Effect of latex material on antioxidant enzymes,lipid peroxidation,DNA damage,and chromosomal aberration

Cell integrity is affected by oxidative stress when the production of active oxidants overwhelms antioxidant defense mechanisms. Latex, a natural polymer obtained from Hevea brasiliensis, is used in medical industry for manufacturing surgical gloves, urinary catheters, and dental dams. The aim of this study was to evaluate the effects of latex material on oxidative stress by in vivo and in vitro methods. In addition, the material was screened for its ability to induce any chromosomal aberrations (CAs) by in vitro method. In vivo studies were carried out with implanted latex material onto subcutaneous tissue of various batches of experimental Wistar rats. At the end of experimental period, animals were anesthetized, blood was collected for serum analysis, and sacrificed. Liver was excised for the determination of antioxidant enzymes and lipid peroxidation (LPO). Subcutaneous tissues were obtained for the extraction of genomic DNA from implanted animals and checked for the presence of 8-hydroxy-2-deoxyguanosine (8-OHdG), considered an indicator of DNA damage. Simultaneously, in vitro studies were carried out using fresh liver and subcutaneous tissue obtained from Swiss albino mice treated with physiological saline extract of latex material. For the estimation of both in vitro and in vivo oxidative stress, 10% liver homogenate was assessed for stress indicators like reduced glutathione, glutathione reductase, glutathione peroxidase, LPO and protein content. The results of both in vivo and in vitro studies indicated that the chemical leachents from the latex material did not significantly affect LPO and the levels of antioxidant enzymes. There was also no significant increase in 8-OHdG content due to the presence of implanted latex material. Finally, the results of in vitro CA test and G banding indicated that extracts of test material did not induce any chromosomal abnormalities.     

海洋双壳类谷胱甘肽S-转移酶活性测定条件的优化

以僧帽牡蛎(Ostrea cucullata)为材料,提取鳃的谷胱甘肽S-转移酶(glutathione s-transferase, GST),测定GST(包括GSTs、GST-π、GST- μ)在不同温度、底物浓度、缓冲液pH条件下的活性,从而确定酶活性测定的最佳条件.结果表明:(1) GSTs活性测定的最佳条件:GSH终浓度2 mmol/L,测定温度31℃,PBS pH 6.5,CDNB终浓度1.5 mmol/L;(2) GST-π活性测定的最佳条件:GSH终浓度2.5 mmol/L,测定温度为29℃,PBS pH7.75,ECA的终浓度2.5 mmol/L;(3) GST-μ活性测定的最佳条件:GSH的终浓度5 mmol/L,外界环境温度为29 ℃,PBS pH 6.0,DCNB的终浓度1.50 mmol/L.     

Catalyzed degradation of disperse dyes by calcium alginate-pectin entrapped bitter gourd （Momordica charantia） peroxidase

Calcium-alginate pectin entrapped bitter gourd peroxidase (BGP) has been employed for the treatment of disperse dyes: Disperse Brown 1 (DB 1) and Disperse Red 17 (DR 17). Peroxidase alone was unable to decolorize DR 17 and DB 1. However, the investigated dyes were decolorized maximally by BGP in the presence of 0.2 mmol/L redox mediator, violuric acid (VA). A slow decrease in percent decolorization was observed when VA concentration was higher than 0.2 mmol/L which could likely be due to the high reactivity of its aminoxyl radical (> N–O.) intermediate, that might undergo chemical reactions with aromatic amino acid side chains of the enzyme thereby inactivating it. Maximum decolorization of the dyes was observed at pH 3.0 and 40°C within 2 hr of incubation. Immobilized peroxidase decolorized 98% DR 17 and 71% DB 1 using 35 U of BGP in batch process in 90 min. Immobilized enzyme decolorized 85% DR 17 and 51% DB 1 whereas soluble enzyme decolorized DR 17 to 48% and DB 1 to 30% at 60°C. UV-visible spectral analysis was used to evaluate the degradation of these dyes and their toxicity was tested by Allium cepa test. The generally observed higher stability of the bioaffinity bound enzymes against various forms of inactivation may be related to the specific and strong binding of enzyme with bioaffinity support which prevents the unfolding/denaturation of enzyme. Thus entrapped peroxidase was found to be effective in the decolorization of the investigated dyes.     

模拟酸雨对龙眼叶绿体的伤害效应

龙眼叶绿体超氧化的歧化酶（SOD）和抗坏血酸过氧化物酶（AsA-POD)活性，谷胱甘肽（GSH）和抗坏血酸含量（AsA)含量随酸雨的pH值下降而降低，膜脂质过氧化产物丙二醛（MDA）的含量则随酸雨的pH下降而增加。pH3.0的酸雨胁迫2h，SOD和AsA-POD活性，GSH和AsA含量有所增加，并随胁迫时间的延长而下降。pH3．5的酸雨处理使类整体膜崩解，叶绿体基粒片层结构混乱；pH3.0的酸雨处理的叶绿体片层结构伤害更为严重。图5图版2参21     

酿酒酵母提取物对废天然橡胶再生的研究

酿酒酵母R1在5L发酵罐中培养36h时,胞内谷胱甘肽(GSH)总量达304mg/L。将酵母菌充分裂解,提取物与废天然橡胶共反应。通过单因素对比实验,确定最适宜的反应条件如下:温度50℃,时间4h,100g胶粉,溶剂用量1.85mol,相转移催化剂用量2.75×10-2mol,酵母菌用量相当于GSH1.0g。处理后最高溶胀度为4.37,是原来的1.49倍。     

Brassinosteroid alleviates phenanthrene and pyrene phytotoxicity by increasing detoxification activity and photosynthesis in tomato

