Expression Of Glutathione S-transferase Research Articles

A Double-Stranded Aptamer for Highly Sensitive Fluorescent Detection of Glutathione S-Transferases.

Aptamer-based biosensors have been widely constructed and applied to detect diverse targets. Glutathione S-transferase (GST), a pivotal phase II metabolic enzyme, plays a critical role in biotransformation in vivo, and aberrant GST expression is associated with various health risks. Herein, aptamers targeting GST were systematically selected from a randomized single-stranded DNA (ssDNA) library of 79 nucleotides (nt) using a biotinylated GST-immobilized streptavidin agarose (SA) bead SELEX technology. Following rigorous screening across eight rounds, four aptamers with strikingly similar secondary structures emerged. Among these, Seq3 exhibited the highest affinity towards GST and was selected for further optimization. A semi-rational post-SELEX truncation strategy was then employed based on base composition analysis, secondary structure analysis and affinity assessment. This strategy enabled the systematic removal of redundant nucleotides in Seq3 without compromising its affinity, ultimately yielding a truncated aptamer, Seq3-3, which retains its specificity with a compact 39nt length. Building upon Seq3-3, a double-stranded fluorescent aptamer probe was ingeniously designed for the in vitro detection of GST. The detection mechanism hinges on the competitive displacement of the complementary chain from the probe, mediated by the target protein, leading to the separation of the antisense oligonucleotide from the double-stranded complex. This process triggers the restoration of the fluorescence signal, enabling sensitive detection, and the probe exhibits excellent response within a linear range of GST activity ranging from 0 to 1500 U/L. The results show that not only an efficient strategy for screening robust and practicable aptamers but also an ultrahighly sensitive detection platform for GST was established.

Transcriptomic Analyses to Unravel Cronobacter sakazakii Resistance Pathways.

The proliferation of antibiotic usage has precipitated the emergence of drug-resistant variants of bacteria, thereby augmenting their capacity to withstand pharmaceutical interventions. Among these variants, Cronobacter sakazakii (C. sakazakii), prevalent in powdered infant formula (PIF), poses a grave threat to the well-being of infants. Presently, global contamination by C. sakazakii is being observed. Consequently, research endeavors have been initiated to explore the strain's drug resistance capabilities, alterations in virulence levels, and resistance mechanisms. The primary objective of this study is to investigate the resistance mechanisms and virulence levels of C. sakazakii induced by five distinct antibiotics, while concurrently conducting transcriptomic analyses. Compared to the susceptible strains prior to induction, the drug-resistant strains exhibited differential gene expression, resulting in modifications in the activity of relevant enzymes and biofilm secretion. Transcriptomic studies have shown that the expression of glutathione S-transferase and other genes were significantly upregulated after induction, leading to a notable enhancement in biofilm formation ability, alongside the existence of antibiotic resistance mechanisms associated with efflux pumps, cationic antimicrobial peptides, and biofilm formation pathways. These alterations significantly influence the strain's resistance profile.

Evaluation of Glutathione S-transferases Expression as Biomarkers by Heavy Metals in Geloina expansa from Sepang Besar River, Selangor, Malaysia

Glutathione S-transferases (GSTs) are enzymes involved in phase II of detoxification metabolism and could be used as biomarkers for water pollution. This study aims to determine heavy metal concentrations in the soft tissue of the mangrove clam Geloina expansa, as well as the expression of GSTs in the species. The acid digestion method was used to digest the samples, followed by a standard USEPA 6010B procedure using inductively coupled plasma optical emission spectrometry (ICP-OES) to measure the heavy metal contents in the samples. GST enzyme activity was measured using 1-chloro-2, 4-dinitrobenzene (CDNB) as substrate. One-way ANOVA was performed to compare the mean values of heavy metal concentration, protein concentration, enzyme activity, and specific activity. There was a significant difference (p<0.05) for Zn, total protein, and specific activity in G. expansa, but no significant difference in Pb, Cu and enzyme activity. GST enzyme activities were estimated at 0.16 ± 0.01 µmol/min, with a protein content of 1.24 ± 0.04 mg. The specific activity for GST was 0.13 ± 0.01 µmol/min/mg, calculated as the ratio of enzyme activity to the total protein. GST-specific activity positively correlates with Pb concentration in the soft tissue of G. expansa. Detailed studies on the effects of pollution on the expression of GST need to be further investigated for the future use of this species as an efficient biomarker model.

