Expression Of Glutathione S-transferase Research Articles

Receptor-Like Cytoplasmic Kinase STK Confers Salt Tolerance in Rice

BackgroundSoil salinization is a major abiotic environmental stress factor threatening crop production throughout the world. Salt stress drastically affects the growth, development, and grain yield of rice (Oryza sativa L.), and the improvement of rice tolerance to salt stress is a desirable approach for meeting increasing food demand. Receptor-like cytoplasmic kinases (RLCKs) play essential roles in plant growth, development and responses to environmental stresses. However, little is known about their functions in salt stress. Previous reports have demonstrated that overexpression of an RLCK gene SALT TOLERANCE KINASE (STK) enhances salt tolerance in rice, and that STK may regulate the expression of GST (Glutathione S-transferase) genes.ResultsThe expression of STK was rapidly induced by ABA. STK was highest expressed in the stem at the heading stage. STK was localized at the plasma membrane. Overexpression of STK in rice increased tolerance to salt stress and oxidative stress by increasing ROS scavenging ability and ABA sensitivity. In contrast, CRISPR/Cas9-mediated knockout of STK increased the sensitivity of rice to salt stress and oxidative stress. Transcriptome sequencing analysis suggested that STK increased the expression of GST genes (LOC_Os03g17480, LOC_Os10g38140 and LOC_Os10g38710) under salt stress. Reverse transcription quantitative PCR (RT-qPCR) suggested that four stress-related genes may be regulated by STK including OsABAR1, Os3BGlu6, OSBZ8 and OsSIK1.ConclusionsThese findings suggest that STK plays a positive regulatory role in salt stress tolerance by inducing antioxidant defense and associated with the ABA signaling pathway in rice.

Transcription Factor AhR Regulates Glutathione S-Transferases Conferring Resistance to lambda-Cyhalothrin in Cydia pomonella.

Aryl hydrocarbon receptor (AhR) enhances insect resistance to insecticides by regulating the detoxification network. Our previous studies have confirmed that overexpressions of cytochrome P450 monooxygenases (P450s) and glutathione S-transferases (GSTs) are involved in lambda-cyhalothrin resistance in Cydia pomonella. Here, we report that CpAhR regulates the expression of GST and P450 genes, thus conferring resistance. Expression patterns indicated that the expression of CpAhR was highly induced by lambda-cyhalothrin exposure and upregulated in a lambda-cyhalothrin-resistant population. RNA interference (RNAi) of CpAhR decreases the expression of key resistance-related genes (CpGSTe3, CpCYP9A121, and CpCYP9A122) and the activity of the GST enzyme, reducing the tolerance to lambda-cyhalothrin. Furthermore, β-naphthoflavone, a novel agonist of AhR, was first proven to be effective in increasing CpAhR expression and larval tolerance to lambda-cyhalothrin. These results demonstrate that CpAhR regulates the expression of key detoxifying genes and GST activity, resulting in the development of resistance to lambda-cyhalothrin in C. pomonella.

Transcriptomic and physiological properties reveal the tolerance mechanism to difenoconazole toxicity in wheat (Triticum aestivum L.)

Difenoconazole (DFZ) is a broad-spectrum fungicide widely applied in wheat production. However, excessive accumulation is linked to phytotoxicity. The effects of DFZ on plants and the response mechanisms to DFZ toxicity are poorly understood. Herein, the uptake, accumulation, and translocation of DFZ and induced changes in the morphology, physiology, and gene expression were investigated under hydroculture of roots treated with 50, 100, and 200 mg/L DFZ concentrations. Compared with the control, DEZ treatment upregulated the expression of genes encoding 4-coumarate-CoA ligase (4CL) and peroxidase (POD) involved in the lignin biosynthesis pathway and enhanced lignin biosynthesis. DFZ accumulated more in older leaves (cotyledons and lower true leaves), with 0.49–5.71 and 0.09–2.14 folds higher than levels in new upper leaves and roots, respectively. The excessive accumulation of DFZ in tissues was rapidly degraded, with a 15.7–69.3% reduction of DFZ content in roots and leaves from 3 DAT to 6 DAT. The genes expression and activity of glutathione S-transferase (GST) were increased. Furthermore, DFZ treatments upregulated genes encoding chalcone synthase (CHS), chalcone isomerase (CHI), and anthocyanidin synthase (ANS) involved in the flavonoid biosynthesis pathway and increased the amount of flavonoid and anthocyanins in leaves. This study provides new insights into the self-protective behaviors exhibited by wheat plants under DFZ stress. The mechanisms included hindering DFZ penetration from roots by enhancing lignin biosynthesis, accumulating more in old leaves, degrading by GST, and alleviating oxidative damage by increasing the content of flavonoids and anthocyanins in leaves.

