Glutathione S-transferase genetic polymorphisms and fluoride-induced reproductive toxicity in men with idiopathic infertility / 亚洲男科学杂志(英文版)

Male infertility caused by idiopathic oligoasthenospermia (OAT) is known as idiopathic male infertility. Glutathione S-transferase (GST) and fluoride may play important roles in idiopathic male infertility, but their effects are still unknown. Our study examined the relationship between GST polymorphisms and fluoride-induced toxicity in idiopathic male infertility and determined the underlying mechanism. Sperm, blood, and urine samples were collected from 560 males. Fluoride levels were measured by a highly selective electrode method, and GST genotypes were identified using polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (PCR-RFLP). Semen parameters, DNA fragmentation index (DFI), mitochondrial membrane potential (MMP), and oxidative stress (OS) biomarkers were statistically assessed at the P < 0.05 level. Compared with healthy fertile group, semen parameters, fluoride levels, OS biomarkers, sex hormone levels, and MMP and DFI levels were lower in the idiopathic male infertility group. For glutathione S-transferase M1 (GSTM1[-]) and glutathione S-transferase T1 (GSTT1[-]) or glutathione S-transferase P1 (GSTP1) mutant genotypes, levels of semen fluoride, OS, MMP, and DFI were considerably higher, and the mean levels of sperm parameters and testosterone were statistically significant in GSTM1(+), GSTT1(+), and GSTP1 wild-type genotypes. Both semen and blood fluoride levels were associated with oxidative stress in idiopathic male infertility patients. Elevated fluoride in semen with the genotypes listed above was linked to reproductive quality in idiopathic male infertility patients. In conclusion, GST polymorphisms and fluorine may have an indicative relationship between reproductive quality and sex hormone levels, and OS participates in the development of idiopathic male infertility.

Correlation between post-transplant non-HLA antibodies and humoral rejection after kidney transplantation / 中华器官移植杂志

Objective:To explore the correlation between post-transplant non-HLA antibodies and humoral rejection(HR)after kidney transplantation(KT).Methods:A retrospective study was conducted for KT recipients with non-HLA antibody level detected from September 2019 to January 2021.The recipients with biopsy confirmed HR and donor-specific HLA antibodies negative or feeble positive at the time of HR were designated as HR group while recipients with stable renal allograft function from 2 weeks post-KT to the time of detecting non-HLA antibody as stable group.The levels of HLA antibody, MHC classⅠchain-related gene A(MICA)antibody and 32 non-HLA antibodies were tested by Luminex single antigen bead and the levels of angiotensin Ⅱ type 1 receptor(AT1R)antibody quantified by enzyme-linked immunosorbent assay (ELISA). Inter-group differences in positive rate of non-HLA antibodies and number of positive non-HLA antibodies were analyzed.Results:Twenty-four recipients had positive non-HLA antibodies while the remainders had no positive non-HLA antibodies.Three HR recipients were positive for actin antibody, collagen Ⅲ antibody, glutathione S-transferase theta-1 antibody or IFN-γ antibody respectively.However, all four non-HLA antibodies of stable recipients were negative.There was significant inter-group difference( P=0.017). Four HR recipients were positive for collagenⅡantibody while only 1 stable recipient was positive for collagenⅡantibody.The positive rate of collagenⅡ antibody was significantly higher in HR recipients than that in stable recipients( P=0.023). HR recipients had an average of 2.36 positive non-HLA antibodies while stable recipients had an average of 0.90.There was significant inter-group difference ( P=0.008). Conclusions:A high level of non-HLA antibodies may elevate the risk of HR after KT.

Avaliação dos níveis da citocina pró-inflamatória fator de necrose tumoral-α em pacientes com diabetes tipo 2 e sua relação com variações genéticas de enzimas antioxidantes e obesidade / Evaluation of proinflammatory cytokine tumor necrosis factor-α levels in obese vs nonobese patients with type 2 diabetes and their relationship to CAT C-262T polymorphism

Introdução: Diabetes tipo 2 (DM2) é um distúrbio multifatorial caracterizado pelo aumento dos níveis de radicais livres. Tanto o estresse oxidativo quanto a obesidade contribuem para um estado inflamatório da doença, principalmente pelo aumento da citocina TNF-α. Sabendo-se que a genética individual pode contribuir para o estresse oxidativo, o estudo avaliou o impacto das variações genéticas de enzimas antioxidantes C262T no gene CAT e polimorfismos nulos dos genes GSTM1 e GSTT1 nos níveis de TNF-α, assim como, avaliou se as variantes genéticas atuariam sinergicamente com a obesidade aumentando os níveis da citocina em diabéticos da Grande Vitória/ES, Brasil.Métodos: O polimorfismo no gene CAT foi avaliado pela técnica PCR/RFLP e nos genes GSTM1 e GSTT1 por PCR multiplex, em 56 pacientes, sendo 28 obesos e 28 não obesos. Níveis de TNF-α foram medidos pela técnica de ELISA sanduíche.Resultados: Frequências das variantes nulas de GSTM1 e GSTT1 foram 44,6% e 17,9%, respectivamente. As frequências genotípicas C262T-CAT foram 73,2%, 25% e 1,8% para homozigoto normal, heterozigoto e homozigoto polimórfico, respectivamente. Não houve associação entre genótipos polimórficos e aumento dos níveis de TNF-α, assim como, não foi demonstrado aumento significante da citocina quando avaliado o sinergismo entre obesidade e genética individual do paciente.Conclusão: Níveis de TNF-α não se elevam em diabéticos tipo 2 na presença dos polimorfismos nos genes CAT, GSTM1 e GSTT1, e a obesidade não atua no aumento dessa citocina na população estudada, separadamente ou em conjunto com a genética individual de variantes nos genes CAT, GSTM1 e GSTT1.

