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https://www.readbyqxmd.com/read/27913299/the-inhibitory-effect-of-beta-lapachone-on-rankl-induced-osteoclastogenesis
#1
Dong Ryun Gu, Joon No Lee, Gi-Su Oh, Hyung Jin Kim, Min Seuk Kim, Seoung Hoon Lee
β-lapachone (β-L) is a substrate of reduced nicotinamide adenine dinucleotide (NADH): quinone oxidoreductase 1 (NQO1). NQO1 reduces quinones to hydroquinones using NADH as an electron donor and consequently increases the intracellular NAD+/NADH ratio. The activation of NQO1 by β-L has beneficial effects on several metabolic syndromes, such as obesity, hypertension, and renal injury. However, the effect of β-L on bone metabolism remains unclear. Here, we show that β-L might be a potent inhibitor of receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclastogenesis...
November 29, 2016: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/27912063/structure-of-mammalian-respiratory-supercomplex-i1iii2iv1
#2
Meng Wu, Jinke Gu, Runyu Guo, Yushen Huang, Maojun Yang
The mammalian respiratory chain complexes assemble into supercomplexes (SCs) and reside in the inner mitochondrial membrane to transfer electrons and establish the proton gradient for complex V to synthesize ATP. The precise arrangement of SCs is largely unknown. Here, we report a 4.0-Å cryo-electron microscopy (cryo-EM) structure of the major SC in porcine heart, the 1.7-MDa SCI1III2IV1. The complex III (CIII) dimer and complex IV (CIV) bind at the same side of the L-shaped complex I (CI). Several accessory or supernumerary subunits of CI, such as NDUFA11, NDUFB4, NDUFB8, and NDUFB9, directly contribute to the oligomerization of CI, CIII, and CIV...
December 1, 2016: Cell
https://www.readbyqxmd.com/read/27909649/in-silico-profiling-of-cell-growth-and-succinate-production-in-escherichia-coli-nzn111
#3
Xingxing Jian, Ningchuan Li, Cheng Zhang, Qiang Hua
BACKGROUND: Succinic acid is a valuable product due to its wide-ranging utilities. To improve succinate production and reduce by-products formation, Escherichia coli NZN111 was constructed by insertional inactivation of lactate dehydrogenase (LDH) and pyruvate formate lyase (PFL) encoded by the genes ldhA and pflB, respectively. However, this double-deletion mutant is incapable of anaerobically growing on glucose in rich or minimal medium even with acetate supplementation. A widespread hold view is that the inactivation of NADH-dependent LDH limits the regeneration of NAD(+) and consequently disables proper growth under anaerobic conditions...
2016: Bioresources and Bioprocessing
https://www.readbyqxmd.com/read/27909264/global-ablation-of-the-mitochondrial-calcium-uniporter-increases-glycolysis-in-cortical-neurons-subjected-to-energetic-stressors
#4
Matthew Nichols, Pia A Elustondo, Jordan Warford, Aruloli Thirumaran, Evgeny V Pavlov, George S Robertson
The effects of global mitochondrial calcium (Ca(2+)) uniporter (MCU) deficiency on hypoxic-ischemic (HI) brain injury, neuronal Ca(2+) handling, bioenergetics and hypoxic preconditioning (HPC) were examined. Forebrain mitochondria isolated from global MCU nulls displayed markedly reduced Ca(2+) uptake and Ca(2+)-induced opening of the membrane permeability transition pore. Despite evidence that these effects should be neuroprotective, global MCU nulls and wild-type (WT) mice suffered comparable HI brain damage...
December 1, 2016: Journal of Cerebral Blood Flow and Metabolism
https://www.readbyqxmd.com/read/27907843/triphenyltin-degradation-and-proteomic-response-by-an-engineered-escherichia-coli-expressing-cytochrome-p450-enzyme
#5
Wenying Yi, Kunliang Yang, Jinshao Ye, Yan Long, Jing Ke, Huase Ou
Although triphenyltin (TPT) degradation pathway has been determined, information about the enzyme and protein networks involved was severely limited. To this end, a cytochrome P450 hydroxylase (CYP450) gene from Bacillus thuringiensis was cloned and expressed in Escherichia coli BL21 (DE3), namely E. coli pET32a-CYP450, whose dosage at 1gL(-1) could degrade 54.6% TPT at 1mgL(-1) within 6 d through attacking the carbon-tin bonds of TPT by CYP450. Sequence analysis verified that the CYP450 gene had a 1214bp open reading frame, encoding a protein with 404 amino acids...
