Progress in the application of three-dimensional cell culture model in toxicity tests of xenobiotic / 中华预防医学杂志

In the process of xenobiotic toxicity prediction and risk assessment, in vitro cell culture models possess high practical application value. With the rapid development of biological technologies such as three-dimensional (3D) bio-printing, organoid culture and organ-on-a-chip systems, in vitro cell culture models have made great progress. Sharing the similarities in structure, function and the physiological environment with tissues or organs in vivo, hazard identification and dose-response analysis based on 3D cell culture models provide access to more accurate toxicity data as a theoretical basis for risk assessment and risk management of chemicals. This review summarizes the establishment of three typical 3D cell culture models, i.e., human cell line-based co-culture model, 3D-printed scaffold-based cell culture model and organoids, and their application in toxicity tests of xenobiotics.

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)

Aplicaciones biotecnológicas de los microorganismos / Biotechnological applications of microorganisms

Resumen La biodiversidad de los microorganismos así como la naturaleza única y las capacidades biosintéticas en condiciones ambientales específicas hacen que los microorganismos sean los probables candidatos para resolver problemas de escases de alimentos, contro de plagas, biodegradación de los xenobióticos, descomposición de la basura, las pilas de desechos producidas, entre otros. Los microorganismos ofrecen un gran potencial para la exploración de moléculas y procesos, y el conocimiento de las especies no convencionales, especialmente dentro del grupo Archaea, ha estimulado la investigación molecular de genes de interés. Estos nuevos genes pueden incorporarse mediante tecnología recombinante en especies biológicamente conocidas, como E. coli y S. cerevisiae, para la síntesis a gran escala de productos. La microbiología tecnológica tiene grandes potenciales para explorar y obstáculos por superar. Por lo tanto, solo la investigación en esta área resulta prometedora para científicos en todo el mundo. En la presente revisión se presentan las aplicaciones más significativas de los microorganismos en la industria de alimentos, la agricultura, compuestos químicos, combustibles, farmacología y materiales.

Abstract The biodiversity of microorganisms as well as the unique nature and biosynthetic capabilities in specific environmental conditions make microorganisms the likely candidates to solve problems of food shortages, pest control, biodegradation of xenobiotics, decomposition of garbage, batteries of produced waste, among others. Microorganisms offer great potential for the exploration of molecules and processes, and knowledge of non-conventional species, especially within the Archaea group, has stimulated the molecular investigation of genes of interest. These new genes can be incorporated by recombinant technology into biologically known species, such as E. coli and S. cerevisiae, for the large-scale synthesis of products. Technological microbiology has great potentials to explore and obstacles to overcome. Therefore, only research in this area is promising for scientists around the world. In this review we present the most significant applications of microorganisms in the food industry, agriculture, chemical compounds, fuels, pharmacology and materials.

Role of pregnane X receptor (PXR) in endobiotic metabolism / 生理学报

As a member of the nuclear receptor superfamily, the pregnane X receptor (PXR) is a ligand-activated transcription factor. PXR is highly expressed in liver and intestinal tissues, and also found in other tissues and organs, such as stomach and kidney. After heterodimerization with retinoid X receptor (RXR), PXR recruits numerous co-activating factors, and binds to specific DNA response elements to perform transcriptional regulation of the downstream target genes. As an acknowledged receptor for xenobiotics, PXR was initially considered as a nuclear receptor regulating drug metabolizing enzymes and transporters. However, nowadays, PXR has also been recognized as an important endobiotic receptor. Recent studies have shown that PXR activation can regulate glucose metabolism, lipid metabolism, steroid endocrine homeostasis, detoxification of cholic acid and bilirubin, bone mineral balance, and immune inflammation in vivo. This review focuses on the role of PXR in metabolism of endogenous substances.

Hepaprotective Effect of Standardized Ecklonia stolonifera Formulation on CCl₄-Induced Liver Injury in Sprague-Dawley Rats

