Fishes; Hazardous Substances; Heavy Metal Poisoning; Humans; Mammals; Metallothionein; Molecular Structure; Oxidative Stress

Bio-organometallics is a research strategy of biology that uses organic-inorganic hybrid molecules. The molecules are expected to exhibit useful bioactivities based on the unique structure formed by interaction between the organic structure and intramolecular metal(s). However, studies on both biology and toxicology of organic-inorganic hybrid molecules have been incompletely performed. There can be two types of toxicological studies of bio-organometallics; one is evaluation of organic-inorganic hybrid molecules and the other is analysis of biological systems from the viewpoint of toxicology using organic-inorganic hybrid molecules. Our recent studies indicate that cytotoxicity of hybrid molecules containing a metal that is nontoxic in inorganic forms can be more toxic than that of hybrid molecules containing a metal that is toxic in inorganic forms when the structure of the ligand is the same. Additionally, it was revealed that organic-inorganic hybrid molecules are useful for analysis of biological systems important for understanding the toxicity of chemical compounds including heavy metals.

Topics: Biology; Endothelial Cells; Heavy Metal Poisoning; Humans; Inorganic Chemicals; Metallothionein; Organic Chemicals; Organometallic Compounds; Poisoning; Toxicology

Metallothionein (MT) is a low molecular weight metal-binding protein induced by endogenous and exogenous stimuli such as cytokines and heavy metals. In 1993 and 1994, two research groups (Choo et al. and Palmiter et al., respectively) produced MT-I/II double-knockout mice (MT-I/II null mice) with null mutations of the MT-I and MT-II genes. Subsequently, MT-I/II null mice have been used to clarify the biological function, physiological role, and pathophysiological relevance of MT by many research groups. Recent studies using MT-I/II null mice to investigate the role of MT in metal toxicity and distribution, oxidative stress, and some disease were reviewed. In addition, several research groups including our laboratory have reported that MT-I/II null mice are highly susceptible to several carcinogenesis caused by 7,12-dimethylbenz[a]anthracene, X-ray, benzo[a]pyrene, N-butyl-N-(4-hydroxybutyl) nitrosamine, lead, and cisplatin. These results suggest that MT is an important protective factor against not only metal toxicity and oxidative stress but also chemical and radiation carcinogenesis. In this review, we present the findings of MT-I/II null mice with regard to the protective role of MT in carcinogenesis and mutagenesis caused by chemical agents and X-ray.

Topics: Animals; Carcinogenesis; Heavy Metal Poisoning; Humans; Metallothionein; Metals, Heavy; Oxidative Stress; Poisoning

Topics: Amino Acid Sequence; Animals; Cadmium; Cadmium Poisoning; Gene Deletion; Heavy Metal Poisoning; Humans; Metallothionein; Metals, Heavy; Molecular Sequence Data; Protein Binding; RNA, Messenger

Metal bioaccumulation and metallothionein (MT) expression were investigated in the gills and liver of the red-blooded Antarctic teleost

Topics: Animals; Cadmium; Copper; Fishes; Gills; Heavy Metal Poisoning; Metallothionein; Metals; Perciformes; Water Pollutants, Chemical

Cadmium (Cd) is toxic non-essential heavy metal that precipitates adverse health effects in humans and animals. Chelation therapy, the typical treatment for cadmium toxicity, has certain safety and efficacy issues to treat long term cadmium toxicity, in particular. Recent studies have shown that essential trace elements can play important roles in obviating experimental Cd toxicity. This study organizes and reviews the prototypical evidences of the protective effects of essential trace elements against Cd toxicity in animals and attempts to point out the underlying mechanisms. Zinc, selenium, iron, and combinations thereof are reported to be active. The major mechanisms elucidated inter alia are-induction of metallothionein (MT) synthesis and Cd-MT binding (for zinc), modulation of oxidative stress and apoptosis, interference in cadmium absorption and accumulation from body-thereby maintenance of essential metal homeostasis and cytoprotection. Based on the findings, essential trace elements can be recommended for the susceptible population. The application of these trace elements appears beneficial for both the prevention and remediation of long-term Cd toxicity operative via multiple mechanisms with no or minimal adverse effects as compared to the conventional chelation therapy.