Introduction: Type 2 diabetes is a multifactorial disorder characterized by increased levels of free radicals. Both oxidative stress and obesity contribute to an inflammatory state of the disease, mainly by increasing the levels of the proinflammatory cytokine tumor necrosis factor-α (TNF-α). Considering that personal genetics may contribute to oxidative stress, this study assessed the impact of CAT C-262T polymorphism and GSTM1 and GSTT1 null polymorphisms on TNF-α levels in patients with type 2. diabetes. The study also evaluated whether the genetic variants act synergistically with obesity to increase TNF-α levels in patients with diabetes from Grande Vitória, Brazil.Methods: Fifty-six patients were included, of whom 28 were obese and 28 were nonobese. The CAT gene polymorphism was assessed using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method, whereas GSTM1 and GSTT1 polymorphisms were assessed using multiplex PCR. TNF-α levels were measured using the sandwich ELISA technique.Results: Frequencies of GSTM1 and GSTT1 null polymorphisms were 44.6% and 17.9%, respectively. The genotype frequencies of CATC-262T polymorphism were 73.2%, 25.0%, and 1.8% for normal homozygote, heterozygote, and polymorphic homozygote, respectively. Polymorphic genotypes were not associated with increased TNF-α levels, and there was no significant increase in TNF-α levels when evaluating the synergism between obesity and personal genetics.Conclusion: The presence of CAT, GSTM1, and GSTT1 gene polymorphisms was not associated with increased TNF-α levels in patients with type 2 diabetes. Obesity alone or combined with personal genetics of CAT, GSTM1, and GSTT1gene polymorphisms did not promote increased TNF-α levels in the study population.

Effects of siRNA interference with Nrf2 on the regulation of glutathione S-transferase and IL-1β in A549 cells exposed to hyperoxia / 中国新生儿科杂志

Objective:To study the gene expression of nuclear factor erythroid-2-related factor 2 (Nrf2), glutathione-S-transferase (GST) and interleukin-1 β (IL-1β) in A549 cells exposed to hyperoxia and cell apoptosis after siRNA interference with Nrf2.Method:Normal A549 cell lines were assigned into normoxia+siRNA group, normoxia+control group, hyperoxia+siRNA group and hyperoxia+control group according to whether siRNA interference was used and the exposure environment (normoxia/hyperoxia). The hyperoxia environment contained 95%O 2 and 5%CO 2. The levels of mRNA expression of Nrf2, GST and IL-1β were detected using quantitative real-time polymerase chain reaction (qRT-PCR). Flow cytometry was used to examine cell apoptosis of the hyperoxia+control group and hyperoxia+siRNA group at different time points. Analysis of variance (ANOVA) was used to test the relative gene expression and apoptosis of A549 cells. Result:(1) Compared with the normoxia+control group, the expression of Nrf2 and GST in the hyperoxia+control group was significantly increased ( P<0.05), and the expression of IL-1β was significantly decreased ( P<0.05); the expression of Nrf2 and GST in the normoxia+siRNA group decreased significantly ( P<0.05), while the expression of IL-1β increased significantly ( P<0.05). (2) Compared with the normoxia+siRNA group, Nrf2 expression in the hyperoxia+siRNA group showed no significant changes ( P=0.230), GST expression increased slightly ( P=0.057), and IL-1β expression decreased slightly ( P=0.112). (3) Compared with the hyperoxia+control group, the expression of Nrf2 and GST in the hyperoxia+siRNA group decreased significantly ( P<0.05), and the expression of IL-1β increased significantly ( P=0.042). (4) Compared with the hyperoxia+control group, the apoptosis of A549 cells in the hyperoxia+siRNA group increased significantly at 24 h, 48 h and 72 h ( P<0.05). Conclusion:After interfering with Nrf2, siRNA may regulate the expression of GST and IL-1β, preventing oxidative stress, reducing inflammatory response and inhibiting apoptosis.