November 28, 2016: Ecotoxicology and Environmental Safety
https://www.readbyqxmd.com/read/27906040/identity-of-fasciola-spp-in-sheep-in-egypt
#6
Said Amer, Ahmed ElKhatam, Shereif Zidan, Yaoyu Feng, Lihua Xiao
BACKGROUND: In Egypt, liver flukes, Fasciola spp. (Digenea: Fasciolidae), have a serious impact on the farming industry and public health. Both Fasciola hepatica and Fasciola gigantica are known to occur in cattle, providing the opportunity for genetic recombination. Little is known on the identity and genetic variability of Fasciola populations in sheep. METHODS: This study was performed to determine the prevalence of liver flukes in sheep in Menofia Province as a representative area of the delta region in Egypt, as measured by postmortem examination of slaughtered animals at three abattoirs...
December 1, 2016: Parasites & Vectors
https://www.readbyqxmd.com/read/27903865/imaging-mitochondrial-dynamics-in-human-skin-reveals-depth-dependent-hypoxia-and-malignant-potential-for-diagnosis
#7
Dimitra Pouli, Mihaela Balu, Carlo A Alonzo, Zhiyi Liu, Kyle P Quinn, Francisca Rius-Diaz, Ronald M Harris, Kristen M Kelly, Bruce J Tromberg, Irene Georgakoudi
Active changes in mitochondrial structure and organization facilitate cellular homeostasis. Because aberrant mitochondrial dynamics are implicated in a variety of human diseases, their assessment is potentially useful for diagnosis, therapy, and disease monitoring. Because current techniques for evaluating mitochondrial morphology are invasive or necessitate mitochondria-specific dyes, their clinical translation is limited. We report that mitochondrial dynamics can be monitored in vivo, within intact human skin by relying entirely on endogenous two-photon-excited fluorescence from the reduced metabolic coenzyme nicotinamide adenine dinucleotide (NADH)...
November 30, 2016: Science Translational Medicine
https://www.readbyqxmd.com/read/27900690/a-microtiter-plate-based-assay-to-screen-for-active-and-stereoselective-hydrolytic-enzymes-in-enzyme-libraries
#8
Dominique Böttcher, Patrick Zägel, Marlen Schmidt, Uwe T Bornscheuer
A procedure for the high-throughput screening (HTS) of esterases is described. This includes a pretest for discrimination of active and inactive clones using an agar plate overlay assay, the enzyme expression in microtiter plates and the measurement of activity and enantioselectivity (E) of the esterase variants using acetates of secondary alcohols as model substrates. Acetic acid released is converted in an enzyme cascade leading to the stoichiometric formation of NADH, which is quantified in a spectrophotometer...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27897348/enzymatic-activity-and-thermodynamic-stability-of-biliverdin-ix%C3%AE-reductase-are-maintained-by-an-active-site-serine
#9
Wen-Ting Chu, Natasha M Nesbitt, Dmitri V Gnatenko, Zongdong Li, Beibei Zhang, Markus A Seeliger, Seamus Browne, Timothy J Mantle, Wadie F Bahou, Jin Wang
Biliverdin reductase IXβ (BLVRB) is a crucial enzyme in heme metabolism. Recent studies in humans have identified a loss-of-function mutation (Ser111Leu) that unmasks a fundamentally important role in hematopoiesis. We have applied experimental and thermodynamic modeling studies to provide further insight into role of the cofactor in substrate accessibility and protein folding properties regulating BLVRB catalytic mechanisms. Site-directed mutagenesis with molecular dynamic (MD) simulations establish the critical role of NAD(P)H-dependent conformational changes on substrate accessibility by forming the "hydrophobic pocket", along with identification of a single key residue (Arg35) modulating NAD(P)H/NADH selectivity...
November 29, 2016: Chemistry: a European Journal
https://www.readbyqxmd.com/read/27896951/hydrogen-peroxide-modulates-energy-metabolism-and-oxidative-stress-in-cultures-of-permanent-human-m%C3%A3-ller-cells-mio-m1
#10
Sven Peters, Max Griebsch, Matthias Klemm, Jens Haueisen, Martin Hammer
In this study the influence of hydrogen peroxide (H2 O2 ) on the redox state, NADH protein binding, and mitochondrial membrane potential in Müller cells is investigated. Cultures of permanent human Müller cells MIO-M1 were exposed to H2 O2 in 75 µM and 150 µM concentration for two hours. Fluorescence emission spectra and lifetimes were measured by two-photon microscopy (excitation wavelength: 740 nm) at the mitochondria which were identified in the microscopic images by their fluorescence properties (spectra and intensity)...
