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ABSTRACT: Heat shock 70kDa protein 12A (HSPA12A) is an atypical member of HSP70 family, and the translationally controlled tumor protein (TCTP) is a novel HSP with chaperone-like activity. They are both involved in protecting organisms against various stressors. In the present study, the cDNAs of HSPA12A and TCTP (called MgHSPA12A and MgTCTP) were identified from Mytilus galloprovincialis by RACE approaches. The full-length cDNA of MgHSPA12A and MgTCTP encoded a peptide of 491 and 171 amino acids, respectively. Real-time PCR was employed to analyze the tissue distribution and temporal expression of these two genes after bacterial challenge and cadmium (Cd) exposure. It was found that the transcripts of MgHSPA12A and MgTCTP were dominantly expressed in gonad and muscle, respectively. The expression level of MgTCTP at 48 h post Vibrio anguillarum challenge was detected to be significantly up-regulated in hepatopancreas (P < 0.05). As concerned to Cd exposure, 2.0-fold increase of MgHSPA12A expression compared to that of the control was observed at 48 h in 5 μg/L Cd(2+)-treated group, while the expression levels of MgTCTP were significantly decreased after exposed to both 5 and 50 μg/L Cd(2+) for 24 h and 96 h. These results suggested the potential involvement of MgHSPA12A and MgTCTP in the mediation of the immune responses and environmental stress in mussels.
Fish & Shellfish Immunology 04/2013; · 3.32 Impact Factor
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ABSTRACT: Suaeda salsa is a native halophyte in saline soils. Salinity is the most important environmental constraint for plant productivity in
the Yellow River Delta. In this work, we investigated the salt-induced effects in root of S. salsa exposed to two environmentally relevant salinities for 1week and 1month using nuclear magnetic resonance-based metabolomics.
Our results indicated that salt stress inhibited the growth of S. salsa and induced significant metabolic responses including decreased amino acids, lactate, 4-aminobutyrate, malate, choline, phosphocholine,
and increased betaine, sucrose, and allantoin in root tissues of S. salsa. In addition, salinity exposures upregulated the activities of superoxide dismutase, glutathione S-transferases, peroxidase,
catalase, and glutathione peroxidase in the aboveground part of seedlings of S. salsa after exposures. Overall, these results demonstrated the osmotic and oxidative stresses, disturbances in protein biosynthesis/degradation,
and energy metabolism in S. salsa exposed to salinities.
Keywords
Suaeda salsa
–Salinity–Metabolic response–NMR–Metabolomics–Antioxidant enzyme activity
Plant Molecular Biology Reporter 04/2012; · 2.45 Impact Factor
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ABSTRACT: Glutathione S-transferases (GSTs) are phase II enzymes that facilitate the detoxification of xenobiotics, and also play important roles in antioxidant defense. In this study, we reported the cloning and molecular characteristics of seven genes of the GST family (VpGSTS1, VpGSTS2, VpGSTS3, VpGSTO, VpGSTMi, VpGSTM and VpGSTR) from Venerupis philippinarum together with mRNA tissue distribution patterns and temporal expression profiles in response to cadmium, copper and benzo[a]pyrene (B[a]P) exposures. The deduced amino acid sequences of VpGSTs showed high similarities to counterparts of other species that clustered into the same clades in the phylogenetic analysis. At basal levels of tissue expression, most VpGSTs were highly expressed in hepatopancreas compared with other tissues. All VpGSTs showed differential response profiles depending on the concentrations of various toxicants and exposure times. More notably, the expressions of VpGSTS2 and VpGSTS3 transcripts were significantly up-regulated in hepatopancreas from Cu and B[a]P-exposed animals, indicating that these two sigma VpGSTs were highly sensitive to Cu and B[a]P exposure. However, the expressions of VpGSTM and VpGSTR were significantly induced by Cu or B[a]P exposure, respectively. These findings suggested the role of VpGSTS2, VpGSTS3, VpGSTM and VpGSTR in defense against oxidative stress and highlighted their potential as biomarkers to Cu or B[a]P exposure.
