Metallothionein-like Proteins Research Articles

Copper-Induced InVivo Gene Amplification in Budding Yeast.

In the biotechnological industry, multicopy gene integration represents an effective strategy to maintain a high-level production of recombinant proteins and to assemble multigene biochemical pathways. In this study, we developed copper-induced invivo gene amplification in budding yeast for multicopy gene expressions. To make copper as an effective selection pressure, we first constructed a copper-sensitive yeast strain by deleting the CUP1 gene encoding a small metallothionein-like protein for copper resistance. Subsequently, the reporter gene fused with a proline-glutamate-serine-threonine-destabilized CUP1 was integrated at the δ sites of retrotransposon (Ty) elements to counter the copper toxicity at 100 μM Cu2+. We further demonstrated the feasibility of modulating chromosomal rearrangements for increased protein expression under higher copper concentrations. In addition, we also demonstrated a simplified design of integrating the expression cassette at the CUP1 locus to achieve tandem duplication under high concentrations of copper. Taken together, we envision that this method of copper-induced invivo gene amplification would serve as a robust and useful method for protein overproduction and metabolic engineering applications in budding yeast.

QTL-seq Identifies Pokkali-Derived QTLs and Candidate Genes for Salt Tolerance at Seedling Stage in Rice (Oryza sativa L.)

Rice is a staple food crop that plays a pivotal role in global food security, feeding more than half of the world’s population. Soil salinity is one of the most important global problems affecting rice productivity. Salt stress at the seedling stage inhibits root growth, impairs nutrient and water uptake, and affects overall plant vigor, resulting in poor establishment and reduced growth. Therefore, acquiring salt tolerance, especially at the seedling stage, is critical for successful rice production in salinity-affected areas. In this study, 160 RILs derived from a cross between Pokkali and KDML105 were evaluated for their salt tolerance at the seedling stage. QTL-seq analysis with this population identified nine QTLs associated with salt tolerance. Through a comprehensive examination of the effects of coding sequence variants of the 360 annotated genes within the QTLs and gene expression under salt stress, 47 candidate genes were prioritized. In particular, Os01g0200700 (metallothionein-like protein) and Os12g0625000 (O-acetylserine (thiol)lyase) were suggested as potential candidates based on annotated functions and expression data. The results provide valuable insights for improving rice productivity and resistance under salt stress conditions during the critical seedling stage.

Cytosolic distribution of copper in the gills of field-collected oysters with different copper bioaccumulation

Oysters can hyper-accumulate copper (Cu) without apparent toxicity, but the mechanism of sequestering excessive cytosolic Cu in oysters remains unclear. We here investigated the Cu distribution in the cytosolic proteins (CPs) in the gills of oysters (Crassostrea hongkongensis) through size-exclusion chromatography coupled to inductively coupled plasma mass spectrometry (SEC-ICP-MS). Oysters collected from the southern coast of China contained a gradient of gill Cu concentrations ranging from 132 to 3540 μg g−1 (dry weight), with 7–41 % of Cu distributed in the CPs fraction. The CPs-Cu concentrations were 8.6 times higher in oysters with high Cu concentrations compared to low concentrations. In the CPs, Cu was dispersed with a broad range of molecular weight, suggesting the involvement of various cytosolic proteins in Cu binding. Among the 10 major Cu peaks, peaks 2 (>600 kDa) and peak 8 (18 kDa) contained substantial Cu and showed obvious differences in response to the variation of CPs-Cu levels. Peak 8 contained metallothionein-like proteins that decreased their role in Cu binding as CPs-Cu concentrations increased. LC-MS/MS analysis revealed that peak 2 contained macromolecular protein complexes (MPCs), which played a critical role in binding excess Cu. The comparison with other bivalve species further suggested that sequestering excess CPs-Cu in MPCs was a special strategy employed by oysters in response to high Cu accumulation. This study provides valuable insights into the mechanism of hyper-accumulation and sequestration of Cu in oysters and helps to better understand Cu biomonitoring by oysters.

