Effects of allelochemicals on activity of nitrate reductase

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Methylenetetrahydrofolate reductase deficiency: Prenatal diagnosis and family studies

Prenatal diagnosis of methylenetetrahydrofolate reductase (MTHFR) deficiency and family studies were performed because of a severely affected first child in this family. The fetus at risk was found to be heterozygous as confirmed by the enzymatic activity assay performed several times after birth. In the father, MTHFR activity was normal in lymphocytes and decreased in fibroblasts, whereas in the asymptomatic mother, the activity was not detectable in fibroblasts and was very low in lymphocytes. The absence of any clinical symptoms in the mother despite a clear MTHFR deficiency and hyperhomocystinemia emphasizes the heterogeneity of this disease.     

Properties of mercuric reductase from a HgCl_2-resistant fungus

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An unusually rapid light-induced nitrate reductase mRNA pulse and circadian oscillations

We thank Dr. G. Gowry for the cDNA clone, and Dr. W. H. Campbell for the antibodies. This project is supported by grants from the Norwegian Research Councils NLVF and NAVF.     

α-萘酚对小球藻谷胱甘肽及其还原酶的影响

研究了--萘酚对2种小球藻谷胱甘肽-包括还型谷胱甘肽GSH与氧化型谷胱甘肽GSSG-含量及其还原酶-GR-活性的影响.结果表明,低浓度--萘酚对藻细胞的谷胱甘肽水平和GR活性有显著的激发作用,随着--萘酚浓度的升高,普通小球藻和蛋白核小球藻的GSH含量、谷胱甘肽总量及GR活性均有所提高,并分别在5mg/L和10mg/L时达到最大值,而GSSG均不断下降且在相同浓度下-2 mg/L-至最小后开始上升.GSSG/GSH先降后升的变化说明藻细胞在--萘酚胁迫下膜脂过氧化加剧,而GSH、GR在清除活性氧、消除过氧化方面起了重要作用.     

a-萘酚对小球藻谷胱甘肽及其还原酶的影响

研究了a-萘酚对2种小球藻谷胱甘肽(包括还型谷胱甘肽GSH与氧化型谷胱甘肽GSSG)含量及其还原酶(GR)活性的影响.结果表明,低浓度a-萘酚对藻细胞的谷胱甘肽水平和GR活性有显著的激发作用,随着a-萘酚浓度的升高,普通小球藻和蛋白核小球藻的GSH含量、谷胱甘肽总量及GR活性均有所提高,并分别在5mg/L和10mg/L时达到最大值,而GSSG均不断下降且在相同浓度下(2mg/L)至最小后开始上升.GSSG/GSH先降后升的变化说明藻细胞在a-萘酚胁迫下膜脂过氧化加剧,而GSH、GR在清除活性氧、消除过氧化方面起了重要作用     

活性污泥中一氧化氮还原酶的提取和测定优化

通过正交实验,研究了超声破碎法和低温高压破碎法提取一氧化氮还原酶（nitric oxide reductase,nor）的活性,分析了超声强度和次数,破碎压力和次数,裂解液的添加量等因素对于胞内可溶性蛋白和核酸（DNA）的释放与提取nor活性的影响.根据活性污泥中酶的催化特性以及其他实验条件,完善了nor催化活性的测定方法.结果表明超声法提取nor的参数应设定为,超声次数为100次,超声强度为500W,裂解液的添加量为0.1mL.低温高压破碎法提取nor的参数应设定为破碎次数为4次,破碎压力为50MPa,裂解液的添加量为0.1mL.低温高压破碎法下的胞内可溶性蛋白和DNA的释放量与nor的最大催化活性要高于超声破碎法.此外,在nor催化活性的测定方法中,nor活性测定终点时间应为15min.     

Prenatal determination of dihydropteridine reductase in a normal fetus at risk for malignant hyperphenylalaninemia

Assay of dihydropteridine reductase in amniotic cells from a fetus at risk for malignant hyperphenylalaninemia is described. A normal result was obtained, this was confirmed after delivery. On two-dimensional polyacrylamide gel electrophoresis the enzyme from amniotic cells showed the same mobility as the mature liver reductase.     

吸入甲醛引起的小鼠肺部GSNO还原酶基因转录上调

为了验证吸入甲醛及气道氧化还原状态变化可在转录水平调控GSNOR表达,研究了甲醛环境暴露和抗氧化剂α-硫辛酸对小鼠肺中GSNOR表达的影响.以昆明系小鼠为实验材料,经吸人甲醛染毒和α-硫辛酸注射后立即脱颈处死,取其肺组织提取总RNA,采用RT-PCR二步法将其中GSNOR及管家基因β-actin扩增后,通过光密度分析染毒前后基因转录水平的变化.实验结果表明,与对照组相比,3.0 mg·m-3浓度的甲醛可使GSNOR基因转录发生显著上调(P＜0.01),而α-硫辛酸的注射可拮抗这种上调作用,表明吸入甲醛可在转录水平诱导GSNOR表达上调,而且这种上调与肺部的氧化还原状态改变有关,这可能是GSNOR在气道表达及其在哮喘发生中的作用基础.     