Comparative Biochemistry and Physiology Part C Toxicology & Pharmacology 04/2012; 155(3):517-27. · 2.62 Impact Factor
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Qingtao Li,
Qing Zhao,
Bo Lu,
Hengbin Zhang,
Song Liu,
Zhipeng Tang,
Lijia Qu,
Rui Zhu,
Jingmin Zhang, Liping You,
Fuhua Yang,
Dapeng Yu
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ABSTRACT: The stability and surface evolution of solid-state nanopores in aqueous solutions are extremely important since they would get immersed in solutions during DNA translocation experiment for DNA analyses. In this work, we systematically studied the size evolution of SiN nanopores in ethanol, deionized water and potassium chloride (KCl) solutions by careful surface characterization and composition analyses using a transmission electron microscope. Surprisingly, we found that nanopores closed up completely in ethanol in an hour and showed a 30% and 20% size decrease in deionized water and KCl solutions, respectively. Strong evidence of surface oxidation was found by composition analyses in the nanopore area. Nanopore size evolution was strongly dependent on initial pore size and solution pH value. In pH = 13 KCl solution, SiN nanopores were observed to increase in size instead of decrease. The results not only provide useful information for DNA detection based on solid-state nanopores, but can also guide design and application of other nanodevices exposed to electrolyte-solvent systems such as cell-on-a-chip devices and biosensors.
Nanoscale 03/2012; 4(5):1572-6. · 5.91 Impact Factor
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ABSTRACT: A poly(methyl methacrylate) assisted dry transfer method was developed to transfer graphene microflake onto a suspended SiN chip in an effective and efficient way for further graphene nanopore drilling for DNA analysis. Graphene microflakes can be patterned by e-beam lithography to a designed shape and size on a large scale of a few thousands simultaneously. Subsequently, individual graphene microflakes can be picked up and transferred to a target hole on a suspended SiN membrane with 1 µm precision via a site-specific transfer-printing method. Nanopores with different diameters from 3 to 20 nm were drilled on the as-transferred graphene membrane in a transmission electron microscope. This method offers a fast and controllable way to fabricate graphene nanopores for DNA analyses.
Nanotechnology 03/2012; 23(8):085301. · 3.98 Impact Factor
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ABSTRACT: Two goose-type lysozymes (designated as MGgLYZ1 and MGgLYZ2) were identified from the mussel Mytilus galloprovincialis. MGgLYZ1 mRNA was widely expressed in the examined tissues and responded sensitively to bacterial challenge in hemocytes, while MGgLYZ2 mRNA was predominately expressed and performed its functions in hepatopancreas. However, immunolocalization analysis showed that both these lysozymes were expressed in all examined tissues with the exception of adductor muscle. Recombinant MGgLYZ1 and MGgLYZ2 could inhibit the growth of several Gram-positive and Gram-negative bacteria, and they both showed the highest activity against Pseudomonas putida with the minimum inhibitory concentration (MIC) of 0.95-1.91 µM and 1.20-2.40 µM, respectively. Protein sequences analysis revealed that MGgLYZ2 had lower isoelectric point and less protease cutting sites than MGgLYZ1. Recombinant MGgLYZ2 exhibited relative high activity at acidic pH of 4-5, while MGgLYZ1 have an optimum pH of 6. These results indicated MGgLYZ2 adapted to acidic environment and perhaps play an important role in digestion. Genomic structure analysis suggested that both MGgLYZ1 and MGgLYZ2 genes are composed of six exons with same length and five introns, indicating these genes were conserved and might originate from gene duplication during the evolution. Selection pressure analysis showed that MGgLYZ1 was under nearly neutral selection while MGgLYZ2 evolved under positive selection pressure with three positively selected amino acid residues (Y(102), L(200) and S(202)) detected in the mature peptide. All these findings suggested MGgLYZ2 perhaps served as a digestive lysozyme under positive selection pressure during the evolution while MGgLYZ1 was mainly involved in innate immune responses.
PLoS ONE 01/2012; 7(9):e45148. · 4.09 Impact Factor
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ABSTRACT: With the objective to identify promising molecular biomarkers for marine pollution monitoring, a new cytochrome P450 gene was identified from Venerupis (Ruditapes) philippinarum and classified as a member of a new subfamily, CYP414A1. Phylogenetic analysis showed that CYP414A1 was closely related to members of the CYP2 family. CYP414A1 mRNA expression was significantly induced by 50 μg/L B[a]P at 96 h, while no significant change was found in 5 μg/L B[a]P-exposed samples. For heavy metals exposure, the expression of CYP414A1 was significantly up-regulated by Cd but sharply depressed by Cu exposure. These results suggested that CYP414A1 responded to various xenobiotics stresses, and could be used as a candidate biomarker of heavy metals and B[a]P.
Environmental toxicology and pharmacology. 01/2012; 33(1):85-91.