Relation among Mercury, Selenium, and Biomarkers of Oxidative Stress in Northern Pike (Esox lucius).

Mercury (Hg) is a toxic environmental contaminant associated with oxidative stress in freshwater fish. A known antagonist to Hg, selenium (Se), may reduce the toxic effects of Hg. In this study, the relation among Se, methylmercury (MeHg), inorganic mercury (IHg), total mercury (THg), and the expression of biomarkers of oxidative stress and metal regulation in livers of northern pike were examined. Livers from northern pike were collected from 12 lakes in Isle Royale National Park, Pictured Rocks National Lakeshore, Sleeping Bear Dunes National Lakeshore, and Voyageurs National Park. The concentrations of MeHg, THg, and Se were measured in liver tissue, and the expression of superoxide dismutase (sod), catalase (cat), glutathione s-transferase (gst), and metallothionein (mt) was assessed. There was a positive relationship between the concentrations of THg and Se, with a Hg:Se molar ratio less than one in all livers examined. There was no significant relation between sod, cat, gst, or mt expression and Hg:Se molar ratios. cat and sod expression were significantly related to increases in percent MeHg, relative to THg; however, gst and mt expression were not significantly altered. This suggests that incorporating biomarkers containing Se may be a better indicator than non-selenium-containing proteins of assessing the long-term effect of Hg and the interactions between Hg and Se in the livers of fish, such as northern pike, especially when molar concentrations of Se are greater than Hg.

Glutathione S-transferase gene diversity and their regulation by Nrf2 in Chinese mitten crab (Eriocheir sinensis) during nitrite stress

Eriocheir sinensis is one of the most important economic aquatic products in China. However, nitrite pollution has become a serious threat to the healthy culture of E. sinensis. Glutathione S-transferase (GST) is an important phase II detoxification enzyme, which plays a leading role in the cellular detoxification of exogenous substances. In this study, we obtained 15 GST genes (designated as EsGST1-15) from E. sinensis, and their expression and regulation in E. sinensis under nitrite stress were studied. EsGST1-15 belonged to different GST subclasses. EsGST1, EsGST2, EsGST3, EsGST4, and EsGST5 belonged to Delta-class GSTs; EsGST6 and EsGST7 are Theta-class GSTs; EsGST8 is a mGST-3-class GST; EsGST9 belonged to mGST-1-class GSTs; EsGST10 and EsGST11 belonged to Sigma-class GSTs; EsGST12, EsGST13, and EsGST14 are Mu-class GSTs; EsGST15 is a Kappa-class GST. Tissue distribution experiments showed that EsGSTs were widely distributed in all detected tissues. The expression level of EsGST1-15 was significantly increased in the hepatopancreas under nitrite stress, indicating that EsGSTs were involved in the detoxification of E. sinensis under nitrite stress. Nuclear factor-erythroid 2 related factor 2 (Nrf2) is a transcription factor that can activate the expression of detoxification enzyme. We detected the expression of EsGST1-15 after interfering with EsNrf2 in the hepatopancreas of E. sinensis with or without nitrite stress. Results showed that EsGST1-15 were all regulated by EsNrf2 with or without nitrite stress. Our study provides new information about the diversity, expression, and regulation of GSTs in E. sinensis under nitrite stress.

Chitosan Nanoparticles Alleviated the Adverse Effects of Sildenafil on the Oxidative Stress Markers and Antioxidant Enzyme Activities in Rats.

Sildenafil (SF) is widely used for erectile dysfunction and other conditions, though with limitations regarding oral absorption and adverse effects. Despite nanotechnological improvements, the effect of nanocarriers on SF hepatotoxicity has not been documented to date. This study aimed at assessing the impact of chitosan nanoparticles either uncoated (CS NPs) or Tween 80-coated (T-CS NPs) on the effects of SF on oxidative stress markers and antioxidant enzyme activities in rats. Test SF-CS NPs prepared by ionic gelation were uniform positively charged nanospheres (diameter 178-215 nm). SF was administered intraperitoneally to male rats (1.5 mg/kg body weight) in free or nanoencapsulated forms as SF-CS NPs and T-SF-CS NPs for 3 weeks. Free SF significantly suppressed the activity of the antioxidant enzymes glutathione S-transferase (GST), glutathione peroxidase (GPx), glutathione reductase (GR), catalase (CAT), and superoxide dismutase (SOD), as well as the levels of glutathione (GSH) and thiobarbituric acid reactive substances (TBARS) as in an indirect measure of free radicals. Interestingly, SF-CS NPs and T-SF-CS-NPs treatments significantly attenuated the inhibitory effects of SF on the activity of these enzymes whereas, GST activity was inhibited. Moreover, the protein expression of GST was downregulated upon treatment of rats with free SF, SF-CS-NPs, and T-SF CS-NPs. In contrast, the activity and protein expression of GPx was induced by SF-CS NPs and T-SF-CS-NPs treatments. The histopathological study showed that SF induced multiple adverse effects on the rat liver architecture which were markedly suppressed particularly by T-SF-CS NPs. In conclusion, chitosan nanoencapsulation of SF counteracted the adverse effects of SF on the activity of antioxidant enzymes and liver architecture. Findings might have significant implications in improving the safety and efficacy of SF treatment of the widely expanding disease conditions.

