Metallothionein Levels Research Articles

ZnT8 Deficiency Protects From APAP-Induced Acute Liver Injury by Reducing Oxidative Stress Through Upregulating Hepatic Zinc and Metallothioneins.

Zinc transporter 8 (ZnT8) is an important zinc transporter highly expressed in pancreatic islets. Deficiency of ZnT8 leads to a marked decrease in islet zinc, which is thought to prevent liver diseases associated with oxidative stress. Herein, we aimed to investigate whether loss of islet zinc affects the antioxidant capacity of the liver and acute drug-induced liver injury. To address this question, we treated ZnT8 knockout (KO) or wild-type control mice with 300 mg/ kg acetaminophen (APAP) or phosphate-buffered saline (PBS). Unexpectedly, we found that loss of ZnT8 in mice ameliorated APAP-induced injury and was accompanied by inhibition of c-Jun N-terminal kinase (JNK) activation, reduced hepatocyte death, and decreased serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST). An increase in hepatic glutathione (GSH) was observed, corresponding to a decrease in malondialdehyde (MDA) and 4-hydroxynonenal (4-HNE) levels. APAP-induced inflammation and glycogen depletion were alleviated. In contrast, no significant changes were observed in cytochrome P450 family 2 subfamily E member 1 (CYP2E1), the main enzyme responsible for drug metabolism. Elevated levels of hepatic zinc and metallothionein (MT) were also observed, which may contribute to the hepatoprotective effect in ZnT8 KO mice. Taken together, these results suggest that ZnT8 deficiency protects the liver from APAP toxicity by attenuating oxidative stress and promoting hepatocyte proliferation. This study provides new insights into the functions of ZnT8 and zinc as key mediators linking pancreatic and hepatic functions.

Antioxidant potential of bulk and nanoparticles of naringenin against cadmium-induced oxidative stress in Nile tilapia, Oreochromis niloticus

Abstract The study assessed the attenuating effect of citrus flavonone, naringenin, and its nanoformulation against cadmium (Cd) toxicity in Nile tilapia (Oreochromis niloticus). Nanoparticles of naringenin (NNn) were synthesized; size 165.1 nm (PDI 0.396) in variable shapes; cluster widespread, spherical accumulated, and tubular bacillary. Parallel, mature male Nile tilapia (n = 120 fish) were used for the exposure study. Group I was negative control. The groups II, III, and IV were exposed to 5 ppm of cadmium chloride monohydrate (CdCl2·H2O) for 21 days. Group III was treated with bulk naringenin (BNn) (3 ppm) and group IV was treated with NNn (3 ppm). Group V was exposed only to NNn (3 ppm). Cd-induced oxidative stress was evident from a significant increase in the hepatic malondialdehyde (MDA) levels and modulation of antioxidant enzymes in the liver and kidney. A significant increase in the hepatic metallothionein and HSP70 levels in the gills was observed. Treatment with both BNn and NNn significantly (p ≤ 0.05) reversed the Cd-induced alterations. However, the protective effect of nano naringenin was more profound on the bioaccumulation of Cd in liver and levels of HSP70 in the gills. These key findings could add to the commercial exploitation of naringenin and its nanosized form as a dietary therapeutic molecule.

The cadmium toxicity in gills of Mytilus coruscus was accentuated by benzo(a)pyrene of higher dose but not lower dose

In natural environment, the existence of interactions of toxic mixtures could induce diverse biochemical pathways and consequently exert different toxicological responses in aquatic organisms. However, little information is available on the effects of combined xenobiotics on lower aquatic invertebrates. Here, we assessed the effects of cadmium (Cd, 0.31 mg/L) as well as the mixture of Cd (0.31 mg/L) and benzo(a)pyrene (Bap, 5 or 50 μg/L) on bioaccumulation, antioxidant, lipid peroxidation (LPO) and metallothionein (MT) responses in gills of thick shell mussel Mytilus coruscus. Upon exposed to single Cd, the metal bioaccumulation, antioxidant enzymes activities, LPO and MT level significantly increased in the gills, suggesting an apparent toxicity to mussels. The interaction of Cd + 5 μg/L Bap did not significantly alter these endpoints compared to single Cd. However, once the dose of Bap elevated to 50 μg/L, the induction of bioaccumulation, antioxidant system and LPO was even more pronounced while the induction of MT was remarkably inhibited, implying an accentuated toxicity. Collectively, the current results demonstrated that 0.31 mg/L Cd exposure resulted in severe toxicity to mussels despite of the induction of MT system to alleviate the metal toxicity. Once the Cd exposure combined with Bap, the lower dose of Bap could not change the Cd toxicity while the higher dose of Bap accentuated the toxicity by inhibiting metallothionein synthesis. These findings might provide some useful clues for elucidation the mechanism of the interaction of combined xenobiotics in molluscs.

