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Nicotinamide Adenine Dinucleotide (nicotinamide + adenine_dinucleotide)
Terms modified by Nicotinamide Adenine Dinucleotide Selected AbstractsElectrocatalysis and Amperometric Detection of the Reduced Form of Nicotinamide Adenine Dinucleotide at Toluidine Blue/Zinc Oxide Coated ElectrodesELECTROANALYSIS, Issue 18 2007Ashok Kumar Abstract Thin toluidine blue (TBO) and zinc oxide (ZnO) hybrid films have been grown on glassy carbon electrode (GCE) and indium tin oxide coated (SnO2) glass electrodes by using cyclic voltammetry (CV). Scanning electron microscopy (SEM) images revealed spherical and beads-like shape of highly oriented TBO/ZnO hybrid films. Energy dispersive spectrometry (EDS) results declared that the films composed mainly of Zn and O. Moreover, TBO/ZnO hybrid films modified electrode is electrochemically active, dye molecules were not easily leached out from the ZnO matrix and the hybrid films can be considered for potential applications as sensor for amperometric determination of reduced nicotinamide adenine dinucleotide (NADH) at 0.0,V. A linear correlation between electrocatalytic current and NADH concentration was found to be in the range between 25,,M and 100,,M in phosphate buffer. In addition, we observed that dopamine, ascorbic acid and uric acid are not interference in amperometric detection of NADH in this proposed method. In addition, TBO/ZnO hybrid film modified electrode was highly stable and its response to the NADH also remained relentless. [source] Synthesis of Nicotinamide Adenine Dinucleotide (NAD) Analogues with a Sugar Modified Nicotinamide MoietyHELVETICA CHIMICA ACTA, Issue 7 2007Natasha Goulioukina Abstract The synthesis of nicotinamide adenine dinucleotide (NAD) analogues in which the ribose unit of the nicotinamide moiety is replaced by a hexitol, altritol, and cyclohexenyl sugar mimic is described. [source] Electroactivity of Polyaniline Multilayer Films in Neutral Solution and Their Electrocatalyzed Oxidation of ,-Nicotinamide Adenine DinucleotideADVANCED FUNCTIONAL MATERIALS, Issue 6 2003S. Tian Abstract In this paper, we report an alternative simple method to shift the electroactivity of polyaniline (PANI) films to neutral pH conditions by forming multilayer assemblies with poly(anions) using the layer-by-layer (LBL) deposition method. A series of self-assembled PANI multilayer films with poly(anions), such as sulfonated polyaniline (SPANI), poly(acrylic acid) (PAA), poly(vinyl sulfonate) (PVS), and poly(styrene sulfonate) (PSS), were prepared by the LBL method. Their electrochemical behavior and catalytic ability for the oxidation of ,-nicotinamide adenine dinucleotide (NADH) in neutral solution were investigated by electrochemistry (EC) combined with surface plasmon spectroscopy (SPS) and the quartz crystal microbalance (QCM) technique. Results indicated that all the films showed very good stability, reversibility, and electroactivity in neutral solution. All the multilayer films can electrocatalyze the oxidation of NADH, with the catalytic ability of PANI/SPANI being higher than that of the other assemblies under the same conditions. The catalytic abilities of the films with the same thickness prepared by the copolymerization method and the LBL method were also compared. [source] Low Potential Detection of NADH at Titanium-Containing MCM-41,Modified Glassy Carbon ElectrodeELECTROANALYSIS, Issue 5 2007Zhihui Dai Abstract Titanium-containing MCM-41 (Ti-MCM-41) modified glassy carbon electrode (GCE) can exhibit an excellent electrocatalytic activity towards the oxidation of ,-Nicotinamide adenine dinucleotide (NADH). A dramatic decrease in the over-voltage of NADH oxidation reaction is observed at 0.28,V (vs. SCE). The modified electrode is found to be stable and reproducible. The electrode shows a linear response for a wide range of 10,1200,,M NADH and the detection limit is 8.0,,M. Ti-MCM-41 mesoporous molecular sieves provide an efficient matrix for development of NADH biosensors and the prepared electrode not only can be used to detect the concentration of NADH in biochemical reaction, but also as the potential matrix of the construction of dehydrogenases biosensor. [source] Restorable Type Conversion of Carbon Nanotube Transistor Using Pyrolytically Controlled Antioxidizing Photosynthesis CoenzymeADVANCED FUNCTIONAL MATERIALS, Issue 16 2009Bo Ram Kang Abstract Here, a pyrolytically controlled antioxidizing photosynthesis coenzyme, , -Nicotinamide adenine dinucleotide, reduced dipotassium salt (NADH) for a stable n-type dopant for carbon nanotube (CNT) transistors is proposed. A strong electron transfer from NADH, mainly nicotinamide, to CNTs takes place during pyrolysis so that not only the type conversion from p-type to n-type is realized with 100% of reproducibility but also the on/off ratio of the transistor is significantly improved by increasing on-current and/or decreasing off-current. The device was stable up to a few months with negligible current changes under ambient conditions. The n-type characteristics were completely recovered to an initial doping level after reheat treatment of the device. [source] Effect of Enzyme and Cofactor Immobilization on the Response of Ethanol Oxidation in Zirconium Phosphate Modified BiosensorsELECTROANALYSIS, Issue 10 2010Mitk'El Abstract Two different self-contained ethanol amperometric biosensors incorporating layered [Ru(phend)2bpy]2+ -intercalated zirconium phosphate (ZrP) as the mediator as well as yeast -alcohol dehydrogenase (y- ADH) and its cofactor nicotinamide adenine dinucleotide (NAD+) were constructed to improve upon a design previously reported where only this mediator was immobilized in the surface of a modified electrode. In the first biosensor, a [Ru(phend)2bpy]2+ -intercalated ZrP modified carbon paste electrode (CPE) was improved by immobilizing in its surface both y- ADH and NAD+ using quaternized Nafion membrane. In the second biosensor, a glassy carbon electrode was modified with [Ru(phend)2bpy]2+ -intercalated ZrP, y- ADH, and NAD+ using Nafion as the holding matrix. Calibration plots for ethanol sensing were constructed in the presence and absence of ZrP. In the absence of ZrP in the surface of the modified glassy carbon electrode, leaching of ADH was observed as detected by UV-vis spectrophotometry. Ethanol sensing was also tested in the presence and absence of ascorbate to measure the selectivity of the sensor for ethanol. These two ethanol biosensors were compared to a previously reported one where the y -ADH and the NAD+ were in solution, not immobilized. [source] Electrocatalysis and Amperometric Detection of the Reduced Form of Nicotinamide Adenine Dinucleotide at Toluidine Blue/Zinc Oxide Coated ElectrodesELECTROANALYSIS, Issue 18 2007Ashok Kumar Abstract Thin toluidine blue (TBO) and zinc oxide (ZnO) hybrid films have been grown on glassy carbon electrode (GCE) and indium tin oxide coated (SnO2) glass electrodes by using cyclic voltammetry (CV). Scanning electron microscopy (SEM) images revealed spherical and beads-like shape of highly oriented TBO/ZnO hybrid films. Energy dispersive spectrometry (EDS) results declared that the films composed mainly of Zn and O. Moreover, TBO/ZnO hybrid films modified electrode is electrochemically active, dye molecules were not easily leached out from the ZnO matrix and the hybrid films can be considered for potential applications as sensor for amperometric determination of reduced nicotinamide adenine dinucleotide (NADH) at 0.0,V. A linear correlation between electrocatalytic current and NADH concentration was found to be in the range between 25,,M and 100,,M in phosphate buffer. In addition, we observed that dopamine, ascorbic acid and uric acid are not interference in amperometric detection of NADH in this proposed method. In addition, TBO/ZnO hybrid film modified electrode was highly stable and its response to the NADH also remained relentless. [source] Electrochemical Preparation of Poly(acriflavine) Film-Modified Electrode and Its Electrolcatalytic Properties Towards NADH, Nitrite and Sulfur OxoanionsELECTROANALYSIS, Issue 9 2007Shen-Ming Chen Abstract Electrochemical polymerization of acriflavine (AF) was carried out onto glassy carbon electrodes (GCE) from the aqueous buffer solution containing 1.5×10,3,M AF monomer (pH,3.5) which produced a thin electrochemically active film. This is noted as poly(AF) film modified electrodes (PAF/GCE). This modified electrode was shown a stable reversible redox couple centered at +0.22,V in pH,3.5 buffer solutions. PAF/GCE was found to be more stable in acidic solutions and its formal potential was found to be pH dependent with a slope close to ,60,mV/pH. The electrochemical deposition kinetics of poly(AF) onto gold coated quartz crystal was studied by using electrochemical quartz crystal microbalance (EQCM) combined with cyclic voltammetry (CV). PAF/GCE was found be good mediator for electrochemical oxidation of reduced nicotinamide adenine dinucleotide (NADH) in pH,5 buffer solutions. The electrocatalytic oxidation of SO and electrocatalytic reduction of NO, SO and S2O were carried out at PAF/GCE electrode in acidic aqueous solutions. The electrocatalytic oxidation of NADH was also investigated by using amperometric method. [source] New Strategy for Dehydrogenase Amperometric Biosensors Using Surfactant to Enhance the Sensitivity of Diaphorase/Ferrocene Modified Carbon Paste Electrodes for Electrocatalytic Oxidation of NADHELECTROANALYSIS, Issue 13 2003César Ramírez-Molina Abstract A carbon paste electrode (CPE) modified with diaphorase (DAP) and ferrocene (FcH) has been developed for determination of NADH at low working potential. The sensitivity and operational stability, towards the detection of the reduced form of the nicotinamide adenine dinucleotide (NADH) in flow injection analysis (FIA), were greatly improved (5 times) upon adding Tween 20 into the electrode matrix. The magnitude of the amperometric signal was dependent on DAP, FcH and surfactant loading, into the modified carbon paste electrode. A rapid and repeatable response was observed to the variation of NADH concentration in the vicinity of the electrode surface. Such advantages of the DAP/FcH/Tween 20 modified carbon paste were successfully used in the construction of L -lactate dehydrogenase modified electrodes. The use of this new approach can be generalized to other dehydrogenases and represents a decisive step for a versatile preparation method of amperometric biosensors. [source] Compound heterozygosity of two missense mutations in the NADH-cytochrome b5 reductase gene of a Polish patient with type I recessive congenital methaemoglobinaemiaEUROPEAN JOURNAL OF HAEMATOLOGY, Issue 6 2003Dorota Grabowska Abstract: A case of type I methaemoglobinaemia observed in a Polish subject with compound heterozygosity for two mutations in the reduced nicotinamide adenine dinucleotide (NADH) cytochrome b5 reductase (b5R) gene is described. One is a novel mutation 647T,C which leads to substitution of isoleucine by threonine at