Salt stress improves the low-temperature tolerance in sugar beet in which carbohydrate metabolism and signal transduction are involved

Low temperatures can inhibit crop growth and yield. To investigate the physiological mechanisms by which salt stress affects low temperature tolerance, sugar beet seedlings subjected to low temperature stress after treated with 0, 200 and 400 mM NaCl were analyzed for physiome, transcriptome and metabolome profiles. Salt stress significantly alleviated the maximum quantum efficiency of PS II (Fv/Fm) and the reduction of relative leaf water content (RWC) caused by low temperature. Salt stress enhanced reactive oxygen species (ROS) metabolism and significantly increased the activities of superoxide dismutase, peroxidase (POX) and dehydroascorbate reductase, and it also increased the gene expression of glutathione S-transferase, POX and L-ascorbate peroxidase. Meanwhile, salt stress also contributed to the increase in carbohydrate enzyme activities (e.g. vacuolar invertase, hexokinase and fructokinase) and gene expression. Salt stress resulted in a decrease in sucrose concentration and an increase in proline concentration. Sucrose concentration was significantly and negatively correlated with the gene expression of the signal transduction pathway. Additionally, the expression of genes related to abscisic acid (protein phosphatase 2 C and receptor), gibberellin signaling (DELLA and F-box protein) and 3-ketoacyl-CoA synthase was significantly increased by salt stress under low temperature. These results suggest that salt stress regulated ROS metabolism, carbohydrate metabolism and hormone signaling, thereby enhancing low temperature tolerance in sugar beet.

Lethal and sub-lethal effects of Cordyceps fumosorosea on the demography and biochemistry of Phenacoccus solenopsis

ABSTRACT The cotton mealybug, Phenacoccus solenopsis Tinsley, is a highly invasive pest of cotton. The use of entomopathogenic fungi as a bio-pesticide could provide an alternative to current chemical control measures that preserve the environment and biodiversity, and ensures good quality of crops. Lethal, sub-lethal and trans-generational studies are needed to assess the effects of myco-insecticides on such pests. The pathogenicity of different spore concentrations of Cordyceps fumosorosea Wize was assessed against second-instar nymphs by immersion while focusing on lethal (LC50) and sub-lethal (LC30) spore concentration treatments of C. fumosorosea on filial (F0) and first filial (F1) generations. After treatment at LC50 and LC30 spore concentrations of C. fumosorosea, the progeny of P. solenopsis showed reduced nymphal duration, adult longevity and fecundity as compared with controls. Also, total pre-oviposition period (TPOP), adult pre-oviposition period (APOP) and oviposition days (Od) were reduced with the increasing spore concentration of C. fumosorosea. In addition, population growth parameters such as net reproductive rates (Ro) and mean generation time (T) were dramatically reduced. The level of expression of acetylcholinesterase (AChE), esterases (EST), acid and alkaline phosphatases (ACP, ALP) and glutathione S-transferases (GST) were also examined. Significant expression of EST, GST, ALP, ACP and AChE (7.15, 4.51, 4.54, 2.80 and 8.32 μmol/min of mg/ml protein), respectively were observed at 8 × 107 conidia/mL of C. fumosorosea. Pathogenicity even at sub-lethal doses as well as the production of detoxifying enzymes indicates that C. fumosorosea is the most effective fungus against P. solenopsis.

Gene expression analysis of antioxidant and DNA methylation on the rat liver after 4-week wood preservative chromated copper arsenate exposure.