Cadmium Accumulation Involves Synthesis of Glutathione and Phytochelatins, and Activation of CDPK, CaMK, CBLPK, and MAPK Signaling Pathways in Ulva compressa.

In order to analyze the effect of cadmium in Ulva compressa (Chlorophyta), the alga was cultivated with 10, 25, and 50 μM of cadmium for 7 days, and the level of intracellular cadmium was determined. Intracellular cadmium showed an increase on day 1, no change until day 5, and an increase on day 7. Then, the alga was cultivated with 10 μM for 7 days, and the level of hydrogen peroxide, superoxide anions, and lipoperoxides; activities of antioxidant enzymes ascorbate peroxidase (AP), dehydroascorbate reductase (DHAR), and glutathione reductase (GR); the level of glutathione (GSH) and ascorbate (ASC); and the level of phytochelatins (PCs) and transcripts encoding metallothioneins (UcMTs) levels were determined. The level of hydrogen peroxide increased at 2 and 12 h, superoxide anions on day 1, and lipoperoxides on days 3 to 5. The activities of AP and GR were increased, but not the DHAR activity. The level of GSH increased on day 1, decreased on day 3, and increased again on day 5, whereas ASC slightly increased on days 3 and 7, and activities of enzymes involved in GSH and ASC synthesis were increased on days 3 to 7. The level of PC2 and PC4 decreased on day 3 but increased again on day 5. The level of transcripts encoding UcMT1 and UcMT2 increased on days 3 to 5, mainly that of UcMT2. Thus, cadmium accumulation induced an oxidative stress condition that was mitigated by the activation of antioxidant enzymes and synthesis of GSH and ASC. Then, the alga cultivated with inhibitors of calcium-dependent protein kinases (CDPKs), calmodulin-dependent protein kinases (CaMKs), calcineurin B-like protein kinases (CBLPKs), and MAPKs and 10 μM of cadmium for 5 days showed a decrease in intracellular cadmium and in the level of GSH and PCs, with the four inhibitors, and in the level of transcripts encoding UcMTs, with two inhibitors. Thus, CDPKs, CaMK, CBLPKS, and MAPKs are involved in cadmium accumulation and GSH and PC synthesis, and GSH and PCs and/or UcMTs may participate in cadmium accumulation.

Effects of Zinc Lactate Supplementation on Growth Performance, Intestinal Morphology, Serum Parameters, and Hepatic Metallothionein of Chinese Yellow-Feathered Broilers

In poultry, organic zinc compounds have higher bioavailability than inorganic zinc sources. However, as an organic zinc source, the application of zinc lactate (ZL) on Chinese yellow-feathered broilers has been rarely reported. Hence, the present study aimed to investigate the effects of ZL supplementation on growth performance, small intestinal morphology, serum biochemical parameters, immune organ index, as well as hepatic metallothionein of Chinese yellow-feathered broilers. A total of 2100 broilers (19 days old) were randomly assigned to 5 treatment groups, including the control (fed basal diet), ZL40 (basal diet plus 40 mg/kg ZL), ZL60 (basal diet plus 60 mg/kg ZL), ZL80 (basal diet plus 80 mg/kg ZL), and ZS80 (basal diet plus 80 mg/kg ZS. Each treatment group had 6 replicates with 70 chickens per replicate. Compared to the control group, the ZL40 and the ZS80 groups had a lower feed to gain ratio (P < 0.05), ZL40 group had higher duodenum and ileum villus heights (P < 0.05), and ZS80 and ZL80 groups had a lower ratio of villus height to crypt depth in the jejunum (P < 0.01). In addition, the ZL60 group had a higher concentration of total protein (P < 0.05) and activity of glutathione peroxidase (GSH-Px) (P < 0.01) compared with the ZS80 and the control groups. Interestingly, the ZL40, ZL60, and ZL80 groups all had higher levels of hepatic metallothionein than the other groups (P < 0.01). In conclusion, zinc lactate had a higher bioavailability and could be used as an alternative to zinc sulfate.