Effect and Mechanism of Banxia Xiexintang on Rats with Chronic Atrophic Gastritis Based on Keap1/Nrf2/ARE Signaling Pathway / 中国实验方剂学杂志

Objective:To explore the mechanism of Banxia Xiexintang （BXXX） in preventing and treating chronic atrophic gastritis （CAG） through Kelch-like ECH-associated protein 1 （Keap1）/nuclear factor erythroid 2-related factor 2 （Nrf2）/antioxidant response element （ARE） signaling pathway. Method:SD rats were divided into a normal group （<italic>n</italic>=12） and an experimental group for CAG model induction. The model rats were then randomly divided into a model group， a vatacoenayme （VG） group （60 mg·kg^-1）， and high- （280 mg·kg^-1）， medium- （140 mg·kg^-1）， and low-dose （70 mg·kg^-1） BXXX groups. The doses in the BXXX groups were equivalent to 28， 14， and 7 g·kg^-1 crude drugs. The rats in the normal group and the model group received distilled water at an equal volume， and those in the VG group and the BXXX groups were treated correspondingly by gavage. After 12 weeks of treatment， hematoxylin-eosin （HE） staining was carried out to observe pathological changes in the gastric mucosa of CAG rats. Western blot and real-time fluorescence-based quantitative PCR was used to detect the protein and mRNA expression levels of Nrf2， glutathione S-transferase （GST）， and NAD （P）H：quinone oxidoreductase 1 （NQO1） in the gastric mucosa of CAG rats. Result:Compared with the normal group， the model group showed increased protein and mRNA expression levels of Nrf2， NQO1， and GST in the gastric mucosa of the rats （<italic>P</italic><0.05）， atrophic gastric mucosa， and even intestinal metaplasia. The protein and mRNA expression levels of Nrf2， NQO1， and GST in the VG group and the high- and medium-dose BXXX groups were lower than those in the model group （<italic>P</italic><0.05）， and gastric mucosa atrophy and intestinal metaplasia were significantly improved， especially in the high-dose BXXX group. However， the effect in the low-dose BXXX group was not significant. Conclusion:BXXX can blunt the transcriptional activity of Nrf2， shut down Nrf2 signaling pathway， and reduce the expression levels of NQO1 and GST to achieve normal oxidation-anti-oxidation balance， which may be one of its action mechanisms in the treatment of CAG.

Novel insights into conjugation of antitumor-active unsymmetrical bisacridine C-2028 with glutathione:Characteristics of non-enzymatic and glutathione S-transferase-mediated reactions / 药物分析学报

Unsymmetrical bisacridines (UAs) are a novel potent class of antitumor-active therapeutics.A significant route of phase II drug metabolism is conjugation with glutathione (GSH),which can be non-enzymatic and/or catalyzed by GSH-dependent enzymes.The aim of this work was to investigate the GSH-mediated metabolic pathway of a representative UA,C-2028.GSH-supplemented incubations of C-2028 with rat,but not with human,liver cytosol led to the formation of a single GSH-related metabolite.Interestingly,it was also revealed with rat liver microsomes.Its formation was NADPH-independent and was not inhibited by co-incubation with the cytochrome P450 (CYP450) inhibitor 1-aminobenzotriazole.Therefore,the direct conjugation pathway occurred without the prior CYP450-catalyzed bioactivation of the substrate.In turn,incubations of C-2028 and GSH with human recombinant glutathione S-transferase(GST) P1-1 or with heat-/ethacrynic acid-inactivated liver cytosolic enzymes resulted in the presence or lack of GSH conjugated form,respectively.These findings proved the necessary participation of GST in the initial activation of the GSH thiol group to enable a nucleophilic attack on the substrate molecule.Another C-2028-GSH S-conjugate was also formed during non-enzymatic reaction.Both GSH S-conju-gates were characterized by combined liquid chromatography/tandem mass spectrometry.Mechanisms for their formation were proposed.The ability of C-2028 to GST-mediated and/or direct GSH conjugation is suspected to be clinically important.This may affect the patient's drug clearance due to GST activity,loss of GSH,or the interactions with GSH-conjugated drugs.Moreover,GST-mediated depletion of cellular GSH may increase tumor cell exposure to reactive products of UA metabolic transformations.

Glutathione S-Transferase Activity in Tissues of Rats Exposed to Fenarimol

Abstract The unconscious use of pesticides causes various adverse effects on non-target organisms, including humans. Enzymes that control metabolism become the target of the pesticide and the organs are damaged due to toxic effects. Glutathione s-transferase (GST, EC 2.5.1.18), an important enzyme of the detoxification mechanism and antioxidant defense system, can be affected by such toxic substances. Therefore, the effect of fenarimol on GST enzyme activity was investigated in our study. For this, 200 mg/kg fenarimol was administered intraperitoneally to male and female rats at different periods (2, 4, 8, 16, 32, 64 and 72 hours). After application, GST enzyme activity was analysed in the liver, kidney, brain and small intestine tissues of the rats. According to our results, activation (liver, kidney, small intestine) or inhibition (brain) of the generally GST enzyme was observed in the tissues of rats exposed to fenarimol. It is thought that the increase and/or decrease in this enzyme activity may be the cause of the toxic effect of fenarimol.