Our previous 4-week repeated dose toxicity study showed that wood preservative chromated copper arsenate (CCA) induced hepatocellular hypertrophy accompanied by biochemical hepatic dysfunction and an increase in oxidative stress marker, 8-hydroxydeoxyguanosine, in female rats. To further explore the molecular mechanisms of CCA hepatotoxicity, we analyzed 10%-buffered formalin-fixed liver samples from female rats for cell proliferation, apoptosis, and protein glutathionylation and conducted microarray analysis on frozen liver samples from female rats treated with 0 or 80 mg/kg/day of CCA. Chemical analysis revealed that dimethylated arsenical was the major metabolite in liver tissues of male and female rats. CCA increase labeling indices of proliferating cell nuclear antigen and decrease terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling accompanied with increased expression of protein glutathionylation, indicating a decrease in glutathione (GSH) in hepatocytes of female rats. Microarray analysis revealed that CCA altered gene expression of antioxidants, glutathione-S-transferase (GST), heat shock proteins and ubiquitin-proteasome pathway, cell proliferation, apoptosis, DNA methylation, cytochrome P450, and glucose and lipid metabolism in female rats. Increased expression of GSTs, including Gsta2, Gsta3, Mgst1, and Cdkn1b (p27), and decreased expression of the antioxidant Mt1, and DNA methylation Dnmt1, Dnmt3a, and Ctcf were confirmed in the liver of female rats in a dose-dependent manner. Methylation status of the promoter region of the Mt1 was not evidently changed between control and treatment groups. The results suggested that CCA decreased GSH and altered the expression of several genes, including antioxidants, GST, and DNA methylation, followed by impaired cell proliferation in the liver of female rats.

Hypoxia Affects the Antioxidant Activity of Glutaredoxin 3 in Scylla paramamosain through Hypoxia Response Elements.

Hypoxia is a major environmental stressor that can damage the oxidation metabolism of crustaceans. Glutaredoxin (Grx) is a key member of the thioredoxin superfamily and plays an important role in the host's defense against oxidative stress. At present, the role of Grx in response to hypoxia in crustaceans remains unclear. In this study, the full-length cDNA of Grx3 (SpGrx3) was obtained from the mud crab Scylla paramamosain, which contains a 129-bp 5' untranslated region, a 981-bp open reading frame, and a 1,183-bp 3' untranslated region. The putative SpGrx3 protein contains an N-terminal thioredoxin domain and two C-terminal Grx domains. SpGrx3 was expressed in all tissues examined, with the highest expression in the anterior gills. After hypoxia, SpGrx3 expression was significantly up-regulated in the anterior gills of mud crabs. The expression of Grx2 and glutathione S-transferases was decreased, while the expression of glutathione peroxidases was increased following hypoxia when SpGrx3 was silenced in vivo. In addition, the total antioxidant capacity of SpGrx3-interfered mud crabs was significantly decreased, and the malondialdehyde content was significantly increased during hypoxia. The subcellular localization data indicated that SpGrx3 was predominantly localized in the nucleus when expressed in Drosophila Schneider 2 (S2) cells. Moreover, overexpression of SpGrx3 reduced the content of reactive oxygen species in S2 cells during hypoxia. To further investigate the transactivation mechanism of SpGrx3 during hypoxia, the promoter region of the SpGrx3 was obtained by Genome Walking and three hypoxia response elements (HREs) were predicted. Dual-luciferase reporter assay results demonstrated that SpGrx3 was likely involved in the hypoxia-inducible factor-1 (HIF-1) pathway during hypoxia, which could be mediated through HREs. The results indicated that SpGrx3 is involved in regulating the antioxidant system of mud crabs and plays a critical role in the response to hypoxia.

Evaluation of Garlic Juice Processing Waste Supplementation in Juvenile Black Rockfish (Sebastes schlegelii) Diets on Growth Performance, Antioxidant and Digestive Enzyme Activity, Growth- and Antioxidant-Related Gene Expression, and Disease Resistance against Streptococcus iniae.

An 8-week feeding trial was conducted to evaluate the effects of various dietary levels of garlic juice processing waste (GJPW) on the growth, feed utilization, digestive and antioxidant enzyme activity, growth- and antioxidant-related gene expression, and resistance to Streptococcus iniae infection of juvenile black rockfish (Sebastes schlegelii). A total of 450 juvenile rockfish were randomly distributed into 30 L rectangular tanks (30 fish per tank). Five experimental diets were prepared in triplicate. The fish were fed experimental diets supplemented with GJPW at concentrations of 0 (GJPW0, control), 2.5 (GJPW2.5), 5 (GJPW5), 7.5 (GJPW7.5), and 10 g kg-1 (GJPW10) diet. All of the GJPW-supplemented treatments (2.5, 5, 7.5, and 10 g kg-1) significantly enhanced weight gain (WG), specific growth rate (SGR), feed efficiency (FE), protein efficiency ratio (PER), and digestive enzyme activity (amylase, trypsin, and lipase). A decreasing trend was seen in plasma aspartate aminotransferase (ALT), alanine aminotransferase (AST), and glucose (GLU) content with increasing dietary levels of GJPW. In contrast, plasma lysozyme and antioxidant enzyme activities were significantly increased with increasing dietary GJPW levels. Furthermore, GJPW administration significantly upregulated the expression of insulin-like growth factor-1 (IGF-1), superoxide dismutase (SOD), catalase (CAT), and glutathione S-transferase (GST) in the liver of rockfish. A challenge test with S. iniae showed significantly higher resistance in the GJPW-supplemented treatments than in the control. In short, dietary supplementation GJPW enhanced growth performance and antioxidant response in juvenile black rockfish, with suitable effects in fish fed with 2.5 g kg-1 GJPW for 8 weeks.

