• Title/Summary/Keyword: nad3

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Sirt1 Promotes DNA Damage Repair and Cellular Survival

  • Song, Seung-Hyun;Lee, Mi-Ok;Lee, Ji-Seon;Oh, Je-Sok;Cho, Sung-Uk;Cha, Hyuk-Jin
    • Biomolecules & Therapeutics
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    • v.19 no.3
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    • pp.282-287
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    • 2011
  • Sirt1, a nicotinamide adenine dinucleotide ($NAD^+$)-dependent histone deacetylase, is known to deacetylate a number of proteins that are involved in various cellular pathways such as the stress response, apoptosis and cell growth. Modulation of the stress response by Sirtuin 1 (Sirt1) is achieved by the deacetylation of key proteins in a cellular pathway, and leads to a delay in the onset of cancer or aging. In particular, Sirt1 is known to play an important role in maintaining genomic stability, which may be strongly associated with a protective effect during tumorigenesis and during the onset of aging. In these studies, Sirt1 was generated in stably expressing cells and during the stimulation of DNA damage to examine whether it promotes survival. Sirt1 expressing cells facilitated the repair of DNA damage induced by either ionizing radiation (IR) or bleomycin (BLM) treatment. Fastened damaged DNA repair in Sirt1 expressing cells corresponded to prompt activation of Chk2 and ${\gamma}$-H2AX foci formation and promoted survival. Inhibition of Sirt1 enzymatic activity by a chemical inhibitor, nicotinamide (NIC), delayed DNA damage repair, indicating that promoted DNA damage repair by Sirt1 functions to induce survival when DNA damage occurs.

Biochemical and Molecular Characterization of Glycerol Dehydrogenase from Klebsiella pneumoniae

  • Ko, Gyeong Soo;Nguyen, Quyet Thang;Kim, Do Hyeon;Yang, Jin Kuk
    • Journal of Microbiology and Biotechnology
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    • v.30 no.2
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    • pp.271-278
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    • 2020
  • Glycerol dehydrogenase (GlyDH) catalyzes the oxidation of glycerol to dihydroxyacetone (DHA), which is the first step in the glycerol metabolism pathway. GlyDH has attracted great interest for its potential industrial applications, since DHA is a precursor for the synthesis of many commercially valuable chemicals and various drugs. In this study, GlyDH from Klebsiella pneumoniae (KpGlyDH) was overexpressed in E. coli and purified to homogeneity for biochemical and molecular characterization. KpGlyDH exhibits an exclusive preference for NAD+ over NADP+. The enzymatic activity of KpGlyDH is maximal at pH 8.6 and pH 10.0. Of the three common polyol substrates, KpGlyDH showed the highest kcat/Km value for glycerol, which is three times higher than for racemic 2,3-butanediol and 32 times higher than for ethylene glycol. The kcat value for glycerol oxidation is notably high at 87.1 ± 11.3 sec-1. KpGlyDH was shown to exist in an equilibrium between two different oligomeric states, octamer and hexadecamer, by size-exclusion chromatography analysis. KpGlyDH is structurally thermostable, with a Tm of 83.4℃, in thermal denaturation experiment using circular dichroism spectroscopy. The biochemical and biophysical characteristics of KpGlyDH revealed in this study should provide the basis for future research on its glycerol metabolism and possible use in industrial applications.

Conservation of metal artifacts excavated from tumulus in Chodang-dong, Kangreung City (강릉시 초당동 고분 출토 금속유물 보존)

  • Yu, Jae-Eun
    • 보존과학연구
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    • s.21
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    • pp.77-99
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    • 2000
  • Tumuli located in Chodang-dong in Kangreung City are sitesexcavated by the Museum of Kangreung University in 1993.Ofthese sites, a gilt bronze crown and gilt bronze sword with ring pommel with a tri-leaf ornament were excavated from the B-16tumulus. Underneath the gilt bronze crown was the gilt bronzes word with ring pommel with tri-leaf ornament, and these artifacts were treated with Polyurethane foams on the spot followed by special measures for conservation. This manuscript describes measures for conservation andanalysis with regard to the gilt bronze crown, gilt bronze sword with ring pommel with tri-leaf ornament and gilt bronze beltornaments. Bronze on the gilt bronze crown was completely corroded, therefore it was consolidated together with soil in the back of the artifact with Paraloid B72 after cleaning. Polyurethanefoams bottom was finished by applying the mixture of Epoxy resinAW106 and HV935K with soil. On the sword with ring pommel, the wooden part of the hilt was coated with Paraloid B72 (in Xylene) and the blade was consolidated with Paraloid NAD-10 30% after desalting with Sodium sesquicarbonate method. The gilt bronzebelt ornaments were treated with 3% Benzotriazole, coated with Paraloid B72 and then joined by using Cyanlacrylate. Specimens for the gilt bronze crown and gilt bronze belt ornaments were produced and gilding layers were examined under a metallographic microscope. The gilt bronze crown and the gilt bronze belt ornament maintained relatively good gilding quality with uniform gilding layers. Analysis of wood for the sheath of the sword with ring pommel revealed it to be Juglans mandshurica. Further more, analysis of constituents for a blue colored grassbead from dissolution of Polyurethane foams found it to be from the $Na_2O$-CaO-$AI_2O_3$-$SiO_2$ family.