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ABSTRACT: Polychlorinated biphenyls (PCBs), dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) binding with the aryl hydrocarbon receptor (AhR) have been correlated with many toxic responses. Hence, it is very necessary to study the interactions between these ligands and AhR for further understanding of the mechanism of toxicity. In this study, an integrated molecular docking and 3D-QSAR approach was employed to investigate the binding interactions between PCBs, PCDDs, PCDFs and AhR. From molecular docking, hydrogen-bonding and hydrophobic interactions were observed to be characteristic interactions between compounds and AhR. Based on the mechanism of interactions, an optimum 3D-QSAR model with good robustness (Q(CUM)(2)=0.907) and predictability (Q(EXT)(2)=0.863) was developed by partial least squares. Additionally, the developed QSAR model indicated that the molecular size, shape profiles, polarizability and electropological states of compounds were related to the binding affinities to AhR.
Environmental toxicology and pharmacology. 11/2011; 32(3):478-85.
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ABSTRACT: Polycyclic aromatic hydrocarbons (PAHs) can be hydroxylated by CYP450-oxidases (1A1 and 1B1 mainly) and may cause DNA damage and cancer. However, the mechanism of such interactions has not been fully understood. In this study, an integrated molecular docking and QSAR approach was employed to further investigate the binding interactions between hydroxylated PAHs (HO-PAHs) and calf thymus DNA (CT-DNA). Molecular docking, hydrogen-bonding, hydrophobic and π-π interactions were observed to be characteristic interactions between HO-PAHs and DNA. An optimum QSAR model with good robustness and predictability was developed based on the molecular structural parameters calculated by the density function theory and partial least squares. Additionally, the developed QSAR model indicated that the molecular size, polarizability and electrostatic potential of HO-PAHs were related to the binding affinities to DNA.
Environmental toxicology and pharmacology. 11/2011; 32(3):373-81.
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Linbao Zhang,
Xiaoli Liu, Liping You,
Di Zhou,
Qing Wang,
Fei Li,
Ming Cong,
Lianzhen Li,
Jianmin Zhao,
Dongyan Liu,
Junbao Yu,
Huifeng Wu
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ABSTRACT: Benzo(a)pyrene is an important polycyclic aromatic hydrocarbon (PAH) which causes carcinogenic, teratogenic and mutagenic effects in various species and the level of contamination of this toxic agent in the marine environment is of great concern. In this study, metabolic responses induced by two doses (0.02 and 0.2μM) of BaP were characterized in the gill tissues of Manila clam Ruditapes philippinarum after exposure for 24, 48 and 96h. The high dose (0.2μM) of BaP induced the disturbances in energy metabolism and osmotic regulation based on the metabolic biomarkers such as succinate, alanine, glucose, glycogen, branched chain amino acids, betaine, taurine, homarine, and dimethylamine in clam gills after 24h of exposure. In addition, hormesis induced by BaP was found in clams exposed to both doses of BaP. Overall, our results demonstrated the applicability of metabolomics for the elucidation of toxicological effects of marine environmental contaminants in a selected bioindicator species such as the Manila clam.
Environmental toxicology and pharmacology. 09/2011; 32(2):218-25.
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ABSTRACT: Polychaetes have often been utilized as indicator species to investigate the impacts of pollutants, such as heavy metals. The uptake of Cd by the polychaete Nereis diversicolor was determined at varying Ca concentrations and with pre-exposure to Ca ion channel blockers and metabolic inhibitors in simulated sea water over 1 week period. The supply of Ca in simulated sea water inhibited Cd uptake and increased Ca concentration in N. diversicolor after 10 μM Cd exposure. Pre-exposure to a Ca-channel blocker (Lanthanum) significantly inhibited Cd uptake, suggesting that the uptake of Cd was exerted at a Ca channel. N-ethylmaleimide, which specifically binds to sulfhydryl groups, inhibited Cd uptake at 10 μM, implying that the transport of Cd is carrier-mediated by proteins or other SH-containing compounds. Subcellular Cd distribution analysis showed that more than 60% of the total Cd associated with the cytosolic fraction. The presence of higher concentration of Ca in simulated sea water did not impact the proportional subcellular distribution of Cd in N. diversicolor. Nevertheless, the supply of Ca could significantly lower Cd concentration in cytosol and cellular debris. The present study provides evidence that Cd transport by N. diversicolor was mediated mainly through lanthanum- sensitive Ca ion channels and accumulated by SH-containing compounds. These results help to understand the uptake mechanism and subcellular distribution of Cd in polychaetes.
