• Title/Summary/Keyword: Diaphorase

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Electrocheimical Evaluation of the Reaction Rafe and Electrochemical Optimization of the Mediated Electrochemical Reduction of NAD$^+$

  • Kang, Young-Wan;Kim, So-Hyoung;Kang, Chan;Yun, Sei-Eok
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2000.10a
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    • pp.181-188
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    • 2000
  • The electrocatalytic reduction of NAD$^{+}$ using diaphorase was studied. methyl viologen (MV$^{2+}$) mediator between an electrode and the enzyme. Steady-state currents could be obtained under the conditions of slow scan rate, low MV$^{2+}$concentration, and high NAD$^{+}$ concentration as the electrode reaction was converted to an electrochemical-catalytic (EC') reaction. The biomecular rate constant for the reaction of the reduced methyl viologen with the oxidized diaphorase was estimated as 7.5$\times$10$^3$M$^{-1}$ s$^{-1}$ from the slope of the current versus [MV$^{2+}$] plot. And the optimal concentrations of diaphorase, MV$^{2+}$ and NAD$^{+}$ in the mediated electrocatalytic reduction of NAD$^{+}$ were decided by applying the cyclic voltammetry. The optimal concentrations of the species were obtained by finding the conditions which gave the highest and steady-state current at a gold-amalgam electrode. The highest and steady-state catalytic current was achieved under the conditions of 1.5 U/ml diaphorase, 0.2 mM MV$^{2+}$, and 4.8 mM NAD$^{+}$ at the scan rate of 2 mV s$^{-1}$ , suggesting that the rate of the electrocatalytic reation is the higest under the former conditions. The electrochemical procedure under the conditions of 1.5 U/ml diaphorase,0.2 mM MV$^{2+}$, and 4.8 mM NAD$^{+}$ was used favorably to drive an enzymatic reduction of pyruvate to D-lactate.

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Mechanism of Action of Anticancer Drug Aziridinylbenzoquinones: Involvement of DT-diaphorase (DNA에 결합하는 항암제의 작용기전)

  • Lee, Chong-Soon-
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1994.11a
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    • pp.147-172
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    • 1994
  • Aziridinylbenzoquinones such as 3, 6-diaziridinyl-1, 4-benzoquinone (DZQ) and its 2, 5-methyl analog (MeDZQ) require bioreductive activation in order to elicit their anticancer activities. To determine the involvement of DTD in the activation of these drugs, we have used a ligation-mediated polymerase chain reaction to map the intracellular alkylation sites in a sing1e copy gene at the nucleotide level. We have performed this analysis in two human colon carcinoma cells, one proficient (HT-29) and one deficient (BE) in DT-diaphorase (DTD) activity. In the DTD proficient HT-29 cell line, DZQ and MeDZQ were found to alkylate both 5'-(A/T)G(C)-3' and 5'-(A/T)A-3' sequences. This is consistent with the nucleotide preferences observed when DZQ and MeDZQ are activated by purified DTD to reactive metabolites capable of alkylating DNA in vitro [Lee, C. -S., Hartley, J. A., Berardini, M. D., Butler, J., Siegel., D., Ross, D., & Gibson, N. W. (1992) Biochemistry, 31: 3019-3025]. Surprisingly in the DTD-deficient BE cell line a pattern of alkylation induced by DZQ and MeDZQ similar to that observed in the DTD-proficient HT-29 cells was observed. This suggests that reductive enzymes other than DTD can be involved in activating DZQ and MeDZQ to DNA reactive species in vivo.

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The Effect of Joint Mobilization on the Expressions of Nadph-Diaphorase and Nitric Oxide Synthase in the Spinal Cord Neurons

  • Kuwaoka, Toshifumi;Jung, HanSuk;Ham, JooHyun;Choi, JinOk;Cho, HyeRin
    • Applied Microscopy
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    • v.45 no.3
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    • pp.135-143
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    • 2015
  • The aim of this study was to investigate effects on joint mobilization in neurochemical changes of nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) and nitric oxide synthase (NOS) of the spinal cord neurons after right knee joint immobilization (RKJI) and in ultrastructural changes of the femoral nerves innervating the muscles acting on RKJI. A total of 15 guinea pigs were used and divided into 5 groups. Immunohistochemistry was performed to detect NADPH-d and NOS. NADPH-d and NOS were not expressed in the ventral horn of control and experimental groups, but were expressed or not in the dorsal horn according to the duration of release after RKJI and the presence or absence of joint mobilization. Ultrastructures of the femoral nerves in experimental groups had partial demyelination and condensed clumps in axon. Effects on manipulative therapy after RKJI were confirmed from expression of NADPH-d and NOS in the dorsal horn of the lumbosacral spinal cord. Manipulative therapy was more effective against a long-term immobilization than a short-term immobilization.

