Tissue Metallothionein Research Articles

Ameliorative effects of Dunaliella salina microalgae on nanoparticle (ZnO NPs)-induced toxicity in fish.

Dunaliella salina (D. salina) is a well-known microalga that contains considerable amounts of nutritious and medicinal bioactive components. This work studied the modulatory role of D. salina against zinc oxide nanoparticle (ZnO NPs)-induced neurotoxic effects in adult zebrafish. Fishes were subjected to 0.69mg L-1 (1/5th 96-h LC50) for 4weeks; then, fishes were supplemented with D. salina in the diet for 2weeks at two levels (15 and 30%). Exposure to ZnO NPs induced a significant increase in the levels of reactive oxygen species (ROS), hydrogen peroxide (H2O2), malondialdehyde (MDA), and 8-hydroxy-2-deoxyguanosine (8-OH-dG) while accompanied with downregulation of antioxidant genes in the brain of exposed fishes. Brain neurochemistry and enzyme activities were also altered following ZnO NP exposure. ZnO NPs significantly reduced the neurotransmitters and acetylcholinesterase (AchE) activity while increasing Alzheimer's disease-related proteins and inflammatory response via upregulation of tumor necrosis factor (TNF-α). Additionally, ZnO NPs increased the indices of brain's DNA oxidative damage, increasing brain tissue's metallothionein (MT) and zinc residues. ZnO NPs upregulated the transcription patterns of apoptosis-related genes (casp3 and p53). D. salina dietary co-supplementation with ZnO NPs alleviated the ZnO NPsZnO NP-induced neuro-oxidative damages by lowering the lipid, DNA damage, and inflammatory biomarkers. Besides, D. salina alleviating responses were linked with increasing the levels of the assessed antioxidants. Conclusively, D. salina dietary supplementation induced potential alleviating effects of the ZnO NP-induced neurotoxicity in adult zebrafish.

Employing molecular, chemical and physiological techniques using Crassostrea virginica to assess ecosystem health along coastal South Carolina and North Carolina, United States

Natural and anthropogenic environmental impacts can introduce contaminants into sensitive habitats, threatening ecosystems and human health. Consistent monitoring of coastal areas provides critical environmental assessment data. Sediments and Eastern Oyster (Crassostrea virginica) tissues were collected at fourteen South Carolina (SC) and four North Carolina (NC) sites as part of the National Oceanic and Atmospheric Administration's Mussel Watch environmental monitoring program. Cellular and molecular techniques were employed to measure C. virginica stress response, specifically, Lipid Peroxidation (LPx), Glutathione (GSH), and qPCR techniques. Gene specific primers targeted for detecting oxidative stress and cellular death were developed in C. virginica to gauge response to current environmental conditions using gill and hepatopancreas (HP) tissue. In order to validate gene specific markers as additional assessment tools, a 96 h zinc (Zn) laboratory exposure was performed. Cellular biomarker data revealed tissue specific responses. Hepatopancreas data showed C. virginica exhibited stress through the lipid peroxidation assay amongst sampling sites, however, response was managed through glutathione detoxification. Gill tissue data had significantly lower levels of cellular biomarker response compared to hepatopancreas. Molecular biomarkers targeting these cellular stress pathways through qPCR analysis show upregulation of Metallothionein in hepatopancreas and gill tissue with a concurrent > 2-fold upregulation in the detoxification marker Superoxide Dismutase (SOD) at three NC sites. SC sites displayed higher stress levels through LPx assays and down-regulation in GPx gene activity. Laboratory zinc exposure revealed no significance in cellular biomarker results, however, molecular data showed gills responding to zinc treatment through upregulation of Metallothionein, SOD and Cathepsin L, indicating an acute response in gills. Collectively, chemical, cellular and molecular methods clarify sentinel stress response of biological impacts and aid in evaluating environmental health in coastal ecosystems. This combined methodological approach provides a detailed analysis of environmental conditions and improves land-use management decisions.

