Glutathione S-transferase Enzyme Activity Research Articles

First report of the effect of lambda-cyhalothrin on the activity of acetylcholinesterase, glutathione S-transferase, and cytochrome C-oxidase enzymes in larvae of Culex quinquefasciatus (Diptera: Culicidae) in East Jakarta

First report of the effect of lambda-cyhalothrin on the activity of acetylcholinesterase, glutathione S-transferase, and cytochrome C-oxidase enzymes in larvae of Culex quinquefasciatus (Diptera: Culicidae) in East Jakarta

Expression and biochemical significance of Piwil2 in stem cell lines

Abstract Introduction P-element induced wimpy testis-like 2 (Piwil2) is in the Piwi gene family. Piwil2 has important roles in the self-renewal mechanism of stem cell induction and progression of numerous types of human malignancies such as lung, breast, colon, prostate, and cervical cancers. Glutathione S-transferase (GST) acts as detoxification in cancer metabolism. This study aimed to investigate the effects of the stem cell protein Piwil2 on MCF10A and MCF-7 at the GST activity levels. Materials/Methods MCF-7/Piwil2 and MCF10A/Piwil2, transfected with a plasmid carrying the Piwil2 gene, and non-transfected MCF-7 and MCF10A were cultured in a complete DMEM/F12 medium. GST A1 and P1 activity was determined in these cell lines using as substrates CDNB, EA respectively. Results According to experimental results, GST P1 activity decreased in the MCF-7/Piwil2 cells as compared with the non-transfected MCF-7 cells, however, MCF-7/Piwil2 cells demonstrated increases in GST A1 (total GST) activity. The statistically significant differences were found for the comparison of non-transfected MCF-7 and MCF-7/Piwil2 (p<0,0001), for GST enzyme activities by using CDNB and EA as substrates. These results were the same for the MCF10A cell line. Discussion It is shown for the first time that transfection studies may affect GST activity at the cellular mechanism level. The study contributes to determining the effect of transfection on GST isoenzymes and also how the Piwil2 gene may affect GST activity in the stem cell line.

BIOCHEMICAL INVESTIGATIONSOF THE EFFECT OF XANAX ON THE CEREBELLAR TISSUES OF MALE MICE

Xanax is an agent with hypnogenic, anxiolytic, anticonvulsant, and muscle relaxant properties and has generally been used as a hypnotic/tranquilizer. The aim of this paper was to investigate the effect of Xanax on acetylcholinesterase and glutathionestransferase enzyme activities on the cerebellar tissues of male mice. Sixty male mice were randomly assigned into four groups (15 mice/each) according to their approximately equal mean body weight. Mice that received orally by gavage 0.5 ml saline solution of 0.9% NaCl were considered as a control mice. Other experimental mice were daily administered orally by gavage with 0.5 ml of different three doses of Xanax (0.5, 1 and 1.5 mg/kg bw), for two months. Biochemical analyses revealed significant decreases in the activities of acetylcholinesterase and glutathionestransferase enzyme in the brain tissues of mice administered with the three doses of Xanax. The major mechanism involved appears to be the result of oxidative stress scavenging action on the neuronal cells of cerebellar cortex of male mice.

The role of Glutathione-S-transferases in phoxim and chlorfenapyr tolerance in a major mulberry pest, Glyphodes pyloalis walker (Lepidoptera: Pyralidae)

Glyphodes pyloalis Walker is a destructive pest on mulberry trees and poses a significant threat to the sericultural industry in China. Phoxim and chlorfenapyr are two commonly used insecticides in mulberry fields. Glutathione-S-transferases (GSTs) comprise a multifunctional protein superfamily that plays important roles in the detoxification of insecticides and xenobiotic compounds in insects. However, whether GSTs participate in the tolerance of phoxim and chlorfenapyr in G. pyloalis is still unknown. To better understand the mechanism of insecticide tolerance in G. pyloalis, the enzymatic activity of GSTs was evaluated under phoxim and chlorfenapyr exposure, respectively. GST enzyme activity was significantly increased after 12, 36 and 48 h of phoxim treatment and 12, 24, 36 and 48 h of chlorfenapyr treatment. Subsequently, eighteen GST genes were identified from the larvae transcriptome of G. pyloalis. Among these, ten GpGSTs had GSH-binding sites and fifteen GpGSTs had variable hydrophobic substrate-binding sites. The expression levels of Delta-GpGST and Epsilon-GpGST genes were significantly influenced by phoxim and chlorfenapyr treatment, and by the time post insecticide application. Furthermore, after silencing GpGST-E4, the mortality rate of G. pyloalis larvae was increased when they were exposed to chlorfenapyr, but it did not significantly alter when the larvae were exposed to phoxim. Our results indicated the vital roles of GpGSTs in the tolerance of insecticides and this action depends on the categories of insecticides. The present study provides a theoretical basis for elucidating insecticide susceptibility and promotes functional research on GST genes in G. pyloalis.