CtrA activates the expression of glutathione S-transferase conferring oxidative stress resistance to Ehrlichia chaffeensis.

Ehrlichia chaffeensis, the causative agent of human monocytic ehrlichiosis (HME), is a Gram-negative obligatory intracellular bacterium, which infects and multiplies in human monocytes and macrophages. Host immune cells produce reactive oxygen species (ROS) to eliminate E. chaffeensis upon infection. E. chaffeensis global transcriptional regulator CtrA activates the expression of GshA and GshB to synthesize glutathione (GSH), the most potent natural antioxidant, upon oxidative stress to combat ROS damage. However, the mechanisms exploited by E. chaffeensis to utilize GSH are still unknown. Here, we found that in E. chaffeensis CtrA activated the expression of glutathione S-transferase (GST) upon oxidative stress, and E. chaffeensis GST utilizes GSH to eliminate ROS and confers the oxidative stress resistance to E. chaffeensis. We found that CtrA bound to the promoter regions of 211 genes, including gst, in E. chaffeensis using chromatin immunoprecipitation coupled to deep sequencing (ChIP-seq). Recombinant E. chaffeensis CtrA directly bound to the gst promoter region determined with electrophoretic mobility shift assay (EMSA), and activated the gst expression determined with reporter assay. Recombinant GST showed GSH conjugation activity towards its typical substrate 2,4-dinitrochlorobenzene (CDNB) in vitro and peptide nucleic acid (PNA) transfection of E. chaffeensis, which can knock down the gst transcription level, reduced bacterial survival upon oxidative stress. Our results demonstrate that E. chaffeensis CtrA regulates GSH utilization, which plays a critical role in resistance to oxidative stress, and aid in the development of new therapeutics for HME.

Nickel translocation, toxicity, and its effect on the expression of genes associated with nickel resistance in red pine (Pinus resinosa)

The main objectives of this study were to 1) investigate metal accumulation and translocation in Pinus spp ., 2) evaluate the level of P. resinosa resistance to nickel; and 3) determine the effects of different concentrations of nickel on gene expression in Pinus resinosa . Seedlings were treated with different concentrations of nickel nitrate (16 mg, 160 mg, and 1,600 mg of nickel per 1 kg of dry soil) and corresponding salt controls in growth chamber screening tests. In general, collected data revealed that P. resinosa is resistant to nickel. The expression of 1-aminocyclopropane-1-carboxylic acid deaminase (ACC deaminase), glutathione-S-transferase (GST), High-affinity Ni transporter (NiCoT or AT2G16800), and natural resistance-associated macrophage protein 3 (NRAMP3) and Serine acetyltransferase (SAT) were analyzed using RT-qPCR. The expression of ACC in samples treated with 1,600 mg/kg of Ni was significantly supressed as it was half of that observed in the water control samples. AT2G16800, and SAT expression were significantly upregulated under 16 mg/kg and 160 mg/kg Ni treatments. But the expression of these genes was repressed in seedlings exposed to 1,600 mg/kg of Ni. The 1,600 mg/kg of Ni induced a GST expression level that was 6.65-fold greater than that of the water control. Overall, low level of nickel ions induce an increase in the expression of ACC, GST, AT2G16800, NRAMP3, and SAT. The levels of bioavailable Ni in soils contaminated with metals was low while Ni concentrations in roots, branches, and needles of P. resinosa exposed to Ni for several years were similar.

Identification of Nrf2 Activators from the Roots of Valeriana officinalis.