The present study was carried out to investigate the effects of exogenously applied 24-epibrassinolide (BR) on growth, gas exchange, chlorophyll fluorescence characteristics, lipid peroxidation and antioxidant systems of tomato seedlings grown under different levels (0, 10, 30, 100 and 300 μM) of phenanthrene (PHE) and pyrene (PYR) in hydroponics. A concentration-dependent decrease in growth, photosynthetic pigment contents, net photosynthetic rate (Pn), stomatal conductance (Gs), maximal quantum yield of PSII (Fv/Fm), effective quantum yield of PSII (Φ_PSII), photochemical quenching coefficient (qP) has been observed following PHE and PYR exposure. By contrast, non-photochemical quenching coefficient (NPQ) was increased. PHE was found to induce higher stress than PYR. However, foliar or root application of BR (50 nM and 5 nM, respectively) alleviated all those depressions with a sharp improvement in the activity of photosynthetic machinery. The activities of guaicol peroxidase (GPOD), catalase (CAT), ascorbate peroxidase (APX) and glutathione reductase (GR) as well as content of malondialdehyde (MDA) were increased in a dose-dependent manner under PHE or PYR treatments. Compared with control the highest increments of GPOD, CAT, APX, GR and MDA by PHE/PYR alone treatments were observed following 300 μM concentration, which were 67%, 87%, 53%, 95% and 74% by PHE and 42%, 53%, 30%, 86% and 62% by PYR, respectively. In addition, both reduced glutathione (GSH) and oxidized glutathione (GSSG) were induced by PHE or PYR. Interestingly, BR application in either form further increased enzymatic and non enzymatic antioxidants in tomato roots treated with PHE or PYR. Our results suggest that BR has an anti-stress effect on tomato seedlings contaminated with PHE or PYR and this effect is mainly attributed by increased detoxification activity.     

Oral administration of potassium bromate, a major water disinfection by-product, induces oxidative stress and impairs the antioxidant power of rat blood

Potassium bromate (KBrO₃) is a widely used food additive, a water disinfection by-product and a known nephrotoxic agent. The effect of KBrO₃ on rat blood, especially on the anti-oxidant defense system, was studied in this work. Animals were given a single oral dose of KBrO₃ (100 mg/kg body weight) and sacrificed 12, 24, 48, 96 and 168 h after this treatment. Blood was collected from the animals and separated into plasma and erythrocytes. KBrO₃ administration resulted in increased lipid peroxidation, protein oxidation, hydrogen peroxide levels and decreased the reduced glutathione content indicating the induction of oxidative stress in blood. Methemoglobin levels and methemoglobin reductase activity were significantly increased while the total anti-oxidant power was greatly reduced upon KBrO₃ treatment. Nitric oxide levels were enhanced while vitamin C concentration decreased in KBrO₃ treated animals. The activities of major anti-oxidant enzymes were also altered upon KBrO₃ treatment. The maximum changes in all these parameters were 48 h after the administration of KBrO₃ and then recovery took place. These results show for the first time that KBrO₃ induces oxidative stress in blood and impairs the anti-oxidant defense system. Thus impairment in the anti-oxidant power and alterations in the activities of major anti-oxidant enzymes may play an important role in mediating the toxic effects of KBrO₃ in the rat blood. The study of such biochemical events in blood will help elucidate the molecular mechanism of action of KBrO₃ and also for devising methods to overcome its toxic effects.     

固定化多环芳烃降解酶——谷胱甘肽S-转移酶的研究

以自制壳聚糖为载体,戊二醛为交联剂固定谷胱甘肽S-转移酶,研究了交联剂浓度、酶用量、pH值、温度及时间等因素对固定化酶活性的影响.结果表明,戊二醛浓度0.5%,pH7.0,温度20℃,时间12h以及液态酶与壳聚糖凝胶1:1配比是谷胱甘肽S-转移酶最佳固定化条件,固定化酶催化菲降解的最佳反应时间是9h.另外,固定化酶与游离酶相比,稳定性和可操作性都有较大的提高.     

谷胱甘肽对甲醛所致Hela细胞毒性的影响

为研究还原型谷胱甘肽(GSH)对甲醛所致Hela细胞毒性的影响,采用体外染毒方式,应用KC1-SDS法和MTT法分别检测GSH对甲醛引起Hela细胞DNA-蛋白质交联(DPC)和细胞活性的影响.结果表明,250μmol·L-1浓度下,DNA-蛋白质交联实验中甲醛染毒组所致的DPC显著高于对照组(p＜0.01),GSH+...     

In vitro Methylation of Arsenite in Flemish giant rabbit Cytosol

The formation of ⁷⁴As‐monomethyIarsonic acid and ⁷⁴As‐dimethylarsinic acid from carrier‐free radiolabelled ⁷⁴As‐arsenite was evaluated in an assay mixture containing 17.6% liver cytosol from the Flemish Giant rabbit. The optimal incubation temperature and pH were respectively, 39°C and 7.6. After a 2h incubation about 90% of ⁷⁴As was protein bound. Protein bound ⁷⁴As was released by hot 2 M HNO₃ (1 min, 110°C). The treatment did not destroy methylated As‐species. Up to 70% of the carrier‐free ⁷⁴As‐arsenite was methylated. Liver cytosol was stored, without loss of activity, in liquid nitrogen in the presence of 2 mM glutathione. The optimal s‐adenosyl‐methionine concentration was 1.7mM. Formation of ⁷⁴As‐monomethylarsonic acid and ⁷⁴As‐dimethylarsinic acid increased up to a glutathione concentration of respectively 10 and 20 mM. Methylation also went through in the presence of other reducing agents and proved to be ATP dependent.