Topics: Animals; Cadmium; Cadmium Poisoning; Chelating Agents; Heavy Metal Poisoning; Metallothionein; Selenium; Trace Elements; Zinc

The VECELL 3-D insert is a new culture scaffold consisting of collagen-coated ePTFE (expanded polytetrafluoroethylene) mesh. We analyzed the effects of VECELL 3-D inserts on the functionality of HepG2, a human hepatocellular carcinoma cell line. HepG2 cells cultured on VECELL 3-D inserts maintained a round shape, while those cultured on a standard culture plate or collagen-coated cell culture plate showed a flattened and cubic epithelial-like shape. HepG2 cells cultured on VECELL 3-D inserts had showed upregulated expression of metallothionein genes and in turn a higher tolerance to toxicity induced by heavy metals. These results suggest that HepG2 cell functions were changed by the cell morphology that is induced by culturing on a VECELL 3-D insert.

Topics: Cell Culture Techniques; Collagen; Drug Tolerance; Gene Expression; Heavy Metal Poisoning; Hep G2 Cells; Humans; Metallothionein; Poisoning; Polytetrafluoroethylene; Tissue Scaffolds; Up-Regulation

In the present study, Gammarus fossarum was used to investigate the bioaccumulation and toxic effects of aquatic pollutants in the real environmental conditions. The novelty of the study is the evaluation of soluble tissue metal concentrations in gammarids as indicators in early assessment of metal exposure. In the Sutla River, industrially/rurally/agriculturally influenced catchment in North-Western Croatia, physico-chemical water properties pointed to disturbed ecological status, which was reflected on population scale as more than 50 times lower gammarid density compared to the reference location, Črnomerec Stream. Significantly higher levels of soluble toxic metals (Al, As, Cd, Pb, Sb, Sn, Sr) were observed in gammarids from the Sutla River compared to the reference site and reflected the data on higher total dissolved metal levels in the river water at that site. The soluble metal estimates were supplemented with the common multibiomarker approach, which showed significant biological responses for decreased acetylcholinesterase activity and increased total soluble protein concentrations, confirming stressed environmental conditions for biota in the Sutla River. Biomarker of metal exposure, metallothionein, was not induced and therefore, toxic effect of metals was not confirmed on molecular level. Comparable between-site pattern of soluble toxic metals in gammarids and total dissolved metal levels in water suggests that prior to biomarker response and observed toxic impact, soluble metals in tissue might be used as early warning signs of metal impact in the aquatic environment and improve the assessment of water quality.

Topics: Amphipoda; Animals; Croatia; Ecology; Environmental Monitoring; Heavy Metal Poisoning; Metallothionein; Metals, Heavy; Poisoning; Rivers; Water Pollutants, Chemical; Water Quality

Exposure to elevated levels of the toxic metals inorganic arsenic (iAs) and cadmium (Cd) represents a major global health problem. These metals often occur as mixtures in the environment, creating the potential for interactive or synergistic biological effects different from those observed in single exposure conditions. In the present study, environmental mixtures collected from two waste sites in China and comparable mixtures prepared in the laboratory were tested for toxicogenomic response in placental JEG-3 cells. These cells serve as a model for evaluating cellular responses to exposures during pregnancy. One of the mixtures was predominated by iAs and one by Cd. Six gene biomarkers were measured in order to evaluate the effects from the metal mixtures using dose and time-course experiments including: heme oxygenase 1 (HO-1) and metallothionein isoforms (MT1A, MT1F and MT1G) previously shown to be preferentially induced by exposure to either iAs or Cd, and metal transporter genes aquaporin-9 (AQP9) and ATPase, Cu(2+) transporting, beta polypeptide (ATP7B). There was a significant increase in the mRNA expression levels of ATP7B, HO-1, MT1A, MT1F, and MT1G in mixture-treated cells compared to the iAs or Cd only-treated cells. Notably, the genomic responses were observed at concentrations significantly lower than levels found at the environmental collection sites. These data demonstrate that metal mixtures increase the expression of gene biomarkers in placental JEG-3 cells in a synergistic manner. Taken together, the data suggest that toxic metals that co-occur may induce detrimental health effects that are currently underestimated when analyzed as single metals.