Influência de polimorfismos nos genes antioxidantes GSTM1, GSTT1 e GSTP1 no desenvolvimento de complicações oftalmológicas em diabéticos tipo 2 / Influence of polymorphisms in the GSTM1, GSTT1, and GSTP1 antioxidant genes on the development of ophthalmic complications in persons with type 2 diabetes

Introdução: A diabetes tipo 2 (DM2) é uma desordem metabólica ocasionada pela disfunção das células beta pancreáticas que interferem na produção de insulina e/ou pela resistência dos órgãos alvos a esse hormônio. Níveis elevados de radicais livres em conjunto com o declínio das defesas antioxidantes presente na DM2 podem ocasionar danos a organelas celulares, promovendo complicações da doença. As glutationas S- transferases (GST) são as principais enzimas antioxidantes que participam da defesa celular contra o estresse oxidativo. Os polimorfismos nos genes que codificam essas enzimas podem acarretar o surgimento de complicações oftalmológicas em diabéticos. Este trabalho avaliou a influência dos polimorfismos nos genes GST no desenvolvimento de doenças como a catarata e o glaucoma em pacientes com DM2 na Grande Vitória (ES). Metodologia: Os polimorfismos dos genes GSTM1 e GSTT1 foram investigados através da técnica de PCR multiplex. Para o gene GSTP1 utilizou-se a técnica PCR- RFLP. A análise estatística foi realizada através do teste exato de Fisher ou do teste do qui-quadrado com P-valor < 0.05. Resultados: Não foi encontrada relação entre os polimorfismos nos genes GSTM1, GSTT1 e GSTP1 e o surgimento de doenças como glaucoma e catarata em pacientes com DM2. Conclusão: Nossos dados sugerem que os polimorfismos nulos nos genes GSTM1 e GSTT1 e o polimorfismo Ile105Val no gene GSTP1 não estão associados com a suscetibilidade individual para o desenvolvimento de complicações oftalmológicas em pacientes com DM2. (AU)

Introduction: Type 2 diabetes mellitus (T2DM) is a metabolic disorder caused by beta cell dysfunction that interferes with insulin production and/or by the resistance of target organs to this hormone. An increase in free radicals together with a decline in antioxidant defenses, present in T2DM, can damage cellular organelles and promote the occurrence of disease complications. Glutathione S-transferases (GSTs) are the main antioxidant enzymes involved in cellular defense against oxidative stress, and polymorphisms in genes encoding GSTs can lead to ophthalmic complications in persons with diabetes. In this study, we evaluated the influence of GST polymorphisms on the development of diseases such as cataract and glaucoma in patients with T2DM in Grande Vitória, Espírito Santo, Brazil. Methods: GSTM1 and GSTT1 polymorphisms were investigated using a multiplex PCR technique. PCR-RFLP was used for the GSTP1 gene. Statistical analysis was performed with Fisher's exact test or the chi-square test, with P-value <0.05. Results: There was no relationship between GSTM1, GSTT1, or GSTP1 polymorphisms and the occurrence of diseases such as glaucoma and cataract in patients with T2DM. Conclusion: Our data suggest that the GSTM1 and GSTT1 null polymorphisms and the ile105Val polymorphism in the GSTP1 gene are not associated with individual susceptibility to the development of ophthalmic complications in persons with T2DM. (AU)

Association between Glutathione-S-Transferase and Gastric Carcinoma: A Case Control Study

Gastric carcinoma is the fourth most common cancer type and the second leading cause of cancer deaths worldwide. Every year, around 1 million new cases and 0.7 million deaths are caused due to gastric carcinoma. Gastrointestinal tract is involved in absorption and metabolism of toxic or potentially carcinogenic compounds which may be present in the food we eat. In this context, digestive tract may be considered as a major site of cancer in humans. Glutathione-S-Transferase (GST) is an important metabolizing enzyme, present in the epithelial cells of human GIT. As nearly all reactive, ultimate carcinogenic forms of chemicals are electrophiles, GST is substantially important as a mechanism for carcinogen detoxification. The present study was conducted to evaluate the role of GST in gastric carcinoma and analyse the level of serum GST in patients suffering from gastric carcinoma. METHODSThis is a case control study, conducted among 50 cases of gastric carcinoma and 50 age sex matched controls. Patients included in this study were diagnosed with gastric carcinoma, after clinical and histological examination. Circulating levels of GST were assayed in the in the serum of control group and in patients with gastric carcinoma, using standardized method. RESULTSMean GST activity in serum was significantly higher (p < 0001) in gastric carcinoma patients (8.24 ± 1.94) as compared to control (5.47 ± 0.52). After chemotherapy (12.34 ± 1.05) the activity of GST was significantly higher (p < 0001) than before chemotherapy (10.23 ± 2.12). The generation of free radicals is as reflected by increased GST and GST-π activity in carcinoma cases. CONCLUSIONSSerum GSTs measurement in plasma may be a useful tumour marker in stomach cancer and serum GSTs activity might be helpful in predicting the response of chemotherapy in advanced stages of cancer. GST values are helpful in predicting the radiation response. Overexpression of GST in neoplasia may be causal, allowing replicative advantage, or casual, accompanying clonal expansion. The major limitation to its widespread use is the time needed for doing the assay and until this is overcome it will remain primarily a research tool.