Elevated metallothionein expression in long-lived species mediates the influence of cadmium accumulation on aging.

Cadmium (Cd) accumulates with aging and is elevated in long-lived species. Metallothioneins (MTs), small cysteine-rich proteins involved in metal homeostasis and Cd detoxification, are known to be related to longevity. However, the relationship between Cd accumulation, the role of MTs, and aging is currently unclear. Specifically, we do not know if long-lived species evolved an efficient metal stress response by upregulating their MT levels to reduce the toxic effects of environmental pollutants, such as Cd, that accumulate over their longer life span. It is also unknown if the number of MT genes, their expression, or both protect the organisms from potentially damaging effects during aging. To address these questions, we reanalyzed several cross-species studies and obtained data on MT expression and Cd accumulation in long-lived mouse models. We confirmed a relationship between species maximum life span in captive mammals and their Cd content in liver and kidney. We found that although the number of MT genes does not affect longevity, gene expression and protein amount of specific MT paralogs are strongly related to life span in mammals. MT expression rather than gene number may influence the high Cd levels and longevity of some species. In support of this, we found that overexpression of MT-1 accelerated Cd accumulation in mice and that tissue Cd was higher in long-lived mouse strains with high MT expression. We conclude that long-lived species have evolved a more efficient stress response by upregulating the expression of MT genes in presence of Cd, which contributes to elevated tissue Cd levels.

Insights on the toxicity of selected rare earth elements in rainbow trout hepatocytes

The increasing extraction of rare earth elements (REEs) for technology applications raised concerns for contamination and toxicity in the environment. The purpose of this study was to examine the toxicity of the following REEs in primary cultures of rainbow trout hepatocytes: yttrium (Y), samarium (Sm), gadolinium (Gd), terbium (Tb) and lutetium (Lu). Hepatocytes were exposed to increasing concentrations of the above elements for 24 h at 15 °C and they were analyzed for viability, metallothioneins (MT), glutathione-S-transferase (GST) and arachidonate cyclooxygenase (COX) as markers of oxidative stress and inflammation. The results revealed that the cytoxicity of REEs were as follows in decreasing order: Y > Sm > Lu > Tb > Gd in concordance with published rainbow trout mortality data. While effects on GST and COX activities were marginal, MT levels were more strongly increased with the 2 most toxic REEs (Y and Sm) and Gd, while MT levels were decreased in the least toxic ones (Tb, Lu). While cell viability followed published trout mortality data, it also followed the redox potential and the glutathione affinity constant (log k). The capacity to induce/decrease MT levels was associated with ionic radius, log k (glutathione) and electronegativity. A proposed mechanism of toxicity for REEs is presented based on the chemical properties of REEs, namely the glutathione binding constant and ionic radius, in light of the observed effects in trout hepatocytes.

Characterization of Brassica rapa metallothionein and phytochelatin synthase genes potentially involved in heavy metal detoxification.

Brassica rapa is an important leafy vegetable that can potentially accumulate high concentrations of cadmium (Cd), posing a risk to human health. The aim of the present study was to identify cadmium detoxifying molecular mechanisms in B. rapa using a functional cloning strategy. A cDNA library constructed from roots of B. rapa plants treated with Cd was transformed into the Cd sensitive yeast mutant strain DTY167 that lacks the yeast cadmium factor (YCF1), and resistant yeast clones were selected on Cd containing media. Two hundred genes potentially conferring cadmium resistance were rescued from the surviving yeast clones and sequenced. Sequencing analysis revealed that genes encoding for metallothionein (MT)1, MT2a, MT2b and MT3, and phytochelatin synthase (PCS)1 and PCS2 accounted for 35.5%, 28.5%, 4%, 11.3%, 18.7% and 2%, respectively of the genes identified. MTs and PCSs expressing DTY167 cells showed resistance to Cd as well as to Zn. PCS1 expressing yeast cells were also more resistant to Pb compared to those expressing MTs or PCS2. RT-PCR results showed that Cd treatment strongly induced the expression levels of MTs in the root and shoot. Furthermore, the different MTs and PCSs exhibited tissue specific expression. The results indicate that MTs and PCS genes potentially play a central role in detoxifying Cd and other toxic metals in B. rapa.

