Lowered Antioxidant Status of Red Blood Cells in Post-Parturient Haemoglobinuria of Buffaloes

The antioxidant status of the red blood cells of buffaloes (n = 20) suffering from post-parturient haemoglobinuria (PPH) was assessed by comparing their tocopherol (vitamin E) and reduced glutathione contents with those of red blood cells from apparently healthy buffaloes (n = 20). The red cell tocopherol content of the diseased buffaloes (1.76±0.11 g/ml) was significantly (p<0.01) lower than that of healthy buffaloes (2.45±0.14 g/ml). This may be the first report comparing the concentration of tocopherol in the red blood cells of buffaloes suffering from PPH and apparently healthy buffaloes.There was a drastic reduction in the reduced glutathione content in the red cells of haemoglobinuric buffaloes (23.74±2.86 mg%) compared to the healthy control buffaloes (73.71±3.87 mg%). The diseased buffaloes also exhibited severe hypophosphataemia. These findings suggest that an impaired or insufficient antioxidant potential of the red blood cells in this disease in buffaloes is associated with the phosphorus deficiency.     

巨尾桉EuGR1基因的克隆及其低温胁迫下的表达特性分析

本研究通过反转录PCR方法克隆了巨尾桉谷胱甘肽还原酶（Glutathione reductase,简称GR）基因,该基因定位于巨尾桉细胞质、长度为1485 bp,在NCBI申请基因注册（GenBank Accession Number KU904639）。构建了pET-EuGR1原核表达载体,酶活检测表明转化菌株GR活性较高。利用实时定量 PCR分析巨尾桉EuGR1的时空表达特性,结果表明：EuGR1的表达量随着巨尾桉叶片的成熟而降低,在幼叶中表达量最大,近根部的叶片表达量最低;在巨尾桉组培苗中,EuGR1在茎中表达量最大,叶中表达量较低,根中表达量最低。通过转基因抗寒巨尾桉温室苗研究EuGR1基因在低温胁迫下的表达模式,发现常温和低温胁迫下耐寒性强的株系其EuGR1的表达量较高（如P40、P41、P52）,耐寒性弱的株系其EuGR1的表达量较低（如P36、F44、F76）,说明巨尾桉EuGR1的表达量与植株的抗寒能力具有相关性。随着低温胁迫时间的延长,EuGR1的表达量在36 h后出现降低趋势,表明EuGR1在桉树耐低温胁迫的前期发挥作用较大。     

Comparison of the effect of Roundup Ultra 360 SL pesticide and its active compound glyphosate on human erythrocytes

The effects of exposure of human erythrocytes to different concentrations of Roundup Ultra 360 SL and its active compound glyphosate were studied. We studied hemolysis after 1, 5, and 24 h incubation; lipid peroxidation, hemoglobin oxidation, the level of reduced glutathione, and the activity of catalase after 1 h. Human erythrocytes were incubated with 100-1500 ppm (100 μg/ml erythrocytes at 5% hematocrite) Roundup Ultra 360 SL and glyphosate. We have found that after 1 h of incubation only Roundup Ultra 360 SL increased the level of methemoglobin, products of lipid peroxidation at 500 ppm and hemolysis at 1500 ppm [Curr. Top. Biophys. 26 (2002) 245], while its active compound glyphosate increased the level of methemoglobin and the level of lipid peroxidation at much higher dose—1000 ppm. At the same time hemolysis was observed to only at the highest dose of glyphosate (1500 ppm) and the longest time of incubation (24 h). Both Roundup Ultra 360 SL and glyphosate did not cause statistically significant changes in the level of GSH, but increased the activity of catalase. Roundup Ultra 360 SL provokes more changes in the function of erythrocytes than its active substance glyphosate, which is probably a result of the properties of additives. Taking into account the limited accumulation of Roundup Ultra 360 SL and glyphosate in the organism as well as the fact that the threshold doses which caused changes in erythrocytes for Roundup Ultra 360 SL were only 500 and 1000 ppm for glyphosate, one may conclude that this pesticide is safe towards human erythrocytes.     

寄主植物对棉铃虫体内解毒酶活性的影响

通过离体酶活性测定 ,研究取食小麦、豌豆等寄主植物和人工饲料对棉铃虫羧酸酯酶、谷胱甘肽 S-转移酶和乙酰胆碱酯酶活性的影响。结果表明取食不同寄主植物的棉铃虫体内羧酸酯酶活性不仅存在量的差异 ,而且存在质的差异 ,寄主植物对棉铃虫谷胱甘肽 S-转移酶、乙酰胆碱酯酶活性有显著影响     

Mapping of genes involved in glutathione, carbohydrate and COR14b cold induced protein accumulation during cold hardening in wheat

Using some of the chromosome substitution lines developed from thecrosses of the donor Cheyenne to Chinese Spring we showed that theaccumulation of water soluble carbohydrates during different stages ofhardening was time dependent. Moreover there was a significantcorrelation between the rate of carbohydrate accumulation and the frosttolerance. The expression and regulation of a wheat gene homologous tothe barley cold regulated cor14b gene was compared in frost sensitiveand frost tolerant wheat genotypes at different temperatures. Studies madewith chromosome substitution lines showed that the threshold inductiontemperature polymorphism of the cor14b wheat homologous genewas controlled by loci located on chromosome 5A of wheat, while cor14b gene was mapped, in Triticum monococcum, onto the longarm of chromosome 2A^m. Our study on the effect of cold hardeningon glutathione (GSH) metabolism showed that chromosome 5A of wheathas an influence on the GSH accumulation and on the ratio of reduced andoxidised glutathione as part of a complex regulatory function during coldhardening. In addition, the level of increase in GSH content duringhardening may indicate the degree of the frost tolerance of wheat.     