RNA-Seq Based Transcriptomic Analysis of Bud Sport Skin Color in Grape Berries

The most common bud sport trait in grapevines is the change in color of grape berry skin, and the color of grapes is mainly developed by the composition and accumulation of anthocyanins. Many studies have shown that MYBA is a key gene regulates the initiation of bud sport color and anthocyanin synthesis in grape peels. In the current study, we used berry skins of ‘Italia’, ‘Benitaka’, ‘Muscat of Alexandria’, ‘Flame Muscat’, ‘Rosario Bianco’, ‘Rosario Rosso’, and ‘Red Rosario’ at the véraison stage (10 weeks post-flowering and 11 weeks post-flowering) as research materials. The relative expressions of genes related to grape berry bud sport skin color were evaluated utilizing RNA-Seq technology. The results revealed that the expressions of the VvMYBA1/A2 gene in the three red grape varieties at the véraison stage were higher than in the three white grape varieties. The VvMYBA1/A2 gene is known to be associated with UFGT in the anthocyanin synthesis pathway. According to the results, VvMYBA1/A2 gene expression could also be associated with the expression of LDOX. In addition, a single gene (gene ID: Vitvi19g01871) displayed the highest expressions in all the samples at the véraison stage for the six varieties. The expression of this gene was much higher in the three green varieties compared to the three red ones. GO molecular function annotation identified it as a putative metallothionein-like protein with the ability to regulate the binding of copper ions to zinc ions and the role of maintaining the internal stable state of copper ions at the cellular level. High expression levels of this screened gene may play an important role in bud sport color of grape berry skin at the véraison stage.

Significance of chemical affinity on metal subcellular distribution in yellow perch (Perca flavescens) livers from Lake Saint-Pierre (QUEBEC, Canada)

The subcellular partitioning approach provides useful information on the location of metals within cells and is often used on organisms with high levels of bioaccumulation to establish relationships between the internal concentration and the potential toxicity of metals. Relatively little is known about the subcellular partitioning of metals in wild fish with low bioaccumulation levels in comparison with those from higher contaminated areas. This study aims to examine the subcellular partitioning of various metals considering their chemical affinity and essentiality at relatively low contamination levels. Class A (Y, Sr), class B (Cu, Cd, MeHg), and borderline (Fe, Mn) metal concentrations were measured in livers and subcellular fractions of yellow perch (n = 21) collected in Lake Saint-Pierre, QC, Canada. The results showed that all metals, apart from MeHg, were distributed among subcellular fractions according to their chemical affinity. More than 60% of Y, Sr, Fe, and Mn were found in the metal-sensitive fractions. Cd and Cu were largely associated with the metallothionein-like proteins and peptides (60% and 67% respectively) whereas MeHg was found mainly in the metal-sensitive fractions (86%). In addition, the difference between the subcellular distribution of Cu and other essential metals like Fe and Mn denotes that, although the essentiality of some metals is a determinant of their subcellular distribution, the chemical affinity of metals is also a key driver. The similarity of the subcellular partitioning results with previous studies on yellow perch and other fish species from higher contaminated areas supports the idea that metals are distributed in the cellular environment according to their chemical properties regardless of the bioaccumulation gradient.

Expression of Metallothionein-Like Protein 2B Gene Induced by Different Concentrations of Copper in White Spruce (Picea glauca)

The alterations to the expressions of metal transporter genes can induce metal ion uptake to maintain cell homeostasis when metal concentrations are low. Metallothioneins (MTs) are a class of low molecular mass (4-8 kDa), cysteine (Cys)-rich proteins that can bind metals via the thiol groups of their Cys residues. Studies on MT gene expressions in higher plants, such as conifers, are limited. White spruce (Picea glauca) is a North American native conifer species that are ecologically and economically important. The objective of the study is to determine the effects of different concentrations of copper on MT2b expression in white spruce (P. glauca). Seedlings were treated with three concentrations of copper sulfate including 1,312, 656, and 130 mg/kg. Potassium sulfate and water were used as controls. Total RNA was extracted from roots and needles. The levels of MT2b gene expression were measured using RT-qPCR. The present study reports for the first time, novel information on the effect of copper ions on the expression of MT2b in P. glauca. The MT2b gene was upregulated in roots when seedlings were exposed to 1,312 mg/kg of copper sulfate but were downregulated in genotypes treated with 656 mg/kg of copper sulfate in both tissues when compared to the water control. The expression of the MT2b gene was significantly downregulated in needles of genotypes treated with 656 mg/kg of copper sulfate when compared to the 1,312 mg/kg concentration. The expression of MT2b was higher in roots compared to needles regardless of the concentrations of copper sulfate or potassium sulfate used. MT2b was highly expressed in the roots of copper-resistant genotypes but was inhibited in the roots of susceptible seedlings when compared to the water control.