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Factors Affecting the Lactate Dehydrogenase Activity of a Spore-forming Lactic Acid Bacteria (포자형성 유산균의 lactate dehydrogenase 역가에 미치는 제요인)

  • ;Hah, Yung Chil;Hong Soon Woo;Lee, Jung Chi
    • Korean Journal of Microbiology
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    • v.15 no.3
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    • pp.103-112
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    • 1977
  • Several strains of spore-forming lacticacid bacteria were isolated from natural sources such as soils, cereals, and foods. The general morphological and physiological characteristics of the strain 6-4 were investigated nad compared with some other industrial strains. The effects of fructose-1,6-diphoshpate (FDP), adenosine triphosphate (ATP), and pH on the lactate dehydrogenase(LDH) activity of the strain were studied, and the changes in LDH activity and spore formation under various cultural conditions were researched. The results were as follows. 1. This strain was identified to Bacillus coagulans Hammer and distributed widely in natural sources. 2. The strain strongly converted various fermentation substrates in to L(+)-lacticacid in anaerobic conditioins, and many spores that were of great advantages to the industrial application were formed easily in the aerobic condition. 3. The LDH activity of this strain was activated by FDP and inhibited by ATP. The optimal pH for the enzyme activity was 6.0-6.5. 4. In the anaerobic culture condifion, the large amount of glucose added in the medium increased the LDH activity, but the cells were not committed to sporulate. 5. When none or a very small amount of glucose (less than 0.5%) was added to culture medium in the aerobic condition, the LDH activity was decreased and many spore were produced with final pH higher than 8.5. 6. The additioin of large amount of glucose (more than 2.0%) in aerobic culture increased the LDH activity and inhibited strongly the spore formation with final pH lower than 6.0.

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The NQO1 rs1800566 Polymorphism and Risk of Bladder Cancer: Evidence from 6,169 Subjects

  • Guo, Zhan-Jing;Feng, Chang-Long
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.12
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    • pp.6343-6348
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    • 2012
  • Objective: The NAD(P)H:quinone oxidoreductase 1 (NQO1) rs1800566 polymorphism, leading to proline-toserine amino-acid and enzyme activity changes, has been implicated in bladder cancer risk, but individually published studies showed inconsistent results. We therefore here conducted a meta-analysis to summarize the possible association. Methods: A systematic literature search up to August 27, 2012 was carried out in PubMed, EMBASE and Wanfang databases, and the references of retrieved articles were screened. Crude odds ratios (ORs) with 95% confidence intervals (CIs) were analyzed for homozygote contrast (TT vs. CC), additive model (T vs. C), dominant model (TT+CT vs. CC), and recessive model (TT vs. CC+CT) to assess the association using fixed- or random-effect models. Results: We identified 12 case-control studies including 3,041 cases and 3,128 controls for the present meta-analysis. Significant association between NQO1 rs1800566 genetic polymorphism and risk of bladder cancer was observed in the additive model (OR = 1.15, 95% CI = 1.01-1.30, p = 0.030). Moreover, in the subgroup analysis stratified by ethnicity, significant associations were observed in Asians (OR = 1.26, 95% CI = 1.08-1.47, p = 0.003 for T vs. C; OR = 1.68, 95% CI = 1.21-2.32, p = 0.002 for TT vs. CC; OR = 1.50, 95% CI = 1.13-1.98, p = 0.005 for TT vs. CT+CC) but not in Caucasians. Conclusions: The results suggest that NQO1 rs1800566 genetic polymorphism may contribute to bladder cancer development, especially in Asians.