A Glutathione S-Transferase from Thinopyrum ponticum Confers Fhb7 Resistance to Fusarium Head Blight in Wheat.

Fusarium head blight (FHB), mainly incited by Fusarium graminearum, has caused great losses in grain yield and quality of wheat globally. Fhb7, a major gene from 7E chromosome of Thinopyrum ponticum, confers broad resistance to multiple Fusarium species in wheat and has recently been cloned and identified as encoding a glutathione S-transferase (GST). However, some recent reports raised doubt about whether GST is the causal gene of Fhb7. To resolve the discrepancy and validate the gene function of GST in wheat, we phenotyped Fhb7 near-isogenic lines (Jimai22-Fhb7 versus Jimai22) and GST overexpressed lines for FHB resistance. Jimai22-Fhb7 showed significantly higher FHB resistance with a lower percentage of symptomatic spikelets, Fusarium-damaged kernels, and deoxynivalenol content than susceptible Jimai22 in three experiments. All the positive GST transgenic lines driven by either the maize ubiquitin promoter or its native promoter with high gene expression in the wheat cultivar 'Fielder' showed high FHB resistance. Only one maize ubiquitin promoter-driven transgenic line showed low GST expression and similar susceptibility to Fielder, suggesting that high GST expression confers Fhb7 resistance to FHB. Knockout of GST in the Jimai22-Fhb7 line using CRISPR-Cas9-based gene editing showed significantly higher FHB susceptibility compared with the nonedited control plants. Therefore, we confirmed GST as the causal gene of Fhb7 for FHB resistance. Considering its major effect on FHB resistance, pyramiding Fhb7 with other quantitative trait loci has a great potential to create highly FHB-resistant wheat cultivars.

Naphthalene and phenanthrene affect differentially two glutathione S-transferases (GSTs) expression, GST activity, and glutathione content in white shrimp P. vannamei

Polycyclic aromatic hydrocarbons (PAHs) are persistent organic pollutants ubiquitous in coastal ecosystems. The white shrimp Penaeus vannamei naturally inhabits in coastal areas and is cultivated in farms located nearby the oceans. PAHs can damage shrimp health, endanger natural populations, and lower shrimp aquaculture productivity. However, crustaceans have enzymes capable of metabolizing organic xenobiotics as PAHs and to neutralize reactive oxygen species (ROS) produced during xenobiotics metabolism. An important superfamily of xenobiotic-metabolizing and antioxidant enzymes are glutathione S-transferases (GSTs). In white shrimp, some GSTs are known, but they have been scarcely studied in response to PAHs. In this study we report the molecular cloning and bioinformatic characterization of two novel nucleotide sequences corresponding to cytosolic GSTs belonging the Delta and Theta classes (GSTD and GSTT). Both proteins genes have tissue-specific patterns of expression under normal conditions, that do not necessarily relate to GST activity and glutathione content. The expression of the GSTD and GSTT, GST activity and glutathione content was analyzed in juvenile P. vannamei exposed to two PAHs, naphthalene (NAP) and phenanthrene (PHE) in sub-lethal concentrations for 96 h. GSTD expression was up-regulated by the two PAHs, while GSTT expression was only induced by NAP. In contrast, GST activity towards CDNB was only up-regulated by PHE, suggesting differential effects of PAHs at gene and protein level. On the other hand, lower reduced glutathione content (GSH) caused by PAHs indicates its utilization for detoxification or antioxidant defenses. However, the GSH/GSSG did not change by PAHs treatment, indicating that shrimp can maintain redox balance during short-term sub-lethal exposure to NAP and PHE. Despite the variations in the responses to NAP and PHE, all these results suggest that the GSTD and GSTT genes could be useful biomarkers for PAH exposure in P. vannamei.

4-Octyl itaconate alleviates dextran sulfate sodium-induced ulcerative colitis in mice via activating the KEAP1-NRF2 pathway.