Metallothionein and Malondialdehyde Correlation in Prostate Cancer Patients

Prostate cancer can be caused by heavy metals exposure that can be identified from increasing levels ofmetallothionein. Exposure to heavy metals is carcinogenic through the formation of ROS in the oxidativestress process. One of the markers of oxidative stress in prostate cancer is malondialdehyde. Purpose ofstudy to determine the relationship between levels of metallothionein and malondialdehyde in prostatecancer. Research design was cross sectional with subjects were 30 patients with prostate cancer at districthospital at Purwokerto, Indonesia. Measurement levels of metallothionein and malondialdehyde usedELISA sandwich method. Data analysis used Spearman correlation test. There was a correlation betweenlevels of metallothionein and malondialdehyde (p &lt;0.05, r = 0.36). The increase of metallothionein levelcause enhancement malondialdehyde level. As a conclusion, study report that there was corelation betweenmetallothionein and malondialdehyde levels illustrates the process of oxidative stress on prostate cancer dueto heavy metals exposure.

ASSESSMENT OF TRACE ELEMENTS UPTAKE ON THE INVASIVE CRAB PORTUNUS SEGNIS HEPATOPANCREAS USING A MULTIVARIATE BIOCHEMICAL APPROACH

In the current investigation, we evaluated the biological consequences of trace elements contamination in the two Tunisian gulfs (Gabes gulf and Tunis gulf) on the blue swimming crabs hepatopancreas (Portunus segnis). The concentrations of three trace elements (cadmium, copper, and lead) in the hepatopancreas P.segnis were evaluated. Additionally, acetylcholinesterase (AChE), metallothioneins (MTs), hydroxide peroxidase (H2O2) and advanced oxidation protein products (AOPP) levels, were chosen as measurements to evaluate the environmental effects on the two crabs’ populations from different gulfs. Macromolecular (lipids, proteins, and DNA) were also determined in P.segnis hepatopancreas. The results of trace elements bioaccumulation in soft P. segnis hepatopancreas showed a high pollution in the Gabes gulf as evidence by significant accumulation of cadmium, cooper, and lead. These findings were confirmed by significant increases of metal pollution index (MPI) and metallothioneins (MTs) levels in the hepatopancreas of P. segnis from Gabes gulf than these from Tunis gulf. Consequently, the trace elements accumulation in P.segnis from Gabes gulf conduct to the generation of lipid peroxidation processes as documented by the high levels of H2O2and LOOH. A significant decrease of AChE activity was recorded in crabs collected from Gabes gulf as compared to these from Tunis gulf. The present study revealed depletion of proteins and lipids contents, while DNA showed significant degradation on crab hepatopancreas collected from Gabes gulf comparing to Tunis gulf. These evidences must be taken in consideration when using P. segnis as an ecological indicator species in the biomonitoring programs.

Apoptosis, oxidative stress and genotoxicity in developing zebrafish after aluminium exposure.

Aluminium is a non-essential metal and potentially toxic to organisms whose environmental concentration increases due to pollution. In our previous studies, the behavioral changes induced by aluminium were already shown on zebrafish, a model organism widely used for ecotoxicology screening. To examine in depth the knowledge about the toxicity mechanism induced by this metal, zebrafish embryos, at 6 hpf, have been exposed to 50, 100 and 200µM of AlCl3 for 72h. Phenotypic alterations, apoptosis and oxidative stress responses have been assessed by evaluations of antioxidant defence and changes in metabolism at the end of treatment. The mRNA expression level of c-fos, appa and appb as marker genes of neural development and function were analyzed by qPCR for the highest used concentration. The data showed that aluminium significantly affected the development of zebrafish inducing morphological alterations and cell death. The oxidative state of larvae was altered, although the formation of reactive oxygen species and the levels of metallothioneins, and the activity of some antioxidant enzymes, decreased at the maximum concentration tested. In addition, at this concentration, the expression of the evaluated genes increased. The comprehensive information obtained gives a realistic snapshot of the aluminium toxicity and provides new information on the mechanism of action of this metal.