An updated meta-analysis of the relationship between glutathione S-transferase T1 null/presence gene polymorphism and the risk of lung cancer

Aim of Study: There were many reports published on the relationship between glutathione S-transferase T1 (GSTT1) null/presence gene polymorphism and the risk of lung cancer in these years. In previous, we conducted a meta-analysis to evaluate the relationship between GSTT1 null/presence gene polymorphism and the risk of lung cancer. This study was conducted to update it. Materials and Methods: The association studies were identified from PubMed and Cochrane Library on March 1, 2016. Results: Sixty-three reports were recruited into this meta-analysis for the association of null genotype of GSTT1 with lung cancer susceptibility, consisting of 21,220 patients with lung cancer and 21,496 controls. There was a marked association between GSTT1 null genotype and lung cancer risk in overall populations and in Asians (overall populations: Odds ratio [OR] = 1.17, 95% confidence interval [95% CI]: 1.07–1.28, P = 0.006; Asians: OR = 1.41, 95% CI: 1.23–1.62, P < 0.00001). However, GSTT1 null genotype was not associated with the risk of lung cancer in Caucasians, Brazilian population, and Africans. Conclusion: GSTT1 null genotype is associated with the lung cancer risk in overall populations and in Asians

Biomarcadores enzimáticos e histológicos em brânquias de Ucides cordatus (Linnaeus, 1763) (Crustacea, Brachyura, Ucididae) indicativos de impactos ambientais em uma região portuária do nordeste do Brasil / [Enzymatic and histological biomarkers in Ucides cordatus (Linnaeus, 1763) (Crustacea, Brachyura, Ucididae) gills indicative of environmental impacts in a port region of northeastern Brazil]

Objetivou-se neste estudo analisar biomarcadores histológicos e bioquímicos em brânquias de U. cordatus indicativos de impactos na Baía de São Marcos. Caranguejos foram coletados em quatro áreas na Baía de São Marcos: A1= Ilha dos Caranguejos (com baixo impacto); A2= Coqueiro, A3= Porto Grande, A4= Cajueiro (áreas potencialmente impactadas). Mediram-se os dados biométricos de cada exemplar de caranguejo. Amostras de brânquias foram submetidas à técnica histológica padrão e homogeneizadas em tampão fosfato, e o sobrenadante foi utilizado para análise das enzimas glutationa-S-transferase (GST) e catalase (CAT). A biometria indicou que os caranguejos de A1 são significativamente (P<0,05) maiores e mais pesados do que os caranguejos das áreas A2, A3 e A4. As alterações branquiais (rompimento das células pilastras, deformação do canal marginal, deslocamento da cutícula e necrose) foram significativamente (P˂0,05) mais frequentes em caranguejos de A2, A3 e A4 do que nos caranguejos de A1. As atividades enzimáticas da GST e CAT nos caranguejos apresentaram diferença significativa (P<0,05) entre as áreas de coletas, com padrão similar ao observado para as alterações branquiais. Os biomarcadores analisados mostraram que os caranguejos estão sob diferentes níveis de impactos (A4>A3>A2>A1) ao longo da Baía de São Marcos.(AU)

The objective of this study was to analyze histological and biochemical biomarkers in U. cordatus gills indicative of impacts in São Marcos Bay. Crabs were collected from four areas in São Marcos Bay: A1=Ilha dos Caranguejos (with low impact); A2=Coqueiro, A3=Porto Grande, A4=Cajueiro (potentially impacted areas). The biometric data of each specimen was measured. Gill samples were submitted to standard histological technique and homogenized in phosphate buffer, and the supernatant was used for analysis of glutathione S-transferase (GST) and catalase (CAT) enzyme activity. Biometric data indicated that crabs found in A1 are significantly (P<0.05) larger and heavier than crabs found in A2, A3 and A4 areas. Gill alterations (rupture of pilaster cells, Dilation of the marginal channel, Cuticle Rupture and necrosis) were significantly (P˂0.05) more frequent in the crabs in A2, A3 and A4 than crabs in A1. The enzymatic activities of GST and CAT showed significant difference (P<0.05) between the sampling areas, similar to that observed for gill alterations. The biomarkers analyzed showed that the crabs are under different impact levels (A4> A3> A2> A1) along the São Marcos Bay.(AU)

Determination of insecticides' lethal concentrations and metabolic enzyme levels in Triatoma dimidiata / Determinación de concentraciones letales y niveles de enzimas metabólicas de insecticidas en Triatoma dimidiata

Abstract: Objective: The feasibility of the use of WHO impregnated paper and biochemical assays to determine lethal concentrations (LC50 and LC99) and insecticide metabolic enzyme levels of Triatoma dimidiata. Materials and methods: LC50 and LC99 were calculated with WHO papers impregnated at different concentrations of malathion, propoxur and deltamethrin; the percentage of insensitive acetylcholinesterase (iAChE); and the levels of esterases, glutathione S-transferases, and monooxygenases in laboratory nymphs of the first stage (5 to 7 days), were undertaken using the WHO biochemical assays. Results: Respectively the LC50 and LC99 µg/cm2 obtained for malathion were 43.83 and 114.38, propoxur 4.71 and 19.29, and deltamethrin 5.80 and 40.46. A 30% of the population had an iAChE, and only a few individuals had high P450 and β-eterase levels. Conclusion: Impregnated papers and biochemical tests developed by WHO for other insects, proved to be feasible methods in monitoring insecticide resistance and metabolic enzymes involved in T. dimidiata.