Relationship between Metallothionein and Mercury (Hg) in the Gill Tissue of the Barbonymus altus in the Brantas River Jombang, East Java

Pollution of river waters is most often the result of human activities in managing agricultural land, industry, and transportation. The entry of pollutants such as heavy metal mercury (Hg) into the waters can interfere with the survival of fish waters. Red Bader Fish (Barbonymus altus) can be used as a biomarker in analyzing aquatic environmental conditions in the Brantas River, Jombang, East Java. This study aimed to analyze the relationship between metallothionein and heavy metal mercury in the gill tissue of Barbonymus altus. Sampling was conducted at three stations with three replications from February to March 2022. Data analysis used regression and correlation methods to determine the relationship between metallothionein and metal mercury in gill tissue. The density and intensity of metallothionein in the gill tissue of Barbonymus altus were analyzed using the immunohistochemical method. The mercury (Hg) content in the gill tissue of Barbonymus altus was analyzed using Atomic Absorption Spectroscopy (AAS). The results showed that the metallothionein density at each station included station 1 of 252.52 × 10-6 – 497.79 × 10-6 MT.µm-2, station 2 of 277.78 × 10-6 – 378.78 × 10-6 MT.µm-2, and station 3 of 303.03 × 10-6 – 404.04 × 10-6 MT.µm-2. The relationship between the metallothionein density value and mercury in the gill tissue has a moderate correlation. In contrast, the metallothionein intensity value with mercury in the gill tissue strongly correlated with a significance value of sig < 0.05. The Hg levels in the gill tissue, density, and intensity have significant values at the all three locations based on the real difference test (sig. 0.00). Keywords: Barbonymus altus, density, intensity, mercury, river.

The effects of mercury exposure on Amazonian fishes: An investigation of potential biomarkers

Exposure to mercury can interfere with the expression of proteins and enzymes, compromise important pathways, such as apoptosis and glucose metabolism, and even induce the expression of metallothioneins. In this study, analytical techniques were used to determine the concentration of total mercury (THg) in muscle and liver tissue, protein pellets, and spots [using graphite furnace atomic absorption spectrometry (GFAAS)], and molecular techniques were used to identify metalloproteins present in mercury-associated protein spots. Thirty individuals from three different fish species, Cichla sp. (n = 10), Brachyplatystoma filamentosum (n = 10), and Semaprochilodus sp. (n = 10) from the Brazilian Amazon were used. Oxidative stress indicators [such as glutathione peroxidase (GSH-Px), catalase (CAT), superoxide dismutase (SOD), a marker of lipid peroxidation (LPO)] and the possible expression of metallothioneins in muscle and liver tissues were investigated. The two piscivorous species, Cichla sp. and B. filamentosum, presented the highest concentrations of mercury in their hepatic tissue, 1219 ± 15.00 and 1044 ± 13.6 μg kg−1, respectively, and in their muscle tissue, 101 ± 1.30 μg kg−1 and 87.4 ± 0.900 μg kg−1, respectively. The non-carnivorous species Semaprochilodus sp. had comparatively low concentrations of mercury in both its hepatic (852 ± 11.1 μg kg−1) and muscle (71.4 ± 0.930 μg kg−1) tissues. The presence of mercury was identified in 24 protein spots using GFAAS; concentrations ranged from 11.5 to 787 μg kg−1, and mass spectrometry identified 21 metal-binding proteins. The activities of GSH-Px, CAT, and SOD, related to oxidative stress, decreased proportionally as tissue Hg concentrations increased, while the levels of LPO markers increased, indicating the presence of stress. Our study results demonstrate possible mercury interference in oxidative stress markers (GSH-Px, CAT, SOD, and LPO), in addition to the identification of 21 metal-binding proteins as possible biomarkers of mercury exposure in fish.

Reducing Endogenous Labile Zn May Help to Reduce Smooth Muscle Cell Injury around Vascular Stents.