Various age-related chronic diseases have been linked to oxidative stress. The cellular antioxidant response pathway is regulated by the transcription factor nuclear erythroid factor 2. Therefore, plant-derived nuclear erythroid factor 2 activators might be useful therapeutics to stimulate the body's defense mechanisms. Our study focused on the discovery of potent nuclear erythroid factor 2 activators from medicinal plants. Initially, a variety of medicinal plant extracts were screened for nuclear erythroid factor 2 activity using a nuclear erythroid factor 2 luciferase reporter cell line. Among these, Valerian (Valeriana officinalis) root was identified as a potent candidate. Sequential extraction and bioassay-guided fractionation led to the isolation of four nuclear erythroid factor 2-active compounds, which were structurally identified by NMR and LC/HRMS as the known compounds isovaltrate, valtrate, jatamanvaltrate-P, and valerenic acid. These four compounds were then tested in relevant biological assays. Firstly, their effects on the expression of glutathione S-transferase, glutamate-cysteine ligase catalytic subunit, glutathione peroxidase, and heme oxygenase 1 were determined in HepG2 cells. Glutathione S-transferase P1 and glutamate-cysteine ligase catalytic subunit were upregulated by isovaltrate, valtrate, and jatamanvaltrate-P, while heme oxygenase 1 was upregulated by isovaltrate, jatamanvaltrate-P, and valerenic acid. The four compounds also increased the levels of glutathione and its metabolite, CysGly. As glutathione aids in the detoxification of hydrogen peroxide, cytoprotective effects of these four nuclear erythroid factor 2 activators against hydrogen peroxide toxicity were investigated, and indeed, the compounds significantly improved cell survival. This study provides evidence that four valepotriates from the roots of V.officinalis are activators of nuclear erythroid factor 2-mediated antioxidant and detoxification pathways. Our data might expand the medical use of this plant beyond its current application as a sleep aid.

Combating Drug Resistance by Exploiting miRNA-200c-Controlled Phase II Detoxification.

Acquired drug resistance constitutes a serious obstacle to the successful therapy of cancer. In the process of therapy resistance, microRNAs can play important roles. In order to combat resistance formation and to improve the efficacy of chemotherapeutics, the mechanisms of the multifaceted hsa-miR-200c on drug resistance were elucidated. Upon knockout of hsa-miR-200c in breast carcinoma cells, a proteomic approach identified altered expression of glutathione S-transferases (GSTs) when cells were treated with the chemotherapeutic drug doxorubicin. In different hsa-miR-200c expression systems, such as knockout, inducible sponge and inducible overexpression, the differential expression of all members of the GST family was evaluated. Expression of hsa-miR-200c in cancer cells led to the repression of a multitude of these GSTs and as consequence, enhanced drug-induced tumor cell death which was evaluated for two chemotherapeutic drugs. Additionally, the influence of hsa-miR-200c on the glutathione pathway, which is part of the phase II detoxification mechanism, was investigated. Finally, the long-term effects of hsa-miR-200c on drug efficacy were studied in vitro and in vivo. Upon doxycycline induction of hsa-miR-200c, MDA-MB 231 xenograft mouse models revealed a strongly reduced tumor growth and an enhanced treatment response to doxorubicin. A combined treatment of these tumors with hsa-miR-200c and doxorubicin resulted in complete regression of the tumor in 60% of the animals. These results identify hsa-miR-200c as an important player regulating the cellular phase II detoxification, thus sensitizing cancer cells not expressing this microRNA to chemotherapeutics and reversing drug resistance through suppression of GSTs.

Alexander Munn

An immunohistochemical study of glutathione S-transferase (GST) expression in hepatocellular carcinoma and cholangiocarcinoma is described. Unlike most animal models of hepatic malignancy pi class GST was not consistently overexpressed in...

Immune Responses of Rhynchophorus ferrugineus to a New Strain of Beauveria bassiana

Evaluating a novel fungal strain’s pathogenicity to important pests and their involved immune responses may give crucial data on a broad scale for future use in pest management strategies. Date palms are mostly destroyed by invading populations of red palm weevils; thus, developing natural biopesticides for them requires a comprehensive screening program of plant secondary metabolites. In this research, we examined the pathogenicity of a new strain of Beauveria bassiana on an important agricultural pest, Rhynchophorus ferrugineus, by measuring the relative activity of defensive enzymes and detoxifying enzymes in certain larval instars. Our findings reveal that the B. bassiana strain may infect the instars of R. ferrugineus, and its pathogenicity to the larvae steadily increases as the spore concentration increases. Seven days after inoculation, the LC50 (the median lethal concentration) of B. bassiana was 490.42 × 105 and 2974.47 × 108 spores/mL for the second and fourth instar R. ferrugineus, respectively, and the LC50 of B. bassiana for each R. ferrugineus instar decreased with infection time, indicating a significant dose effect. Infected R. ferrugineus larvae of the second instars showed considerable changes in the activity of both protecting and detoxifying enzymes (peroxidase, catalase, superoxide dismutase, Cytochrome P450, glutathione S transferase (GST), and esterase) as infection time progressed. In addition, R. ferrugineus larvae that were infected with B. bassiana had enzyme activity that persisted from 24 to 48 h, which was much longer than in the control group. Lethality of B. bassiana resulted in elevated expressions of GST, Esterase, and Cytochrome P450 responsive genes. In conclusion, the results of this research indicate that B. bassiana may be utilized as a bio-insecticide to suppress young larvae of R. ferrugineus in an integrated pest management program.