Ulcerative colitis (UC) is a chronic idiopathic inflammatory bowel disease with a relapsing-remitting course. Although its etiology remains unknown, excessive oxidative stress in colon is a major intermediate factor that can promote the progression of UC. In the present study, we investigated the effect and the underlying mechanisms of 4-Octyl itaconate (OI) on dextran sulfate sodium (DSS)-induced UC in mice. Our work identified that OI alleviated the colitis by reducing the oxidative stress and the apoptosis in colon tissue, then increasing the tight junction proteins expression and in turn enhancing the intestinal barrier function, thereby creating less severe inflammatory responses. Moreover, our results demonstrated that OI reduced the Kelch-like ECH-associated protein 1 (KEAP1) expression and subsequent upregulated nuclear factor E2-related factor (NRF2) expression and its nuclear translocation which in turn induced the expression of glutathione S-transferase (GST) and NAD(P)H: quinone oxidoreductase 1 (NQO1). In addition, ML385, a NRF2 antagonist, can inhibit the protective effects of OI on UC, indicating that the role of OI in this colitis model could be dependent on the activation of KEAP1-NRF2 pathway. Notably, OI co-administration significantly enhanced the therapeutic effects of mesalazine or 1400W on UC. Collectively, itaconate may have a great potential for use in the treatment of IBD.

Role of glutathione S-transferases in the mode of action of herbicides that inhibit amino acid synthesis in Amaranthus palmeri

Acetolactate synthase inhibitors (ALS inhibitors) and glyphosate are two classes of herbicides that act by inhibiting an enzyme in the biosynthetic pathway of branched-chain or aromatic amino acids, respectively. Besides amino acid synthesis inhibition, both herbicides trigger similar physiological effects in plants. The main aim of this study was to evaluate the role of glutathione metabolism, with special emphasis on glutathione S-transferases (GSTs), in the mode of action of glyphosate and ALS inhibitors in Amaranthus palmeri. For that purpose, plants belonging to a glyphosate-sensitive (GLS) and a glyphosate-resistant (GLR) population were treated with different doses of glyphosate, and plants belonging to an ALS-inhibitor sensitive (AIS) and an ALS-inhibitor resistant (AIR) population were treated with different doses of the ALS inhibitor nicosulfuron. Glutathione-related contents, GST activity, and related gene expressions (glutamate-cysteine ligase, glutathione reductase, Phi GST and Tau GST) were analysed in leaves. According to the results of the analytical determinations, there were virtually no basal differences between GLS and GLR plants or between AIS and AIR plants. Glutathione synthesis and turnover did not follow a clear pattern in response to herbicides, but GST activity and gene expression (especially Phi GSTs) increased with both herbicides in treated sensitive plants, possibly related to the rocketing H2O2 accumulation. As GSTs offered the clearest results, these were further investigated with a multiple resistant (MR) population, compressing target-site resistance to both glyphosate and the ALS inhibitor pyrithiobac. As in single-resistant plants, measured parameters in the MR population were unaffected by herbicides, meaning that the increase in GST activity and expression occurs due to herbicide interactions with the target enzymes.

Glutathione S-transferase activity facilitates rice tolerance to the barnyard grass root exudate DIMBOA

BackgroundIn paddy fields, the noxious weed barnyard grass secretes 2,4-dihydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one (DIMBOA) to interfere with rice growth. Rice is unable to synthesize DIMBOA. Rice cultivars with high or low levels of allelopathy may respond differently to DIMBOA.ResultsIn this study, we found that low concentrations of DIMBOA (≤ 0.06 mM) promoted seedling growth in allelopathic rice PI312777, while DIMBOA (≤ 0.08 mM) had no significant influence on the nonallelopathic rice Lemont. DIMBOA treatment caused changes in the expression of a large number of glutathione S-transferase (GST) proteins, which resulting in enrichment of the glutathione metabolic pathway. This pathway facilitates plant detoxification of heterologous substances. The basal levels of GST activity in Lemont were significantly higher than those in PI312777, while GST activity in PI312777 was slightly induced by increasing DIMBOA concentrations. Overexpression of GST genes (Os09g0367700 and Os01g0949800) in these two cultivars enhanced rice resistance to DIMBOA.ConclusionsTaken together, our results indicated that different rice accessions with different levels of allelopathy have variable tolerance to DIMBOA. Lemont had higher GST activity, which helped it tolerate DIMBOA, while PI312777 had lower GST activity that was more inducible. The enhancement of GST expression facilitates rice tolerance to DIMBOA toxins from barnyard grass root exudates.