Vascular stent service involves complex service environments and performance requirements, among which the histocompatibility of the stent could seriously affect the therapeutic effect. In the pathology of vascular disease, the thin fiber cap is easily ruptured, exposing the necrotic core below, and triggering a series of dangerous biochemical reactions. In contrast, the thin neointima, considered an essential structure growing on the stent, may evolve into vulnerable plaque structures due to lesions induced by the stent. Therefore, the reduction of necrosis around the stent below the thin neointima is indispensable. In this work, different cell model experiments suggested that the content of endogenous labile Zn positively correlated with cell injury. Zinquin-Zn fluorescence experiments and zinc ion channels research suggested that the change in the content of endogenous labile Zn in smooth muscle cells is affected by different stent coatings. The content of endogenous labile Zn in cells negatively correlated with cell viability. Animal experiments indirectly verified the increase in endogenous labile Zn by detecting the expression of Zn regulatory protein (metallothionein) in the necrotic tissues. Reducing the content of endogenous labile Zn may favor a reduction in smooth muscle cell injury and necrosis. This biochemical mechanism is effective in improving the therapeutic effect of vascular stents.

Metallothionein and Cadmium Toxicology—Historical Review and Commentary

More than one and a half centuries ago, adverse human health effects were reported after use of a cadmium-containing silver polishing agent. Long-term cadmium exposure gives rise to kidney or bone disease, reproductive toxicity and cancer in animals and humans. At present, high human exposures to cadmium occur in small-scale mining, underlining the need for preventive measures. This is particularly urgent in view of the growing demand for minerals and metals in global climate change mitigation. This review deals with a specific part of cadmium toxicology that is important for understanding when toxic effects appear and, thus, is crucial for risk assessment. The discovery of the low-molecular-weight protein metallothionein (MT) in 1957 was an important milestone because, when this protein binds cadmium, it modifies cellular cadmium toxicity. The present authors contributed evidence in the 1970s concerning cadmium binding to MT and synthesis of the protein in tissues. We showed that binding of cadmium to metallothionein in tissues prevented some toxic effects, but that metallothionein can increase the transport of cadmium to the kidneys. Special studies showed the importance of the Cd/Zn ratio in MT for expression of toxicity in the kidneys. We also developed models of cadmium toxicokinetics based on our MT-related findings. This model combined with estimates of tissue levels giving rise to toxicity, made it possible to calculate expected risks in relation to exposure. Other scientists developed these models further and international organizations have successfully used these amended models in recent publications. Our contributions in recent decades included studies in humans of MT-related biomarkers showing the importance of MT gene expression in lymphocytes and MT autoantibodies for risks of Cd-related adverse effects in cadmium-exposed population groups. In a study of the impact of zinc status on the risk of kidney dysfunction in a cadmium-exposed group, the risks were low when zinc status was good and high when zinc status was poor. The present review summarizes this evidence in a risk assessment context and calls for its application in order to improve preventive measures against adverse effects of cadmium exposures in humans and animals.

METALLOTHIONEIN EXPRESSION IN A PARASITIC CRUSTACEAN, LAMPROGLENA CLARIAE (CRUSTACEA: COPEPODA), ON CLARIAS GARIEPINUS (TELOESTEI: CLARIIDAE) CORRESPONDS TO WATER QUALITY.