Regulation of the molecular repertoires of oxidative stress response in the gills and olfactory organ of Atlantic salmon following infection and treatment of the parasite Neoparameoba perurans

The present study investigated the involvement of key molecular regulators of oxidative stress in amoebic gill disease (AGD), a parasitic infestation in Atlantic salmon. In addition, the study evaluated how these molecular biomarkers responded when AGD-affected fish were exposed to a candidate chemotherapeutic peracetic acid (PAA). Atlantic salmon were experimentally infected with the parasite Neoparameoba perurans, the causative agent of AGD, by bath exposure and after 2 weeks, the fish were treated with three commercial PAA products (i.e., Perfectoxid, AquaDes and ADDIAqua) at a dose of 5 ppm. Two exposure durations were evaluated – 30 min and 60 min. Sampling was performed 24 h and 2 weeks after PAA treatment (equivalent to 2- and 4-weeks post infection). At each sampling point, the following parameters were evaluated: gross gill pathology, gill parasitic load, plasma reactive oxygen species (ROS) and total antioxidant capacity (TAC), histopathology and gene expression profiling of genes with key involvement in oxidative stress in the gills and olfactory organ. AGD did not result in systemic oxidative stress as ROS and TAC levels remained unchanged. There were no clear patterns of AGD-mediated regulation of the oxidative stress biomarkers in both the gills and olfactory organ; significant changes in the expression were mostly related to time rather than infection status. However, the expression profiles of the oxidative stress biomarkers in AGD-affected salmon, following treatment with PAA, revealed that gills and olfactory organ responded differently – upregulation was prominent in the gills while downregulation was more frequent in the olfactory organ. The expression of catalase, glutathione S-transferase and thioredoxin reductase 2 was significantly affected by the treatments, both in the gills and olfactory organ, and these alterations were influenced by the duration of exposure and PAA product type. Parasitic load in the gills did significantly increase after treatment regardless of the product and exposure duration; the parasite was undetectable in some fish treated with AquaDes for 30 mins. However, PAA treated groups for 30 min showed lower macroscopic gill scores than the infected-untreated fish. Histology disclosed the classic pathological findings such as multifocal hyperplasia and increased number of mucous cells in AGD-affected fish. Microscopic scoring of gill injuries showed that AGD-infected-PAA-treated fish had lower scores, however, an overall trend could not be established. The morphology and structural integrity of the olfactory organ were not significantly altered by parasitism or PAA treatment. Collectively, the results indicate that AGD did not affect the systemic and mucosal oxidative status of Atlantic salmon. However, such a striking profile was changed when AGD-affected fish were exposed to oxidative chemotherapeutics. Moreover, the gills and olfactory organ demonstrated distinct patterns of gene expression of oxidative stress biomarkers in AGD-infected-PAA-treated fish. Lastly, PAA treatment did not fully resolve the infection, but appeared not to worsen the mucosal health either.

Effects of Exogenous Linoleic Acid on Barley (Hordeum vulgare L.) Seedlings Under Salinity

Salt stress adversely affects plants and causes different levels of morphological, physiological, biochemical, and molecular changes at different growth stages. Polyunsaturated fatty acids (PUFAs), such as linoleic acid, are main components of membrane lipids and determine the fluidity and stability of the cell membrane. In addition, PUFAs have a crucial role in maintaining the structure and function of the cell membrane which is damaged by salinity. There may be a relationship between level of PUFAs in membrane lipids and salinity tolerance. The present study was carried out to examine the effects of exogenous application of 0.5 mM linoleic acid (LA) on barley seedlings (Hordeum vulgare L. cv. Martı) grown in hydroponic conditions under 160 mM NaCl. The treatment with LA ameliorated the stress generated by NaCl by increasing osmolyte level and decreasing ion leakage percentage and H2O2 content within hours. Besides, LA significantly enhanced expression of salt-responsive transcription factor HvDRF2 and ROS scavenger gene HvMT2 as 105- and 40-fold, respectively, in the leaves of barley seedlings under salinity conditions. While LA slightly increased the gene expression of ascorbate peroxidase (HvAPX), glutathione S-transferase (HvGST6) and copper zinc superoxide dismutase (HvCu/ZnSOD) in the roots of barley seedlings, the expression of these genes was not changed in the leaves under salinity compared to salt-stressed samples. This study provides novel insights for effects of LA on improvement of salinity tolerance in barley.