Metallothionein-like 5 expression is correlated with poor prognosis and promotes proliferation of cervical squamous cell carcinoma

ABSTRACT Cervical cancer represents one of the most important female genital cancers. Cervical squamous cell carcinoma (CESC) accounts for about 90% of all cervical malignancies and the prognosis are unsatisfied. Here we aimed to investigate the clinical relevance of metallothionein-like 5 (MTL5), a novel metallothionein-like protein, in CESC. RT-qPCR and immunohistochemistry staining showed that MTL5 was upregulated in CESC tissues than nontumorous cervix tissues, which is consistent with the data from TCGA database. Kaplan–Meier survival analysis revealed that higher MTL5 can help predict worse prognosis. In addition, Cox hazard regression analysis verified an independent predictive role of MTL5 in CESC. To further investigate the involvement of MTL5 in CESC, we conducted knockdown experiments in two CESC cell lines. As a result, silencing MTL5g significantly inhibited proliferation of CESC cells. Finally, we validated that silencing MTL5 can suppress CESC tumor growth in vivo using the mice subcutaneous xenografts model. Taken together, higher MTL5 indicates worse survival of CESC after surgical resection. Targeting MTL5 represents a potential therapy of CESC by inhibiting tumor growth, which deserves further investigations.

ScMT10, a metallothionein-like gene from sugarcane, enhances freezing tolerance in Nicotiana tabacum transgenic plants

Sugarcane (Saccharum spp. hybrid) is of tropical origin, yet generally cultivated in subtropical areas. Cold stress is a major environmental factor that impairs the production of sugarcane in that area. Metallothioneins, a family of small, cysteine-rich proteins, are proved to be involved in metal homeostasis and detoxification and thus alleviate stress-induced ROS accumulation. Here, a cDNA clone deducing a single Cys-rich domain type 2 metallothionein-like protein, designated ScMT10, was isolated from sugarcane. The level of ScMT10 expression was up-regulated by salicylic acid (SA), cold, and oxidative stress (H2O2) in sugarcane seedlings, but it was down-regulated in response to abscisic acid (ABA), methyl jasmonate (MeJA), heavy metal ions (Cu2+, Cd2+), and salinity. Transgenic tobacco (Nicotiana tabacum) plants (ScMT10-OE) showed a remarkable enhancement in freezing tolerance compared with wild-type plants (WT). Furthermore, ScMT10-OE tobacco showed a lower electrolyte leakage, reduced malondialdehyde contents, and slower ROS (including H2O2) accumulation under freezing stress and retained higher proline, soluble sugar, and relative chlorophyll content compared to WT plants. The expression profiles of ScMT10 in the transgenic tobacco were similar to those in sugarcane seedlings under different stresses. Besides, the level of H2O2 in the ScMT10-OE plants was comparable to that observed in WT plants under heavy metal, salt, and drought stress. Therefore, the transgenic plants did not perform better than WT plants under those stresses. Taken together, we present a hypothesis that ScMT10 was exclusively and positively involved in the cold stress-induced accumulation of reactive oxygen species (ROS). The over-expression of ScMT10 in tobacco decreased the contents of endogenous ROS, MDA, and relative electric conductivity. It increased the accumulation of proline, soluble sugar, and relative chlorophyll under freezing stress, which alleviated the oxidative damage to the cell membrane during freezing exposure. ScMT10 can be considered as a candidate gene for cold-tolerance trait improvement.

Monitoring metallothionein-like protein concentrations and cholinesterase activity in tropical cup oysters as biomarkers of exposure to metals and pesticides in the southern Caribbean, Colombia.