Influence of salinity on the toxicity of copper and cadmium to Zebrafish embryos.

Salinization has become a serious worldwide environmental perturbation in freshwater ecosystems. Concomitantly, many of such ecosystems are already impacted by other toxicants, which together with increased salinity may result in synergistic, antagonistic or additive toxic effects to biota. This work intended to assess the influence of increasing salinity (by using NaCl) on the lethal and sublethal toxicity of two metallic elements (copper and cadmium) in embryos of the fish species Danio rerio. This goal was achieved by exposing zebrafish embryos to seven concentrations of NaCl, individually or combined with each metal, using a full factorial design. The following endpoints were monitored in the test organisms: mortality, hatching, malformations and the enzymatic activity of glutathione S-transferase (GST) and cholinesterase (ChE). Overall, moderate salinity levels alleviated the lethal toxicity of both copper and cadmium although this effect was stronger in the copper assay. This effect was also influenced, as expected, by the concentrations of the metals indicating that the protective effect of salt only reaches some levels, after what is overwhelmed by the high metal toxicity, especially with the non-essential metal cadmium. At sub-lethal concentrations, the interactive effect resulting from NaCl and metals was not consistent and varied with the endpoint analyzed and the metal tested. Overall, the interactions between the salt and metals seem complex and with more drastic effects (positive or negative) on lethal endpoints than sub-lethal.

Biological Effects Assessmentof Antibiofouling EDCs: Gaeta Harbor (South Italy) Benthic Communities' Analysis by Biodiversity Indices and Quantitative gpx4 Expression.

The most representative organisms of the Harbor of Gaeta Gulf in South Italy were analyzed for biofouling by visual census and confirmed later by molecular approach on an artificial Conatex panel dipped 3 m into a eutrophic area during the Covid-19 pandemic. Mitochondrial Cytochrome C oxidase subunit 1(COI) gene was sequenced from 20 different marine species (flora: 2 families, 2 orders; fauna: 16 families, 11 orders) to test whether the morphology-based assignment of the most common biofouling member was supported by DNA-based species identification. Twelve months of submersion resulted in generation of sufficient data to obtain a facies climax represented mainly by the bivalve mollusk, Mytilus galloprovincialis. Specific diversity and variations of the biofouling biomasses were analyzed using two different anti-biofouling paints: an endocrine disrupting chemical (EDC)-containing metal biocide, and a biocide-free paint. Also, their effects on detoxification and reproductive health of M. galloprovincialis were evaluated using glutathione S-transferase enzymatic activity and RTqPCR expression of the fertility antioxidant gene glutathione peroxidase 4 (gpx4). The obtained data provide useful indications on which future investigations may be focused and may become a potential management tool for a harbor biofouling database to assist local administrations in EDCs protection of autochthonous benthic communities and their fertility using innovative antifouling paints.

