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排序方式: 共有149条查询结果,搜索用时 15 毫秒
1.
Robin Wijngaard Meritxell Perramn Marina Parra-Robert Susana Hidalgo Gina Butrico Manuel Morales-Ruiz Muling Zeng Eudald Casals Wladimiro Jimnez Guillermo Fernndez-Varo Gerald I. Shulman Gary W. Cline Gregori Casals 《International journal of molecular sciences》2021,22(9)
The metabolic ratios lactate/pyruvate and β-hydroxybutyrate/acetoacetate are considered valuable tools to evaluate the in vivo redox cellular state by estimating the free NAD+/NADH in cytoplasm and mitochondria, respectively. The aim of the current study was to validate a gas-chromatography mass spectrometry method for simultaneous determination of the four metabolites in plasma and liver tissue. The procedure included an o-phenylenediamine microwave-assisted derivatization, followed by liquid-liquid extraction with ethyl acetate and silylation with bis(trimethylsilyl)trifluoroacetamide:trimethylchlorosilane 99:1. The calibration curves presented acceptable linearity, with a limit of quantification of 0.001 mM for pyruvate, β-hydroxybutyrate and acetoacetate and of 0.01 mM for lactate. The intra-day and inter-day accuracy and precision were within the European Medicines Agency’s Guideline specifications. No significant differences were observed in the slope coefficient of three-point standard metabolite-spiked curves in plasma or liver and water, and acceptable recoveries were obtained in the metabolite-spiked samples. Applicability of the method was tested in precision-cut liver rat slices and also in HepG2 cells incubated under different experimental conditions challenging the redox state. In conclusion, the validated method presented good sensitivity, specificity and reproducibility in the quantification of lactate/pyruvate and β-hydroxybutyrate/acetate metabolites and may be useful in the evaluation of in vivo redox states. 相似文献
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This study investigated cellular Nicotinamide Adenine Dinucleotide Phosphate (NADPH) fluorescence as a potential indicator of biohydrogen production in Chlamydomonas reinhardtii and a β-NADPH standard. NADPH fluorescence profiles of cultures grown in TAP-S (Tris-acetate phosphate minus sulphur) media, TAP (Tris-acetate phosphate) media and TAP + 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) were subsequently compared. Hydrogen production induced from sulphur depletion was found to correlate directly (r = 0.941) with NADPH over the ten day period. The addition of leachate was used to increase hydrogen yields, and subsequently increased the NADPH concentration by 50%–70%. A direct correlation was observed (r = 0.929) between NADPH and hydrogen when the leachate supplemented media was used. As NADPH is the terminal electron acceptor in the photosynthetic chain, results show that NADPH has a pivotal role in hydrogen production as a carrier molecule. Under sulphur depletion, cellular NADPH fluorescence can be used as an indicator of hydrogen production. 相似文献
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Minimizing photodecomposition of flavin adenine dinucleotide fluorescence by the use of pulsed LEDs 下载免费PDF全文
Dynamic alterations in flavin adenine dinucleotide (FAD) fluorescence permit insight into energy metabolism‐dependent changes of intramitochondrial redox potential. Monitoring FAD fluorescence in living tissue is impeded by photobleaching, restricting the length of microfluorimetric recordings. In addition, photodecomposition of these essential electron carriers negatively interferes with energy metabolism and viability of the biological specimen. Taking advantage of pulsed LED illumination, here we determined the optimal excitation settings giving the largest fluorescence yield with the lowest photobleaching and interference with metabolism in hippocampal brain slices. The effects of FAD bleaching on energy metabolism and viability were studied by monitoring tissue pO2, field potentials and changes in extracellular potassium concentration ([K+]o). Photobleaching with continuous illumination consisted of an initial exponential decrease followed by a nearly linear decay. The exponential decay was significantly decelerated with pulsed illumination. Pulse length of 5 ms was sufficient to reach a fluorescence output comparable to continuous illumination, whereas further increasing duration increased photobleaching. Similarly, photobleaching increased with shortening of the interpulse interval. Photobleaching was partially reversible indicating the existence of a transient nonfluorescent flavin derivative. Pulsed illumination decreased FAD photodecomposition, improved slice viability and reproducibility of stimulus‐induced FAD, field potential, [K+]o and pO2 changes as compared to continuous illumination. 相似文献
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Lutein Prevents High Fat Diet‐Induced Atherosclerosis in ApoE‐Deficient Mice by Inhibiting NADPH Oxidase and Increasing PPAR Expression 下载免费PDF全文