Effects of Moringa oleifera leaves extract, vitamin C, and taurine co-exposures on calcium and metallothionein levels, oxidative stress, and gill histopathological changes in Clarias gariepinus exposed to sub-lethal cadmium

Cadmium aquatic environmental pollution poses great threats to fish and their would-be consumers. The present work investigated the effects of ethyl acetate extract of Moringa oleifera leaves (EAEMOL), vitamin C, and taurine co-exposures on calcium and metallothionein levels, oxidative stress, and gill histopathological changes in Clarias gariepinus exposed to sub-lethal cadmium (CdCl2) for 28 days. Fish were exposed to CdCl2 only (1.048 mg/L) as well as co-exposed with EAEMOL (20 mg/L), vitamin C (5 mg/L), and taurine (5 mg/L) separately. There was significant (p < 0.05) deterioration in fish water quality with increasing exposure period but no significant (p > 0.05) changes occurred between the exposed groups. However, the co-exposure of EAEMOL, vitamin C, and taurine did not significantly (p > 0.05) improve the CdCl2-induced fish water quality deterioration. Sub-lethal exposure to CdCl2 only caused significant (p < 0.05) increase in the serum malondialdehyde (MDA) and liver metallothionein (MT) levels with significant (p < 0.05) decrease in serum catalase activity only. However, EAEMOL, vitamin C, and taurine co-exposures did not significantly (p > 0.05) improve the MDA, superoxide dismutase, catalase, and glutathione activities, as well as MT and calcium (Ca2+) levels, condition factor (CF), hepatosomatic index (HSI), and gill histopathological changes induced by the CdCl2 exposure. Similarly, none of the present exposures, CdCl2 only or its co-exposures with EAEMOL, vitamin C, and taurine significantly (p > 0.05) altered the normal functioning of the gills despite the observed histopathological changes based on the degree of tissue change protocol. Therefore, EAEMOL, vitamin C, and taurine co-exposures, as administered in the present case, did not considerably alter the physicochemical parameters of the experimental fish water. However, outside significantly (p < 0.05) increasing MDA level, EAEMOL, vitamin C, and taurine co-exposures did not significantly (p > 0.05) improve the CdCl2-induced Ca2+, MT, CF, HSI, and gill histopathological alterations by sub-lethal CdCl2 exposure of C. gariepinus.

Density and intensity of metallothionein of Crassostrea sp. as biomarkers of heavy metal contamination in the Northern coast of East Java, Indonesia

The present study aimed to observe the expression level of metallothionein by analyzing its density and intensity in the gill and stomach of Crassostrea glomerata and Crassostrea iredalei. The mussels were collected from three sampling locations in the Northern coast of East Java, Indonesia: Mayangan, Probolinggo (Station 1), Kenjeran beach, Surabaya (Station 2), and Gresik Ports (Station 3). The density and intensity of metallothionein were analyzed by using immunohistochemistry technique. The results showed that the highest density of metallothionein in Crassostrea iredalei was at Station 3 (Gresik) i.e. 47.8 × 10−3 (MT/µm2) in gill and 42.2 × 10−3 (MT/µm2) in stomach. While, the highest density value in Crassostrea glomerata was observed at Station 3 (Gresik) i.e. 64 × 10−3 (MT/µm2) in gill and 45.6 × 10−3 (MT/µm2) in stomach. Furthermore, the correlation analysis of heavy metals concentration with the density and intensity of metallothionein showed a significant correlation. This result indicates that the concentrations of the investigated heavy metals (i.e. Pb, Cd and Hg) in seawater correlated with the density and intensity of metallothionein in the gill and stomach of mussels (Crassostrea glomerata and Crassostrea iredalei), which can be used as biomarkers of heavy metal contamination.

Systemic Deletion of Atp7b Modifies the Hepatocytes’ Response to Copper Overload in the Mouse Models of Wilson Disease

Wilson disease (WD) is a genetic disorder caused by inactivation of the copper (Cu) transporter Atp7b and Cu overload, especially in the liver. Mice with Atp7b deleted either globally (systemic inactivation) or only in hepatocyte recapitulate various aspects of human WD. However, their phenotypes vary, and neither the common response to Cu overload nor factors responsible for phenotypes variability are well defined. Using metabolic, histologic, and proteome analyses in three mouse strains with mutated Atp7b, we show that global inactivation of Atp7b enhances and specifically modifies the hepatocyte response. The loss of Atp7b only in hepatocytes dysregulates lipid and nucleic acid metabolisms and increases the abundance of respiratory chain components and redox balancing enzymes. In global knockouts, independently of the background strain, the metabolism of lipid, nucleic acid, and amino acid is inhibited, respiratory chain components are down-regulated, inflammatory response and regulation of chromosomal replication are enhanced. Decrease in glucokinase and lathosterol oxidase and elevation of mucin-13 and S100A10 are observed in all Atp7b mutant strains and reflects the extent of liver injury. The magnitude of proteome changes in different Atp7b-/- strains and in individual animals inversely correlates with their metallothioneins levels rather than Cu content. These results augment the current model of WD pathogenesis, facilitate the identification of WD-specific markers, and assist the development of new therapies