position 215 (I215T). This maternal mutation was found in several family members. A previously known mutation, 757G,A, leads to the replacement of valine by methionine at position 252 (V252M). The latter mutation was found also in the father and one of the two brothers. The effects of these mutations were analysed on a model of the human b5R protein obtained by homology modelling. Although both amino acid substitutions are located in the NADH-binding domain, the whole protein structure, especially the region between the flavin adenine dinucleotide and NADH-binding domains, is disturbed. The structural changes in the I215T mutant are less prominent than those in the V252M mutant. We presume that the 647T,C mutation is a type I mutation, however, it has not been observed in the homozygous state. [source] Is Helicobacter pylori a True Microaerophile?HELICOBACTER, Issue 4 2006Stephanie Bury-Moné Abstract Background:, There is no general consensus about the specific oxygen and carbon dioxide requirements of the human pathogen Helicobacter pylori. This bacterium is considered a microaerophile and consequently, it is grown under atmospheres at oxygen tensions 5,19% and carbon dioxide tensions 5,10%, both for clinical and basic and applied research purposes. The current study compared the growth of H. pylori in vitro, under various gas atmospheres, and determined some specific changes in the physiology of bacteria grown under different oxygen partial pressures. Methods:, Measurements of bacterial growth under various conditions were carried out employing classical solid and liquid culture techniques. Enzymatic activities were measured using spectrophotometric assays. Results:,H. pylori and all the other Helicobacter spp. tested had an absolute requirement for elevated carbon dioxide concentrations in the growth atmosphere. In contrast with other Helicobacter spp., H. pylori can tolerate elevated oxygen tensions when grown at high bacterial concentrations. Under 5% CO2, the bacterium showed similar growth in liquid cultures under oxygen tensions from microaerobic (< 5%) to fully aerobic (21%) at cell densities higher than 5 × 105 cfu/ml for media supplemented with horse serum and 5 × 107 cfu/ml for media supplemented with ,-cyclodextrin. Evidence that changes occurred in the physiology of H. pylori was obtained by comparing the activities of ferredoxin:NADH (nicotinamide adenine dinucleotide) oxidoreductases of bacteria grown under microaerobic and aerobic atmospheres. Conclusions:,H. pylori is a capnophile able to grow equally well in vitro under microaerobic or aerobic conditions at high bacterial concentrations, and behaved like oxygen-sensitive microaerophiles at low cell densities. Some characteristics of H. pylori cells grown in vitro under microaerobic conditions appeared to mimic better the physiology of organisms grown in their natural niche in the human stomach. [source] Synthesis of Nicotinamide Adenine Dinucleotide (NAD) Analogues with a Sugar Modified Nicotinamide MoietyHELVETICA CHIMICA ACTA, Issue 7 2007Natasha Goulioukina Abstract The synthesis of nicotinamide adenine dinucleotide (NAD) analogues in which the ribose unit of the nicotinamide moiety is replaced by a hexitol, altritol, and cyclohexenyl sugar mimic is described. [source] Electroactivity of Polyaniline Multilayer Films in Neutral Solution and Their Electrocatalyzed Oxidation of ,-Nicotinamide Adenine DinucleotideADVANCED FUNCTIONAL MATERIALS, Issue 6 2003S. Tian Abstract In this paper, we report an alternative simple method to shift the electroactivity of polyaniline (PANI) films to neutral pH conditions by forming multilayer assemblies with poly(anions) using the layer-by-layer (LBL) deposition method. A series of self-assembled PANI multilayer films with poly(anions), such as sulfonated polyaniline (SPANI), poly(acrylic acid) (PAA), poly(vinyl sulfonate) (PVS), and poly(styrene sulfonate) (PSS), were prepared by the LBL method. Their electrochemical behavior and catalytic ability for the oxidation of ,-nicotinamide adenine dinucleotide (NADH) in neutral solution were investigated by electrochemistry (EC) combined with surface plasmon spectroscopy (SPS) and the quartz crystal microbalance (QCM) technique. Results indicated that all the films showed very good stability, reversibility, and electroactivity in neutral solution. All the multilayer films can electrocatalyze the oxidation of NADH, with the catalytic ability of PANI/SPANI being higher than that of the other assemblies under the same conditions. The catalytic abilities of the films with the same thickness prepared by the copolymerization method and the LBL method were also compared. [source] Indoleamine 2,3-dioxygenase in T-cell tolerance and tumoral immune escapeIMMUNOLOGICAL REVIEWS, Issue 1 2008Jessica B. Katz Summary: Indoleamine 2, 3-dioxygenase (IDO) degrades the essential amino acid tryptophan in mammals, catalyzing the initial and rate-limiting step in the de novo biosynthesis nicotinamide adenine dinucleotide (NAD). Broad evidence implicates IDO and the tryptophan catabolic pathway in generation of immune tolerance to foreign antigens in tissue microenvironments. In particular, recent findings have established that IDO is overexpressed in both tumor cells and antigen-presenting cells in tumor-draining lymph nodes, where it promotes the establishment of peripheral immune tolerance to tumor antigens. In the normal physiologic state, IDO is important in creating an environment that limits damage to tissues due to an overactive immune