November 29, 2016: Journal of Biophotonics
https://www.readbyqxmd.com/read/27896650/neuroprotection-by-chlorpromazine-and-promethazine-in-severe-transient-and-permanent-ischemic-stroke
#11
Xiaokun Geng, Fengwu Li, James Yip, Changya Peng, Omar Elmadhoun, Jiamei Shen, Xunming Ji, Yuchuan Ding
Previous studies have demonstrated depressive or hibernation-like roles of phenothiazine neuroleptics [combined chlorpromazine and promethazine (C + P)] in brain activity. This ischemic stroke study aimed to establish neuroprotection by reducing oxidative stress and improving brain metabolism with post-ischemic C + P administration. Sprague-Dawley rats were subjected to transient (2 or 4 h) middle cerebral artery occlusion (MCAO) followed by 6 or 24 h reperfusion, or permanent (28 h) MCAO without reperfusion...
November 28, 2016: Molecular Neurobiology
https://www.readbyqxmd.com/read/27896382/the-mechanism-of-improved-intracellular-organic-selenium-and-glutathione-contents-in-selenium-enriched-candida-utilis-by-acid-stress
#12
Gao-Chuan Zhang, Da-Hui Wang, Dong-Hua Wang, Gong-Yuan Wei
Batch culture of Candida utilis CCTCC M 209298 for the preparation of selenium (Se)-enriched yeast was carried out under different pH conditions, and maximal intracellular organic Se and glutathione (GSH) contents were obtained in a moderate acid stress environment (pH 3.5). In order to elucidate the physiological mechanism of improved performance of Se-enriched yeast by acid stress, assays of the key enzymes involved in GSH biosynthesis and determinations of energy supply and regeneration were performed. The results indicated that moderate acid stress increased the activity of γ-glutamylcysteine synthetase and the ratios of NADH/NAD(+) and ATP/ADP, although no significant changes in intracellular pH were observed...
November 28, 2016: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/27895979/optical-redox-ratio-identifies-metastatic-potential-dependent-changes-in-breast-cancer-cell-metabolism
#13
Kinan Alhallak, Lisa G Rebello, Timothy J Muldoon, Kyle P Quinn, Narasimhan Rajaram
The development of prognostic indicators of breast cancer metastatic risk could reduce the number of patients receiving chemotherapy for tumors with low metastatic potential. Recent evidence points to a critical role for cell metabolism in driving breast cancer metastasis. Endogenous fluorescence intensity of nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FAD) can provide a label-free method for assessing cell metabolism. We report the optical redox ratio of FAD/(FAD + NADH) of four isogenic triple-negative breast cancer cell lines with varying metastatic potential...
November 1, 2016: Biomedical Optics Express
https://www.readbyqxmd.com/read/27892932/reciprocal-regulation-of-carbon-monoxide-metabolism-and-the-circadian-clock
#14
Roman Klemz, Silke Reischl, Thomas Wallach, Nicole Witte, Karsten Jürchott, Sabrina Klemz, Veronika Lang, Stephan Lorenzen, Miriam Knauer, Steffi Heidenreich, Min Xu, Jürgen A Ripperger, Michael Schupp, Ralf Stanewsky, Achim Kramer
Circadian clocks are cell-autonomous oscillators regulating daily rhythms in a wide range of physiological, metabolic and behavioral processes. Feedback of metabolic signals, such as redox state, NAD(+)/NADH and AMP/ADP ratios, or heme, modulate circadian rhythms and thereby optimize energy utilization across the 24-h cycle. We show that rhythmic heme degradation, which generates the signaling molecule carbon monoxide (CO), is required for normal circadian rhythms as well as circadian metabolic outputs. CO suppresses circadian transcription by attenuating CLOCK-BMAL1 binding to target promoters...
November 28, 2016: Nature Structural & Molecular Biology
https://www.readbyqxmd.com/read/27888691/redox-imbalance-and-mitochondrial-abnormalities-in-the-diabetic-lung
#15
Jinzi Wu, Zhen Jin, Liang-Jun Yan
Although the lung is one of the least studied organs in diabetes, increasing evidence indicates that it is an inevitable target of diabetic complications. Nevertheless, the underlying biochemical mechanisms of lung injury in diabetes remain largely unexplored. Given that redox imbalance, oxidative stress, and mitochondrial dysfunction have been implicated in diabetic tissue injury, we set out to investigate mechanisms of lung injury in diabetes. The objective of this study was to evaluate NADH/NAD(+) redox status, oxidative stress, and mitochondrial abnormalities in the diabetic lung...