Investigation of the effect of isoxazole derivatives on glutathione-dependent enzymes associated with cancer

Globally, cancer is currently the second-most common cause of death. As such, it is a crucial problem that affects society's well-being all around the world. Chemotherapy is the most commonly used treatment for cancer. It is a technique that uses drugs to kill cancerous cells or the cancerous organism itself. Chemotherapy has been used to treat metastatic cancer for the past 50 years, with varying degrees of effectiveness. One of the main problems with many chemotherapeutic drugs is that they might damage healthy cells and organs. These drugs frequently have a low therapeutic index, a restricted ability to selectively target cancerous cells, and a high propensity to develop drug resistance after extended use. Furthermore, a rise in the expression of glutathione reductase (GR; EC 1.8.1.7) and glutathione S-transferase (GST; EC 2.5.1.18) can be seen with time. Extensive study on proteins and their functions in the development of cancer cells has been conducted in recent years. Glutathione-S-transferases (GSTs) are a superfamily of enzymes that play a critical role in the detoxification of cells by protecting them against reactive electrophiles such as chemotherapeutic drugs and reactive oxygen species. Glutathione reductase (GR), on the other hand, is an essential antioxidant enzyme that supports cellular protection against oxidative stress. In addition to its antioxidant role, GR has become a target for the synthesis of antimalarial and anticancer medications. In the current investigation, affinity chromatography was used to purificate the GST and GR enzymes from human erythrocytes. Through the calculation of IC50 and KI values for these derivatives, the effects of isoxazole derivatives on these purified enzymes were examined, with an emphasis on identifying the forms of inhibition.

Rapid discovery of a novel “green” and natural GST inhibitor for sensitizing hepatocellular carcinoma to Cisplatin by visual screening strategy

Over-expression of glutathione S-transferase (GST) can promote Cisplatin resistance in hepatocellular carcinoma (HCC) treatment. Hence, inhibiting GST is an attractive strategy to improve Cisplatin sensitivity in HCC therapy. Although several synthesized GST inhibitors have been developed, the side effects and narrow spectrum for anticancer seriously limit their clinical application. Considering the abundance of natural compounds with anticancer activity, this study developed a rapid fluorescence technique to screen “green” natural GST inhibitors with high specificity. The fluorescence assay demonstrated that schisanlactone B (hereafter abbreviated as C1) isolated from Xue tong significantly down-regulated GST levels in Cisplatin-resistant HCC cells in vitro and in vivo. Importantly, C1 can selectively kill HCC cells from normal liver cells, effectively improving the therapeutic effect of Cisplatin on HCC mice by down-regulating GST expression. Considering the high GST levels in HCC patients, this compound demonstrated the high potential for sensitizing HCC therapy in clinical practice by down-regulating GST levels.

Phytochelatin-Mediated Cultivar-Dependent Cd Accumulations of Lactuca sativa and Implication for Cd Pollution-Safe Cultivars Screening.

Cd pollution-safe cultivar (Cd-PSC) is a feasible strategy to minimize Cd contamination in leafy vegetables. The shoot Cd concentrations of 23 Lactuca sativa cultivars under Cd stress ranged from 0.124 to 2.155 mg·kg-1 with a maximum cultivar difference of 8 folds. Typical Cd-PSC C16 (L) and high-Cd-accumulating cultivar C13 (H) were screened to investigate the mechanisms of Cd accumulations in L. sativa through determining Cd concentrations, Cd subcellular distributions, phytochelatin profiles, and phytochelatin biosynthesis-related genes' expressions. Higher Cd distribution in a heat stable fraction in C13 (H) indicated that the high Cd accumulation trait of C13 (H) mainly depended on the Cd-phytochelatin complexes. Root phytochelatin concentrations were significantly elevated in C13 (H) (5.83 folds) than in C16 (L) (2.69 folds) (p < 0.05) under Cd stress. Significantly downregulated expressions of glutathione S-transferase rather than the regulation of phytochelatin synthesis genes in the root of C13 (H) might be responsible for sufficient glutathione supply for phytochelatins synthesis. These findings suggested that phytochelatin elevation in C13 (H) would favor the Cd root to shoot transportation, which provides new insights into the phytochelatin-related cultivar-dependent Cd accumulating characteristic in L. sativa.