The liver is an essential organ for the detoxification of exogenous xenobiotics, drugs and toxic substances. The incidence rate of non-alcoholic liver injury increases due to dietary habit change and drug use increase. Our previous study demonstrated that Ecklonia stolonifera (ES) formulation has hepatoprotective effect against alcohol-induced liver injury in rat and tacrine-induced hepatotoxicity in HepG2 cells. This present study was designated to elucidate hepatoprotective effects of ES formulation against carbon tetrachloride (CCl₄)-induced liver injury in Sprague Dawley rat. Sixty rats were randomly divided into six groups. The rats were treated orally with ES formulation and silymarin (served as positive control, only 100 mg/kg/day) at a dose of 50, 100, or 200 mg/kg/day for 21 days. Seven days after treatment, liver injury was induced by intraperitoneal injection of CCl₄ (1.5 ml/kg, twice a week for 14 days). The administration of CCl₄ exhibited significant elevation of hepatic enzymes (like AST and ALT), and decrease of antioxidant related enzymes (superoxide dismutase, glutathione peroxidase and catalase) and glutathione. Then, it leaded to DNA damages (8-oxo-2′-deoxyguanosine) and lipid peroxidation (malondialdehyde). Administration of ES formulation inhibited imbalance of above factors compared to CCl₄ induced rat in a dose dependent manner. Real time PCR analysis indicates that CYP2E1 was upregulated in CCl₄ induced rat. However, increased gene expression was compromised by ES formulation treatment. These findings suggests that ES formulation could protect hepatotoxicity caused by CCl₄ via two pathways: elevation of antioxidant enzymes and normalization of CYP2E1 enzyme.

Estudo da expressão gênica dos principais metabolizadores de fase II de xenobióticos: GSTM1, GSTP1 e GSTT1 em carcinoma de células escamosas bucal / Study of the gene expression of the main xenobiotic phase II metabolizers: GSTM1, GSTP1 and GSTT1 in squamous cell carcinoma of the oral cavity

Tabaco e álcool são considerados os principais fatores de risco para o carcinoma de células escamosas (CCE) bucal contribuindo de maneira desfavorável para o tratamento e desfecho clínico. Seus carcinógenos são metabolizados em duas fases, sendo a segunda fase realizada pelas Glutationa S-transferases (GSTs). O objetivo do presente estudo foi avaliar a expressão gênica da forma selvagem dos genes GSTM1, GSTP1 e GSTT1 por qPCR em 33 amostras de CCE bucal de fumantes, ex-fumantes e não fumantes, e 15 controles em busca de uma correlação clínica com consumo de tabaco, álcool e estadiamento clínico. A dependência nicotínica foi avaliada pelo Teste de Fagerström pra Dependência a Cigarros (TFDC) e para consumo de etílicos o Teste AUDIT. Foi observado aumento da expressão de GSTM1 no Grupo CCE fumante em relação ao Grupo Controle (p=0,0161). Contrariamente, foi encontrada uma menor expressão de GSTT1 no Grupo CCE fumante em relação ao Grupo Controle fumante (p=0,0183). No grupo CCE fumante não foi encontrada uma correlação entre a expressão dos genes estudados e fatores ligados ao tabagismo, etilismo e estadiamento clinico. No grupo Controle fumante, houve correlação entre teste AUDIT e a expressão de GSTM1 (p=0,0000). Para GSTP1 e GSTT1 houve correlação entre a expressão quando comparada a idade do paciente (p=0,0008; p=0,0095), idade de inicio do tabagismo (p=0,0033; p=0,0081), TFDC (p=0,0102; p=0,0085) e AUDIT (p=0,0052; p=0,0219) respectivamente. Para GSTT1 foi encontrada uma correlação entre a expressão e número de cigarros/dia (p=0,0175). Concluímos que as formas selvagens das GSTs estudadas apresentaram uma alta expressão nas amostras de CCE bucal, entretanto, quantitativamente essa expressão foi baixa, com grande variabilidade interindividual. Outrossim, não houve uma correlação direta entre níveis de expressão, carga tabágica, TFDC, teste AUDIT e estadiamento clínico. O aumento da expressão de GSTM1 e GSTP1 parece não ter tido um efeito protetor. A baixa expressão de GSTT1 em pacientes fumantes com CCE bucal se mostrou um potencial marcador a ser avaliado em pacientes fumantes que ainda não desenvolveram uma neoplasia maligna(AU)