Globally, parasites are sensitive toward environmental changes, and, in some cases, they are even more sensitive than their hosts. However, there is limited knowledge on the physiological responses of parasites and their effects on their hosts in relation to environmental degradation. In this study, metallothioneins (MTs) were isolated and compared between the ectoparasite Lamproglena clariae and its host fish Clarias gariepinus. Differences in the levels of MTs in the parasite and host were compared to physicochemical water quality variables and metals to determine if MT expression was linked with changes in water quality. Clarias gariepinus individuals were sampled from 2 sites of differing water quality along the Vaal River using gill nets and assessed for L. clariae. Gill, muscle, and liver tissue of the host and L. clariae were collected and stored in liquid nitrogen for analysis of MT. Water and sediment samples were collected for metal analysis by inductively coupled plasma-optical emission spectrometry and inductively coupled plasma-mass spectrometry. Nutrient levels and water hardness in water samples were assessed using spectrophotometry. MTs were quantified using spectrophotometry and size exclusion chromatography in the host and parasite, respectively. Infections by L. clariae differed between sites, with higher parasite intensity at the unpolluted Vaal Dam site. Concentrations of MT in host tissues and L. clariae were significantly higher at the polluted site, below the Vaal River Barrage, compared to the Vaal Dam site. Parasite MT concentrations were significantly lower compared to concentrations in the liver and gill tissue of C. gariepinus individuals. In conclusion, differences in the concentrations of MT and infection biology of L. clariae reflected the state of the environment and support the usefulness of this parasite and other Lamproglena spp. as bioindicators.

Edible bird's nest protects histomorphology of rat's uterus against cadmium (Cd) toxicity through a reduction of Cd deposition and enhanced antioxidant activity.

Cadmium (Cd) is often associated with reproductive disorders of mammals. Edible bird’s nest (EBN) is a natural food product made of swiftlet's salivary secretion used to make their nests and it has been consumed as a tonic food for decades. This research aimed to study the protective effects of EBN against Cd-induced uterine toxicity in Sprague Dawley rats. Thirty (30) female Sprague Dawley rats were assigned into five groups as follows: group 1- negative control (NC) received distilled water; group 2 - positive control (PC) administered with CdCl2, 5 mg/kg BW; while groups EBN-1, EBN-2, and EBN-3 received CdCl2 (5 mg/kg BW) plus graded concentrations of 60, 90 and 120 mg/kg BW of EBN, respectively. After four weeks of daily oral treatment, rats were euthanized to collect the uterus for evluations of histopathological changes, Cd concentrations and Metallothionein (MT) expressions using H&E stain, inductive coupled plasma mass spectrometry (ICP-MS) and immunohistochemistry, respectively. Blood samples were collected for superoxide dismutase (SOD) analysis using SOD assay kit. Results revealed that the CdCl2 without EBN supplement (PC) group had elevated levels of Cd in the uterus along with increased MT expressions and decreased SOD enzyme activity as compared to the NC group. Moreover, uterine histopathological changes, including glandular cysts and loss of normal structure of luminal epithelium (LE) and glandular epithelium (GE) were found in the PC group. Interestingly, groups treated with CdCl2 along with EBN (EBN1, EBN2, EBN3) showed lower levels of uterine tissue Cd deposition and MT expression, lower degenerative changes with normal histomorphology of glands, and increased SOD activity as compared to the PC group. Overall, the findings revealed that oral exposure to Cd at a dose of 5 mg/kg BW resulted in significant alterations in the rat's uterus. However, the toxicity effect was averted by EBN treatment in a dose dependant manner; highest protection achieved with EBN 120 mg/kg BW, through a possible detoxification mechanism and prevention of Cd deposition.

The Herbal Constituents in An-Gong-Niu-Huang Wan (AGNH) Protect against Cinnabar- and Realgar-Induced Hepatorenal Toxicity and Accumulations of Mercury and Arsenic in Mice.