Resumen: Objetivo: La factibilidad de usar los papeles impregnados y ensayos bioquímicos según la OMS para determinar concentraciones letales (CL50 y CL99) y niveles enzimáticos en la resistencia a insecticidas en Triatoma dimidiata. Material y métodos: Se calcularon la CL50 y CL99 con papeles impregnados según la OMS a diferentes concentraciones de malatión, propoxur y deltametrina; el porcentaje de acetilcolinesterasa insensible (iAChE); y los niveles de esterasas, glutatión S-transferasas, y monooxigenasas en ninfas de laboratorio del estadio I (5-7 días) se determinaron usando los ensayos bioquímicos según la OMS. Resultados: Se obtuvieron las CL50 y CL99 µg / cm2 respectivamente para malatión 43.83 y 114.38, propoxur 4.71 y 19.29, y deltametrina 5.80 y 40.46. Un 30% de las chinches tuvo iAChE, y sólo pocos individuos tuvieron niveles superiores de P450 y β-eterasas. Conclusión: Los papeles impregnados y ensayos bioquímicos que describe la OMS para otros insectos demostraron ser métodos factibles para monitorear la resistencia a insecticidas y las enzimas metabólicas involucradas en T. dimidiata.

Yeast glutaredoxin, GRX4, functions as a glutathione S-transferase required for red ade pigment formation in Saccharomyces cerevisiae

As a tumor suppressor, p53 preserves genomic integrity in eukaryotes. However, limited evidence is availablefor the p53 shuttling between the cytoplasm and nucleus. Previous studies have shown that b-actin polymerization negatively regulates p53 nuclear import through its interaction with p53. In this study, we found thatDNA damage induces both b-actin and p53 accumulation in the nucleus. b-actin knockdown impaired thenuclear transport of p53. Additionally, b-actin could interact with p53 which was enhanced in response togenotoxic stress. Furthermore, N terminal deletion mutants of p53 shows reduced levels of association with bactin. We further identified Ser15, Thr18 and Ser20 of p53 are critical to the b-actin: p53 interaction, whichupon mutation into alanine abrogates the binding. Taken together, this study reveals that b-actin regulates thenuclear import of p53 through protein–protein interaction.

Cáncer de próstata y polimorfismos de la glutation s-transferasa en una población venezolana / Prostate cancer and polymorphisms of the glutathione s-transferase in a population Venezuelan

En vista de la alta prevalencia del cáncer de próstata en la población venezolana y la ausencia de un patrón genético conocido en relación a la expresión de las enzimas Glutatión S-transferasas, se estudió la relación entre la expresión de un polimorfismo nulo de estas enzimas y la presencia de cáncer de adenocarcinoma prostática Métodos: Se incluyen 100 individuos para el muestreo no probabilístico, 50 pacientes con diagnóstico de adenocarcinoma de próstata comprobado mediante biopsia y 50 controles con hiperplasia prostática benigna demostrada mediante tacto y corroborada por ultrasonido transrectal, provenientes de los principales hospitales del país, se procedió a tomar muestra de sangre y mediante reacción de cadena de polimerasa, se determinó la presencia o ausencia de los genes para las enzimas Glutatión S-transferasa Mu 1 (GST M) y glutatión S-transferasa theta 1 (GST T1). Resultados: se logró evidenciar que el genotipo nulo se encontró en 40 y 24% de los pacientes mientras que para los controles fue de 38% y 22% respectivamente, demostrando que en la población venezolana estudiada no existen diferencias significativas entre casos y controles. Conclusiones: No se pudo demostrar una diferencia significativa entre los dos grupos estudiados. Recomendaciones: A pesar de nuestros hallazgos, se necesitan estudios futuros con muestras de mayor tamaño para dilucidar la posible asociación entre este patrón enzimático con el riesgo de presentar cáncer de próstata(AU)

Prostate cancer presents with a high incidence in the Venezuelan population. there is no known genetic pattern related to the expression of drug metabolizing enzymes Glutathione S-transferases. Methods: We proceeded to study the possible correlation between null polymorphism for these enzymes and prostate adenocarcinoma. the sample included 100 patients recruited from the Urology Department of three University Hospitals in Caracas, Venezuela, 50 cancer patients and 50 cancer free controls. Blood samples were drawn from each patient and polymorphisms for Glutathione S-transferase Mu 1 (GST M1) and Glutathione-sS-transferase theta 1(GST T1) were determined by polymerase chain reaction from lymphocytes. Results: Null genotype was found in 40% and 24% of cancer patients whereas the percentage in controls was 38 and 22% respectively, showing no statistically significant differences between the two groups. Conclusions: It was not possible to show a significant difference between the two groups. Recommendations: Due to the small size of the sample, it would be necessary to explore further in a larger population sample to determine whether there is an association between the expression of these enzymes and prostate cancer(AU)