Amperometric Enzyme Electrode for the Determination of $NH_4^+$ ($NH_4^+$ 정량을 위한 Amperometric Enzyme Electrode)

  • Moo Lyong Seo;Jae Sang Kim;Shim Sung Lee;Zun Ung Bae;Heung Lark Lee;Tae Myung Park
    • Journal of the Korean Chemical Society
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    • v.37 no.11
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    • pp.937-942
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    • 1993
  • Enzyme electrodes for amperometric measurement of ammonia was prepared by immobilization of L-glutamate dehydrogenase on an Immobilon-AV Affinity membrane and attachment to a glassy carbon electrode. Reduced nicotinamide adenine dinucleotide (NADH) was used as the electroactive species. The electrochemical oxidation of NADH was monitored at +1.0 volt vs. Ag/AgCl. Response was linear from $4.0\;{\times}\;10^{-5}\;to\;4.0\;{\times}\;10^{-4}$ M. The detection limit was 2.0 ${\times}\;10^{-6}$ M. Response time, the optimum pH and life time of enzyme immobilized membrane were 2 min, pH 7.3∼7.6 (Dulbecco's buffer solution) and about 25 days respectively. When the enzyme electrode was applied to the $NH_4^+$ determination with amperometric method, other physiological materials had no interference.

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Harmal Extract Induces Apoptosis of HCT116 Human Colon Cancer Cells, Mediated by Inhibition of Nuclear Factor-κB and Activator Protein-1 Signaling Pathways and Induction of Cytoprotective Genes

  • Elkady, Ayman I;Hussein, Rania A;El-Assouli, Sufian M
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.4
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    • pp.1947-1959
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    • 2016
  • Background: Colorectal cancer (CRC) is a major cause of morbidity and mortality, being the second most common type of cancer worldwide in both men and women. It accounts yearly for approximately 9% of all new cases of cancers. Furthermore, the current chemotherapeutic regimens seem unsatisfactory, so that exploration of novel therapeutic modalities is needed. The present study was undertaken to investigate the inhibitory effects of a crude alkaloid extract (CAERS) of a medicinal herb, Rhazya stricta, on proliferation of CRC HCT116 cells and to elucidate mechanisms of action. To achieve these aims, we utilized MTT, comet, DNA laddering and gene reporter assays, along with Western blot and RT-PCR analyses. Results: We found that CAERS inhibited cell proliferation and induced apoptotic cell death in HCT116 cells. Hallmarks of morphological and biochemical signs of apoptosis were clearly evident. CAERS down-regulated DNA-binding and transcriptional activities of NF-${\kappa}B$ and AP-1 proteins, while up-regulating expression of the Nrf-2 protein. It also down-regulated expression levels of the ERK MAPK, Bcl-2, cyclin D1, CDK-4, survivin and VEGF and up-regulated levels of Bax, caspase-3/7 and -9, p53, p21, Nrf-2. Markedly, it promoted mRNA expression levels of cytoprotective genes including the hemeoxygenase-1, NAD(P)H quinine oxidoreductase 1 and UDP-glucuronyltransferase. Conclusions: These findings indicate that CAERS exerts antiproliferative action on CRC cells through induction of apoptotic mechanisms, and suggest CAERS could be a promising agent for studying and developing novel chemotherapeutic agents aimed at novel molecular targets for the treatment of CRC.

An Analysis on Electrical Double Layers at the Silicon Semiconductor Interfaces Using the Zeta Potential (Zeta전위에 의한 Silicon 반도체 계면의 전기이중층 해석)

  • Chun, Jang-Ho
    • Journal of the Korean Institute of Telematics and Electronics
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    • v.24 no.2
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    • pp.242-247
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    • 1987
  • Electrophysical phenomena at the silicon semiconductor-electrolyte solution interfaces were analyzed based on the zeta potential of the electrical double layer and microelectrophoresis. The suspensions were composed of the p or n-type silicon particles suspended in the KCI or pH buffer solutions. The approximate diameter of the prepared and sampled sioicon semiconductor pardticles was 1.5\ulcorner. The sign of the zeta poetntials of the p and n-type silicon particles in the KCl and pH buffer solution was positive. A range of electrophoretic mobilities of the p and n-type silicons in the KCl solutions was 5.5-8.9x10**-4 cm\ulcornerV-sec and 4.2-7.9x10**-4cm\ulcornerV-sec, respectively. The range of zeta potentials corresponding to the electrophoretic mobilities is 70.4-114.0mV nad 53.9-101.2mV, respectively. On the other hand, a range of electrophoretic mobilities of the p and n-type silicons in the pH buffer solutions was 1.1x10**-4-2.2x10**-3cm\ulcornerV-sec and 0-2.1x10**-3cm\ulcornerV-sec, respectively. The range of zeta potentials corresponding to the electrophoretic mobilities is 14.1-281.6mV and 0-268.8mV, respectively. The zeta potentials and electrical double layers of the doped silicon semiconductors are decisively influenced by the positively charged ions in the solutions. The maximum values of the zeta potentials in the KCl solutions appeared at a concentration of about 10-\ulcorner. The isoelectric point of the n-type silicon semiconductors appeared at about a pH 7. The effect of the space charge of the doped silicon semiconductors can be neglected compare with the effect of the surface charge.