Tobacco and alcohol are considered to be the main risk factors for oral squamous cell carcinoma (SCC), contributing to treatment and clinical outcome. Its carcinogens are metabolized in two phases, being the second phase carried out by Glutathione Stransferases (GSTs). The objective of the present study was to evaluate the wild-type gene expression of the GSTM1, GSTP1 and GSTT1 genes by qPCR in 33 samples of oral SCC from smokers, former smokers and nonsmokers, and 15 controls looking for a clinical correlation with tobacco and alcohol consumption and clinical staging. Nicotinic dependence was assessed by the Fagerström Test for Cigarette Dependence (TFCD) and alcohol consumption by the AUDIT Test. Increased expression of GSTM1 in the Smoker SCC Group was observed in relation to the Control Group (p=0.0161). Conversely, a lower expression of GSTT1 was found in the smoker SCC group compared to the Smoker Control Group (p=0.0183). In the smoker SCC group, no correlation was found between the genes expression studied and factors related to smoking, alcoholism and clinical staging. In the Smoker Control Group, there was a correlation between the AUDIT test and the GSTM1 expression (p=0.0000). For GSTP1 and GSTT1, there was a correlation between the expression compared with the patient's age (p=0.0008, p=0.0095), age of starting smoking (p=0.0033, p=0.0081), FTCD (p=0.0102, p=0.0085) and AUDIT (p=0.0052, p=0.0219) respectively. For GSTT1 a correlation was found between expression and number of cigarettes/day (p=0.0175). We concluded that the wild forms of the GSTs studied presented a high expression in the samples of oral SCC; however, quantitatively this expression was low, with great interindividual variability. Also, there was no direct correlation between levels of expression, pack-years, FTCD, AUDIT Test and clinical stage. Increased expression of GSTM1 and GSTP1 appears to have had no protective effect. The low GSTT1 expression in smokers with oral SCC was shown to be a potential marker to be evaluated in smoker patients who have not yet developed a malignant neoplasm(AU)

Ecotoxicidad de nanopartículas metálicas y superparamagnéticas de óxido de hierro en dos especies / Ecotoxicity of metalic and superparamagnetics iron oxide nanoparticles in two species

Introducción: la nanotecnología y el empleo de materiales a nano escala son un área relativamente nueva de la ciencia y la tecnología con un gran crecimiento en el mercado global. Muchos de los productos no cuentan con estudios que garanticen su uso seguro, tanto para el hombre como para los ecosistemas. Los estudios ecotoxicológicos permiten evaluar los efectos de un determinado xenobiótico sobre especies representativas de los diferentes compartimentos ambientales. Objetivo: evaluar los efectos tóxicos de nanopartículas de Ag, Au, Ag/Ag y superparamagnéticas de óxido de hierro, en dos especies bioindicadoras de los ecosistemas terrestre y acuático. Métodos: como parte de los estudios de seguridad se realizaron ensayos de toxicidad aguda por contacto en lombriz de tierra de la especie Eisenia andrei, con una duración de 96 horas y estudios en anfibios de la especie Osteopillus septentrionales en diferentes etapas del desarrollo (embrionario y larval). Se evaluó la ocurrencia de mortalidad y de efectos tóxicos, en el caso del ensayo en lombriz de tierra; se determinó además la viabilidad celular. Resultados: los efectos tóxicos más significativos en el caso de la lombriz de tierra fueron, la ocurrencia de alteraciones fisiológicas y conductuales al ser expuesta a NPs de Ag de 3 nm y superparamagnéticas de óxido de hierro, estas últimas provocaron citotoxicidad a la concentración 1,38 mg/mL. En el caso de los anfibios se evidenció toxicidad en NPs de Ag 3 nm y superparamagnéticas de óxido de hierro. Conclusiones: todas las nanopartículas mostraron efectos tóxicos en las especies bioindicadoras evaluadas(AU)

Introduction: Nanotechnology and the use of nanoscale materials are a relatively new area of science and technology with big growth in the global market. Many of these products don't have studies that guarantee their safe use, both for man and for ecosystems. Ecotoxicological studies allow the evaluation of the effects of a particular xenobiotic on representative species of the different environmental compartments. Objective: To evaluate the toxic effects of nanoparticles of Ag, Au, Ag / Ag and super paramagnetic iron oxide in two bioindicators of terrestrial and aquatic ecosystems. Methods: Acute contact toxicity tests were carried out on ground worm of the Eisenia andrei species, with a duration of 96 hours and studies on amphibians of the species Osteopillus septentrionales at different stages of development (embryonic and larval). The occurrence of mortality and toxic effects was evaluated in the case of earthworm test; cell viability was also determined. Results: The most significant toxic effects in the case of earthworms were the occurrence of physiological and behavioral alterations when exposed to 3 nm Ag of superparamagnetic iron oxide nanoparticles, where the latter caused cytotoxicity at concentration of 1.38 mg / mL. In the case of amphibians, toxicity was evidenced in Ag 3 nm nanoparticles and superparamagnetic iron oxide. Conclusions: All nanoparticles showed toxic effects in the evaluated bioindicator species(AU)

Tank mixture of pesticides for Spodoptera frugiperda control in maize with triflumuron / Mistura em tanque de produtos fitossanitários no controle de Spodoptera frugiperda na cultura do milho com triflumuron