Hao Han Wei Cui Linzhi Wang Yufang Xiong Liegang Liu Xiufa Sun Liping Hao 《Lipids》2015,50(3):261-273
Epidemiological and experimental studies provide supportive evidence that lutein, a major carotenoid, may act as a chemopreventive agent against atherosclerosis, although the underlying molecular mechanisms are not well understood. The main aim of this study was to investigate the effects of lutein on the alleviation of atherosclerosis and its molecular mechanisms involved in oxidative stress and lipid metabolism. Male apolipoprotein E knockout mice (n = 55) were fed either a normal chow diet or a high fat diet (HFD) supplemented with or without lutein for 24 weeks. The results showed that a HFD induced atherosclerosis formation, lipid metabolism disorders and oxidative stress, but noticeable improvements were observed in the lutein treated group. Additionally, lutein supplementation reversed the decreased protein expression of aortic heme oxygenase‐1 and increased the mRNA and protein expressions of aortic nicotinamide‐adenine dinucleotide phosphate oxidase stimulated by a HFD. Furthermore, the decreased mRNA and protein expression levels of hepatic peroxisome proliferator‐activated receptor‐α, carnitine palmitoyltransferase 1A, acyl CoA oxidase 1, low density lipoprotein receptors and scavenger receptor class B type I observed in mice with atherosclerosis were markedly enhanced after treatment with lutein. Taken together, these data add new evidence supporting the anti‐atherogenic properties of lutein and describing its mechanisms of action in atherosclerosis prevention, including oxidative stress and lipid metabolism improvements. 相似文献
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Ateek Shah Yashwant Kumar S. Rohan Amrita B. Hazra 《Chembiochem : a European journal of chemical biology》2023,24(11):e202300055
Flavin adenine dinucleotide (FAD) is an essential redox cofactor in cellular metabolism. The organic synthesis of FAD typically involves coupling flavin mononucleotide (FMN) with adenosine monophosphate, however, existing synthesis routes present limitations such as multiple steps, low yields, and/or difficult-to-obtain starting materials. In this study, we report the synthesis of FAD nucleobase analogues with guanine/cytosine/uracil in place of adenine and deoxyadenosine in place of adenosine using chemical and enzymatic approaches with readily available starting materials, achieved in 1–3 steps with moderate yields (10–57 %). We find that the enzymatic route using Methanocaldococcus jannaschii FMN adenylyltransferase (MjFMNAT) is versatile and can produce these FAD analogues in high yields. Further, we demonstrate that Escherichia coli glutathione reductase is capable of binding and using these analogues as cofactors. Finally, we show that FAD nucleobase analogues can be synthesized inside a cell from cellular substrates FMN and nucleoside triphosphates by the heterologous expression of MjFMNAT. This lays the foundation for their use in studying the molecular role of FAD in cellular metabolism and as biorthogonal reagents in biotechnology and synthetic biology. 相似文献
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为了提高核糖核酸(RNA)单元应用于棉织物阻燃整理的耐久性以及降低生物阻燃成本,采用烯丙基溴对RNA单元衍生物5'-腺嘌呤核苷酸(AMP)进行改性制备生物阻燃单体。通过紫外光接枝法将AMP单体和AMP单体/丙烯酰胺分别接枝到纯棉织物上,制备了光接枝AMP单体和光接枝AMP单体/丙烯酰胺阻燃棉织物。对阻燃单体化学结构进行了表征。测试了纯棉织物及阻燃棉织物的阻燃性能。研究结果表明:AMP单体中成功引入不饱和双键;2种光接枝阻燃棉织物的分解温度均低于纯棉织物,阻燃棉织物的失重率降低;光接枝AMP单体/丙烯酰胺阻燃棉织物的续燃时间及阴燃时间可降低至0 s,AMP单体与丙烯酰胺之间存在一定的协效阻燃作用且可降低成本。 相似文献
9.
提出了测定烟酰胺含量的新方法 ,它是基于在碱性介质中烟酰胺与羟胺反应生成异羟肟酸 ,反应产物与 Fe3+反应生成紫红色配合物 ,在 496 nm波长处有最大吸收 ,其配合比为3∶ 1 .该方法的线性范围为 1 0 .0~ 2 0 0 μg/m L,相关系数为 0 .9995 ,摩尔吸光系数为 5 .11× 10 4L·mol-1·cm-1 ,平均回收率为 10 1 .2 % 相似文献
10.
Scope: Phenethyl isothiocyanate (PEITC) is a constituent of cruciferous vegetables that has demonstrated cancer preventive activity in a number of cancer models including lung, prostate, and breast cancer. Our objective was to examine the effects of the oral administration of PEITC for 7 days on the hepatic expression of genes important in drug metabolism and toxicity in Sprague Dawley rats. The liver is the major site for the metabolism of various xenobiotics and carcinogens, and determining the effects of PEITC on the gene expression of hepatic enzymes may provide insight into mechanisms underlying the cancer preventive activity of PEITC. Methods and results: Using a microarray containing 282 genes, we observed that PEITC significantly up‐regulated UDP‐glucuronosyltransferase UGT1A6 and strongly down‐regulated nicotinamide N‐methyltransferase (NNMT). We also confirmed the down‐regulation of NNMT by real‐time quantitative RT‐PCR. Other genes that were significantly up‐regulated were the drug metabolizing enzyme cyp2b15, the anti‐apoptotic gene bcl2l2, and the stress regulators Gadd45b, Dnajb9, Dnajb5 and Hspb1. Conclusion: Our results indicate new targets that may be important in the mechanisms of the anticancer effects of PEITC. Of particular significance was the down‐regulation of NNMT which may represent a new target for the treatment of a variety of cancers. 相似文献