Evaluation of phytochemical properties and larvicidal activities of Cynodon dactylon, Clerodendrum viscosum, Spilanthes acmella and Terminalia chebula against Aedes aegypti

Objective: To investigate the phytochemical, antioxidant and larvicidal property of Cynodon dactylon, Clerodendrum viscosum, Spilanthes acmella and Terminalia chebula against Aedes aegypti. Methods: Antioxidant capacity of methanolic extract of the plants was studied by 2,2- Diphenyl-1-picryl-hydrazyl-hydrate (DPPH) assay, ferric reducing antioxidant power assay, 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonate) assay (ABTS), thiobarbituric acid reactive substance (TBARS) assay, superoxide anion scavenging activity and total antioxidant activity assay following standard protocol. Total phenolic content, total flavonoid content, carbohydrate, and plant protein were also estimated following standard protocols. Larvicidal property of plant extracts were determined following World Health Organization standard protocol. Additionally, glutathione-s-transferase (GST) and acetylcholinesterase (AchE) inhibitory property was also tested biochemically. Results: Phytochemically, high protein, carbohydrate and phenolic were found in Terminalia chebula, while Cynodon dactylon showed high flavonoid contents. Similarly, high antioxidant activity was found in Terminalia chebula with IC50 values at 13.7, 2.9, 45.2 and 46.0 μg/mL in DPPH, ABTS, TBARS and superoxide anion scavenging activity, respectively. Larvicidal study showed strongest activity in Spilanthes acmella followed by Cynodon dactylon, and Clerodendrum viscosum and Terminalia chebula. GST and AchE of Aedes aegypti larvae showed reduced enzyme activity when pre-incubated with Cynadon dactylon and Spilanthes acmella. Conclusions: The methanolic crude extracts of Cynodon dactylon, Clerodendrum viscosum, Spilanthes acmella and Terminalia chebula possess strong antioxidant and larvicidal property against Aedes aegypti and therefore, may be further investigated for the molecular mode of action.

Analysis of different proteins in primary open angle glaucoma based on aqueous proteome / 中华实验眼科杂志

Objective@#To analyze the changes of protein expression in aqueous humor of patients with primary open angle glaucoma (POAG), and to explore the biological markers and potential therapeutic targets associated with disease occurrence.@*Methods@#A retrospective case-control study was adopted.Ten patients with age-related cataract and 10 patients with POAG combined with cataract.were collected at the Tianjin Medical University Eye Hospital from October 2016 to December 2017.In the process of surgery, 100 μl of aqueous humor were collected with 1 ml syringe and No.1 needle through the surgical access.Those proteins extracted from aqueous humor were analyzed by quantitative proteomic mass spectrometry.The differential significance test was performed by Maxquant significances A. The differential proteins of the two groups were screened and determined with the conditions of P<0.05 and difference multiple>2.The biological big data was annotated by the function of GO and KEGG.The study was approved by the Ethics Committee of the Tianjin Medical University Eye Hospital (No.2013KY[L]-10). Patients and their guardians all signed the informed consent.@*Results@#Ninty-seven differential proteins were detected in this proteomic analysis, including 48 up-regulated proteins and 49 down-regulated proteins.GO analysis significantly differs in protein function involved in many aspects, some different proteins including lipopolysaccharide-binding protein (LBP), cluster of Differentiation 163(CD163), C-reactive protein (CRP), annexin A1 (ANXA1) were involved in inflammation; redox-related proteins were glutathione S-transferase P (GSTP1), thioredoxin (TXN), some different proteins related with cell adhesion movement were cartilage oligomeric matrix protein (COMP), desmocollin-2 (DSC2) and laminin subunit beta-2 (LAMB2); procollagen-lysine, 2-oxoglutarate 5-dioxygenase 1 (PLOD1), solid protein (tenascin, TNC) are associated with fibrosis; some different proteins related to nerve growth were reelin (RELN), semaphorin-3F(SEMA3F), semaphorin-4B(SEMA4B). Metabolism-related proteins included pyruvate kinase (PKM), carb oxypeptidase N subunit 2 (CPN2) and so on.KEGG analysis indicated that the expression of NrF2/ERK signaling pathway and TGF-β/NF-κB signaling pathway were different between the two groups.@*Conclusions@#The expression of GSTP1 and TXN in the aqueous humor of POAG is significantly decreased, which may regulate cell adhesion and activity and expression of extracellular matrix by regulating NrF2/ERK1/2 and TGF-β/NF-κB signaling pathways.GSTP1 and TXN may serve as a potential biomarker and therapeutic target of POAG.