Metallothionein-like protein concentrations (MT) and three functionally defined fractions of cholinesterase activity (ChE) were quantified in gill and digestive gland homogenates of tropical cup oysters from 5 nearshore locations in the Colombian Caribbean and correlated with sediment and tissue metal (9 metals) and pesticide (22 organophosphates, OPs, and 20 organochlorines-OCPs), as well as water physical-chemical parameters (salinity, pH, temperature, and dissolved oxygen). Tissue and sediment pesticide concentrations were below detection limits in all samples, whereas sediment and tissue metal concentrations exceeded environmental thresholds at several locations. Tissue MT and ChE biomarkers varied by a factor of 5-6 between locations. Inhibition of cholinesterase activity was negligible for all 5 sites, despite spatial-temporal variation in ChE activity, consistent with below-detection OP concentrations. Tissue MT and ChE biomarkers correlated with tissue and metal sediment concentrations, yet, statistically significant covariance between biomarkers and water chemistry parameters was also observed, indicating that both, metal concentrations and physical-chemical variables, are likely to be responsible for generating the observed spatial-temporal variations in biomarker patterns.

Response of biomarkers to metals, hydrocarbons and organochlorine pesticides contamination in crabs (Callinectes ornatus and C. bocourti) from two tropical estuaries (São José and São Marcos bays) of the Maranhão State (northeastern Brazil)

Response of biomarkers to chemical contamination was evaluated in crabs of the Callinectes genus (Callinectes ornatus and C. bocourti) from two tropical estuaries (São José and São Marcos bays) of the Maranhão State (northeastern Brazil). Biomarkers evaluated included hepatopancreatic metallothionein-like proteins (MTLP) and lipid peroxidation (LPO), as well as muscle acetylcholinesterase (AChE). Tissue concentrations of metals (pereiopod muscle and hepatopancreas), hydrocarbons (hepatopancreas) and organochlorine pesticides (hepatopancreas) were also evaluated. Crab samples were collected in three sites of each estuary (São Marcos Bay and São José Bay). Sampling was performed in August/2012 (dry season), January/2013 (rainy season), August/2013 (dry season), and January/2014 (rainy season). Concentrations of chemical contaminants and responses of biomarkers showed significant spatial (São Marcos Bay and São José Bay) and/or seasonal (dry and rainy seasons) and annual (2012–2014) variability. However, a general higher Zn concentration was observed in hepatopancreas of crabs from São José Bay. In turn, a general higher Cd concentration paralleled by oxidative damage (LPO) was observed in hepatopancreas of crabs from São Marcos Bay. As expected, these findings support the idea that this bay is more intensively or chronically impacted by industrial activities while the São José Bay is likely more affected by domestic activities. Interestingly, LPO level in crab hepatopancreas showed to be the most reliable and adequate biomarker to distinguish the two bays.

Bioaccumulation and biotransformation of inorganic arsenic in zhikong scallop (Chlamys farreri) after waterborne exposure

Arsenic (As) and As speciation in marine bivalves have been widely investigated. However, little is known about the bioaccumulation and biotransformation of inorganic As in different tissues of scallops. Therefore, the tissue-specific accumulation, biotransformation and subcellular partitioning of As were investigated in Chlamys farreri following 12 d inorganic As [arsenite (AsⅢ) and arsenate (AsⅤ)] exposure and 30 d depuration. Total As levels were highest in the kidneys and lowest in the adductor muscle after 12 d exposure for both As (Ⅲ) and As (Ⅴ) treatment groups, and the bioavailability of As (Ⅲ) was significantly higher than that of As (Ⅴ) for C. farreri. After 30 d elimination, total As levels were significantly decreased to the control levels. The subcellular fate of As in five different tissues was similar for different inorganic As treatment groups. The greatest proportion of As was found in the metallothionein-like protein fraction (MTLP) and the second was the cellular debris (CD). A little part of As (Ⅲ) could be oxidized to As (Ⅴ) in the gill and digestive gland for As (Ⅲ) treatment groups, and the reduction of As (Ⅴ) to As (Ⅲ) happened in the gill and kidney under As (Ⅴ) exposure. Although a high methylation activity was found in C. farreri, it varied in different tissues with different inorganic As species exposure. The present results indicated that exposure to As (Ⅲ) and As (Ⅴ) could induce different responses in bioaccumulation and biotransformation in five tissues of C. farreri.