Regulatory T cells induce polarization of pro-repair macrophages by secreting sFGL2 into the endometriotic milieu

An increased number of highly active regulatory T cells (Tregs) and macrophages has been found in peritoneal fluid from women with endometriosis. Here, we show that the level of Tregs-derived soluble fibrinogen-like protein 2 (sFGL2) increases in the peritoneal fluid of women with endometriosis. Higher expression of FGL2 and its receptor CD32B is observed in eutopic endometrium and ectopic tissues. The production of sFGL2 in Tregs may be enhanced by several cytokines. sFGL2 selectively induces pro-repair macrophage polarization mainly through the activation of the SHP2-ERK1/2-STAT3 signaling pathway, and the suppression of the NF-κB signaling pathway. Furthermore, sFGL2 induces a much higher level of metallothionein (MT) expression that in turn facilitates pro-repair macrophages polarization. sFGL2-induced pro-repair macrophages promote Th2 and Tregs differentiation, creating a positive feedback loop. These findings suggest that sFGL2 secreted by Tregs skews macrophages toward a pro-repair phenotype via SHP2-ERK1/2-STAT3 signaling pathway, which is involved in the progression of endometriosis.

Assessment spermatogenic cell apoptosis and the transcript levels of metallothionein and p53 in Meretrix meretrix induced by cadmium

Cadmium (Cd) has been widely used in industry and can accumulate in the water, soil, and food. Meretrix meretrix is one of the marine shellfishes cultivated for economic purpose in China. The increasing Cd levels in coastal marine water could adversely affect the economic benefits of shellfish cultivation. In the present study, M. meretrix were exposed to different Cd2+ concentrations (0, 1.5, 3, 6, and 12 mg L-1) for 5 d to evaluate the effects of Cd on spermatogenic cell. The Cd accumulation, survival rate and the indices of oxidative stress and apoptosis were determined in the spermatogenic cells of M. meretrix. The expression levels of p53 and metallothionein (MT) mRNA were also measured in the spermatogenic cells. Cd accumulation and the mortality rate of spermatogenic cells were found to increase in a dose-response manner with Cd2+ concentrations. Histopathology changes, especially the damage of membranous structure, were more severe as the Cd2+ levels in the testis became higher. The indexes of oxidative stress, including reactive oxygen species, malondialdehyde, protein carbonyl derivates and DNA-protein crosslinks all increased after exposure to Cd2+. However, the total antioxidant capacity gradually decreased with the increasing Cd2+ concentration. In addition, exposure to Cd2+ increased the apoptotic rate and caspase-3 and 9 activities but decreased the level of mitochondrial membrane potential and cytochrome C oxidase in the spermatogenic cells. MT mRNA expression increased in lower Cd2+ concentration treated groups whereas decreased in higher groups, while the p53 mRNA expression increased in a dose-response manner with Cd2+ and was positively correlated with the oxidative damage indices. These results indicated that Cd2+ caused oxidative stress and p53 induced apoptosis in the spermatogenic cells, and thus decreased the survival rate of sperm cells. This finding highlights that Cd can reduce the reproductive capacity of M. meretrix, thus threatening to wild shellfish populations and reducing the efficiency of shellfish farming.

