运用三维荧光光谱解析高温厌氧产氢反应过程对温度变化的响应

为建立反应器运行状态的荧光光谱快速监测方法,应用三维荧光光谱表征了高温厌氧反应器的出水,采用平行因子分析方法分析后得到3种荧光组分——蛋白质、辅酶NADH和核黄素,并获得它们的激发发射光谱图及相对浓度.结果表明,当温度从50 ℃上升到60 ℃的过程中,3种荧光物质的相对浓度均呈现相应的变化,它们与反应器的运行参数呈现明显相关性.进一步研究表明,核黄素是高温厌氧产氢反应中的特有荧光物质,采用标准加入法和二阶校正方法可以测定其实际浓度,出水中核黄素浓度变化范围为0.04~0.13 mg·L^-1.该研究为厌氧产氢反应器的运行监控提供了理论基础.     

Prenatal diagnosis of NADH:ubiquinone oxidoreductase deficiency

NADH:ubiquinone oxidoreductase (complex I of the mitochondrial respiratory chain) deficiency is a severe disorder with an often early fatal outcome. Prenatal diagnosis for complex I defects currently relies mainly on biochemical assays of complex I in fetal tissues such as chorionic villi (CV), and is only in a minority of cases possible by means of mutational analysis of nuclear-encoded genes of complex I. We report on our experience to date with prenatal diagnosis in pregnancies at risk for complex I deficiency. We measured complex I activity in native CV and/or cultured CV in 23 pregnancies in 15 families. In accordance with the results of the investigations in CV, 15 children were born clinically unaffected. Two prenatally diagnosed unaffected fetuses and two prenatally diagnosed affected fetuses were lost prematurely with spontaneous or provoked abortions, respectively. Two affected children were born (prenatally found to be affected). In two pregnancies a discrepancy between native and cultured cells was found. We conclude that prenatal diagnosis for complex I deficiency can be reliably performed. Pitfalls were encountered in using cultured CV as a result of maternal cell contamination (MCC). Future research on pathogenic nuclear mutations underlying complex I deficiency will extend the possibilities for prenatal diagnosis at the molecular level. Copyright © 2001 John Wiley & Sons, Ltd.     

Electropolymerized poly（Toluidine Blue）-modified carbon felt for highly sensitive amperometric determination of NADH in flow injection analysis

Poly(pheniothiazine) films were prepared on a porous carbon felt (CF) electrode surface by an electrooxidative polymerization of three phenothiazine derivatives (i.e.,Tthionine (TN), Toluidine Blue (TB) and Methylene Blue (MB)) from 0.1 mol/L phosphate buffer solution (pH 7.0). Among the three phenothiazies, the poly(TB) film-modified CF exhibited an excellent electrocatalytic activity for the oxidation of nicotinamide adenine dinucleotide reduced form (NADH) at +0.2 V vs. Ag/AgCl. The poly(TB) film-modified CF was successfully used as working electrode unit of highly sensitive amperometric flow-through detector for NADH. The peak currents (peak heights) were almost unchanged, irrespective of a carrier flow rate ranging from 2.0 to 4.1 mL/min, resulting in the measurement of NADH (ca. 30 samples/hr) at 4.1 mL/min. The peak current responses of NADH showed linear relationship over the concentration range from 1 to 30 mol/L (sensitivity: 0.318 A/( mol/L); correlation coefficient: 0.997). The lower detection limit was found to be 0.3 mol/L (S/N = 3).     

Terpenylnaphthoquinones are reductively activated by NADH/NADH dehydrogenase

Novel terpenylnaphthoquinones were found to enhance the rate of oxygen consumption in the presence of NADH/NADH dehydrogenase in 1?:?1?v/v mixtures of 40?mM phosphate buffer (pH 7.4) and DMSO. Initial rates of oxygen consumption increase with an increase in the half-wave reduction potentials of the quinones. The amounts of spin trapped methyl radicals in the presence of DMSO, produced through the Haber–Weiss reaction of the oxygen consumption process, and of the corresponding semiquinones, generated under anaerobic conditions, also correlate with the quinone redox potential. Since this enzymatic system is found in mitochondria, a possible pathway in the cytotoxic activity of these terpenylnaphthoquinones could be the interference or inhibition of mitochondrial respiration by NADH depletion or by mitochondrial degradation due to reactive oxygen species generation.     