Accumulation, transformation and subcellular distribution of arsenite associated with five carbon nanomaterials in freshwater zebrafish specific-tissues

Although carbon nanomaterials (CNMs) commonly exist throughout the aquatic environment, their effect on arsenic (As) distribution and toxicity is unclear. In this study, arsenite accumulation, transformation, subcellular distribution, and enzyme activity were assessed in adult zebrafish (Danio rerio) intestines, heads and muscles, following co-exposure to arsenite and CNMs with different structures (single-walled carbon nanotubes (SWCNTs), multi-walled carbon nanotubes (MWCNTs), fullerene (C60), graphene oxide (GO), and graphene (GN)). Results show that GN and GO promoted As toxicity in D. rerio, as carriers increasing total As accumulation in the intestine, resulting in arsenite adsorbed by GO and GN being released and transformed mainly into moderately-toxic monomethylarsonic acid (MMA), which was mostly distributed in organelles and metallothionein-like proteins (MTLPs). Moreover, GO and GN influenced As species distribution in D. rerio due to the excellent electron transfer ability. However, the effect was marginal for SWCNT, MWCNT and C60, because of the different structure and suspension stability in fish-culture water. In addition, in the muscle and head tissues, As was mainly distributed in cellular debris in the forms of dimethylarsinic acid (DMA) and arsenobetaine (AsB). These findings help better understand the influence of CNMs on the mechanism of As toxicity in natural aquatic environments.

The countryside or the city: Which environment is better for the honeybee?

For a number of years, the decline of honeybee (Apis mellifera) in North America and Europe has been the subject of much debate. Among the many factors proposed by hundreds of studies to explain this phenomenon is the hypothesis that agricultural activities using pesticides contribute to the weakness of bee colonies. Moreover, while urban beekeeping is presently booming in several cities, we do not know if this environment is more beneficial for bees than the typical, rural area. In the summer of 2018, we sampled honeybees (foragers and larvae) in rural (Laurentians) and urban (city of Montreal) areas and compared them using the following biomarkers: carotenoids, retinoids, α-tocopherol, metallothionein-like proteins (MTLPs), lipid peroxidation, triglycerides, acetylcholinesterase activity (AChE) and proteins. Pesticides, pharmaceuticals and personal care products (PPCPs) and metals were also quantified in honeybees' tissues. Our result revealed that, globally, urban foragers had higher levels of insecticides and PPCPs and that metals were in greater concentrations in urban larvae. Compared to rural foragers, urban foragers had higher concentrations of MTLPs, triglycerides, protein and AChE activity. The multifactorial analysis confirmed that insecticides, some metals and PPCPs were the most influential components in the contaminant‒biomarker relationships for both foragers and larvae.

Detecting the effects of chronic metal exposure on benthic systems: Importance of biomarker and endpoint selection

Understanding metal toxicity to benthic systems is still an ecotoxicological priority and, although numerous biomarkers exist, a multi-biomarker and endpoint approach with sediment as the delivery matrix combined with life-history relevant exposure timescales is missing. Here we assess potential toxicity by measuring a suite of biomarkers and endpoints after exposing the ecologically important polychaete Alitta(Nereis)virens to sediment spiked with environmentally relevant concentrations of copper and zinc (and in combination) for 3, 6 and 9 months. We compared biomarker and endpoint sensitivity providing a guide to select the appropriate endpoints for the chosen time frame (exposure period) and concentration (relevant to Sediment Quality Guidelines) needed to identify effects for benthic polychaetes such as A. virens. Target bioavailable sediment and subsequent porewater concentrations reflect the global contamination range, whilst tissue concentrations, although elevated, were comparable with other polychaetes. Survival reduced as concentrations increased, but growth was not significantly different between treatments. Metabolic changes were restricted to significant reductions in protein after 9 months exposure across all copper concentrations, and reductions in lipid at high copper concentrations (3 months). Significant changes in feeding behaviour and increases in metallothionein-like protein concentration were limited to the medium and high copper and zinc concentrations, respectively, both after 6 months exposure. Despite data highlighting A. virens′ metal tolerance, DNA damage and protein concentrations are the most sensitive biomarkers. Copper and zinc cause biomarker responses at concentrations routinely found in coastal sediments that are characterised as low contamination, suggesting a reappraisal of the current input sources (especially copper) is required.