Topics: Arsenic; Cadmium; Cell Line, Tumor; China; Dose-Response Relationship, Drug; Female; Heavy Metal Poisoning; Humans; Metallothionein; Metals, Heavy; Oxidative Stress; Placenta; Poisoning; Pregnancy; Water Pollutants, Chemical

The general population is exposed to metals as trace amounts of metallic compounds are present in air, water, and food. Information on background exposures and biomarker concentrations of environmental chemicals in the general Portuguese population is limited. Therefore, the purpose of this study was to determine the levels of important nonessential metals with recognized toxicity cadmium (Cd) and lead (Pb) and essential metals copper (Cu), nickel (Ni), chromium (Cr), and zinc (Zn) in placentas of mothers living in south Portugal (Algarve). Due to the difficulty in establishing the effects of chemicals in a complex and variable environment, this study also aimed to examine the response of biomarkers, such as biochemical changes that occurs at subcellular levels in the presence of contaminants. The investigated biomarkers in placentas indicative of metal exposure or damage included the metallothioneins (MT), delta-aminolevulinic acid dehydratase (ALAD) (specific for Pb), and lipid peroxidation (LPO) as an index of oxidative stress damage. Moreover, HJ-BIPLOT was applied in order to identify and categorize mothers vulnerable to environmental contamination in this region. Metal concentrations in the placenta were not excessive but within the range found in most European studies. In general, the biomarkers MT and LPO were positively correlated with metal levels, while with ALAD the opposite occurred, indicating the selected battery of biomarkers were suitable to study the effects of metals on human placenta. Further, the application of multivariate analysis with HJ-BIPLOT showed that most significant factors contributing to maternal and fetal exposures via placenta were dietary and smoking habits.

Topics: Adolescent; Adult; Biomarkers; Cross-Sectional Studies; Diet; Environmental Monitoring; Epidemiological Monitoring; Female; Heavy Metal Poisoning; Hospitals, Public; Humans; Lipid Peroxidation; Maternal Exposure; Maternal Nutritional Physiological Phenomena; Metallothionein; Metals, Heavy; Placenta; Poisoning; Porphobilinogen Synthase; Portugal; Pregnancy; Smoking; Trace Elements; Young Adult

Metallothioneins (MTs) are low molecular weight, cysteine-rich, metal-binding proteins. Because of their rich thiol groups, MTs bind to the biologically essential metals and perform these metals' homeostatic regulations; absorb the heavy metals and assist with their transportation and extraction. The aim of this study was to investigate the association between the metallothionein 2A (MT2A) core promoter region -5 A/G single nucleotide polymorphism (SNP) and Cd, Pb, Zn and Cu levels in the blood samples. MT2A polymorphism was determined by the standard polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique using the 616 blood samples and the genotype frequencies were found as 86.6% homozygote typical (AA), 12.8% heterozygote (AG) and 0.6% homozygote atypical (GG). Metal levels were analyzed by dual atomic absorption spectrophotometer system and the average levels of Cd, Pb, Zn and Cu in the blood samples were 1.69±1.57 ppb, 30.62±14.13 ppb, 0.98±0.49 ppm and 1.04±0.45 ppm, respectively. As a result; highly statistically significant associations were detected between the -5 A/G core promoter region SNP in the MT2A gene and Cd, Pb and Zn levels (p=0.004, p=0.012 and p=0.002, respectively), but no association was found with Cu level (p=0.595). Individuals with the GG genotype had statistically lower Zn level and higher Cd and Pb levels in the blood samples than individuals with AA and AG genotypes. This study suggests that having the GG genotype individuals may be more sensitive for the metal toxicity and they should be more careful about protecting their health against the toxic effects of the heavy metals.