Nowadays, few studies have been conducted to evaluate the effect of tank mixture of pesticides on pest chemical control in maize crops, however the farmers are often using these mixtures, without any technical support. The current study focused on assessing some pesticide mixtures to control Spodoptera frugiperda with triflumuron in a conventional corn hybrid. Moreover, this experiment aimed to assess likely plant physiological effects, as well as physical-chemical characteristics of these tank mixes. To this end, four treatments were carried out in a randomized block design with five replications, being repeated in two seasons. Treatments consisted of different pesticide mixtures, such as insecticide (INS) + adjuvant (ADJ); INS + ADJ + fungicide (FUN); INS + ADJ + FUN + foliar fertilizers (FF); control (Ctrl). The treatments underwent analyses of mixture physical-chemical characteristics, sampled caterpillar number and size, leaf gas exchange (IRGA), and amount of chlorophyll a and b. Spray liquid characteristics changed with used mixture. Fertilizer addition caused major changes in spray liquid properties as increasing pH and electrical conductivity. No effect was observed by mixing azoxystrobin, cyproconazole, zinc oxide and manganese sulfate to triflumuron concerning control effectiveness of Spodoptera frugiperda in the corn plants. Conversely, each pesticide mixture had a distinct effect on plant physiological variables.

Têm-se atualmente poucos estudos sobre os efeitos de diferentes misturas de produtos fitossanitários na cultura do milho. A mistura em tanque é prática frequente a campo, mas sem o devido respaldo técnico. Os objetivos deste trabalho foram avaliar a eficácia de diferentes misturas em tanque no controle de Spodoptera frugiperda em um híbrido de milho convencional e os possíveis efeitos fisiológicos na planta, assim como as características físico-químicas da calda. O delineamento experimental utilizado foi o de blocos ao acaso (DBC), com quatro tratamentos e cinco repetições. O experimento foi repetido em duas épocas. Os tratamentos foram constituídos de diferentes misturas de produtos fitossanitários: 1- Inseticida (in) + Adjuvante (adj); 2 ­ in + adj + Fungicida (fg); 3 ­ in + adj + fg + fertilizantes foliares; 4 ­ testemunha. Foram avaliadas características físico-químicas da calda e número e tamanho de lagartas. Também foram realizadas avaliações de trocas gasosas com analisador de gás IRGA e de quantidade de clorofilas a e b com um clorofilômetro. As características físico-químicas de calda foram alteradas pelo uso de misturas de diferentes produtos fitossanitários. A adição de fertilizantes causou as maiores mudanças, aumentando o pH e a condutividade elétrica da calda. Não houve efeito da mistura de azoxistrobina, ciproconazol, oxido de zinco e sulfato de manganês ao triflumuron na eficácia de controle de Spodoptera Frugiperda na cultura do milho. As avaliações fisiológicas (trocas gasosas e clorofila) foram afetadas de forma distinta pelas misturas de produtos fitossanitários.

Platelet Shape Changes and Cytoskeleton Dynamics as Novel Therapeutic Targets for Anti-Thrombotic Drugs

Platelets play an essential role in hemostasis through aggregation and adhesion to vascular injury sites but their unnecessary activation can often lead to thrombotic diseases. Upon exposure to physical or biochemical stimuli, remarkable platelet shape changes precede aggregation or adhesion. Platelets shape changes facilitate the formation and adhesion of platelet aggregates, but are readily reversible in contrast to the irrevocable characteristics of aggregation and adhesion. In this dynamic phenomenon, complex molecular signaling pathways and a host of diverse cytoskeleton proteins are involved. Platelet shape change is easily primed by diverse pro-thrombotic xenobiotics and stimuli, and its inhibition can modulate thrombosis, which can ultimately contribute to the development or prevention of thrombotic diseases. In this review, we discussed the current knowledge on the mechanisms of platelet shape change and also pathological implications and therapeutic opportunities for regulating the related cytoskeleton dynamics.

Low-Dose Bisphenol A Increases Bile Duct Proliferation in Juvenile Rats: A Possible Evidence for Risk of Liver Cancer in the Exposed Population?

Increasing concern is being given to the association between risk of cancer and exposure to low-dose bisphenol A (BPA), especially in young-aged population. In this study, we investigated the effects of repeated oral treatment of low to high dose BPA in juvenile Sprague-Dawley rats. Exposing juvenile rats to BPA (0, 0.5, 5, 50, and 250 mg/kg oral gavage) from post-natal day 9 for 90 days resulted in higher food intakes and increased body weights in biphasic dose-effect relationship. Male mammary glands were atrophied at high dose, which coincided with sexual pre-maturation of females. Notably, proliferative changes with altered cell foci and focal inflammation were observed around bile ducts in the liver of all BPA-dosed groups in males, which achieved statistical significance from 0.5 mg/kg (ANOVA, Dunnett’s test, p<0.05). Toxicokinetic analysis revealed that systemic exposure to BPA was greater at early age (e.g., 210-fold in C(max), and 26-fold in AUC at 50 mg/kg in male on day 1 over day 90) and in females (e.g., 4-fold in C(max) and 1.6-fold in AUC at 50 mg/kg vs. male on day 1), which might have stemmed from either age- or gender-dependent differences in metabolic capacity. These results may serve as evidence for the association between risk of cancer and exposure to low-dose BPA, especially in young children, as well as for varying toxicity of xenobiotics in different age and gender groups.