system. However, by fostering immune suppression, IDO can facilitate the survival and growth of tumor cells expressing unique antigens that would be recognized normally as foreign. In preclinical studies, small-molecule inhibitors of IDO can reverse this mechanism of immunosuppression, complementing classical cytotoxic cancer chemotherapeutic agents' ability to trigger regression of treatment-resistant tumors. These results have encouraged the clinical translation of IDO inhibitors, the first of which entered phase I clinical trials in the fall of 2007. In this article, we survey the work defining IDO as an important mediator of peripheral tolerance, review evidence of IDO dysregulation in cancer cells, and provide an overview of the development of IDO inhibitors as a new immunoregulatory treatment modality for clinical trials. [source] Nicotinamide , biologic actions of an emerging cosmetic ingredientINTERNATIONAL JOURNAL OF COSMETIC SCIENCE, Issue 5 2005N. Otte Synopsis Nicotinamide, the water-soluble amide of nicotinic acid, is a component of the two most important coenzymes , nicotinamide adenine dinucleotide and nicotinamide adenine dinucleotide phosphate. Thus nicotinamide is involved in numerous oxidation,reduction reactions in mammalian biological systems. Nicotinamide essentially acts as an antioxidant. Most effects are exerted via poly-adenosine diphosphate-ribose polymerase inhibition. Thus nicotinamide increasingly gains interest in the prevention and treatment of several skin diseases. It is well established in the systemic therapy of pellagra, a deficiency disease linked to nicotinic acid, but with respect to topical use there is still a need for further evidence with respect to its manifold potential uses. Currently, its local use is established in the care of acne-prone skin. Résumé Le nicotinamide, l,amide hydrosoluble de l'acide nicotinique est un composant des deux plus importants co-enzymes NAD et NADP. Le nicotinamide est impliqué dans de nombreuses réactions d'oxydo-réduction dans les systèmes biologiques des mammifères. Le nicotinamide agit essentiellement en tant qu'anti-oxydant. La plupart des effets sont exercés via l'inhibition de la poly (ADP-ribose) polymerase (PARP). Ainsi, l'intérêt du nicotinamide croît dans la prévention et le traitement de nombreuses maladies cutanées. Son rôle est bien établi dans la thérapie systémique de la pellagre, une maladie déficiente liée à l'acide nicotinique, mais en ce qui concerne son utilisation topique, du fait de ses multiples applications potentielles, il est encore nécessaire d'accumuler davantage de preuves. Son utilisation locale est couramment admise dans le traitement des peaux sujettes à l'acné. [source] Coimmobilization of malic enzyme and alanine dehydrogenase on organic,inorganic hybrid gel fibers and the production of L -alanine from malic acid using the fibers with coenzyme regenerationJOURNAL OF APPLIED POLYMER SCIENCE, Issue 5 2010Koji Nakane Abstract Malic enzyme (EC 1.1.1.39) and alanine dehydrogenase (EC 1.4.1.1) were entrap-immobilized on hybrid gel fibers of cellulose acetate (CA) and zirconium (Zr) alkoxide by air-gap wet spinning. The production of L -alanine from malic acid with coenzyme regeneration was examined with the enzymes immobilized on the fibers. The productivity of L -alanine of the immobilized enzymes decreased to approximately one-fifth of that of free enzymes, but the CA,Zr-fiber-immobilized enzymes retained a high level of productivity after repeated use. Reduced form of nicotinamide adenine dinucleotide (NADH) recycling also occurred effectively for the enzymes immobilized on the fiber. © 2010 Wiley Periodicals, Inc. J Appl Polym Sci, 2010 [source] Enhancement of poly-adenosine diphosphate-ribosylation in human hepatocellular carcinomaJOURNAL OF GASTROENTEROLOGY AND HEPATOLOGY, Issue 5 2000Fumio Nomura Abstract Background: Poly-adenosine diphosphate (ADP)-ribosylation, catalysed by poly(ADP-ribose) polymerase (PARP), is a post-translational modification of nuclear proteins and is involved in a wide range of biological processes including DNA repair, cell proliferation and malignant transformation. Alteration of this reaction in human hepatocellular carcinoma (HCC) is of interest, but has not yet been explored. The aim of this study was to evaluate poly-ADP-ribosylation and to compare the expression of PARP in HCC and adjacent non-tumour tissues. Methods: Tumorous and adjacent non-tumorous tissues were obtained from five consecutive patients with HCC during surgery for tumour resection. Tissue homogenates were subjected to ADP-ribosylation with [32P]-nicotinamide adenine dinucleotide. The ADP-ribosylated proteins were separated by sodium dodecylsulfate,polyacrylamide gel electrophoresis, followed by autoradiography. Expression of PARP was also evaluated by western blotting. Results: Several proteins were ADP-ribosylated in human HCC tissues. Notably, the radiolabelling of a 116-kDa protein was remarkably greater than that in adjacent non-tumorous tissues (86.5 ± 35.2 arbitrary units by densitometry vs 12.2 ± 9.9, mean± SD, n = 5, P < 0.02). The radiolabelling of the 116-kDa protein was decreased in the presence of PARP inhibitors in a concentration-dependent manner. Immunoblot analyses revealed that the radiolabelled protein was PARP and that its expression was significantly greater in HCC than in adjacent non-tumorous tissues (333 ± 204% of non-tumorous tissue, P < 0.05). Conclusions: We found that poly-ADP-ribosylation and PARP expression were significantly increased in