Characterization of hepatic and extrahepatic glutathione S-transferases in rainbow trout (Oncorhynchus mykiss) and their induction by 3,3′,4,4′-tetrachlorobiphenyl

Liver, kidney, gill and olfactory epithelium cytosolic fractions of rainbow trout (Oncorhynchus mykiss) were examined for glutathione S-transferase (GST) contents. Proteins retained on a glutathione (GSH)-affinity matrix were separated as monomers by reversed-phase HPLC and characterized by immunoblotting, mass spectrometry and partial amino acid sequence. For each organ concerned, a specific pattern of these proteins was determined and appeared similar for liver and kidney on one hand, and for gill and olfactory epithelium on the other hand. It was confirmed that the prominent hepatic GST is a class enzyme, also constitutively expressed as a major isoform in the four organs studied. Moreover, a class variant and two new class GST subunits were characterized in minor fractions. An unknown protein, which was found major in gills and olfactory epithelium, exhibited some characteristics of class GSTs. Occurrence of possible GSH-adduct formation observed on two distinct monomers in specific experimental conditions is discussed. These results and methods were used to investigate the effect of 3,3,4,4-tetrachlorobiphenyl (TCB), a polychlorinated biphenyl (PCB), on GST expression in trout liver. From HPLC-profiling, significant co-induction of the major class and the two minor class GST subunits was observed in trout after waterborne exposure to TCB which was followed by a slight increase in 1-chloro-2,4-dinitrobenzene (CDNB) activity. The present work allows qualitative evaluation of the specific detoxification potential of rainbow trout. The use of HPLC-profiling of GSTs as a possible tool for the biomonitoring of polluted aquatic environment is suggested.     

Sulphur accumulation after Verticillium dahliae infection of two pepper cultivars differing in degree of resistance

Elemental sulphur levels, sulphur localization in stems, and levels of sulphate, glutathione and cysteine were studied in pepper ( Capsicum annuum ) cvs Yolo Wonder (higher resistance) and Luesia (lower resistance) after inoculation with Verticillium dahliae , the cause of vascular wilt. Accumulation of elemental sulphur (S⁰) was first detected 10 days after inoculation in Yolo Wonder (mean S⁰ level 7·3  µ g g⁻¹ DW), and 15 days after inoculation in Luesia (mean S⁰ level 3·3  µ g g⁻¹ DW). The maximum level was reached 21 days after inoculation in Yolo Wonder (14·1  µ g g⁻¹ DW). In control plants, elemental sulphur was not detected. SEM-EDX (scanning electron microscopy-energy dispersive X-ray microanalysis) indicated that the sulphur was not restricted to a specific location, but was dispersed throughout the vascular tissue. Sulphate levels showed a decline at the end of the experiment in inoculated plants, possibly related to the increase in sulphur levels seen in the two cultivars. The differences in sulphate levels between the two cultivars may be due to faster sulphate breakdown in cv. Yolo Wonder.     

The effects of dietary oxidized fat and selenium source on performance,glutathione peroxidase,and glutathione reductase activity in broiler chickens

Normal or elevated selenium status of broilers, which is influenced by dietary selenium sources, improves the bird’s ability to overcome the adverse effects of reactive oxygen metabolites. The objective of this study was to evaluate the effects of feeding graded levels of peroxidized poultry fat on blood and hepatic glutathione peroxidase (GSH-Px), and hepatic glutathione reductase activity in broiler chickens fed either inorganic sodium selenite (SEL) or organic selenium enriched in the organic selenium yeast product Sel-Plex (SP). Nine starter diets, varying in levels of oxidized fat (0, 3, and 6 mEq/kg) and dietary selenium sources, were fed to 360 male chicks from hatch to 21 d of age. Sel-Plex or SEL was added to the basal diet to provide either 0 or 0.2 ppm of supplemental selenium in the diets. Blood and hepatic samples were obtained for each treatment group at 21 d of age. Neither peroxidized fat nor selenium source significantly altered the activity of hepatic glutathione reductase (P ≤ 0.05). Blood GSH-Px was influenced significantly by both fat and selenium source (P ≤ 0.05), but the fat × selenium source interaction was not significant (P ≥ 0.3). A selenium source effect on the hepatic GSH-Px activity (P ≤ 0.05) was evidenced by higher GSH-Px activity, even in the basal diet with no added peroxidized fat. An increase in GSH-Px activity was seen in the erythrocyte and hepatic samples in both the SEL and SP treatments when peroxidized fat was given at 3 mEq/kg, but in the erythrocytes and in the hepatic tissues from SEL-supplemented birds, there was an apparent inhibition of GSH-Px activity. This inhibition was not seen in the hepatic tissue samples from SP-fed birds. Because elevated GSH-Px activity is indicative of oxidative stress, it was concluded that dietary SP supplementation resulted in better selenium and redox status in broilers than did SEL. These results indicate that the dietary selenium supplied in an organic form (selenium yeast as SP) improved the selenium and redox status in broilers, leading to greater resistance to oxidative stress than when the inorganic form of selenium (SEL) was fed.