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Cytoprotective Effect of Makgeolli Lees on Paraquat Induced Oxidative Stress in A549 Cells via Activation of NRF2 and Antioxidant Genes

  • Jeon, Miso;Rahman, Naimur;Kim, Yong-Sik
    • Journal of Microbiology and Biotechnology
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    • v.26 no.2
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    • pp.277-286
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    • 2016
  • Makgeolli lees (ML) has several physiological effects such as antioxidant, antidiabetic, and anticancer properties, but its biological functions have not been determined definitively. Here, we tested whether ML has a cytoprotective effect on paraquat (PQ)-induced oxidative stress in the human lung carcinoma cell line A549. At 0.1 mg/ml ML, viability of PQ-exposed A549 cells was restored by 12.4%, 18.5%, and 48.6% after 24, 48, and 72 h, respectively. ML also reduced production of the intracellular reactive oxygen species (ROS) that were generated by PQ treatment. Further experiments revealed that ML treatment enhanced the expression and nuclear translocation of nuclear factor erythroid 2-related factor 2 (NRF2) as well as ARE-GFP reporter activity. ML treatment also effectively increased the expression of NRF2's target genes NAD(P)H dehydrogenase quinone 1 (NQO1) and heme oxygenase 1 (HO-1). Moreover, we found that expression of cytoprotective genes, including glutathione peroxidases (GPXs), superoxide dismutase (SOD1), catalase (CAT), peroxiredoxin 3 (PRDX3), and peroxiredoxin 4 (PRDX4), was greatly enhanced by treatment with ML during PQ exposure. Taken together, the data suggest that treatment of PQ-exposed A549 cells with ML ameliorates cytotoxicity through induction of NRF2 expression and its target genes HO-1, NQO1, and other antioxidant genes. Thus, ML may serve as a functional food applicable to ROS-mediated human diseases.

Echinostoma mekongi n. sp. (Digenea: Echinostomatidae) from Riparian People along the Mekong River in Cambodia

  • Cho, Jaeeun;Jung, Bong-Kwang;Chang, Taehee;Sohn, Woon-Mok;Sinuon, Muth;Chai, Jong-Yil
    • Parasites, Hosts and Diseases
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    • v.58 no.4
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    • pp.431-443
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    • 2020
  • Echinostoma mekongi n. sp. (Digenea: Echinostomatidae) is described based on adult flukes collected from humans residing along the Mekong River in Cambodia. Total 256 flukes were collected from the diarrheic stool of 6 echinostome egg positive villagers in Kratie and Takeo Province after praziquantel treatment and purging. Adults of the new species were 9.0-13.1 (av. 11.3) mm in length and 1.3-2.5 (1.9) mm in maximum width and characterized by having a head collar armed with 37 collar spines (dorsal spines arranged in 2 alternative rows), including 5 end group spines. The eggs in feces and worm uterus were 98-132 (117) ㎛ long and 62-90 (75) ㎛ wide. These morphological features closely resembled those of Echinostoma revolutum, E. miyagawai, and several other 37-collar-spined Echinostoma species. However, sequencing of the nuclear ITS (ITS1-5.8S rRNA-ITS2) and 2 mitochondrial genes, cox1 and nad1, revealed unique features distinct from E. revolutum and also from other 37-collar-spined Echinostoma group available in GenBank (E. bolschewense, E. caproni, E. cinetorchis, E. deserticum, E. miyagawai, E. nasincovae, E. novaezealandense, E. paraensei, E. paraulum, E. robustum, E. trivolvis, and Echinostoma sp. IG). Thus, we assigned our flukes as a new species, E. mekongi. The new species revealed marked variation in the morphology of testes (globular or lobulated), and smaller head collar, collar spines, oral and ventral suckers, and cirrus sac compared to E. revolutum and E. miyagawai. Epidemiological studies regarding the geographical distribution and its life history, including the source of human infections, remain to be performed.