Heavy metal toxicity in Buriganga river alters the immunology of Nile tilapia (Oreochromis niloticus L)

The objective of the current study was to evaluate the biochemical and immunological responses of tilapia, Oreochromis niloticus due to heavy metals pollution. Histomorphological alterations in the liver and kidney suggested tissue damages due to this polluted water exposure. The brain acetylcholinesterase (AChE) as an indicator of neurotoxicity was significantly (P < 0.01) decreased after 10 days exposure of fish to heavy metal contained river water, while plasma glutamate oxalacetate transaminase and plasma glutamate pyruvate transaminase were significantly increased (P < 0.01). Moreover, superoxide dismutase, catalase, glutathione peroxidase, glutathione S-transferase enzyme activities, as well as reduced glutathione and malondialdehyde levels were significantly increased in heavy metals contained river water treated fish compared to the control. Additionally, glucose level and blood serum Ca2+ concentrations were significantly (P < 0.01) decreased in fish exposed to heavy metal contained river water compared to the control. Hematological indices such as Hemoglobin, RBC, WBC, MCV etc. of polluted river water treated fish were significantly (P < 0.01) different in comparison to that of control fish. The cytokines i.e. IL-1β, IL-6, and TNF-α level were significantly (P < 0.01) increased in the fish exposed to heavy metals contained river water in comparison to that of control fish. The present findings explored the detrimental effects of heavy metal contained river water on fish at biochemical and immunological levels.

Sublethal Effects of Abamectin on the Development, Reproduction, Detoxification Enzyme Activity, and Related Gene Expression of the Oriental Fruit Moth (Lepidoptera: Tortricidae).

Grapholita molesta is one of the most important fruit pests worldwide. Abamectin is a biological pesticide frequently used to control fruit borers like G. molesta in part owing to its translaminar properties. In this study, we characterized the toxicity of abamectin to G. molesta larvae using the diet incorporation method. The sublethal effects of abamectin on the development, reproduction, detoxification enzyme activity, and related gene expression of G. molesta were assessed. The results showed that the LC20 and LC50 values of the insecticide against G. molesta 72 h post-treatment were 1.17 mg L-1 and 5.85 mg L-1, whereas the LC20 and LC50 values 96 h post-treatment were 0.34 mg L-1 and 3.63 mg L-1. When compared to the control, sublethal concentrations of abamectin 1) significantly increased the mortality of the larvae, prepupae, and pupae of G. molesta, 2) prolonged the duration of 3rd to 5th instar larva, prepupal and pupal periods, 3) shortened the longevity of adults, and 4) reduced female fecundity. The enzymatic activity of glutathione S-transferase (GST) varied significantly after exposure to sublethal concentrations of abamectin, but the cytochrome P450 monooxygenases and carboxylesterase activity were not significantly affected. Thirteen of the 25 GST genes were significantly upregulated under different sublethal concentrations of abamectin. The combined findings increase our understanding of the effects of abamectin on G. molesta and the potential role of GSTs in the metabolic interactions of abamectin in this pest, and have applications for more rational and effective use of abamectin to control G. molesta.

Role of Glutathione-S-Transferase M1 (GSTM1) and T1 (GSTT1) Genes on Aluminum Concentration and Oxidative Markers among Autistic Children

Autism Spectrum Disorder (ASD) is considered a multifaceted neurodevelopmental disorder. The last two decades showed an increase in its prevalence until reached about 1 in 54 children. Autistic symptoms may be exacerbated when the interaction of the genetic and the environmental risk factors occur, suggesting that gene-environment interaction could be a mechanism underlying the etiology of ASD. Aluminum is a known neurotoxic metal that has known health effects in humans. Glutathione-S-transferase (GST) genes and their enzymes play a major role in the detoxification of many toxic metals.Data were collected from 76 children aged 2-8 years diagnosed with ASD and 30 sex and age matched healthy children. The aim of this study was to investigate the association of polymorphisms in the two GST genes (GSTM1 and GSTT1) with mean aluminum concentrations (as, gene-environment interaction) and oxidative status markers (GST enzyme, malondialdehyde and nitric oxide) among the studied groups. The study started at December 2019 and last for one year at the clinics of National Research Centre, Egypt.The results of this study showed that the null GSTM1 and GSTT1 genotype is the most common type in ASD and that genotype may predispose ASD children to decreased antioxidant status (GST enzyme activity) which in term lead to mal detoxification of aluminum. There is marked increase in aluminum concentrations in hair of ASD children and oxidative markers (increase in MDA and NO) leading to oxidative damage that may play an important role in children autistic status. The study recommends adding antioxidant supplements to daily diet of ASD children to improve their antioxidant status and in term improving management of patients with autism spectrum disorders. Further studies are needed to describe other GST gene polymorphisms.