human HCC compared with those in adjacent non-tumorous tissues in surgically obtained specimens. [source] Sirtuins, melatonin and circadian rhythms: building a bridge between aging and cancerJOURNAL OF PINEAL RESEARCH, Issue 1 2010Brittney Jung-Hynes Abstract:, Histone deacetylases (HDAC) have been under intense scientific investigation for a number of years. However, only recently the unique class III HDAC, sirtuins, have gained increasing investigational momentum. Originally linked to longevity in yeast, sirtuins and more specifically, SIRT1 have been implicated in numerous biological processes having both protective and/or detrimental effects. SIRT1 appears to play a critical role in the process of carcinogenesis, especially in age-related neoplasms. Similarly, alterations in circadian rhythms as well as production of the pineal hormone melatonin have been linked to aging and cancer risk. Melatonin has been found act as a differentiating agent in some cancer cells and to lower their invasive and metastatic status. In addition, melatonin synthesis and release occurs in a circadian rhythm fashion and it has been linked to the core circadian machinery genes (Clock, Bmal1, Periods, and Cryptochromes). Melatonin has also been associated with chronotherapy, the timely administration of chemotherapy agents to optimize trends in biological cycles. Interestingly, a recent set of studies have linked SIRT1 to the circadian rhythm machinery through direct deacetylation activity as well as through the nicotinamide adenine dinucleotide (NAD+) salvage pathway. In this review, we provide evidence for a possible connection between sirtuins, melatonin, and the circadian rhythm circuitry and their implications in aging, chronomodulation, and cancer. [source] Interactions between melatonin and nicotinamide nucleotide: NADH preservation in cells and in cell-free systems by melatoninJOURNAL OF PINEAL RESEARCH, Issue 2 2005Dun-Xian Tan Abstract:, Interactions of melatonin and nicotinamide adenine dinucleotide (NADH) have been studied in different experimental models including NADH-promoted oxyhemoglobin oxidation, vanadate-induced NADH oxidation and paraquat-induced NADH depletion in cultured PC12 cells. Our findings indicate that melatonin preserves NADH levels under oxidative stress both in cell-free systems and in cultured PC12 cells. These interactions likely involve electron donation by melatonin and reduction of the NAD radical. As a result, the NAD radical is recycled to NADH and melatonin is oxidized to N1 -acetyl- N2 -formyl-5-methoxykynuramine (AFMK). NADH is a central molecule at the crossroads between energy metabolism and the antioxidant defense system in organisms. Recycling of NADH by melatonin might improve the efficiency of NADH as an energy carrier and as an antioxidant. Interactions between melatonin and NADH may be implicated in mitochondrial metabolism. [source] Melatonin protects against streptozotocin, but not interleukin-1,-induced damage of rodent pancreatic ,-cellsJOURNAL OF PINEAL RESEARCH, Issue 3 2001Annika K. Andersson In the present study, we examined whether melatonin can protect rodent pancreatic islets against streptozotocin (STZ) and interleukin-1, (IL-1,)-induced suppression of ,-cell function. Formation of free radicals, DNA damage and extensive DNA repair leading to depletion of intracellular nicotinamide adenine dinucleotide (NAD) may mediate STZ toxicity. Activation of inducible nitric oxide synthase and nitric oxide (NO) formation may cause IL-1,-induced ,-cell impairment. We also studied the effect of melatonin against STZ-induced hyperglycemia in C57BL/Ks mice. For in vitro studies, cultured rat islets were exposed to melatonin (100 ,M,1 mM) 30 min prior to STZ (0.5 mM) or IL-1, (25 U/mL) addition. After an additional 30 min incubation with STZ, islet function and NAD content were analyzed either acutely or after 18 hr of recovery in fresh culture medium. For IL-1, experiments, islets were incubated for 48 hr with the cytokine before evaluation of islet function. We found that melatonin counteracted STZ-induced inhibition of glucose metabolism and insulin release in cultured rat islets after 18 hr of recovery. Moreover, NAD levels were higher in the melatonin-treated group at this time point. Melatonin had no effect on IL-1,-induced islet inhibition of glucose oxidation or NO formation. Diabetes induced by STZ (140 mg/kg body weight; i.v.) was effectively prevented by administration of melatonin (100 mg/kg body weight; i.p.) 30 min before STZ injection. We conclude that the protective effects of melatonin against ,-cell damage may be related to interference with DNA damage and poly(ADP-ribose) polymerase (PARP) activation rather than through effects on NO generation pathways. [source] Contribution of NADH Increases to Ethanol's Inhibition of Retinol Oxidation by Human ADH IsoformsALCOHOLISM, Issue 4 2009Jennifer R. Chase Background:, A decrease in retinoic acid levels due to alcohol consumption has been proposed as a contributor to such conditions as fetal alcohol spectrum diseases and ethanol-induced cancers. One molecular mechanism, competitive inhibition by ethanol of the catalytic activity of human alcohol dehydrogenase (EC 1.1.1.1) (ADH) on all-trans-retinol oxidation has been shown for the ADH7 isoform. Ethanol metabolism also causes an increase in the free reduced nicotinamide adenine dinucleotide (NADH) in cells, which might reasonably be expected to decrease the retinol oxidation rate by product inhibition of ADH isoforms. Methods:, To