An-Gong-Niu-Huang Wan (AGNH) has been a well-known cinnabar- and realgar-containing compound recipe for cerebral diseases. Unfortunately, its clinical practice is often restrained by the specific hepatorenal toxicity of cinnabar and realgar (C + R). In previous research studies, we have found that the antioxidative and anti-inflammatory effects of its herbal constituents could mitigate the risks from the toxicity. The underlying detoxification mechanisms are still unsolved. The present study investigated the protective effects of AGNH's herbal constituents on hepatorenal injury induced by C + R. For the mice treated with C + R, the increased expression levels of sensitive biomarkers of metal exposure and hepatorenal toxicity, including metallothionein (MT) in both hepatorenal tissues and kidney induced molecule-1 (KIM-1) in the kidney, were simultaneously reduced when C + R coadministered with other herbal medicines. In addition, the contents of trivalent As (AsIII), pentavalent As (Asv), and mercury (Hg) in hepatorenal tissues of mice were also significantly reduced benefiting from the herbal constituents in AGNH. Further mechanism studies showed that the herbal constituents in AGNH could downregulate the expressions of uptake transporters (AQP9 and OAT1) and upregulate the expressions of efflux transporters (P-gp, MRP2, and MRP4) in mice intoxicated by C + R. Our results suggested that AGNH's herbal constituents protect the body against C + R-induced hepatorenal toxicity and accumulations of Hg and As, which could be associated with the reestablishment of heavy metal homeostasis and the detoxification system.

Comparative responses of Sinopotamon henanense to acute and sub-chronic Cd exposure.

Studies on the freshwater crab Sinopotamon henanense have shown that acute and sub-chronic Cd2+ exposure induced differential alterations in the respiratory physiology and gill morphology. To elucidate Cd2+ toxicity under these two exposure conditions, crabs were acutely exposed to 7.14, 14.28, and 28.55 mg/L Cd2+ for 96 h and sub-chronically exposed to 0.71, 1.43, and 2.86 mg/L Cd2+ for 3 weeks. The Cd2+ accumulation, total metallothionein (MT), superoxide dismutase, and malondialdehyde (MDA) contents in the gill tissues were detected. Moreover, the glucose-6-phosphate dehydrogenase (G6PDH) activity, NADPH content, reduced glutathione (GSH), oxidized glutathione (GSSG), and GSH/GSSG ratio in the hepatopancreas were determined. The morphology of the X-organ-sinus gland complex was also observed. The results showed that sub-chronical Cd2+ exposure induced lower MT content and higher MDA level in the gills than in the acute exposure. In the hepatopancreas, acute Cd2+ exposure decreased the pentose phosphate pathway activity and NADPH content; however, an increased G6PDH activity and NADPH content were detected in sub-chronic Cd2+ exposure (2.86 mg/L). Morphological changes occurred in the sinus gland in crabs exposed to 2.86 mg/L Cd2+ for 3 weeks. The tightly packed structure composed by the axons, enlarged terminals, and glial cells, became loose and porous. Ultra-structurally, a large number of vacuoles and few neurosecretory granules were observed in the axon terminal. These effects added to our understanding of the toxic effects of Cd2+ and provide biochemical and histopathological evidence for S. henanense as a biomarker of acute or long-term waterborne Cd2+ pollution.

Coal mine effluent-induced metal bioaccumulation, biochemical, oxidative stress, metallothionein, and histopathological alterations in vital tissues of the catfish, Clarias batrachus.

In the present study, a multi-biomarker approach was used to assess the toxicity of the coal mine effluent (CME) generated at the Rajrappa coal mine on the catfish Clarias batrachus. A core of biomarkers indicative of nutritional value, oxidative stress, and histopathology was selected to illustrate the toxic effects of CME-containing different heavy metals and other toxicants. The results of metal bioaccumulation in CME-exposed fish tissues revealed the highest metal concentration in liver (1.34-297.68mg/kg) while lowest in muscles (1.47-23.26mg/kg) as compared to other tissues and so was the metallothionein level. The high value of bioaccumulation observed in liver, kidney, and gills reflects their affinity for metals. In addition, the values of metal pollution index (MPI) of different fish tissues further affirmed that liver followed by kidney and gills are at greater risk than brain, skin, and muscles. Significant alterations in the activity of certain enzymes (aspartate amino transferase, alanine amino transferase, alkaline phosphatase) as well as oxidative stress markers (superoxide dismutase, catalase and lipid peroxidation) were detected in the tissues of CME-exposed fish. The tissue-specific metal accumulation and increased metallothionein levels may be associated with the biochemical and physiological activity of an organ and its constitutive antioxidant defenses. The histopathological changes in the various tissues of the CME-exposed fish justify the high metal accumulation and biochemical alterations. Overall results indicate that the Rajrappa coal mine effluent is very toxic having adverse health impact on the fish and might also affect the human health when consumed.