Investigation of oxidative stress status in cumulus cells in patıents with in vitro fertilization

The negative impact of oxidative stress on oocytes obtained from in vitro fertilization (IVF) patients is a challenge for the optimization of live birth rates. In this study, it is aimed to investigate whether oxidant/antioxidant parameters have a predictive value in terms of determining the count and quality of oocytes. Catalase (CAT), glutathione-S-transferase (GST), arylesterase (ARE) enzyme activities, and malondialdehyde (MDA) levels were analysed in cumulus cells of poor responder (n = 28, oocyte count ≤ 4), normo responder (n = 48, 5 ≤ oocyte count ≤ 14), and high responder (n = 26, oocyte count ≥ 15) patient groups continuing IVF treatment. The cumulus cell GST enzyme activity were statistically significantly increased in the high responders group compared to the poor responder and the normo responder’s groups (p < 0.001 and p = 0.002, respectively). The cumulus cell MDA levels were significantly decreased in the high responder group compared to the poor responder group (p = 0.008). The cumulus cell CAT (p = 0.175) and ARE (p = 0.124) enzyme activities were examined but no statistically significant difference found between the groups. The significant increase in GST enzyme activity and significant decrease in MDA levels in the high responder group indicate that oxidative stress has an effect oocyte status and quality.

Differential regulation of antioxidant enzymes in Frankliniella occidentalis (Thysanoptera: Thripidae) exposed to thermal stress

Frankliniella occidentalis is an invasive insect pest that incites damage to ornamental and agronomic crops on a global scale. In this study, the effects of temperature on gene expression and enzyme activity were studied for superoxide dismutase (SOD), peroxidase (POD), and glutathione-S-transferase (GST) in F. occidentalis. SOD, POD and GST enzyme activity increased significantly at 35–37 °C but declined as the temperature increased to 41 °C. In a time course study at 35 °C, SOD, POD and GST activities were significantly elevated at 0.5, 1 and 2 h in comparison to the control at 26 °C. Expression patterns were evaluated for the three antioxidant genes under high and low temperature stress. In a time course study at –4 °C, SOD, POD and GST expression peaked at 1 h and declined at 2 h of exposure. In contrast, when transcription was monitored at 35 °C, expression was lowest at 1 h and increased at 2 h. The results provide data that will be useful in deciphering the role of antioxidant enzymes in the adaptation of F. occidentalis to climate change.

DCOIT unbalances the antioxidant defense system in juvenile and adults of the marine bivalve Amarilladesma mactroides (Mollusca: Bivalvia)

DCOIT is a co-biocide that is part of the formulation of the commercial antifouling Sea-Nine 211® and although it is “safe to use”, negative effects have been reported on the antioxidant defense system of non-target organisms. Therefore, the objective of this research was to verify and compare the response of antioxidant enzymes of juveniles and adults of Amarilladesma mactroides exposed to DCOIT. The animals were exposed to solvent control (DMSO 0.01%) and DCOIT (measured concentration 0.01 mg/L and 0.13 mg/L) for 96 h, then gills, digestive gland and mantle were collected for analysis of the enzymatic activity of glutathione S-transferase (GST), superoxide dismutase (SOD) and catalase (CAT). The results revealed that adults, in relation to juveniles, have low basal activity of GST and SOD enzymes in the gills and digestive gland and high basal activity of SOD and CAT in the mantle. DCOIT did not alter GST activity in the gills of any life stage, while both concentrations decreased SOD and CAT in adults. In the digestive gland, it was observed that DCOIT (0.13 mg/L) decreased the GST activity in adults and CAT in juveniles, and both concentrations of the co-biocide decreased the SOD and CAT in adults. In the mantle, DCOIT (0.13 mg/L) increased CAT in juveniles. We conclude that juveniles have greater basal activity of antioxidant enzymes than adults and, in addition, DCOIT negatively affected the adults of A. mactroides, mainly decreasing the activity of GST, SOD and CAT in the gills and digestive gland of these organisms.