Analysis of different proteins in primary open angle glaucoma based on aqueous proteome / 中华实验眼科杂志

Objective To analyze the changes of protein expression in aqueous humor of patients with primary open angle glaucoma ( POAG ) , and to explore the biological markers and potential therapeutic targets associated with disease occurrence. Methods A retrospective case-control study was adopted. Ten patients with age-related cataract and 10 patients with POAG combined with cataract. were collected at the Tianjin Medical University Eye Hospital from October 2016 to December 2017. In the process of surgery,100 μl of aqueous humor were collected with 1 ml syringe and No. 1 needle through the surgical access. Those proteins extracted from aqueous humor were analyzed by quantitative proteomic mass spectrometry. The differential significance test was performed by Maxquant significances A. The differential proteins of the two groups were screened and determined with the conditions of P<0. 05 and difference multiple>2. The biological big data was annotated by the function of GO and KEGG. The study was approved by the Ethics Committee of the Tianjin Medical University Eye Hospital (No. 2013KY[L]-10). Patients and their guardians all signed the informed consent. Results Ninty-seven differential proteins were detected in this proteomic analysis,including 48 up-regulated proteins and 49 down-regulated proteins. GO analysis significantly differs in protein function involved in many aspects,some different proteins including lipopolysaccharide-binding protein (LBP),cluster of Differentiation 163(CD163),C-reactive protein (CRP),annexin A1 (ANXA1) were involved in inflammation;redox-related proteins were glutathione S-transferase P (GSTP1),thioredoxin (TXN), some different proteins related with cell adhesion movement were cartilage oligomeric matrix protein ( COMP ) , desmocollin-2 ( DSC2 ) and laminin subunit beta-2 ( LAMB2 );procollagen-lysine, 2-oxoglutarate 5-dioxygenase 1 (PLOD1),solid protein (tenascin,TNC) are associated with fibrosis;some different proteins related to nerve growth were reelin (RELN),semaphorin-3F(SEMA3F),semaphorin-4B(SEMA4B). Metabolism-related proteins included pyruvate kinase ( PKM ) , carb oxypeptidase N subunit 2 ( CPN2 ) and so on. KEGG analysis indicated that the expression of NrF2/ERK signaling pathway and TGF-β/NF-κB signaling pathway were different between the two groups. Conclusions The expression of GSTP1 and TXN in the aqueous humor of POAG is significantly decreased,which may regulate cell adhesion and activity and expression of extracellular matrix by regulating NrF2/ERK1/2 and TGF-β/NF-κB signaling pathways. GSTP1 and TXN may serve as a potential biomarker and therapeutic target of POAG.

Evaluation of phytochemical properties and larvicidal activities of Cynodon dactylon, Clerodendrum viscosum, Spilanthes acmella and Terminalia chebula against Aedes aegypti

Objective: To investigate the phytochemical, antioxidant and larvicidal property of Cynodon dactylon, Clerodendrum viscosum, Spilanthes acmella and Terminalia chebula against Aedes aegypti. Methods: Antioxidant capacity of methanolic extract of the plants was studied by 2,2- Diphenyl-1-picryl-hydrazyl-hydrate (DPPH) assay, ferric reducing antioxidant power assay, 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonate) assay (ABTS), thiobarbituric acid reactive substance (TBARS) assay, superoxide anion scavenging activity and total antioxidant activity assay following standard protocol. Total phenolic content, total flavonoid content, carbohydrate, and plant protein were also estimated following standard protocols. Larvicidal property of plant extracts were determined following World Health Organization standard protocol. Additionally, glutathione-s-transferase (GST) and acetylcholinesterase (AchE) inhibitory property was also tested biochemically. Results: Phytochemically, high protein, carbohydrate and phenolic were found in Terminalia chebula, while Cynodon dactylon showed high flavonoid contents. Similarly, high antioxidant activity was found in Terminalia chebula with IC

Evaluation of hepatoprotective activity of Syringa oblata leaves ethanol extract with the indicator of glutathione S-transferase A1

Abstract The leaves of Syringa oblata Lindl., Oleaceae, had been extensively used as a folk medicine to treat various infections, heal inflammations, icteric hepatitis and acute mastitis. The study was designed to evaluate the hepatoprotective activity of S. oblata leaves ethanol extract against CCl4-induced hepatotoxicity in primary hepatocytes and mice with the indicator of glutathione S-transferase alpha 1. The hepatoprotective effects of S. oblata leaves ethanol extract were evaluated on the basis of liver histopathology and biochemical parameters as well as hepatic oxidative stress markers. The results showed that CCl4 negatively modulated biochemical parameters and liver antioxidant activities. However, the use of S. oblata leaves ethanol extract restored altered-serum biochemical parameters and liver antioxidant activities in a dose-dependent manner. Importantly, the trends in S-transferase alpha 1 were similar to alanine aminotransferase and aspartate aminotransferase level, and S-transferase alpha 1 was suggested to be a marker for the evaluation of hepatoprotective activity of S. oblata leaves ethanol extract. Histopathological examination showed that CCl4 causes significant hepatic injury relative to control group. The above findings suggested that S. oblata leaves ethanol extract has hepatoprotective effects against CCl4-induced hepatic injury and S-transferase alpha 1 may be an indicator to evaluate the protective effects of S. oblata leaves ethanol extract.