Female-specific pancreatic cancer survival from CT imaging of visceral fat implicates glutathione metabolism in solid tumors

To identify if body composition, assessed with preoperative CT-based visceral fat ratio quantification as well as tumor metabolic gene expression, predicts sex-dependent overall survival (OS) in patients with pancreatic ductal adenocarcinoma (PDAC). This was a retrospective analysis of preoperative CT in 98 male and 107 female patients with PDAC. Relative visceral fat (rVFA; visceral fat normalized to total fat) was measured automatically using software and corrected manually. Median and optimized rVFA thresholds were determined according to published methods. Kaplan Meier and log-rank tests were used to estimate OS. Multivariate models were developed to identify interactions between sex, rVFA, and OS. Unsupervised gene expression analysis of PDAC tumors from The Cancer Genome Atlas (TCGA) was performed to identify metabolic pathways with similar survival patterns to rVFA. Optimized preoperative rVFA threshold of 38.9% predicted significantly different OS in females with a median OS of 15 months (above threshold) vs 24 months (below threshold; p=0.004). No significant threshold was identified in males. This female-specific significance was independent of age, stage, and presence of chronic pancreatitis (p=0.02). Tumor gene expression analysis identified female-specific stratification from a five-gene signature of glutathione S-transferases. This was observed for PDAC as well as clear cell renal carcinoma and glioblastoma. CT-based assessments of visceral fat can predict pancreatic cancer OS in females. Glutathione S-transferase expression in tumors predicts female-specific OS in a similar fashion.

Busulfan induces steatosis in HepaRG cells but not in primary human hepatocytes: Possible explanations and implication for the prediction of drug-induced liver injury.

The antineoplastic drug busulfan can induce different hepatic lesions including cholestasis and sinusoidal obstruction syndrome. However, hepatic steatosis has never been reported in patients. This study aimed to determine whether busulfan could induce steatosis in primary human hepatocytes (PHH) and differentiated HepaRG cells. Neutral lipids were determined in PHH and HepaRG cells. Mechanistic investigations were performed in HepaRG cells by measuring metabolic fluxes linked to lipid homeostasis, reduced glutathione (GSH) levels, and expression of genes involved in lipid metabolism and endoplasmic reticulum (ER) stress. Analysis of two previous transcriptomic datasets was carried out. Busulfan induced lipid accumulation in HepaRG cells but not in six different batches of PHH. In HepaRG cells, busulfan impaired VLDL secretion, increased fatty acid uptake, and induced ER stress. Transcriptomic data analysis and decreased GSH levels suggested that busulfan-induced steatosis might be linked to the high expression of glutathione S-transferase (GST) isoenzyme A1, which is responsible for the formation of the hepatotoxic sulfonium cation conjugate. In keeping with this, the GST inhibitor ethacrynic acid and the chemical chaperone tauroursodeoxycholic acid alleviated busulfan-induced lipid accumulation in HepaRG cells supporting the role of the sulfonium cation conjugate and ER stress in steatosis. While the HepaRG cell line is an invaluable tool for pharmacotoxicological studies, it might not be always an appropriate model to predict and mechanistically investigate drug-induced liver injury. Hence, we recommend carrying out toxicological investigations in both HepaRG cells and PHH to avoid drawing wrong conclusions on the potential hepatotoxicity of drugs and other xenobiotics.

Validation of biomarkers for neonicotinoid exposure in Folsomia candida under mutual exposure to diethyl maleate.