Identification of glutathione-S-transferase m19 and m34 among responsive GST genes against 1-chloro-2,4-dinitrobenzene treatment of Tetrahymena thermophila

Industrial xenobiotic pollutants have toxic effects on diverse organisms in their natural environments. This study aims to identify the Glutathione-S-transferases (GST) from Tetrahymena thermophila that are highly responsive to the treatment of synthetic substrate 1-chloro-2,4-dinitrobenzene (CDNB). The LD50 value of CDNB was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test as 0.079 mM at 9 h exposure. The glutathione affinity-purified 22 kDa and 23 kDa GSTs from CDNB-treated cells were identified as GSTm19 and GSTm34 with 2D-gel electrophoresis coupled MALDI-Tof MS/MS analysis. The specific activitiy of the affinity-purified GSTs was upregulated upon the treatment of 0.072 mM CDNB with the decreased cell survival. GSTm19 and GSTm34 had also upregulated the mRNA expression under the highest dose treatment. The high cell survival and elevated total GST enzyme activity at 9 h under CDNB doses could be the result of both transcriptional upregulations as well as post-translational modifications. As a result, the cell survival of Tetrahymena thermophila was significantly affected by CDNB exposure in a concentration-dependent manner with the effect of low-dose stimulation and high-dose inhibition.

Difference in toxicity of Pd (II) and mechanism of action before and after reduction by Bacillus wiedmannii MSM.

At present, there are many studies on microbial reduction of Pd (II), although few of these studies assess the bacterial toxicity of Pd (II) before and after reduction by microorganisms. In this study, the toxicity of Pd (II) to Bacillus wiedmannii MSM (B. wiedmannii MSM) was assessed before and after reduction by live B. wiedmannii MSM cells (referred to as "Pd (0)-loaded cells") and after biosorption by dead B. wiedmannii MSM cells (referred to as "Pd (II)-loaded cells"). B. wiedmannii MSM is a widely occurring, nonpathogenic aerobic bacterium. Compared with Pd (II), the EC20, EC50, and EC80 of Pd (0)-loaded cells increased by 77.73-, 112.75-, and 163.91-fold, respectively, while the EC20, EC50, and EC80 of Pd (II)-loaded cells increased by only 11.55-, 9.77-, and 8.29-fold, respectively. The sterilization contribution rates of the mechanisms of action of Pd (0)-loaded cells were ranked in the order of: remaining Pd (II) > oxidative stress > physical puncture. Pd (II) was found to increase cell membrane permeability, glutathione S-transferase (GST) enzyme activity, and reactive oxygen species levels in cells and decrease the cell membrane potential. XPS results indicated that Pd (II) increased the content of polysaccharides and peptides and decreased the content of hydrocarbons in cells. These findings reveal the bactericidal mechanism of toxicity of Pd (II) and Pd (0)-loaded cells on Bacillus wiedmannii MSM and provide an environmentally friendly and inexpensive method for Pd (II) detoxification.

Biomarkers of oxidative stress in the post-embryonic characterization of the neotropical annual killifish.

Annual killifish are among the most remarkable extremophile species with the shortest vertebrate life span. Few studies have reported on the oxidative balance throughout their life cycle and its association to the natural aging process of these neotropical animals in a natural environment. We standardized and analyzed physiological markers related to the redox balance of the annual killifish (Cynopoecilus fulgens) throughout the post-embryonic life cycle (enzyme activity of Superoxide Dismutase, Catalase, Glutathione Peroxidase, and Glutathione S-transferase, as well as the determination of the levels of Lipoperoxidation, Carbonylated Proteins, and Total Proteins). We tested the influence of environmental variables on these biomarkers. Individuals were collected, including juveniles, adults, and seniles, in three sampling units around the Parque Nacional da Lagoa do Peixe, located in the Coastal Plain of Rio Grande do Sul. We observed that males and females used different physiological strategies of their redox balance during their life cycle, and their oxidative balance was influenced by their reproductive period and environmental variables (water temperature, abundance of predators, abundance of another sympatric annual killifish species, and abundance of C. fulgens). The population of each temporary pond presented different physiological responses to the adaptation of their life cycle, and there was an influence of environmental component as a modulator of this cycle. Our study offers reference values that will be useful for comparison in future research with short-lived organisms.

Toxicological Impacts on Antioxidant Responses, Stress Protein, and Genotoxicity Parameters of Aluminum Oxide Nanoparticles in the Liver of Oreochromis niloticus.