Subcellular metal distribution in two deep-sea mollusks: Insight of metal adaptation and detoxification near hydrothermal vents

In this study, we determined the concentrations of Cu, Zn, Ni, Cd, Pb and As and their subcellular distributions within the tissues of mussels (Bathymodiolus marisindicus) and snails (Gigantopelta aegis) from two hydrothermal vent regions, i.e., Tiancheng and Longqi, at Southwest Indian Ridge. Mussels collected from the two venting regions showed comparable concentrations for Ni and Pb, but Cu, Zn, Cd and As concentrations were significantly different in mussel gills between the two vent regions. Similar ranges of metal concentrations were found in the snails as those in the mussels, but most of the metals were mainly accumulated in the viscera, except for Ni. Similar subcellular partitioning of Cu, Zn and Cd was documented in different mussel tissues, with cellular debris (50%) being the predominant fraction, followed by equivalent values in other fractions. Lead was distributed in both cellular debris and metal-rich granules (MRG) fraction, whereas Ni was predominantly distributed in MRG (90%). Arsenic was mainly partitioned in cellular debris and metallothionein-like protein. However, deep-sea snails displayed elevated subcellular partitioning of Cu in the organelles (up to 60%) and may be more susceptible to Cu stress than the mussels. Our results demonstrated the metal-specificity of detoxification strategies in these deep-sea hydrothermal vent mollusks, and the mussels may be more adaptable to high metal exposures than the snails at hydrothermal vent.

Biomarkers of oxidative stress and cell damage in freshwater bivalves Diplodon parodizi exposed to landfill leachate.

Landfill is a public and environmental health problem; establishing and understanding methodologies to decrease its toxicity are thus necessary. Leachate samples were collected, at a sanitary landfill, immediately after the exit from the landfill, i.e. raw leachate (collection point A), after conventional treatment (point B) and after treatment by wetlands (point C). D. parodizi specimens were exposed to 3%, 10% and control (0%) dilutions of leachate from these collection points for 7days. Markers of antioxidant defences and cell damage were analysed. At point B, the gills of D. parodizi showed higher glutathione-S-transferase (GST) and glutathione reductase (GR) activity; the latter is a supplier of glutathione reductase (GSH). The low GST activity at point A was associated with the hormesis effect. Higher levels of superoxide dismutase (SOD), ethoxyresorufin-O-deethylase (EROD) and glutathione peroxidase (GPx) occurred at point A. Glucose-6-phosphate dehydrogenase (G6PDH) was inhibited at the points with the highest pollutant load and at the highest leachate dilutions. Higher levels of markers at point A may be related to the high pollutant charge and specific compounds present in the untreated leachate. The multi-xenobiotic resistance mechanism (MXR), metallothionein-like proteins (MT) and lipid peroxidation (LPO) did not vary among treatments. The biomarker responses showed negative effects of the leachate on the freshwater bivalve and simultaneously showed that the wetland treatment employed at the Caximba sanitary landfill is effective.

Integrated assessment of the acclimation capacity of the marine bivalve Crenomytilus grayanus under naturally highly contaminated conditions: Subcellular distribution of trace metals and structural alterations of nephrocytes

The aim of our work was to assess whether the cellular processes in the nephrocytes of the long-lived mussel Crenomytilus grayanus tend to acclimation or destruction under trace metal contamination. Mussels were collected from three sites in the north-western Pacific Ocean: reference site, upwelling site, and a site highly contaminated with trace metals. Concentration, subcellular distribution of trace metals (Cd, Cu, Zn, and Pb) in the mussel kidneys, and ultrastructural alterations of the nephrocytes were studied. To assess the total load of accumulated trace metals, the total concentration coefficient (∑СС) was determined.In the kidneys of the reference C. grayanus, trace metals were eliminated from cell metabolism mainly by lysosomal granules or residue bodies. Under high levels of contamination, the defense mechanisms of C. grayanus are practically suppressed (no metallothionein-like protein peak, decreased content of granules) by the total effect of accumulated pollutants that leads to the destruction of cellular structures. Under natural conditions (upwelling site), increased accumulation of trace metals in the mussel kidneys did not lead to an increase in the number or size of lysosomal granules. However, abnormal high Cd accumulation in the kidneys caused the synthesis of high levels of metallothionein-like proteins that sequester most of the studied trace metals. To quickly lower the metal levels in nephrocytes under these conditions, a unique long-term acclimatory response — apocrine-like secretion in nephrocytes, which provides rapid elimination of me—MTLP complexes from the cell arose. Thus, our integrated study of the subcellular distribution of trace metals and ultrastructural alterations in nephrocytes allowed us to characterize the features of the structural and functional alterations in mussel cells under the field conditions tested.