High-dose dietary supplementation with zinc prevents gut inflammation: Investigation of the role of metallothioneins and beyond by transcriptomic and metagenomic studies.

Although it is known that zinc has several beneficial roles in the context of gut inflammation, the underlying mechanisms have not been extensively characterized. Zinc (Zn) is known to be the primary physiological inducer of the expression of the metallothionein (MT) superfamily of small stress-responsive proteins. The expression of MTs in various tissues is induced or enhanced (including the gastrointestinal tract (GIT)) by a variety of stimuli, including infection and inflammation. However, the MTs' exact role in inflammation is still subject to debate. In order to establish whether or not MTs are the sole vectors in the Zn-based modulation of intestinal inflammation, we used transcriptomic and metagenomic approaches to assess the potential effect of dietary Zn, the mechanisms underlying the MTs' beneficial effects, and the induction of previously unidentified mediators. We found that the expression of endogenous MTs in the mouse GIT was stimulated by an optimized dietary supplementation with Zn. The protective effects of dietary supplementation with Zn were then evaluated in mouse models of chemically induced colitis. The potential contribution of MTs and other pathways was explored via transcriptomic analyses of the ileum and colon in Zn-treated mice. The microbiota's role was also assessed via fecal 16S rRNA sequencing. We found that high-dose dietary supplementation with Zn induced the expression of MT-encoding genes in the colon of healthy mice. We next demonstrated that the Zn diet significantly protected mice in the two models of induced colitis. When comparing Zn-treated and control mice, various genes were found to be differentially expressed in the colon and the ileum. Finally, we found that Zn supplementation did not modify the overall structure of the fecal microbiota, with the exception of (i) a significant increase in endogenous Clostridiaceae, and (ii) some subtle but specific changes at the family and genus levels. Our results emphasize the beneficial effects of excess dietary Zn on the prevention of colitis and inflammatory events in mouse models. The main underlying mechanisms were driven by the multifaceted roles of MTs and the other potential molecular mediators highlighted by our transcriptomic analyses although we cannot rule out contributions by other factors from the host and/or the microbiota.

Heavy metal toxicity has an immunomodulatory effect on metallothionein and glutathione peroxidase gene expression in Cyprinus carpio inhabiting a wetland lake and a culture pond.

The study provides cumulative data on the status of the two water bodies. The study designed revealed physicochemical properties (temperature, dissolved oxygen, pH, total dissolved solids and conductivity) to be in the desirable range, however, amongst the heavy metals excepting for Cd all were found to be higher than the permissible limits set by WHO and USEPA. It was observed that these elements cast their impact on bioindices (hepatosomatic index, condition factor, spleenosomatic index and kidney somatic index), renal marker enzyme (creatine kinase), hepatic marker enzymes (aspartate aminotransferase, alanine aminotransferase and alkaline phosphatase), histology of immune organs (liver, spleen, head-kidney and thymus) and level of serum immunoglobulin (IgM). Further, expression levels of Metallothionein (MT) and Glutathione peroxidase (GPX) genes in immune-related tissues (liver, spleen, head-kidney, thymus and blood) observed indicates metal pollution and abiotic stresses. These alterations are reliable indicators of the cellular and humoral immune response in Cyprinus carpio.