understand the relative importance of these two mechanisms by which ethanol decreases the retinol oxidation in vivo we need to assess them quantitatively. We have built a model system of 4 reactions: (1) ADH oxidation of ethanol and NAD+, (2) ADH oxidation of retinol and NAD+, (3) oxidation of ethanol by a generalized Ethanoloxidase that uses NAD+, (4) NADHoxidase which carries out NADH turnover. Results:, Using the metabolic modeling package ScrumPy, we have shown that the ethanol-induced increase in NADH contributes from 0% to 90% of the inhibition by ethanol, depending on (ethanol) and ADH isoform. Furthermore, while the majority of flux control of retinaldehyde production is exerted by ADH, Ethanoloxidase and the NADHoxidase contribute as well. Conclusions:, Our results show that the ethanol-induced increase in NADH makes a contribution of comparable importance to the ethanol competitive inhibition throughout the range of conditions likely to occur in vivo, and must be considered in the assessment of the in vivo mechanism of ethanol interference with fetal development and other diseases. [source] Single sample extraction protocol for the quantification of NAD and NADH redox states in Saccharomyces cerevisiaeJOURNAL OF SEPARATION SCIENCE, JSS, Issue 18 2008Jennifer L. Sporty Abstract A robust redox extraction protocol for quantitative and reproducible metabolite isolation and recovery has been developed for simultaneous measurement of nicotinamide adenine dinucleotide (NAD) and its reduced form, NADH, from Saccharomyces cerevisiae. Following culture in liquid media, yeast cells were harvested by centrifugation and then lysed under nonoxidizing conditions by bead blasting in ice-cold, nitrogen-saturated 50 mM ammonium acetate. To enable protein denaturation, ice cold nitrogen-saturated CH3CN/50 mM ammonium acetate (3:1 v/v) was added to the cell lysates. Chloroform extractions were performed on supernatants to remove organic solvent. Samples were lyophilized and resuspended in 50 mM ammonium acetate. NAD and NADH were separated by HPLC and quantified using UV,Vis absorbance detection. NAD and NADH levels were evaluated in yeast grown under normal (2% glucose) and calorie restricted (0.5% glucose) conditions. Results demonstrate that it is possible to perform a single preparation to reliably and robustly quantitate both NAD and NADH contents in the same sample. Robustness of the protocol suggests it will be (i) applicable to quantification of these metabolites in other cell cultures; and (ii) amenable to isotope labeling strategies to determine the relative contribution of specific metabolic pathways to total NAD and NADH levels in cell cultures. [source] Sodium acetate enhances hydrogen peroxide production in Weissella cibariaLETTERS IN APPLIED MICROBIOLOGY, Issue 1 2009A. Endo Abstract Aims:, To investigate hydrogen peroxide production by lactic acid bacteria (LAB) and to determine the key factors involved. Methods and Results:, Six strains of Weissella cibaria produced large amounts (2·2,3·2 mmol l,1) of hydrogen peroxide in GYP broth supplemented with sodium acetate, but very low accumulations in glucose yeast peptone broth without sodium acetate. Increased production of hydrogen peroxide was also recorded when strains of W. cibaria were cultured in the presence of potassium acetate, sodium isocitrate and sodium citrate. Oxidases and peroxidases were not detected, or were present at low levels in W. cibaria. However, strong nicotinamide adenine dinucleotide (NADH) oxidase activity was recorded, suggesting that the enzyme plays a key role in production of hydrogen peroxide by W. cibaria. Conclusions:,Weissella cibaria produces large quantities of hydrogen peroxide in aerated cultures, in a process that is dependent on the presence of acetate in the culture medium. NADH oxidase is likely the key enzyme in this process. Significance and Impact of the Study:, This is the first study showing that sodium acetate, normally present in culture media of LAB, is a key factor for hydrogen peroxide production by W. cibaria. The exact mechanisms involved are not known. [source] Assimilation of NAD+ precursors in Candida glabrataMOLECULAR MICROBIOLOGY, Issue 1 2007Biao Ma Summary The yeast pathogen Candida glabrata is a nicotinamide adenine dinucleotide (NAD+) auxotroph and its growth depends on the environmental supply of vitamin precursors of NAD+. C. glabrata salvage pathways defined in this article allow NAD+ to be synthesized from three compounds , nicotinic acid (NA), nicotinamide (NAM) and nicotinamide riboside (NR). NA is salvaged through a functional Preiss,Handler pathway. NAM is first converted to NA by nicotinamidase and then salvaged by the Preiss,Handler pathway. Salvage of NR in C. glabrata occurs via two routes. The first, in which NR is phosphorylated by the NR kinase Nrk1, is independent of the Preiss,Handler pathway. The second is a novel pathway in which NR is degraded by the nucleosidases Pnp1 and Urh1, with a minor role for Meu1, and ultimately converted to NAD+ via the nicotinamidase Pnc1 and the Preiss,Handler pathway. Using C. glabrata mutants whose growth depends exclusively on the external NA or NR supply, we also show that C. glabrata utilizes NR and to a lesser extent NA as NAD+ sources during disseminated infection. [source] Reduction in DNA Synthesis During Two-photon Microscopy of Intrinsic Reduced Nicotinamide Adenine Dinucleotide Fluorescence,PHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 2 2005Michael G. Nichols ABSTRACT Two-photon laser