Effects of waterborne exposure to cadmium on biochemical responses in the freshwater gastropod, Bellamya aeruginosa.

The biochemical responses of Bellamya aeruginosa as a dominant and widespread freshwater gastropod throughout China to waterborne cadmium (Cd) were investigated to explore the impacts of exposure concentration and duration in this potential sentinel species. After the 7 days' test of dosage-mortality relationship, gastropods were exposed for either 7 days at the LC50 (1.7mg/L), the LC10 (0.7mg/L) and 0.02mg/L Cd, or 28 days at 0.02mg/L Cd. A suite of biochemical indicators including metallothionein-like protein (MTLP), reduced glutathione (GSH), catalase (CAT), contents of tissue metal (Cd, Fe, Mn, Cu, Zn), and the compartments of these metals bound to MTLP were examined. The treatment of 0.02mg/L Cd led to the increase of Cd bound to MTLP (Cd-MTLP) levels, the decrease of GSH content, and the upregulation of CAT activity, but no induction of MTLP, indicating that the intrinsic MTLP and GSH worked together for the detoxification of Cd at the low exposure. When the exposure concentration increased, GSH was depleted severely and synthesis of MTLP was triggered, leading to a strong and significant relationship between MTLP level and Cd accumulation. At the lethal concentrations (1.7mg/L), both MTLP induction and CAT activity were inhibited while the proportion of Cd-MTLP to total Cd were increased, suggesting more intrinsic MTLP were utilized to sequester free Cd ions. Therefore, the content of Cd-MTLP in digestive glands of B. aeruginosa was recommended as a reliable biomarker for Cd contamination.

The role of tesmin in the physiology and pathogenesis of human diseases

Tesmin (Testis-specific metallothionein-like protein) was first observed during meiotic division of spermatocytes in mouse testis. Expression of tesmin was also found in somatic cells of various organs. In addition to the suggested function of regulating the meiotic division of germ cells, the cysteine-rich domains in tesmin structure allow to bind heavy metal ions. In addition, this protein acts as a co-activator of the nuclear mineralocorticoid receptor, which may affect the expression of many other genes. Tesmin could also potentially play a role in disease states such as: chronic obstructive pulmonary disease, osteoporosis, AIDS encephalopathy, as well as neoplasms: acute lymphoblastic leukemia, prostate cancer or non-small cell lung cancer. The above reports on the multifaceted role of tesmin in various disease processes suggest that it could be an interesting direction for further research, what this paper aims to present.

Identification of Differentially Up-regulated Genes in Apple with White Rot Disease.

Fuji, a major apple cultivar in Korea, is susceptible to white rot. Apple white rot disease appears on the stem and fruit; the development of which deteriorates fruit quality, resulting in decreases in farmers' income. Thus, it is necessary to characterize molecular markers related to apple white rot resistance. In this study, we screened for differentially expressed genes between uninfected apple fruits and those infected with Botryosphaeria dothidea, the fungal pathogen that causes white rot. Antimicrobial tests suggest that a gene expression involved in the synthesis of the substance inhibiting the growth of B. dothidea in apples was induced by pathogen infection. We identified seven transcripts induced by the infection. The seven transcripts were homologous to genes encoding a flavonoid glucosyltransferase, a metallothionein-like protein, a senescence-induced protein, a chitinase, a wound-induced protein, and proteins of unknown function. These genes have functions related to responses to environmental stresses, including pathogen infections. Our results can be useful for the development of molecular markers for early detection of the disease or for use in breeding white rotresistant cultivars.