November 17, 2016: Redox Biology
https://www.readbyqxmd.com/read/27887026/chaetomium-thermophilum-formate-dehydrogenase-has-high-activity-in-the-reduction-of-hydrogen-carbonate-hco3-to-formate
#16
Aşkın Sevinç Aslan, Jarkko Valjakka, Jouni Ruupunen, Deniz Yildirim, Nicholas J Turner, Ossi Turunen, Barış Binay
While formate dehydrogenases (FDHs) have been used for cofactor recycling in chemoenzymatic synthesis, the ability of FDH to reduce CO2 could also be utilized in the conversion of CO2 to useful products via formate (HCOO(-)). In this study, we investigated the reduction of CO2 in the form of hydrogen carbonate (HCO3(-)) to formate by FDHs from Candida methylica (CmFDH) and Chaetomium thermophilum (CtFDH) in a NADH-dependent reaction. The catalytic performance with HCO3(-) as a substrate was evaluated by measuring the kinetic rates and conducting productivity assays...
November 24, 2016: Protein Engineering, Design & Selection: PEDS
https://www.readbyqxmd.com/read/27886975/in-vitro-metabolic-engineering-of-bioelectricity-generation-by-the-complete-oxidation-of-glucose
#17
Zhiguang Zhu, Y-H Percival Zhang
The direct generation of electricity from the most abundant renewable sugar, glucose, is an appealing alternative to the production of liquid biofuels and biohydrogen. However, enzyme-catalyzed bioelectricity generation from glucose suffers from low yields due to the incomplete oxidation of the six-carbon compound glucose via one or few enzymes. Here, we demonstrate a synthetic ATP- and CoA-free 12-enzyme pathway to implement the complete oxidation of glucose in vitro. This pathway is comprised of glucose phosphorylation via polyphosphate glucokinase, NADH generation catalyzed by glucose 6-phosphate dehydrogenase (G6PDH) and 6-phosphogluconate dehydrogenase (6PGDH), electron transfer from NADH to the anode, and glucose 6-phosphate regeneration via the non-oxidative pentose phosphate pathway and gluconeogenesis...
November 23, 2016: Metabolic Engineering
https://www.readbyqxmd.com/read/27886267/in-vivo-characterization-of-hair-and-skin-derived-carbon-quantum-dots-with-high-quantum-yield-as-long-term-bioprobes-in-zebrafish
#18
Jing-Hui Zhang, Aping Niu, Jing Li, Jian-Wei Fu, Qun Xu, De-Sheng Pei
Carbon quantum dots (CDs) were widely investigated because of their tunable fluorescence properties and low toxicity. However, so far there have been no reports on in vivo functional studies of hair and skin derived CDs. Here, hair derived CDs (HCDs) and skin derived CDs (SCDs) were produced by using human hair and pig skin as precursors. The quantum yields (QYs) of HCDs and SCDs were quite high, compared to citric acid derived CDs (CCDs). HCDs and SCDs possess optimal photostability, hypotoxicity and biocompatibility in zebrafish, indicating that HCDs and SCDs possess the capacity of being used as fluorescence probes for in vivo biological imaging...
November 25, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27885835/sequential-enzymatic-conversion-of-%C3%AE-angelica-lactone-to-%C3%AE-valerolactone-through-hydride-independent-c-c-bond-isomerization
#19
Nikolaus G Turrini, Elisabeth Eger, Tamara C Reiter, Kurt Faber, Mélanie Hall
A case of hydride-independent reaction catalyzed by flavin-dependent ene-reductases from the Old Yellow Enzyme (OYE) family was identified. α-Angelica lactone was isomerized to the conjugated β-isomer in a nicotinamide-free and hydride-independent process. The catalytic cycle of C=C bond isomerization appears to be flavin-independent and to rely solely on a deprotonation-reprotonation sequence through acid-base catalysis. Key residues in the enzyme active site were mutated and provided insight on important mechanistic features...
November 25, 2016: ChemSusChem
https://www.readbyqxmd.com/read/27885267/contributory-roles-of-two-l-lactate-dehydrogenases-for-l-lactic-acid-production-in-thermotolerant-bacillus-coagulans
#20
Lifan Sun, Caili Zhang, Pengcheng Lyu, Yanping Wang, Limin Wang, Bo Yu
Thermotolerant Bacillus coagulans is considered to be a more promising producer for bio-chemicals, due to its capacity to withstand harsh conditions. Two L-lactate dehydrogenase (LDH) encoding genes (ldhL1 and ldhL2) and one D-LDH encoding gene (ldhD) were annotated from the B. coagulans DSM1 genome. Transcriptional analysis revealed that the expression of ldhL2 was undetectable while the ldhL1 transcription level was much higher than that of ldhD at all growth phases. Deletion of the ldhL2 gene revealed no difference in fermentation profile compared to the wild-type strain, while ldhL1 single deletion or ldhL1ldhL2 double deletion completely blocked L-lactic acid production...
November 25, 2016: Scientific Reports
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