Topics: Adenine Nucleotides; Adolescent; Adult; Aged; Cadmium; Copper; Female; Genetic Markers; Genotype; Guanine Nucleotides; Heavy Metal Poisoning; Humans; Lead; Male; Metallothionein; Metals, Heavy; Middle Aged; Poisoning; Polymorphism, Single Nucleotide; Promoter Regions, Genetic; Young Adult; Zinc

Organisms are able to detoxify accumulated metals by, e.g. binding them to metallothionein (MT) and/or sequestering them in metal-rich granules (MRG). The different factors involved in determining the capacity or efficiency with which metals are detoxified are not yet known. In this work we studied how the sub-cellular distribution pattern of cadmium, copper and zinc in whole tissue of zebra mussels from clean and polluted surface waters is influenced by the total accumulated metal concentration and by its physiological condition. Additionally we measured the metallothionein concentration in the mussel tissue. Metal concentration increased gradually in the metal-sensitive and detoxified sub-cellular fractions with increasing whole tissue concentrations. However, metal concentrations in the sensitive fractions did not increase to the same extent as metal concentrations in whole tissues. In more polluted mussels the contribution of MRG and MT became more important. Nevertheless, metal detoxification was not sufficient to prevent metal binding to heat-sensitive low molecular weight proteins (HDP fraction). Finally we found an indication that metal detoxification was influenced by the condition of the zebra mussels. MT content could be explained for up to 83% by variations in Zn concentration and physiological condition of the mussels.

Topics: Animals; Cadmium Poisoning; Cell Fractionation; Copper; Dreissena; Environmental Monitoring; Heavy Metal Poisoning; Inactivation, Metabolic; Metallothionein; Metals, Heavy; Statistics, Nonparametric; Subcellular Fractions; Water Pollutants, Chemical; Zinc

Remobilization of chemicals from contaminated sediments is a major risk associated with dredging and disposal operations in harbour areas. In this work caged mussels, Mytilus galloprovincialis, were chosen as bioindicator organisms to reveal the impact and recovery of organisms from these activities in the harbour of Piombino (Tuscany, Italy) where approximately 100,000 m(3) of sediments were removed and disposed in a local confined disposal facility (CDF). Organisms were deployed before, during and after the end of operations, selecting sites differently impacted by these activities. Temporal changes in environmental bioavailability and biological effects of pollutants were assessed by integrating analyses of trace metals and polycyclic aromatic hydrocarbons (PAHs) accumulated in tissues of caged mussels with a wide array of biomarkers reflecting exposure to specific classes of pollutants and different levels of cellular unbalance or toxicity. Such biological responses included levels of metallothioneins, activity of acyl CoA oxidase (AOX) as a marker of peroxisome proliferation, oxidative stress biomarkers (content of glutathione, enzymatic activities of catalase, glutathione S-transferases, glutathione reductase, glutathione peroxidases), total oxyradical scavenging capacity (TOSC) toward peroxyl and hydroxyl radicals, lysosomal membrane stability and genotoxic effects measured as DNA strand breaks and frequency of micronuclei. Obtained results indicated that a general disturbance was already present in the whole harbour area and especially in the inner site before the beginning of operations, when caged mussels exhibited a significant accumulation of PAHs and Pb, lower TOSC values and higher levels of both lysosomal and genotoxic damages. Bioavailability of trace metals and PAHs markedly increased during dredging activities with values up to 40 microg/g for Pb and up to 2200 ng/g for PAHs in tissues of caged mussels, a significant inhibition of antioxidant efficiency and increase of oxidative damages. While bioavailability of trace metals returned to the pre-dredging values after the end of operations, the accumulation of PAHs, oxidative effects and genotoxic damages remained elevated in mussels caged in the inner area and in front of CDF. Overall this study confirmed the utility of caged mussels to assess the remobilization of chemicals from dredged sediments and the onset of potentially harmful biological effects.