Neonicotinoid insecticides are harmful to non-target soil invertebrates, which are crucial for sustainable agriculture. Gene expression biomarkers could provide economic and high-throughput metrics of neonicotinoid exposure and toxicity to non-target invertebrates. Thereby, biomarkers can help guide remediation efforts or policy enforcement. Gene expression of Glutathione S-Transferase 3 (GST3) has previously been proposed as a biomarker for the neonicotinoid imidacloprid in the soil ecotoxicological model species Folsomia candida (Collembola). However, it remains unclear how reliably gene expression of neonicotinoid biomarkers, such as GST3, can indicate the exposure to the broader neonicotinoid family under putative GST enzymatic inhibition. In this work, we exposed springtails to two neonicotinoids, thiacloprid and imidacloprid, alongside diethyl maleate (DEM), a known GST metabolic inhibitor that imposes oxidative stress. First, we determined the influence of DEM on neonicotinoid toxicity to springtail fecundity. Second, we surveyed the gene expression of four biomarkers, including GST3, under mutual exposure to neonicotinoids and DEM. We observed no effect of DEM on springtail fecundity. Moreover, the expression of GST3 was only influenced by DEM under mutual exposure with thiacloprid but not with imidacloprid. The results indicate that GST3 is not a robust indicator of neonicotinoid exposure and that probable GST enzymatic inhibition mediates the toxicity of imidacloprid and thiacloprid differentially. Future research should investigate biomarker reliability under shifting metabolic conditions such as provided by DEM exposure.

Expression comparisons of defense-related genes in resistant and susceptible chickpea cultivars in response to Ascochyta rabiei

Ascochyta blight caused by Ascochyta rabiei is a destructive disease affecting chickpea crops worldwide. Numerous studies have sought to identify resistance genes involved in the defense response against Ascochyta blight, but the function of potential resistant genes across different chickpea genetic backgrounds remains poorly understood. This study evaluated the potential role of 16 genes involved in defense responses in partially resistant (ILC482) and susceptible (Sarı98) chickpea cultivars. The leaf samples were collected from 2-week-old seedlings at 6, 12, 24, 48, and 72 h post-inoculation (hpi) with A. rabiei and analyzed for differential expression of defense genes using qRT-PCR. Twelve out of 16 genes were differentially regulated between chickpea cultivars. The qRT-PCR analysis indicated that the expression of defense genes was significantly higher in the partially resistant cultivar than that in the susceptible cultivar. The time point of the highest upregulation ratio for defense genes was variable between the cultivars. The expression of glutathione S-transferase, serine/threonine-protein kinase, WRKY gene (CaWRKY16), and wall associated-receptor kinase genes reached maximum levels in the resistant cultivar at 6 hpi, while antimicrobial peptide precursor (SNAKIN2), polymorphic antigen membrane protein, ethylene receptor-like sequences (CaETR1), and two nucleotide-binding site-leucine-rich repeat genes showed the highest activity at 24 hpi. In particular, CaWRKY16, CaWRKY50, glutathione S-transferase, CaETR1, and wall-associated-receptor kinase genes provided valuable information for assessing chickpea cultivars with different resistance levels. The results indicated that the expression changes of these genes involved in defense pathways play a crucial role in enhanced resistance of chickpea plants against Ascochyta blight and, therefore could be potential candidate genes for future breeding studies.

Effect of hexavalent chromium exposure on the reproductive status and biomarker responses of female Geloina erosa.

Hexavalent chromium (Cr (VI)) is widely distributed in the marine environment of Hainan Province, China and poses a potential threat to its mangrove ecosystems. However, the mechanisms underlying Cr-induced stress and reproductive toxicity in clams remain largely unknown. In this study, the clams, Geloina erosa, were exposed to 4.34, 8.69, 17.38 and 34.76 mg/L Cr (VI) for 24, 48 and 72 h. The gonad-somatic index (GSI) was determined and histological alterations of the ovaries were quantified by light microscopy. The micronucleus test was performed which quantifies the genotoxic presence of small cytoplasmic bodies in eukaryotic cells. Enzymatic assays for catalase (CAT), glutathione reductase (GR), and malondialdehyde (MDA) activities were done. Quantitative real-time PCR (qRT-PCR) was used to quantify the expression of glutathione-S-transferase (GST), heat shock protein 70 (HSP70) and vitellogenin (Vtg) in ovaries of G. erosa. The results showed that the micronucleus frequency was significantly increased when clams were exposed to Cr (VI). Cr (VI) exposure induced the accumulation of MDA and affected CAT and GR enzyme activities. The high Cr (VI) concentration of 34.76 mg/L significantly increased the levels of GR activity, GST expression and HSP70 expression and inhibited Vtg expression and CAT activity. MDA content was significantly increased after 72 h at the high Cr (VI) exposure (34.76 mg/L). Therefore, Cr (VI) exposure may be toxic to the development of ovaries of G. erosa.