Tanshinone IIA Protects Endothelial Cells from H₂O₂-Induced Injuries via PXR Activation

Tanshinone IIA (Tan IIA) is a pharmacologically active substance extracted from the rhizome of Salvia miltiorrhiza Bunge (also known as the Chinese herb Danshen), and is widely used to treat atherosclerosis. The pregnane X receptor (PXR) is a nuclear receptor that is a key regulator of xenobiotic and endobiotic detoxification. Tan IIA is an efficacious PXR agonist that has a potential protective effect on endothelial injuries induced by xenobiotics and endobiotics via PXR activation. Previously numerous studies have demonstrated the possible effects of Tan IIA on human umbilical vein endothelial cells, but the further mechanism for its exerts the protective effect is not well established. To study the protective effects of Tan IIA against hydrogen peroxide (H₂O₂) in human umbilical vein endothelial cells (HUVECs), we pretreated cells with or without different concentrations of Tan IIA for 24 h, then exposed the cells to 400 μM H₂O₂ for another 3 h. Therefore, our data strongly suggests that Tan IIA may lead to increased regeneration of glutathione (GSH) from the glutathione disulfide (GSSG) produced during the GSH peroxidase-catalyzed decomposition of H₂O₂ in HUVECs, and the PXR plays a significant role in this process. Tan IIA may also exert protective effects against H₂O₂-induced apoptosis through the mitochondrial apoptosis pathway associated with the participation of PXR. Tan IIA protected HUVECs from inflammatory mediators triggered by H₂O₂ via PXR activation. In conclusion, Tan IIA protected HUVECs against H₂O₂-induced cell injury through PXR-dependent mechanisms.

Lymphocyte DNA damage and plasma antioxidant status in Korean subclinical hypertensive patients by glutathione S-transferase polymorphism

BACKGROUND/OBJECTIVES: Glutathione S-transferase (GST) forms a multigene family of phase II detoxification enzymes which are involved in the detoxification of xenobiotics by conjugating substances with glutathione. The aim of this study is to assess the antioxidative status and the degree of DNA damage in the subclinical hypertensive patients in Korea using glutathione S-transferase polymorphisms. SUBJECTS/METHODS: We examined whether DNA damage and antioxidative status show a difference between GSTM1 or GSTT1 genotype in 227 newly diagnosed, untreated (systolic blood pressure (BP) ≥ 130 mmHg or diastolic BP ≥ 85 mmHg) subclinical hypertensive patients and 130 normotensive subjects (systolic BP < 120 mmHg and diastolic BP < 80 mmHg). From the blood of the subjects, the degree of the DNA damage in lymphocyte, the activities of erythrocyte superoxide dismutase, the catalase, and the glutathione peroxidase, the level of glutathione, plasma total radical-trapping antioxidant potential (TRAP), anti-oxidative vitamins, as well as plasma lipid profiles and conjugated diene (CD) were analyzed. RESULTS: Of the 227 subjects studied, 68.3% were GSTM1 null genotype and 66.5% were GSTT1 null genotype. GSTM1 null genotype had an increased risk of hypertension (OR: 2.104, CI: 1.38-3.35), but no significant association in GSTT1 null genotype (OR 0.982, CI: 0.62-1.55). No difference in erythrocyte activities of superoxide dismutase, catalase, or glutathione peroxidase, and plasma TRAP, CD, lipid profiles, and GSH levels were observed between GSTM1 or GSTT1 genotype. Plasma levels of α-tocopherol increased significantly in GSTT1 wild genotype (P < 0.05); however, plasma level of β-carotene increased significantly in GSTT1 null genotype (P < 0.01). DNA damage assessed by the Comet assay was significantly higher in GSTM1 null genotype than wild genotype (P < 0.05). CONCLUSIONS: These results confirm the association between GSTM1 null genotype and risk of hypertension as they suggest that GSTM1 null genotype leads to an increased oxidative stress compared with wild genotype.