Environmental quality assessment by multiple biogeochemical indicators of an intertidal flat under anthropogenic influence from the southwest of Buenos Aires (Argentina)

Chemical measures combined with biological data (biomarkers) are recommended in monitoring programs of marine environments. In the present research, the use of multiples biogeochemical indicators allowed interrelating different environmental measurements to deepen the knowledge of the quality/status of the intertidal flat from Puerto Rosales (Bahia Blanca estuary, Argentina) subjected to anthropogenic pressure. The sediments from this site presented a eutrophic status and high organic matter concentration with high nutritional value for the benthic community. The TPR/TCH ratio (~ 10) would evidence the untreated sewage discharge contribution and increments in protein content due to the complexation of nitrogen during phytodetritus accumulation and degradation. Trace metals levels in the fine sediments were lower than those recommended by international guides for uncontaminated sites. In the sediments, except Cu, all metals analyzed were rarely associated with adverse biological effects. Finally, metallothioneins levels in the burrowing crab Neohelice granulata were lower than those found in the literature and significantly higher in females than in males. Even though the different analyses and indices performed indicate that this intertidal flat is in good environmental condition, the wastewater discharge influenced it. Thus, it is advisable to continue with this kind of study by applying multi-combined proxies in this and other impacted tidal flats.

Exposure to aluminium causes behavioural alterations and oxidative stress in the brain of adult zebrafish

Aluminium (Al) water pollution is an increasing environmental problem. Accordingly, this study aimed to find out more about its toxic effects on aquatic organisms. Adult zebrafish were exposed to 11 mg/L of Al and the behavioural responses and its correlation with brain oxidative stress, antioxidant-defences, changes in metabolism and neurotransmission were assessed at 10, 15 and 20 days of exposure. The behavioural and locomotory responses, suggest an increase in the anxiety state, especially observed in animals exposed to Al for 15 days. The reactive oxygen species increased in a time-dependent trend, while the oxidative damage varied over exposure time. The activity of antioxidant enzymes, as superoxide dismutase, glutathione peroxidase and glutathione S-transferases, and the metallothioneins levels increased after short-term exposures and tended to decrease or stabilize at longer times. The results contribute to understand the toxic mechanisms activated by Al highlighting correlations like behavioural disorders and oxidative state.

Systemic deletion of Atp7b modifies the hepatocytes\u2019 response to copper overload in the mouse models of Wilson disease

Wilson disease (WD) is caused by inactivation of the copper transporter Atp7b and copper overload in tissues. Mice with Atp7b deleted either globally (systemic inactivation) or only in hepatocyte recapitulate various aspects of human disease. However, their phenotypes vary, and neither the common response to copper overload nor factors contributing to variability are well defined. Using metabolic, histologic, and proteome analyses in three Atp7b-deficient mouse strains, we show that global inactivation of Atp7b enhances and specifically modifies the hepatocyte response to Cu overload. The loss of Atp7b only in hepatocytes dysregulates lipid and nucleic acid metabolisms and increases the abundance of respiratory chain components and redox balancing enzymes. In global knockouts, independently of their background, the metabolism of lipid, nucleic acid, and amino acids is inhibited, respiratory chain components are down-regulated, inflammatory response and regulation of chromosomal replication are enhanced. Decrease in glucokinase and lathosterol oxidase and elevation of mucin-13 and S100A10 are observed in all Atp7b mutant strains and reflect the extent of liver injury. The magnitude of proteomic changes in Atp7b−/− animals inversely correlates with the metallothioneins levels rather than liver Cu content. These findings facilitate identification of WD-specific metabolic and proteomic changes for diagnostic and treatment.

Metal load and oxidative stress driven by organotin compounds on rainbow trout

Tributyltin-based (TBT) antifouling paints, widely used for the treatment of flooded surfaces, have been banned in 2008 for their high environmental persistence and bioaccumulation in aquatic organisms. Although it is still present in aquatic ecosystems, oxidative stress driven by TBT has been still poorly investigated in fish. The aim of the study was to examine the time-course stress responses in liver of rainbow trout that received a single intraperitoneal injection of tributyltin chloride (TBTC) or tributyltin ethoxide (TBTE), both at a dose of 0.05 and 0.5 mg/kg. Levels of metallothioneins, total glutathione, malondialdehyde, superoxide dismutase, catalase, glutathione peroxidase and glutathione S-transferase were evaluated at 3 and 6 days post-injection. Tin load was measured in the muscle of the same fish. Differences were observed in the time-course accumulation of tin with a clear dose-response relationship. Although individual oxidative stress biomarkers varied, the biomarker profile indicated different stress mechanisms caused by both TBTC and TBTE. The weak induction of metal-trapping metallothioneins and the changes of oxidative stress biomarkers suggested a stress-pressure in both TBT-treated trout, advising for an ecotoxicological risk for freshwater ecosystems.