Ecological effects and molecular mechanisms of single and coexisting PFOS and Cu exposure on submerged macrophytes and periphytic biofilms in aquatic environments

Perfluorooctane sulfonate (PFOS) is an accumulative emerging persistent organic pollutant that coexists with copper (Cu) in aquatic environments and is a severe threat to human health. PFOS has genotoxicity, male reproductive toxicity, neurotoxicity, developmental toxicity and endocrine disrupting effects, and is considered to be a kind of environmental pollutant with systemic multi-organ toxicity. However, the combined effects of PFOS and Cu on aquatic organisms have not been analyzed previously. In this study, we investigate the response of Vallisneria Natans (V. natans) and biofilms to single and combined treatments of PFOS and Cu at environmental concentrations (1.0, 10.0, and 100.0 μg·L−1). Results indicate that single and combined treatment triggered the reactive oxygen species (ROS) metabolism with increased activities of GSH, MDA, CAT, TSOD and an antagonistic joint toxicity. Meanwhile, transcriptomic analyses showed that the peroxidase (PER), 9 glutaredoxin (GRX), and 39 glutathione S-transferase (GST) expression of genes related to ROS detoxification was up-regulated. In addition, single and combined exposure induced the up-regulation of the transport channel genes which encode aquaporins, rapid-type anion channels, and copper-exporting P-type ATPase transporters, promoting the uptake of PFOS and Cu in V. natans, Enhancing detoxification and mitigating the effects of Cu and PFOS exposure. Moreover, PFOS and Cu changed the structural characteristics of extracellular polysaccharides in biofilms, synthesizing large amounts of β-D-glucopyranose PS, α-D-glucopyranose PS. And the community diversity and species evenness changed, with Rhizobiaceae, Chloroplast, and Weeksellaceae being the dominant phylum. This study provides insights into the molecular mechanism of joint toxicity of PFOS and Cu by Transcriptomics and PCR analysis on the submerged macrophyte and their periphyton biofilms.

Nomilin and its analogue obacunone alleviate NASH and hepatic fibrosis in mice via enhancing antioxidant and anti-inflammation capacity.

Nonalcoholic steatohepatitis (NASH) and hepatic fibrosis are leading causes of cirrhosis with rising morbidity and mortality worldwide. Currently, there is no appropriate treatment for NASH and hepatic fibrosis. Many studies have shown that oxidative stress is a main factor inducing NASH. Nomilin (NML) and obacunone (OBA) are limonoid compounds naturally occurring in citrus fruits with various biological properties. However, whether OBA and NML have beneficial effects on NASH remains unclear. Here, we demonstrated that OBA and NML inhibited hepatic tissue necrosis, inflammatory infiltration and liver fibrosis progression in methionine and choline-deficient (MCD) diet, carbon tetrachloride (CCl4 )-treated and bile duct ligation (BDL) NASH and hepatic fibrosis mouse models. Mechanistic studies showed that NML and OBA enhanced anti-oxidative effects, including reduction of malondialdehyde (MDA) level, increase of catalase (CAT) activity and the gene expression of glutathione S-transferases (GSTs) and Nrf2-keap1 signaling. Additional, NML and OBA inhibited the expression of inflammatory gene interleukin 6 (Il-6), and regulated the bile acid metabolism genes Cyp3a11, Cyp7a1, multidrug resistance-associated protein 3 (Mrp3). Overall, these findings indicate that NML and OBA may alleviate NASH and liver fibrosis in mice via enhancing antioxidant and anti-inflammation capacity. Our study proposed that NML and OBA may be potential strategies for NASH treatment.

The mechanisms of metabolic resistance to pyrethroids and neonicotinoids fade away without selection pressure in the tarnished plant bug Lygus lineolaris.