The aim of this study was to determine the toxic effects of aluminum oxide nanoparticles (Al2O3 NPs) on oxidative stress, stress protein, and genotoxicity parameters in Oreochromis niloticus. Ninety-six-hour LC50 value of Al2O3 NPs was found as 52.4 ppm for O. niloticus. The fish were exposed to 2.6 ppm (5% of the 96-h LC50) and 5.2 ppm (10% of the 96-h LC50) for 3 days and 7 days. Various biochemical parameters, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione-S-transferase (GST) activities, glutathione (GSH), thiobarbituric acid reactive substance (TBARS), heat shock protein 70 (HSP70; stress protein), and genotoxicity biomarker 8-hydroxy-2-deoxyguanosine (8-OHdG) levels, were determined. Results showed that antioxidant enzymes were significantly decreased in SOD, CAT, and GPx enzyme activity, but GST enzyme activity was significantly increased in 7 days. The oxidative stress parameters, GSH levels, were significantly decreased while 8-OHdG and TBARS levels were increased in 3 and 7 days. HSP70 levels were decreased in the concentrations of Al2O3 NPs and exposure times. Our results showed that as a result of changes in oxidative stress parameters, stress protein, and genotoxicity parameters, O. niloticus liver tissue is highly sensitive and toxic to aluminum oxide nanoparticle exposure.

Protective effect of glutathione S-transferase enzyme activity against aflatoxin B1 in poultry species: relationship between glutathione S-transferase enzyme kinetic parameters, and resistance to aflatoxin B1

Comparative studies designed to investigate the role of glutathione S-transferase (GST) activity on the enzyme catalyzed trapping of aflatoxin B1-8,9-epoxide (AFBO) with glutathione, and the relationship with aflatoxin B1 (AFB1) resistance have not been conducted in poultry. Hepatic cytosolic fractions of chickens, quail, turkeys and ducks were used to measure in vitro the enzymatic parameters maximal velocity (Vmax), Michaelis-Menten constant (Km) and intrinsic clearance (CLint) for GST activity. AFB1 used ranged from 2.0 to 157.5 µM and the AFB1-GSH produced was identified and quantitated by HPLC. Significant differences were found in GST Vmax values, being the highest in chickens, followed by quail, ducks and turkeys. The Km values were also significantly different, with chickens < ducks < turkeys < quail. Chickens had the higher CLint value in contrast to ducks. Differences by sex showed that duck females had a higher CLint value than the turkey and quail, whereas duck males had a CLint close to that of turkey. The ratio “AFBO production /AFB1-GSH production” follows the order duck>turkey>quail>chicken, in agreement with the known poultry sensitivity. The extremely high “AFB1 epoxidation activity/ GST activity” ratio observed in ducks might be the explanation for the development of hepatocellular carcinoma in this species.

Comparative transcriptome combined with biochemical and physiological analyses provide new insights toward cadmium accumulation with two contrasting Nicotiana species.

Cadmium (Cd) is known as one of the most hazardous elements in the environment and a persistent soil constraint toxic to all flora and fauna. In this study, we conducted physiological, biochemical, and transcriptomic analyses of Nicotiana rustica (N. rustica) and Nicotiana tabacum (N. tabacum) treated with CdCl2 to know the underlying molecular mechanisms of Cd accumulation. As a result, N. rustica had more dry weight than N. tabacum. Additionally, N. rustica accumulated higher Cd concentration (69.65 times), Cd2+ influx (1.32-fold), glutathione S-transferases (GST) enzyme activity (2.54 times), GSH/GSSG (oxidized form of GSH) ratio, increase of superoxide dismutase and CAT and a lower H2 O2 and superoxide (O2 •- ) accumulation in their roots than N. tabacum. Cd mainly distributed in the cytoplasm of both species and N. rustica had a significant proportion in the cell wall. Furthermore, the transcriptomic analysis revealed 173 and 710 differentially expressed genes (DEGs) between control and Cd-stressed plants in the leaves and roots of N. rustica, while 576 and 1543 DEGs were found in the leaves and roots of N. tabacum, respectively. In N. rustica, phenylpropanoid biosynthesis and phenylalanine metabolism were the most enriched pathways, while GSH metabolism, ATP-binding cassette transporters and phenylpropanoid biosynthesis were the most enriched in N. tabacum. Finally, we found that DEGs related to metal influx, sequestration, remobilization, and chelation were responsible for Cd accumulation. These results indicated that N. rustica accumulated higher Cd content than N. tabacum, suggesting that each species utilized different response mechanism under the same Cd stress conditions. The DEGs identified in this study might lead to the identification of genes or pathways related to Cd regulation. This study identifies important regulators related to Cd accumulation.