Interactive effects of cadmium and hypoxia on metabolic responses and bacterial loads of eastern oysters Crassostrea virginica Gmelin

Pollution by toxic metals including cadmium (Cd) and hypoxia are important stressors in estuaries and coastal waters which may interactively affect sessile benthic organisms, such as oysters. We studied metabolic responses to prolonged hypoxic acclimation (2 weeks at 5% O₂) in control and Cd-exposed (30 d at 50 μg L⁻¹ Cd) oysters Crassostrea virginica, and analyzed the effects of these stressors on abundance of Vibrio spp. in oysters. Hypoxia-acclimated oysters retained normal standard metabolic rates (SMR) at 5% O₂, in contrast to a decline of SMR observed during acute hypoxia. However, oysters spent more time actively ventilating in hypoxia than normoxia resulting in enhanced Cd uptake and 2.7-fold higher tissue Cd burdens in hypoxia. Cd exposure led to a significant decrease in tissue glycogen stores, increase in free glucose levels and elevated activity of glycolytic enzymes (hexokinase and aldolase) indicating a greater dependence on carbohydrate catabolism. A compensatory increase in activities of two key mitochondrial enzymes (citrate synthase and cytochrome c oxidase) was found during prolonged hypoxia in control oysters but suppressed in Cd-exposed ones. Cd exposure also resulted in a significant increase in abundance of Vibrio parahaemolyticus and Vibrio vulnificus levels during normoxia and hypoxia, respectively. Overall, Cd- and hypoxia-induced changes in metabolic profile, Cd accumulation and bacterial flora of oysters indicate that these stressors can synergistically impact energy homeostasis, performance and survival of oysters in polluted estuaries and have significant consequences for transfer of Cd and bacterial pathogens to the higher levels of the food chain.     

共价功能化对单壁碳纳米管产生DNA损伤和活性氧的影响研究

以质粒DNA评价法表征了不同程度共价功能化单壁碳纳米管(SWNTs)的生物毒性,并在近红外-可见光光谱、拉曼光谱和活性氧(ROS)表征的基础上研究了共价功能化对其物理结构和化学活性的影响.结果表明,共价功能化增强了碳纳米管对质粒DNA的损伤程度,3种不同类型的碳纳米管对DNA损伤的强度为羧基功能化SWNTs(SWNT-COOH)>聚乙二醇功能化SWNTs(SWNT-PEG)>非共价功能化SWNTs(uSWNTs).随着共价功能化程度的加深,碳纳米管表面石墨晶格的sp2结构受损程度逐渐增高,以致使其反应活性增大.共价功能化后SWNTs可作为电子转移中间体将NADH中的电子转移至O2并生成ROS从而对DNA造成损伤.     

Effects of the inhibitor of pyruvate dehydrogenase multi-enzyme complex on hydrogen production by fermentative microbes

The redox state of carbon sources directly affected the ratio of NADH/NAD⁺ which was coupled to the hydrogen production by Bacillus sp. FS2011. The addition of the inhibitor of pyruvate dehydrogenase multi-enzyme complex (PDHc)E1 could regulate hydrogen production by FS2011 or pretreated compost in batch cultivation. With the addition of appropriate amount of inhibitor, hydrogen production via the NADH pathway was increased, leading to the higher overall hydrogen production. The maximum hydrogen yields of 307.6 ± 13.21 mL/g by FS2011 with the inhibitor of 80 ppm and 362.1 ± 10.1 mL/g by pretreated compost with the inhibitor of 60 ppm were observed, which were increased by 8.7% and 17.8% compared with the controls, respectively. Meanwhile the production of soluble metabolic byproducts such as butyrate, acetate and so on were decreased, resulting in reducing the difficulty of wastewater treatment.