The mechanism of Yinchenhao decoction in treating obstructive-jaundice-induced liver injury based on Nrf2 signaling pathway.

BACKGROUNDObstructive jaundice (OJ) is caused by bile excretion disorder after partial or complete bile duct obstruction. It may cause liver injury through various mechanisms. Traditional Chinese medicine (TCM) has a lot of advantages in treating OJ. The recovery of liver function can be accelerated by combining Chinese medicine treatment with existing clinical practice. Yinchenhao decoction (YCHD), a TCM formula, has been used to treat jaundice. Although much progress has been made in recent years in understanding the mechanism of YCHD in treating OJ-induced liver injury, it is still not clear.AIMTo investigate chemical components of YCHD that are effective in the treatment of OJ and predict the mechanism of YCHD.METHODSThe active components and putative targets of YCHD were predicted using a network pharmacology approach. Gene Ontology biological process and Kyoto Encyclopedia of Genes and Genomes path enrichment analysis were carried out by cluster profile. We predicted the biological processes, possible targets, and associated signaling pathways that YCHD may involve in the treatment of OJ. Thirty male Sprague–Dawley rats were randomly divided into three groups, each consisting of 10 rats: the sham group (Group S), the OJ model group (Group M), and the YCHD-treated group (Group Y). The sham group only received laparotomy. The OJ model was established by ligating the common bile duct twice in Groups M and Y. For 1 wk, rats in Group Y were given a gavage of YCHD (3.6 mL/kg) twice daily, whereas rats in Groups S and M were given the same amount of physiological saline after intragastric administration daily. After 7 d, all rats were killed, and the liver and blood samples were collected for histopathological and biochemical examinations. Total bilirubin (TBIL), direct bilirubin (DBIL), alanine aminotransferase (ALT), and aspartate transaminase (AST) levels in the blood samples were detected. The gene expression levels of inducible nitric oxide synthase (iNOS) and endothelial nitric oxide synthase (eNOS), and the nucleus positive rate of NF-E2 related factor 2 (Nrf2) protein were measured. Western blot analyses were used to detect the protein and gene expression levels of Nrf2, Kelch-like ECH-associated protein 1, NAD(P)H quinone dehydrogenase 1 (NQO1), and glutathione-S-transferase (GST) in the liver tissues. One-way analysis of variance was used to evaluate the statistical differences using the statistical package for the social sciences 23.0 software. Intergroup comparisons were followed by the least significant difference test and Dunnett’s test.RESULTSThe effects of YCHD on OJ involve biological processes such as DNA transcription factor binding, RNA polymerase II specific regulation, DNA binding transcriptional activator activity, and nuclear receptor activity. The protective effects of YCHD against OJ were closely related to 20 pathways, including the hepatitis-B, the mitogen-activated protein kinase, the phosphatidylinositol 3-kinase/protein kinase B, and tumor necrosis factor signaling pathways. YCHD alleviated the swelling and necrosis of hepatocytes. Following YCHD treatment, the serum levels of TBIL (176.39 ± 17.03 μmol/L vs 132.23 ± 13.88 μmol/L, P < 0.01), DBIL (141.41 ± 14.66 μmol/L vs 106.43 ± 10.88 μmol/L, P < 0.01), ALT (332.07 ± 34.34 U/L vs 269.97 ± 24.78 U/L, P < 0.05), and AST (411.44 ± 47.64 U/L vs 305.47 ± 29.36 U/L, P < 0.01) decreased. YCHD promoted the translocation of Nrf2 into the nucleus (12.78 ± 0.99 % vs 60.77 ± 1.90 %, P < 0.001). After YCHD treatment, we found a decrease in iNOS (0.30 ± 0.02 vs 0.20 ± 0.02, P < 0.001) and an increase in eNOS (0.18 ± 0.02 vs 0.32 ± 0.02, P < 0.001). Meanwhile, in OJ rats, YCHD increased the expressions of Nrf2 (0.57 ± 0.03 vs 1.18 ± 0.10, P < 0.001), NQO1 (0.13 ± 0.09 vs 1.19 ± 0.07, P < 0.001), and GST (0.12 ± 0.02 vs 0.50 ± 0.05, P < 0.001), implying that the potential mechanism of YCHD against OJ-induced liver injury was the upregulation of the Nrf2 signaling pathway.CONCLUSIONOJ-induced liver injury is associated with the Nrf2 signaling pathway. YCHD can reduce liver injury and oxidative damage by upregulating the Nrf2 pathway.