Metallothioneins enhance chromium detoxification through scavenging ROS and stimulating metal chelation in Oryza sativa

Metallothioneins (MTs) is a metal ion binding protein to detoxify heavy metal stress in plant cells. This study examines involvement of MTs in metal chelation and ROS scavenging in rice seedling under Cr induction either Cr(VI) or Cr(III) at three different effective concentrations using Agilent 44K rice microarray and real-time PCR technology. Results showed that the concentration of Cr was higher in roots than in shoots in both Cr treatments. Accumulation of both H2O2 and O2− in rice tissues was evident, but the fluctuation of H2O2 was more remarkable than O2−. Both Cr exposures resulted in enhancement of MTs in plant tissues. Results from PCR analysis confirmed that ten specific OsMT genes responsible for regulating ROS removal were expressed differentially in plant tissues as well as in Cr variants, suggesting that their different regulation and responsiveness strategies. Expression patterns of metal chelation-related OsMT genes, after Cr exposure were also inconsistent in rice tissues. Longer exposure periods caused more transcriptional changes in both Cr treatments. We also noticed that OsMT1b might carry more weight during Cr chelation in roots rather than in shoots, while OsMT2c had more important role in eliminating H2O2 accumulation in shoots than roots. These results suggest that different speciation of Cr in rice tissues resulted in inconsistent transcriptional changes of OsMT genes, which functioned in different regulation and responsiveness pathways responsible for metal ions chelating and ROS scavenging during Cr detoxification.

Mass spectrometric imaging of cysteine rich proteins in human skin

Looking insight pathological processes, metallothioneins (MTs) are considered to be potential biomarkers for monitoring of a development of various types of diseases, such as cancer. The early identification of the MTs in biological tissues could be important tool for the estimation of appropriate clinical therapy. Therefore, here we investigated the application of matrix assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI) together with immunohistochemical analyses (IHC) using MT-1/2 antibody for MT detection in formalin-fixed paraffin-embedded (FFPE) biopsy specimens of human skin. Principal component analyses revealed differences in the peptide/protein profiles separating healthy skin from the carcinoma specimens. Statistically significant ion peaks at m/z 6038, 6300, 6676, and 7026 were more frequently detected in squamous cell carcinoma (SCC), basal cell carcinoma (BCC) and melanoma. Using IHC, we found that MT-1/2 was significantly higher in SCC and melanoma compared to healthy skin. Surprisingly, significantly low levels of MT-1/2 were found in BCC. On one side, the results indicate important role of MTs in melanoma occurrence and progression, as on the second side, there are hidden processes associated with MTs based on differences of the occurrence of the MS peaks, which could be associated with cycling of MTs isoforms.

Preliminary analysis of the interactions between CdTe quantum dots and human metallothionein.

Metallothionein (MT) plays the important role in the detoxification of heavy metals, protection against oxidative compounds and as a prognostic marker in the development of tumors. It is important to find selective, stable and sensitive tools and probes to evaluate the presence of MT in biological fluids or tissues. QDs linked with ligands such as peptides or small molecules are a promising tool for selective, fast, and sensitive tagging and imaging in medicine. In previous findings, the authors proved the possibility of interaction with QDs (particularly with CdTe) and analyzed the stability of the formed complexes between CdTe and MT during incubation over time. Following that, an initial analysis of the interactions between CdTe quantum dots (QDs) and human metallothionein (MT) was performed. Complexes of mercaptosuccinic acid-covered CdTe QDs + MT were investigated using fluorescence intensity changes along a timeline, quenching analysis, stability interpretation based on zeta potential, and quenching intensity. Based on the preliminary results, it appears as though the possible interactions depend on the size of the CdTe QDs. Additionally, the formation of complexes between CdTe and human MT likely depends mostly on structural changes and conformational reorganization rather than on electrostatic interactions. Both types of interactions are responsible for complex creation and stabilization.