Topics: Acyl-CoA Oxidase; Animals; Biological Availability; Catalase; Comet Assay; DNA Damage; Ecotoxicology; Geologic Sediments; Glutathione; Glutathione Peroxidase; Heavy Metal Poisoning; Metallothionein; Metals, Heavy; Micronucleus Tests; Mytilus; Polycyclic Aromatic Hydrocarbons; Principal Component Analysis; Water Pollutants, Chemical

Distribution of Cu, Zn, Cd, Ag, Hg, and Se were determined in hepatocytosol of northern fur seals (Callorhinus ursinus), black-footed albatrosses (Diomedea nigripes), and Dall's porpoises (Phocoenoides dalli). Copper, Zn, and Cd were accumulated preferentially in metallothionein (MT) fraction and their contents in MT fraction increased with the amounts in the hepatocytosol. Silver was bound to both high-molecular-weight substances (HMWS) and MT in the hepatocytosol for all three species, whereas the distribution of Ag in the cytosol was different among the three species. In northern fur seals, Ag mainly was bound to MT, whereas it mainly was associated with HMWS in Dall's porpoises. In contrast, Ag was distributed almost equally in both HMWS and MT for black-footed albatrosses. Mercury content in HMWS and Se content in HMWS and low-molecular-weight substances (LMWS) increased with their contents in hepatocytosol for all the three species. A significant positive correlation was found between Se and Hg contents in high-molecular weight (HMW) fraction in cytosol. The molar ratio of Hg and Se was close to unity in HMW fraction of the specimens with high Hg concentration in cytosol, implying that the Hg-Se complex was bound to the HMWS. Analysis of metals in the hepatocytosol by high-performance liquid chromatography/inductively coupled plasma-mass spectrometry (HPLC/ICP-MS) suggests that multiple isoforms of MT are present in hepatocytosol of the three species and that the metal profiles in hepatocytosols are different among the species. To our knowledge, this is the first report on the association of Ag with HMWS and MT in hepatocytosol of marine mammals and seabirds. Also, distribution and interaction of Hg and Se were investigated for the first time in hepatocytosol of the higher trophic marine animals.

Topics: Animals; Birds; Cytosol; Environmental Exposure; Female; Heavy Metal Poisoning; Liver; Male; Metallothionein; Porpoises; Seals, Earless; Tissue Distribution; Water Pollutants

An active biomonitoring experiment was performed using mussels collected at a clean site, Fier d'Ars, and transplanted to two locations, outside the harbor of La Rochelle and in the Baie de L'Aiguillon along the coast of Charentes (French Atlantic coast) beginning in April for several months. Mussels were collected in June and October. The cadmium, copper, and zinc concentrations of all resident and transplanted mussel samples and the polycyclic aromatic hydrocarbon and polychlorinated biphenyl concentrations in some mussel samples and in the sediment samples were determined. Mussel response was evaluated for several biochemical biomarkers: concentrations of metallothionein, activities of glutathione S-transferase and acetylcholinesterase (AChE) and levels of thiobarbituric reactive substance (TBARS). The physiological status of the animals was assessed using the condition index. A principal component analysis performed with the chemical and biochemical results of the evaluations of the resident and transplanted mussels collected in June allowed them to be separated into three groups: resident mussels from la Rochelle with high metal and TBARS levels, resident mussels from Baie de L'Aiguillon with a very high condition index, and resident mussels from Fier d'Ars and transplanted mussels at La Rochelle and Baie de L'Aiguillon with low TBARS and AChE activities. Strong seasonal variation from June to October of all parameters was noted. Mussels transplanted to La Rochelle appeared to be the most "polluted" in their pollutant concentrations and biochemical responses; moreover, the La Rochelle site had the highest concentration of organics in sediments of all the sites. The choice of Fier d'Ars as a reference site may be questionable because some of the biomarker responses of the mussels were higher than expected there, although these pollutants in mussels and sediment were present at the lowest concentrations measured.

Topics: Acetylcholinesterase; Animals; Biomarkers; Bivalvia; Environmental Monitoring; Environmental Pollutants; France; Geologic Sediments; Glutathione Transferase; Heavy Metal Poisoning; Metallothionein; Metals, Heavy; Polychlorinated Biphenyls; Polycyclic Aromatic Hydrocarbons; Water Pollutants