Ecotoxicology 08/2011; 21(1):104-10. · 2.36 Impact Factor
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Xiaoli Liu,
Linbao Zhang, Liping You,
Junbao Yu,
Ming Cong,
Qing Wang,
Fei Li,
Lianzhen Li,
Jianmin Zhao,
Chenghua Li,
Huifeng Wu
CLEAN - Soil Air Water 08/2011; 39(8):759 - 766. · 2.18 Impact Factor
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Linbao Zhang,
Xiaoli Liu,
Leilei Chen, Liping You,
Dong Pei,
Ming Cong,
Jianmin Zhao,
Chenghua Li,
Dongyan Liu,
Junbao Yu,
Huifeng Wu
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ABSTRACT: Glutathione peroxidases (GPx) are key enzymes in the antioxidant systems of living organisms by catalyzing the reduction of peroxides to non-reactive products. In the present study, the full-length cDNA encoding a selenium-dependent GPx was identified from Venerupis philippinarum (designated as VpSe-GPx), and the spatial and temporal expression patterns post-Vibrio anguillarum, heavy metals and benzo[a]pyrene (B[a]P) challenge were also investigated. VpSe-GPx possessed all the conserved features critical for the fundamental structure and function of glutathione peroxidase. The VpSe-GPx mRNA was found to be most abundantly expressed in hepatopancreas. Vibrio challenge could significantly up-regulate the mRNA expression of VpSe-GPx, and the highest expression level was detected at 24 h post-infection with 6.5-fold increase compared with that in the control group. For heavy metals exposure, the expression of VpSe-GPx was significantly induced by 20, 40 μg L(-1) Cd and 10, 20 μg L(-1) Cu but depressed by 10 μg L(-1) Cd and 40 μg L(-1) Cu. With regards to B[a]P exposure, the expression of VpSe-GPx mRNA was significantly induced at 48 and 96 h post challenge. All these results suggested that VpSe-GPx was potentially involved in mediating the immune response and antioxidant defense in V. Philippinarum.
Fish & Shellfish Immunology 07/2011; 31(6):831-7. · 3.32 Impact Factor
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Xiaoli Liu,
Cuiyun Yang,
Linbao Zhang,
Lianzhen Li,
Sujing Liu,
Junbao Yu, Liping You,
Di Zhou,
Chuanhai Xia,
Jianmin Zhao,
Huifeng Wu
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ABSTRACT: Cadmium is a non-essential element to living organisms and has become the severe contaminant in both seawater and sediment in the intertidal zones of the Bohai Sea. The halophyte, Suaeda salsa is the pioneer plant in the intertidal zones of Bohai Sea and has been widely applied in environmental sciences. In this study, the dose- and time-dependent effects induced by environmentally relevant concentrations (2, 10 and 50 μg l(-1)) of cadmium were characterized in S. salsa using NMR-based metabolomics. The levels of amino acids (valine, leucine, glutamate, tyrosine, etc.), carbohydrates (glucose, sucrose and fructose), intermediates of tricarboxylic acid cycle (succinate, citrate, etc.) and osmolyte (betaine) were altered in the S. salsa samples after cadmium exposures. These metabolic biomarkers indicated the elevated protein degradation and disturbances in the osmotic regulation and energy metabolism caused by cadmium in S. salsa. Overall, our results demonstrated the applicability of NMR-based metabolomics for the detection of metabolic biomarkers that could be used for the interpretation of toxicological effects induced by contaminants in the pioneer plant S. salsa in the intertidal zones. In addition, the metabolic biomarkers could be potentially useful for the bio-monitoring of contaminants in the intertidal zones.
Ecotoxicology 05/2011; 20(6):1422-31. · 2.36 Impact Factor
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ABSTRACT: Copper is an important heavy metal contaminant with high ecological risk in the Bohai Sea. In this study, the metabolic responses in the bioindicator, Manila clam (Ruditapes philippinarum), to the environmentally relevant copper exposures were characterized using NMR-based metabolomics. The significant metabolic changes corresponding to copper exposures were related to osmolytes, intermediates of the Krebs cycle and amino acids, such as the increase in homarine, branched chain amino acids and decrease in succinate, alanine and dimethylamine in the copper-exposed clam gills during 96 h exposure period. Overall, Cu may lead to the disturbances in osmotic regulation and energy metabolism in clams during 96 h experimental period. These results demonstrate that NMR-based metabolomics is applicable for the discovery of metabolic biomarkers which could be used to elucidate the toxicological mechanisms of marine heavy metal contaminants.