scanning microscopy (TPLSM) of endogenous reduced nicotinamide adenine dinucleotide (NAD(P)H) provides important information regarding the cellular metabolic state. When imaging the punctate mitochondrial fluorescence originating from NAD(P)H in a rat basophilic leukemia (RBL) cell at low laser powers, no morphological changes are evident, and photobleaching is not observed when many images are taken. At higher powers, mitochondrial NAD(P)H fluorescence bleaches rapidly. To assess the limitations of this technique and to quantify the extent of photodamage, we have measured the effect of TPLSM on DNA synthesis. Although previous reports have indicated a threshold power for "safe" two-photon imaging, we find the laser power to be an insufficient indicator of photodamage. A more meaningful metric is a two-photon-absorbed dose that is proportional to the number of absorbed photon paris. A temporary reduction of DNA synthesis in RBL cells occurs whenever a threshold dose of approximately 2 × 1053 photon2 cm,4 s,1 is exceeded. This threshold is independent of laser intensity when imaging with average powers ranging from 5 to 17 mW at 740 nm. Beyond this threshold, the extent of the reduction is intensity dependent. DNA synthesis returns to control levels after a recovery period of several hours. [source] Alterations in Mitochondrial and Apoptosis-regulating Gene Expression in Photodynamic Therapy-resistant Variants of HT29 Colon Carcinoma Cells,PHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 2 2005Xiao Yun Shen ABSTRACT Photodynamic therapy (PDT) is a novel cancer therapy inducing irreversible photodamage to tumor tissue via photosensitizer-mediated oxidative cytotoxicity. The cellular and molecular responses associated with PDT are only partially understood. We have reported previously the generation of several photosensitizer-specific PDT-resistant cell variants of HT29 human colon adenocarcinoma cells by selecting cells from sequential PDT treatment using different photosensitizers. In this report, we describe the use of messenger RNA (mRNA) differential display to identify genes that were differentially expressed in the parental HT29 cells compared with their resistant variants. In comparison with parental HT29 cells, mRNA expression was increased in the PDT-resistant cell variants for BNIP3, estrogen receptor-binding fragmentassociated gene 9, Myh-1c, cytoplasmic dynein light chain 1, small membrane protein I and differential dependent protein. In contrast, expression in the PDT-resistant variants was downregulated for NNX3, human HepG2 3,region Mbol complementary DNA, glutamate dehydrogenase, hepatomaderived growth factor and the mitochondrial genes coding for 16S ribosomal RNA (rRNA) and nicotinamide adenine dinucleotide (NADH) dehydrogenase subunit 4. The reduction for mitochondrial 16S rRNA in the PDT-resistant variants was confirmed by Northern blotting, and the elevated expression of the proapoptotic BNIP3 in the PDT-resistant variants was confirmed by Northern and Western blotting analysis. We also examined the expression of some additional apoptosis-regulating genes using Western blotting. We show an increased expression of Bcl-2 and heat shock protein 27 and a downregulation of Bax in the PDT-resistant variants. In addition, the mutant p53 levels in the parental HT29 cells were reduced substantially in the PDT-resistant variants. We suggest that the altered expression in several mitochondria1 and apoptosisregulating genes contributes to PDT resistance. [source] Endogenous Fluorescence Spectroscopy of Cell Suspensions for Chemopreventive Drug Monitoring,PHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 1 2005Nathaniel D. Kirkpatrick ABSTRACT Cancer chemopreventive agents such as N -4-(hydroxyphenyl)-retinamide (4HPR) are thought to prevent cancers by suppressing growth or inducing apoptosis in precancerous cells. Mechanisms by which these drugs affect cells are often not known, and the means to monitor their effects is not available. In this study endogenous fluorescence spectroscopy was used to measure metabolic changes in response to treatment with 4HPR in ovarian and bladder cancer cell lines. Fluorescence signals consistent with nicotinamide adenine dinucleotide (NADH), flavin adenine dinucleotide (FAD) and tryptophan were measured to monitor cellular activity through redox status and protein content. Cells were treated with varying concentrations of 4HPR and measured in a stable environment with a sensitive fluorescence spectrometer. Results suggest that redox signal of all cells changed in a similar dose-dependant manner but started at different baseline levels. Redox signal changes depended primarily on changes consistent with NADH fluorescence, whereas the FAD fluorescence remained relatively constant. Similarly, tryptophan fluorescence decreased with increased drug treatment, suggesting a decrease in protein production. Given that each cell line has been shown to have a different apoptotic response to 4HPR, fluorescence redox values along with changes in tryptophan fluorescence may be a response as well as an endpoint marker for chemopreventive drugs. [source] Knockout of major leaf ferredoxin reveals new redox-regulatory adaptations in Arabidopsis thalianaPHYSIOLOGIA PLANTARUM, Issue 3 2008Ingo Voss Ferredoxins are the major distributors for electrons to the various acceptor systems in plastids. In green tissues, ferredoxins are reduced by photosynthetic electron flow in the light, while in heterotrophic tissues, nicotinamide adenine