Heavy selection pressure prompted the development of resistance in a serious cotton pest tarnished plant bug (TPB), Lygus Lineolaris in the mid-southern USA. Conversely, a laboratory resistant TPB strain lost its resistance to five pyrethroids and two neonicotinoids after 36 generations without exposure to any insecticide. It is worthwhile to examine why the resistance diminished in this population and determine whether the resistance fade away has practical value for insecticide resistance management in TPB populations. A field-collected resistant TPB population in July (Field-R1) exhibited 3.90-14.37-fold resistance to five pyrethroids and two neonicotinoids, while another field-collected TPB population in April (Field-R2) showed much lower levels of resistance (0.84-3.78-fold) due to the absence of selection pressure. Interestingly, after 36 generations without exposure to insecticide, the resistance levels in the same population (lab-R) significantly decreased to 0.80-2.09-fold. The use of detoxification enzyme inhibitors had synergistic effects on permethrin, bifenthrin and imidacloprid in resistant populations of L. lineolaris. The synergism was more pronounced in field-R2 than lab-S and lab-R TPB population. Moreover, esterase, GST, and P450 enzyme activities increased significantly by approximately 1.92-, 1.43-, and 1.44- fold in field-R1, respectively, and 1.38-fold increased P450 enzyme activities in field-R2 TPB population, compared to the lab susceptible (lab-S) TPB. In contrast, the three enzyme activities in the lab-R strain were not significantly elevated anymore relative to the lab-S population. Additionally, field-R1 TPB showed elevated expression levels of certain esterase, GST and P450 genes, respectively, while field-R2 TPB overexpressed only P450 genes. The elevation of these gene expression levels in lab-R expectedly diminished to levels close to those of the lab susceptible (lab-S) TPB populations. Our results indicated that the major mechanism of resistance in TPB populations was metabolic detoxification, and the resistance development was likely conferred by increased gene expressions of esterase, GST, and P450 genes, the fadeaway of the resistance may be caused by reversing the overexpression of esterase, GST and P450. Without pesticide selection, resistant gene (esterase, GST, P450s) frequencies declined, and detoxification enzyme activities returned to lab-S level, which resulted in the recovery of the susceptibility in the resistant TPB populations. Therefore, pest's self-purging of insecticide resistance becomes strategically desirable for managing resistance in pest populations. This article is protected by copyright. All rights reserved.

Abstract 73: Specific Activity of GSTs and Protein Expression of GSTT1, GSTP1 and CYP1A1 in Head and Neck Cancer

Abstract Purpose: Rising incidence of head and neck cancer have been observed in developing countries over last decade. It has been postulated that carcinogen detoxifying genes may be involved directly or indirectly in pathogenesis of head and neck cancer. Three genes (CYP1A1, GSTM1 and GSTP1) were studied for their association with head and neck cancer. Cytochrome P450 1A1 (CYP1A1) is phase I enzyme while glutathione S transferases (GSTs) constitute the phase II detoxification enzymes. Abnormal protein expression of CYP1A1 and GSTs lead to cancer initiation. Methods: The current case control study was designed to evaluate the possible associations of these enzymes with head and neck cancer. Serum GSTs activity was determined by using ELISA and CYP1A1, GSTT1, GSTP1 expression in tissue by immunohistochemistry with specific antibodies. Results: Mean serum GSTs specific activity was 7.7 (+11.5)U/L and control was 11.4 (+7.5)U/L. Significant decrease (P&amp;lt;0.05) in GSTs specific activity was observed in head and neck cancer patients compared to controls (P&amp;lt;0.001). Moreover, CYP1A1 and GSTT1 was down expressed whereas GSTP1 was over expressed in head and neck cancer tissues compared with adjacent normal control tissue. Conclusion: Conclusion: Decreased activity of GSTs, reduced expression of GSTs, CYP1A1, GSTT1 and increased expression of GSTP1 may contribute to head and neck cancer development in Pakistani population. Citation Format: Nosheen Masood, Azra Yasmin. Specific Activity of GSTs and Protein Expression of GSTT1, GSTP1 and CYP1A1 in Head and Neck Cancer [abstract]. In: Proceedings of the 11th Annual Symposium on Global Cancer Research; Closing the Research-to-Implementation Gap; 2023 Apr 4-6. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2023;32(6_Suppl):Abstract nr 73.