Marine environmental research 05/2011; 72(1-2):33-9. · 1.76 Impact Factor
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ABSTRACT: The Manila clam (Ruditapes philippinarum) has been considered a good sentinel species for metal pollution monitoring in estuarine tidal flats. Along the Bohai coast of China, there are dominantly distributed three species of clams (White, Liangdao Red and Zebra in Yantai population) endowed with distinct tolerances to environmental stressors. In this study, adductor muscle samples were collected from both control and acute mercury exposed White, Liangdao Red and Zebra clams, and the extracts were analyzed by NMR-based metabolomics to compare the metabolic profiles and responses to the acute mercury exposure to determine the most sensitive clam species capable of acting as abioindicator for heavy metal pollution monitoring. The major abundant metabolites in the White clam sample were branched-chain amino acids (leucine, isoleucine and valine), lactate, arginine, aspartate, acetylcholine, homarine and ATP/ADP, while the metabolite profile of Zebra clam sample comprised high levels of glutamine, acetoacetate, betaine, taurine and one unidentified metabolite. For the Liangdao Red clam sample, the metabolite profile relatively exhibited high amount of branched-chain amino acids, arginine, glutamate, succinate, acetylcholine, homarine and two unassigned metabolites. After 48h exposure of 20μgL(-1) Hg(2+), the metabolic profiles showed significant differences between three clam species, which included increased lactate, succinate, taurine, acetylcholine, betaine and homarine and decreased alanine, arginine, glutamine, glutamate, acetoacetate, glycine and ATP/ADP in White clam samples, and elevated succinate, taurine and acetylcholine, and declined glutamine, glycine, and aspartate in Liangdao Red clam samples, while the increased branched-chain amino acids, lactate, succinate, acetylcholine and homarine, and reduced alanine, acetoacetate, glycine and taurine were observed in the Zebra clam samples. Overall, our findings showed that White clams could be a preferable bioindicator for the metal pollution monitoring based on the more sensitive metabolic changes in the adductor muscle compared with other two (Liangdao Red and Zebra) clam species.
Environmental toxicology and pharmacology. 03/2011; 31(2):323-32.
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ABSTRACT: The controllable deformation of nanopores was realized by moving a convergent electron beam in a high-resolution transmission electron microscope. Nanostructures with the desired geometries were successfully fabricated from the original nanopores in 100 nm-thick and 260 nm-thick Si(3)N(4) membranes. The formation dynamics is a competition process between the knock-on effect of the high-energy electron beam and surface tension driven shrinkage. This approach can be used to finely tune critical dimensions and deform nanopores to particular desired geometries with single-nanometer precision, which offers substantial opportunities in flexibly fabricating nanostructures for various applications such as nanoelectronics and nanofluidics.
Nanotechnology 03/2011; 22(11):115302. · 3.98 Impact Factor
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Xiaoli Liu,
Linbao Zhang, Liping You,
Junbao Yu,
Jianmin Zhao,
Lianzhen Li,
Qing Wang,
Fei Li,
Chenghua Li,
Dongyan Liu,
Huifeng Wu
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ABSTRACT: Mercury is a hazardous pollutant in the Bohai marine environments due to its high toxicity to the marine organisms and subsequent ecological risk. Manila clam Ruditapes philippinarum is one of important sentinel organisms in 'Mussel Watch Program' launched in China and therefore used as a bioindicator in marine and coastal ecotoxicology. There are dominantly distributed three pedigrees of clam (White, Liangdao Red and Zebra) in Yantai population endowed with different tolerances to environmental stressors. In this study, gill tissues were collected from both untreated and mercury exposed White, Liangdao Red and Zebra clams, and the extracts were analyzed by NMR-based metabolomics to compare the original metabolomes and the toxicological effects induced by mercury exposure in three pedigrees. The major abundant metabolites in White clam sample were branched-chain amino acids, lactate, alanine, arginine, acetoacetate, glutamate, succinate, citrate, malonate and taurine, while the metabolite profile of Liangdao Red clam sample comprises relative high levels of alanine, arginine, glutamate, succinate and glycogen. For Zebra clam sample, the metabolite profile exhibited relatively high amount of aspartate, acetylcholine and homarine. After 48 h exposure of 20 μg l(-1) Hg(2+), the metabolic profiles from all the three pedigrees of clams commonly showed significant increases in alanine, arginine, glutamate, aspartate, α-ketoglutarate, glycine and ATP/ADP, and decreases in citrate, taurine and homarine. The unique metabolic differences between the metabolomes of gill tissues from Hg(2+)-exposed White, Liangdao Red and Zebra clams were found, including elevated acetylcholine and branched-chain amino acids in White clams, and the declined succinate in both White and Liangdao Red samples as well as the declined betaine in Zebra and White clams. Overall, our findings showed the differential toxicological responses to mercury exposure and that White clams could be a preferable bioindicator for the metal pollution monitoring based on the metabolic changes from gill compared with other two (Liangdao Red and Zebra) pedigrees of clams.
Ecotoxicology 11/2010; 20(1):177-86. · 2.36 Impact Factor