Characterization of arsenate transformation and identification of arsenate reductase in a green alga Chlamydomonas reinhardtii

Arsenic (As) is a pervasive and ubiquitous environmental toxin that has created catastrophic human health problems world-wide. Chlamydomonas reinhardtii is a unicellular green alga, which exists ubiquitously in freshwater aquatic systems. Arsenic metabolism processes of this alga through arsenate reduction and sequent store and efflux were investigated. When supplied with 10 mol/L arsenate, arsenic speciation analysis showed that arsenite concentration increased from 5.7 to 15.7 mg/kg dry weight during a 7-day period, accounting for 18%–24% of the total As in alga. When treated with different levels of arsenate (10, 20, 30, 40, 50 mol/L) for 7 days, the arsenite concentration increased with increasing external arsenate concentrations, the proportion of arsenite was up to 23%–28% of the total As in alga. In efflux experiments, both arsenate and arsenite could be found in the efflux solutions. Additionally, the efflux of arsenate was more than that of arsenite. Furthermore, two arsenate reductase genes of C. reinhardtii (CrACR2s) were cloned and expressed in Escherichia coli strain WC3110 (ΔarsC) for the first time. The abilities of both CrACR2s genes to complement the arsenatesensitive strain were examined. CrACR2.1 restored arsenate resistance at 0.8 mmol/L. However, CrACR2.2 showed much less ability to complement. The gene products were demonstrated to reduce arsenate to arsenite in vivo. In agreement with the complementation results, CrACR2.1 showed higher reduction ability than CrACR2.2, when treated with 0.4 mmol/L arsenate for 16 hr incubation.     

东湖沉积物中dNaR活性和硝酸盐还原菌的垂向分布

测定了武汉东湖污染负荷不同的3个点位(污染负荷为3号>2号>1号)柱状沉积物中异化性硝酸还原酶(dNaR)活性以及硝酸盐还原菌数量、TOC、TN、NO3--N的垂向变化,分析了dNaR活性与硝酸盐还原菌数量、TOC、TN、NO3--N的相关性.结果表明,不同点位沉积物中dNaR活性不同,污染负荷最高的3号点位dNaR活性最高,污染负荷最低的1号点活性最低.不同点位沉积物中dNaR活性在垂向分布上具有相似特征:0~15cm沉积物中活性较大,15cm以下随着深度的增加,酶活性迅速降低.硝酸盐还原菌数量在垂向分布上随着深度的增加而减小. dNaR活性与TOC、TN不具有相关性,而与NO3--N呈正相关(P<0.05);dNaR活性与硝酸盐还原菌数量呈显著性相关(P<0.01).     

不同氮源对海洋卡盾藻生长和硝酸还原酶活性的影响

在实验室条件下,研究了不同氮源对海洋卡顿藻(Chattonella marina)生长和藻细胞硝酸还原酶活性(NRA)的影响.结果表明,海洋卡盾藻可以有效利用无机氮源,如NH4Cl、NaNO3、NaNO2,对有机氮源如尿素、甘氨酸和1,4-丁二胺盐酸盐也有一定的利用能力,但不能有效利用丙氨酸.海洋卡顿藻的生长速率与营养盐的同化速率不一致,存在一定的滞后效应,最大比生长率可达到0.7 d-1.藻细胞硝酸还原酶活性在以NaNO3为氮源时,藻细胞酶活性(NRAmax)最大,为20.6 fmol/(min·cell).     

Evaluation of a fetus at risk for dihydropteridine reductase deficiency by direct mutation analysis using denaturing gradient gel electrophoresis

Dihydropteridine reductase (DHPR) is an enzyme involved in the recycling of tetrahydrobiopterin (BH₄), which is an obligate co-factor of the aromatic amino acid hydroxylases. DHPR deficiency is a rare, autosomal recessive disorder caused by mutations in the QDPR gene. DHPR-deficient patients are diagnosed by a lack of response to a low phenylalanine diet and by severe neurological symptoms. Final diagnosis is made by measurements of neurotransmitters and pterin metabolites in cerebrospinal fluid (CSF) and urine, in addition to DHPR enzyme activity, which can be assessed in whole red blood cells. Treatment of DHPR deficiency can be difficult and the outcome is not always satisfying, even if all treatment strategies are followed. Therefore prenatal diagnosis is of great importance in affected families. Prenatal diagnosis is possible by measuring DHPR activity in different cell types but this is time consuming. More than 25 different mutations have to date been identified in the QDPR gene and direct identification of a mutation in a fetus would be easy and rapid. We have developed a method based on denaturing gradient gel electrophoresis (DGGE) for the analysis of the QDPR gene. The method is useful for rapid and simultaneous scanning of all exons and flanking intronic sequences of the QDPR gene. We describe the first prenatal diagnosis conducted using this method. Copyright © 2001 John Wiley & Sons, Ltd.     

NR途径参与调节SO2胁迫下拟南芥含硫抗氧化物水平

以拟南芥为材料,研究了硝酸还原酶(NR)途径在植株响应SO_2过程中对含硫抗氧化物水平的影响.结果发现,野生型拟南芥植株中谷胱甘肽过氧化物酶(GPX)和谷胱甘肽硫转移酶(GST)活性高于NR缺失突变体(nia1nia2);SO_2暴露后,野生型和突变体nia1nia2植株中半胱氨酸(Cys)和谷胱甘肽(GSH)含量显著增加,GPX和GST的活性诱导性增高,且野生型植株中上述4项指标的增幅高于突变体;拟南芥逆境响应基因GSTU24和GPX7在野生型和突变体nia1nia2中差异表达,且对SO_2的响应程度不同.研究表明,植株体内NR途径与拟南芥细胞中含硫抗氧化物水平有关,NR缺失影响逆境应答中含硫抗氧化物水平的提高及抗逆基因转录,说明NR途径参与调节了植株逆境响应中含硫抗氧化物的合成及相关氧化还原反应与解毒过程.