Synthesis, Cytotoxicity, Antioxidant and Antimicrobial Activity of Indole Based Novel Small Molecules

Aim: In this study experiments were carried out to explore antioxidant, antimicrobial, cytotoxic properties of novel indole derivative 1-ethyl-2-phenyl-3-phenylethyl-3-thiophen-2-yl-1Hindole (EPI) together with its effect on glutathione S-transferases (GST) activities in human liver carcinoma (HepG2) cells. Background:: Indoles probably represent one of the most important heterocyclic structures that have been attracting the interest of many scientists in drug discovery. Objective: The present study was carried out to explore antioxidant, antimicrobial, cytotoxic properties of novel indole derivative 1-ethyl-2-phenyl-3-phenylethyl-3-thiophen-2-yl-1H-indole (EPI) and its effect on glutathione S-transferases (GST) activities in human liver carcinoma (HepG2) cells. Materials and Methods: Pd-catalyst Sonogashira coupling reactions, MTT Assay, Antioxidant capacity test, Antimicrobial test, GST enzyme activity test. Results: 1-ethyl-2-phenyl-3-(phenylethynyl)-1H-indole had antioxidant and antimicrobial properties. It displayed significant induction in glutathione S-transferases (GST) enzyme activity in human liver cancer cell lines (HepG2), but cytotoxic effect on all tested cancer cell lines could not be observed. Conclusion:: All of these results showed that 1-ethyl-2-phenyl-3-(phenylethynyl)-1H-indole had antioxidant and antimicrobial properties without cytotoxic effect, which could make it a promising active component with further studies.

Effects of Different Ammonia Levels on Tribenuron Methyl Toxicity in Daphnia magna.

The present study investigates the toxicity of the herbicide tribenuron methyl (TBM) as an anthropogenic agent and ammonia as an abiotic factor on Daphnia magna at environmentally relevant concentrations. These stressors may coexist in surface waters in agricultural regions. To achieve this objective, D. magna were exposed to TBM at a nominal concentration of 0.81μg/L in association with a low ammonia (LA) concentration of 0.65mg/L and a high ammonia (HA) concentration of 1.61mg/L in acute toxicity tests of 96-h duration and chronic toxicity tests of 21-day duration. The D. magna also were exposed to TBM, HA, and LA singly. The D. magna were analysed for various biomarkers of sublethal toxicity. Glutathione peroxidase (GPx), glutathione S-transferase (GST), cholinesterase (ChE) enzyme activities, and levels of thiobarbituric acid reactive substances (TBARS) and total protein were determined spectrophotometrically. Mitochondrial membrane potential (MMP) was analysed by microscopy with fluorescence staining. Cytochrome c and 5' AMP-activated protein kinase (AMPK) were analysed by Western blotting. Morphometric properties were examined microscopically. This is the first study in which AMPK, an indicator of intracellular energy, was measured in D. magna. GST and ChE enzyme activities and TBARS and total protein levels did not change during acute exposures (i.e., 96h) in all treatments. GPx activity increased in D. magna from the HA + TBM treatment compared with single-exposure groups. The level of cytochrome c protein was elevated in D. magna from the LA and LA + TBM treatments. AMPK protein levels increased in all treatments with daphnids, except in the LA group. MMP was depolarised in D. magna from all treatments, whereas the most notable change was observed in HA + TBM mixture group in chronic exposures. The results show that GST and ChE may not be sensitive biomarkers for evaluating the sublethal toxic effects to D. magna exposed to environmentally relevant concentrations of ammonia and TBM. Acute and chronic exposure to ammonia and TBM probably caused an energetic crisis in D. magna. Therefore, AMPK and MMP are promising biomarkers for these toxicants.