Natural carriers in bioremediation: a review

Bioremediation of contaminated groundwater or soil is currently the cheapest and the least harmful method of removing xenobiotics from the environment. Immobilization of microorganisms capable of degrading specific contaminants significantly promotes bioremediation processes, reduces their costs, and also allows for the multiple use of biocatalysts. Among the developed methods of immobilization, adsorption on the surface is the most common method in bioremediation, due to the simplicity of the procedure and its non-toxicity. The choice of carrier is an essential element for successful bioremediation. It is also important to consider the type of process ( n s tu or ex s tu), type of pollution, and properties of immobilized microorganisms. For these reasons, the article summarizes recent scientific reports about the use of natural carriers in bioremediation, including efficiency, the impact of the carrier on microorganisms and contamination, and the nature of the conducted research.

Ethanol Extract of Cirsium japonicum var. ussuriense Kitamura Exhibits the Activation of Nuclear Factor Erythroid 2-Related Factor 2-dependent Antioxidant Response Element and Protects Human Keratinocyte HaCaT Cells Against Oxidative DNA Damage

Keratinocytes are constantly exposed to extracellular insults, such as ultraviolet B, toxic chemicals and mechanical stress, all of which can facilitate the aging of keratinocytes via the generation of intracellular reactive oxygen species (ROS). Nuclear factor erythroid 2-related factor 2 (Nrf2) is a transcription factor that plays a critical role in protecting keratinocytes against oxidants and xenobiotics by binding to the antioxidant response element (ARE), a cis-acting element existing in the promoter of most phase II cytoprotective genes. In the present study, we have attempted to find novel ethanol extract(s) of indigenous plants of Jeju island, Korea that can activate the Nrf2/ARE-dependent gene expression in human keratinocyte HaCaT cells. As a result, we identified that ethanol extract of Cirsium japonicum var. ussuriense Kitamura (ECJUK) elicited strong stimulatory effect on the ARE-dependent gene expression. Supporting this observation, we found that ECJUK induced the expression of Nrf2, hemoxygenase-1, and NAD(P)H:quinone oxidoreductase-1 and this event was correlated with Akt1 phosphorylation. We also found that ECJUK increased the intracellular reduced glutathione level and suppressed 12-O-tetradecanoylphorbol acetate-induced 8-hydroxyguanosine formation without affecting the overall viability. Collectively, our results provide evidence that ECJUK can protect against oxidative stress-mediated damages through the activation of Nrf2/ARE-dependent phase II cytoprotective gene expression.

Microbiome-Linked Crosstalk in the Gastrointestinal Exposome towards Host Health and Disease / 대한소아소화기영양학회지

The gastrointestinal exposome represents the integration of all xenobiotic components and host-derived endogenous components affecting the host health, disease progression and ultimately clinical outcomes during the lifespan. The human gut microbiome as a dynamic exposome of commensalism continuously interacts with other exogenous exposome as well as host sentineling components including the immune and neuroendocrine circuit. The composition and diversity of the microbiome are established on the basis of the luminal environment (physical, chemical and biological exposome) and host surveillance at each part of the gastrointestinal lining. Whereas the chemical exposome derived from nutrients and other xenobiotics can influence the dynamics of microbiome community (the stability, diversity, or resilience), the microbiomes reciprocally alter the bioavailability and activities of the chemical exposome in the mucosa. In particular, xenobiotic metabolites by the gut microbial enzymes can be either beneficial or detrimental to the host health although xenobiotics can alter the composition and diversity of the gut microbiome. The integration of the mucosal crosstalk in the exposome determines the fate of microbiome community and host response to the etiologic factors of disease. Therefore, the network between microbiome and other mucosal exposome would provide new insights into the clinical intervention against the mucosal or systemic disorders via regulation of the gut-associated immunological, metabolic, or neuroendocrine system.

Estudo do papel das células de kupffer na modulação dos citocromos P450 hepáticos por estímulos inflamatórios

É conhecido que processos inflamatórios podem modular a expressão e atividade deenzimas CYPs. Não é claro, entretanto, o modo pelo qual estímulos inflamatórios regulam aexpressão dessas enzimas. Neste trabalho investigamos a hipótese de que as células deKupffer do fígado exerceriam um papel na modulação dos CYPs hepáticos em resposta àinflamação exacerbada ou sepse. O cloreto de gadolínio, é um inibidor seletivo das células deKupffer, conhecido por atenuar o quadro de inflamação exacerbada, quandoadministradopreviamente ao estímulo inflamatório, em diferentes modelos animais. Algunsautores sugeriram que as células de Kupffer atuariam como intermediários na modulação daatividade de CYPs hepáticos desencadeada por estímulos inflamatórios. Há estudos quesugerem que a diminuição da população das células de Kupfferatenua ou elimina a regulaçãodas CYPs hepáticas por estímulos inflamatórios.Além disso, estudos em culturas dehepatócitos in vitro, na ausência de células de Kupffer, tem constatado a regulação negativa daexpressão de CYPs hepáticas após estimulação com LPS. Nessa linha, o objetivo destepresente trabalho é investigar o papel das células de Kupffer na regulação da atividade deenzimas hepáticas de biotransformação de xenobióticos (CYPs) após estimulação inflamatóriacom LPS. Para isso, os níveis séricos de transaminases, e a histopatologia foram empregadospara avaliar o efeito do tratamento com diferentes doses de GdCl3sobre o tecido hepático.Noexperimento principal para investigar o papel das células de Kupffer, os ratos foram alocadosao acaso em quatro grupos. Foram quantificadosmarcadores bioquímicos no soro dos animaispara evidenciar danos ao tecido hepático causados pelos tratamentos e realizado o examehistopatológico...