dinucleotide (reduced) (NADPH) generated in the oxidative pentose-phosphate pathway (OPP) is the reductant. We have used a Ds -T-DNA insertion line of Arabidopsis thaliana for the gene encoding the major leaf ferredoxin (Fd2, At1g60950) to create a situation of high electron pressure in the thylakoids. Although these plants (Fd2-KO) possess only the minor fraction of leaf Fd1 (At1g10960), they grow photoautotrophically on soil, but with a lower growth rate and less chlorophyll. The more oxidized conditions in the stroma due to the formation of reactive oxygen species are causing a re-adjustment of the redox state in these plants that helps them to survive even under high light. Redox homeostasis is achieved by regulation at both, the post-translational and the transcriptional level. Over-reduction of the electron transport chain leads to increased transcription of the malate-valve enzyme NADP-malate dehydrogenase (MDH), and the oxidized stroma leads to an increased transcription of the OPP enzyme glucose-6-P dehydrogenase. In isolated spinach chloroplasts, oxidized conditions give rise to a decreased activation state of NADP-MDH and an activation of glucose-6-P dehydrogenase even in the light. In Fd2-KO plants, NADPH-requiring antioxidant systems are upregulated. These adjustments must be caused by plastid signals, and they prevent oxidative damage under rather severe conditions. [source] Molecular data reveals California as the potential source of an invasive leafhopper species, Macrosteles sp. nr. severini, transmitting the aster yellows phytoplasma in HawaiiANNALS OF APPLIED BIOLOGY, Issue 3 2009J.J. Le Roux Abstract A species of aster leafhopper (Macrosteles sp.) became established in 2001 on Oahu, Hawaii, and through the transmission of the aster yellows phytoplasma, caused devastating losses to the island's watercress industry. DNA sequence data were analysed from two mitochondrial genes [cytochrome oxidase subunit 1(CO1) and nicotinamide adenine dinucleotide 1 (NADH1)] and one nuclear gene (wingless, Wg) (combined total of 1874 bp) to reconstruct phylogenetic relationships between putative US mainland source populations of aster leafhoppers and those introduced to Hawaii. These data were applied to elucidate the origin(s) and identity of Hawaiian infestations and the amount of genetic diversity within introduced invasive populations. Both phylogenetic search criteria (Bayesian and maximum likelihood models) converged onto similar tree topologies for all three gene regions and suggested that Hawaii infestations represent a single undescribed leafhopper species unrelated to the common aster leafhopper, Macrosteles quadrilineatus. An exact haplotype match was found from a specimen intercepted from watercress shipped to Hawaii from Los Angeles, California, suggesting this region as the potential source for Hawaiian infestations. Two mitochondrial haplotypes were identified in Hawaii suggesting two or perhaps just a single introduction of more than one female. [source] IMPACT OF BLOOD FLOW OCCLUSION ON LIVER NECROSIS FOLLOWING THERMAL ABLATIONANZ JOURNAL OF SURGERY, Issue 1-2 2006Mehrdad Nikfarjam Background: Laser, radiofrequency and microwave are common techniques for local destruction of liver tumours by thermal ablation. The main limitation of thermal ablation treatment is the volume of necrosis that can be achieved. Blood flow occlusion is commonly advocated as an adjunct to thermal ablation to increase the volume of tissue necrosis based on macroscopic and histological assessment of immediate or direct thermal injury. This study examines the impact of blood flow occlusion on direct and indirect laser induced thermal liver injury in a murine model using histochemical methods to assess tissue vitality. Methods: Thermal ablation produced by neodymium yttrium-aluminium-garnet laser (wavelength 1064 nm) was applied to the liver of inbred male CBA strain mice at 2 W for 50 s (100 J). Treatment was performed with and without temporary portal vein and hepatic artery blood flow occlusion. Animals were killed upon completion of the procedure to assess direct thermal injury or at 24, 48 and 72 h to assess the progression of tissue damage. The maximum diameter of necrosis was assessed by vital staining for nicotinamide adenine dinucleotide (NADH) diaphorase. Microvascular changes were assessed by laser Doppler flowmetry, confocal in vivo microscopy and scanning electron microscopy. Results: The direct thermal injury (mean SE) assessed by NADH diaphorase staining was significantly greater following thermal ablation treatment without blood flow occlusion than with blood flow occlusion (3.3 (0.4) mm vs 2.9 (0.3) mm; P = 0.005). Tissue disruption, cracking and vacuolization was more pronounced adjacent to the fibre insertion site in the group treated with thermal ablation combined with blood flow occlusion. There was an equivalent increase in the extent of injury following therapy in both groups that reached a peak at 48 h. The maximum diameter of necrosis in the thermal ablation alone group at 48 h was significantly greater than the thermal ablation combined with blood flow occlusion group (5.8 (0.4) mm vs 5.3 (0.3) mm; P = 0.011). The patterns of microvascular injury were similar in both groups, varying in extent. Conclusion: Temporary blood flow inflow occlusion appears to decrease the extent of initial injury measured by vital staining techniques and does not alter the time sequence of progressive tissue injury following thermal ablation therapy. [source] | |||||||||||||||||||