Assessing lead toxicity in the clam Ruditapes philippinarum: Bioaccumulation and biochemical responses

Lead (Pb) is a non-essential metal. Its occurrence in the environment is related principally to anthropogenic contamination. Pb is toxic to aquatic organisms and can provoke damage to membranes and inhibit the activity of essential enzymes. The filter-feeding, Manila clam Ruditapes philippinarum is widely used as a biomonitor organism to assess metal toxicity. Among biomarkers related to the Pb toxicity, the enzymatic activity of δ-aminolevulinic acid dehydratase (δ-ALAD) has been adopted as a specific tool. Metallothionein (MT), lipid peroxidation (LPO) and antioxidant enzymes activities, such as catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR) and glutathione S-transferase (GST) and superoxidase dismutase (SOD) have also been employed to assess the toxic effect of metals. Two target tissues, the gills and the digestive gland, were selected to examine biomarker responses. In order to assess the effects of Pb accumulation and the mechanisms involved in the recovery from it, clams were exposed at two Pb levels (10 and 100 µg/L) for 7 days and were later maintained in clean water for 7 days as a depuration period. Pb accumulation was dependent on the exposure concentration and higher Pb levels were observed in the gills compared to the digestive gland. Inhibition of δ-ALAD, GST and SOD and the induction of MT and LPO over the exposure period were observed in the gills and the digestive gland of R. philippinarum. The depuration period showed a continuous inhibition of the δ-ALAD activity and induction of MT and LPO in both tissues. These results demonstrate that lead induced an exposure effect and the 7 days of depuration were not sufficient to recover the basal health status of the clams.

Influence of SR-90 on the Morphometric Indices and the Level of Proteins of Metallothioneins in the Soft Tissues of Terrestrial Mollusks Bradybaena Fruticum in the Area of Location of the Regional Radioactive Waste Storage Facility

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Mercury and metallothioneins in blood fractions and tissues of captive Morelet's crocodiles in Quintana Roo, Mexico

Even though studies of heavy metals and their relation with metallothioneins (MTs) in reptile tissues have been conducted, research on heavy metals and MTs in organs and blood fractions of crocodylians is limited. To date there are no studies on the distribution of MTs in organs and their relation with mercury (Hg), or the concentration of MTs in plasma and erythrocytes of crocodylians. In order to understand the role of MTs in crocodiles, our aim was to assess the detoxification mechanisms for exposure to metals in Morelet's crocodile (Crocodylus moreletii) by using two biomarkers (Hg and MTs) in blood fractions and tissues, and comparing them with concentrations between two populations of crocodiles, one previously wild and currently captive (Theme Park) and another raised in a Wildlife Management Unit (WMU). The caudal scutes from the Theme Park showed higher concentrations of Hg than those from the skin in the WMU samples, and significant negative relationships were observed between the total length (TL) and Hg in the scutes. The significant negative relationship between Hg and hemoglobin (Hb) may be due to disorders in the oxidation process and even denature of this protein, while the positive trend observed between MTs and Hb is consistent with the detoxifying functions and the protection against oxidative damage. This study is the first to report Hg in the erythrocytes of crocodylians and the use of MTs for testing the potential of these biomarkers as a tool to assess Hg exposure in crocodile's habitats.

The effect of ozone on the expression of metallothionein in tissues of rats chronically exposed to cadmium.

Our aims were to evaluate the expression of metallothionein (MT) in an experimental rat model which experienced chronic exposure to cadmium (Cd) and to measure its expression after ozone therapy (OT) or oxygen (Ox) in the same model, as compared to the control group, which was exposed to neither cadmium nor ozone. Forty male Wistar rats were divided into 5 groups: control, Cd, Cd and Ox, Cd and Oz, and Oz. During our research, Cd concentration (ASA) and MT concentration (ELISA) were determined in supernatants of the kidneys, liver and pancreas. SDS-PAGE analyses and immunohistochemical localization were used to evaluate the level of MT expression in the tissue. In rats intoxicated with Cd, the highest concentration of both Cd and MT was observed in the kidneys and liver, with a significantly lower concentration measured in the pancreas. Ozone therapy reduces the accumulation of cadmium in the liver and kidneys, resulting in a reduced expression of metallothionein in those tissues.