It is known that inflammatory processes may modulate the expression and activity ofCYP enzymes. The mode by which inflammatory stimuli regulate CYP expression andactivity, however, remains unclear. Kupffer cells are resident macrophages in the liver andthus play an important role in a systemic inflammatory process or in sepsis. Gadoliniumchloride (GdCl3) has been reported to selectively kill an/or inhibit the activity of Kupffercells. Along this line, it has also been described that Gd decreases exacerbated inflammatoryresponses when it is administered prior to inflammatory stimuli in various animal models.Therole of Kupffer cells in the modulation of CYPs activity triggered by inflammatory stimuli,however, is not entirely clear in the literature. There are studies suggesting that a reduction inthe population of Kupffer cells attenuates the down-regulation of hepatic CYPs induced byinflammatory stimuli. However, GdCl3 was also described to decrease liver CYP activityirrespective of whether it depletes or not Kupffer cells. Moreover, in vitro studies showed thatLPS down-regulates the expression of CYP forms in hepatocyte cell lines in culture (in theabsence of Kupffer cells). To investigate whether Kupffer cells play a role in the regulation ofthe activity of liver xenobiotic biotransformation enzymes (CYPs) by inflammatorystimulation with LPS. The activity of transaminases in the blood serum (a marker for liverinjury) was determined and liver histopathology was evaluated in female Wistar rats. In a setof preliminary tests, rats were treated with different doses of GdCl3 or with LPS for selectingdoses and euthanasia time in the main experiment. In the main study experimental groups.Treatment associated liver injury was evaluated by levels of transaminases and alkalinephosphatase in the blood serum and by liver histopathology examination...

Estudo experimental da exposição ao PCB126 sobre a indução de Diabetes mellitus tipo II / Experimental study of PCB126 exposure on induction of Diabetes mellitus type II

A exposição ambiental aos poluentes orgânicos persistentes tem recebido amplo destaque na literatura recentemente devido à extensa associação entre o desenvolvimento de doenças metabólicas, obesidade e/ou diabetes mellitus, e a presença destes poluentes, principalmente os organoclorados, como as bifenilas policloradas (PCBs), no organismo. Por outro lado, os mecanismos de ação destes poluentes é controverso devido à elevada quantidade de representantes destas classes, gerando diversidade de protocolos de exposição e escassez de estudos experimentais. Por isto, foi objetivo deste trabalho elucidar os mecanismos de ação tóxica do PCB126, nas doses de 0,1; 1 ou 10 µg/kg de massa corpórea, em ratos Wistar machos, durante quinze dias, expostos por instilação intranasal. O procotolo de exposição empregado foi caracterizado e considerado suficiente para causar toxicidade, uma vez que foram observadas alterações no sistema imune, metabolismo e em parâmetros relacionados à gênese do diabetes mellitus. A caracterização da exposição foi determinada pela quantificação da concentração de PCB126 no fígado e pulmão (CG/MS) e pelo aumento da expressão do receptor aril hidrocarboneto (AhR) no rim, fígado, pulmão e tecido adiposo (Western Blot). O efeito imunossupressor do PCB126 foi evidenciado pelo comprometimento da produção de células na medula óssea e, consequentemente, no número de células totais no sangue circulante. Adicionalmente, foi evidenciada a interferência do poluente na via de ativação mediada por receptores acoplados à proteína G (GPCRs), principalmente em neutrófilos, alterando importantes funções destas células, como a expressão de moléculas de adesão, geração de espécies reativas de oxigênio e migração. Entre as alterações metabólicas observadas, destacamos o aumento dos níveis de triglicerídeos e colesterol sérico, aumento da liberação de ácidos graxos livres; aumento da atividade da enzima hepática gama glutamil transferase; aumento da resistência à insulina e aumento da geração de óxido nítrico pelas ilhotas de Langerhans, dados estes, possivelmente relacionados ao comprometimento das células beta (ß) pancreáticas, confirmados pelo aumento da expressão de GLUT4 no tecido adiposo, aumento da concentração de insulina sérica e aumento do estresse oxidativo nas ilhotas de Langerhans. Em conjunto, os dados obtidos destacam importantes alterações causadas pela exposição intranasal ao PCB126, evidenciando a participação do poluente na gênese do diabetes mellitus do tipo II

The environmental exposure to persistent organic pollutants has been widely highlighted in recent literature due to the extensive association between the development of metabolic diseases, obesity and/or diabetes mellitus, and presence of these pollutants, especially organochlorines such as polychlorinated biphenyls (PCBs) in organism. Moreover, the mechanisms of action of these pollutants are controversial due to the high number of PCBs congeners, diversity of exposure protocols and lack of experimental studies. Therefore, the aim of this study was to elucidate the mechanisms of PCB126's toxic action at doses of 0.1; 1 or 10 µg/kg body weight in male Wistar rats exposed by intranasal instillation for 15 days. The established exposure procotol was characterized and considered sufficient to cause toxicity since changes were observed in the immune system, metabolism and in parameters related to the pathogenesis of diabetes mellitus. Characterization of exposure was determined by quantifying the concentration of PCB126 in liver and lung (GC-MS) and by the increased expression of aryl hydrocarbon receptor (AhR) in kidney, liver, lung, and adipose tissue (Western blot). The immunosuppressive effect of PCB126 was evidenced by impairment of cell production in the bone marrow and thus the total number of cells in the circulation. In addition, the interference of the pollutant in the activation pathway mediated by G-protein coupled receptors (GPCRs), in particular in neutrophils, was observed by changing important functions of these cells such as the expression of adhesion molecules, reactive oxygen species generation, and migration. Among the metabolic changes observed, we highlight the increased levels of triglycerides and serum cholesterol, increased release of free fatty acids; increased gamma glutamyl transferase hepatic enzyme activity; increased insulin resistance and increased generation of nitric oxide by the islets of Langerhans, these data possibly related to the impairment of beta cells (ß) pancreatic function, suggested by the increased expression of GLUT4 in adipose tissue, increased serum insulin concentration and increased oxidative stress in the islets of Langerhans. Altogether, these results highlight important changes caused by intranasal exposure to PCB126, suggesting participation of the pollutant in the genesis of diabetes mellitus type II

Biodegradation of kerosene: Study of growth optimization and metabolic fate of P. janthinellum SDX7

Penicillum janthinellum SDX7 was isolated from aged petroleum hydrocarbon-affected soil at the site of Anand, Gujarat, India, and was tested for different pH, temperature, agitation and concentrations for optimal growth of the isolate that was capable of degrading upto 95%, 63% and 58% of 1%, 3% and 5% kerosene, respectively, after a period of 16 days, at optimal growth conditions of pH 6.0, 30 °C and 180 rpm agitation. The GC/MS chromatograms revealed that then-alkane fractions are easily degraded; however, the rate might be lower for branched alkanes, n-alkylaromatics, cyclic alkanes and polynuclear aromatics. The test doses caused a concentration-dependent depletion of carbohydrates of P. janthinellum SDX7 by 3% to 80%, proteins by 4% to 81% and amino acids by 8% to 95% upto 16 days of treatment. The optimal concentration of 3% kerosene resulted in the least reduction of the metabolites of P. janthinellum such as carbohydrates, proteins and amino acids with optimal growth compared to 5% and 1% (v/v) kerosene doses on the 12th and 16th day of exposure. Phenols were found to be mounted by 43% to 66% at lower and higher concentrations during the experimental period. Fungal isolate P. janthinellum SDX7 was also tested for growth on various xenobiotic compounds.

Regulation of organic anion transporting polypeptides expression and activity / 药学学报

Organic anion transporting polypeptides (OATP), a member of solute carrier (SLC) superfamily, is considered as an important transmembrane uptake transporters. OATP is involved in the transport of a variety of endo- and xenobiotics (bile acids, bilirubin, prostaglandin, thyroid hormones, steroid hormone conjugates), drugs and toxins in a Na+ and ATP independent manner. Multiple factors (eg. hormones, proinflammatory cytokines, drugs) can affect the distribution, expression and activity of OATPs, leading to an altered accumulation of OATP substrates and related food-drug and drug-drug interactions. Changes in the distribution and expression of OATPs in malignant tissues may be related to the pathological process of cancer, while the modulation epigenetic mechanism also contributes to its distribution patterns. This review describes the factors that can affect the expression or function of OATPs, which may provide a valuable reference for drug development and the clarification of pathogenesis.