ASSOCIATION OF RED CELL X-PROTEIN, NADH-DIAPHORASE 1, MALIC ENZYME AND SERUM ARYLESTERASE PHENOTYPES WITH ECONOMIC TRAITS IN SHEEP

  • Lee, S.L.;Mukherjee, T.K.;Agamuthu, P.;Panandam, J.M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.8 no.4
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    • pp.353-356
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    • 1995
  • The red cell X-protein, NADH-diaphorase 1, malic enzyme and serum arylesterase phenotypes of 50 Thai Longtail and 53 Cameroon X Thai Longtail ($F_1$) crossbred sheep were determined by horizontal starch gel electrophoresis. None of the economic traits was influenced by DIA1, ME and EsA phenotypes. However, XP phenotypes showed a highly significant association with body weight, body height, heart girth and back girth, with mean values of XP+ve phenotype greater than XP-ve. The $XP^+$ allele was associated with greater body weight, body height, heart girth and back girth.

Basic Studies on the Preparation of Diagnostic Membranes by Using Multi-Layered Gelatin Films to Measure Blood Glucose Level of Diabetics (당뇨병 환자의 혈당치 측정을 위한 다층 젤라틴 필름을 이용한 진단막의 제조에 관한 기초연구)

  • 권석기;음성진
    • Membrane Journal
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    • v.8 no.1
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    • pp.22-28
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    • 1998
  • Multi-layered gelatin films were used for the preparation of diagnostic membranes to measure blood glucose concentration. Diffusion rates of glucose through multi-layer gelatin films were monitored to obtain blood-glucose level by using glucose dehydrogenase and diaphorase. The effects of coating edge distance, enzyme quantities, film thickness, and outside temperature on the maximum diffusion rates of glucose were examined.

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Effects of the Bee Venom Herbal Acupuncture on the Neurotransmitters of the Rat Brain Cortex

  • Yun, Hyoung-Seok;Lee, Jae-Dong
    • Journal of Pharmacopuncture
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    • v.4 no.1
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    • pp.125-139
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    • 2001
  • In order to study the effects of bee venom herbal acupuncture on the neurotransmitters of the rat brain cortex, herbal acupuncture with the bee venom group and normal saline group was performed bilaterally on the point corresponding to LI 4 of the rat. The average optical density of the neurotransmitters from the cerebral cortex was analyzed 30 minutes after the herbal acupuncture with immunohistochemical methods. The results were as follows: 1. The density of NADPH-diaphorase in the bee venom group was increased significantly at the motor cortex, visual cortex, auditory cortex, cingulate cortex, retrosplenial cortex, and perirhinal cortex, compared to the normal saline group. 2. The average optical density of vasoactive intestinal peptide in the bee venom group had significant changes at the insular cortex, retrosplenial cortex, and perirhinal cortex, compared to the normal saline group. 3. The average optical density of neuropeptide-Y in the bee venom group increased significantly at the visual cortex and cingulate cortex, compared to the normal saline group.

Effect of Low Frequency Electroacupuncture on Nicotidamide Adenine Dinucleotide Phosphate-diaphorase(NADPH-d) Positive Neurons in the Brain Cortex of Rat with Adjuvant Induced Rheumatoid Arthritis (저빈도 전침자극이 류마토이드 관절염 유발 흰쥐 대뇌피질 Nicotidamide Adenine Dinucleotide Phosphate-diaphorase(NADPH-d) 양성세포 발현에 미치는 영향)

  • Jung, Ki-Hoon;Roh, Jeong-Du;Kim, E-Hwa;Lee, Eun-Yong
    • Journal of Acupuncture Research
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    • v.25 no.3
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    • pp.179-187
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    • 2008
  • Objectives & Methods : This study was to investigate effect of low frequency electroacupuncture on NADPH-d positive neurons in the brain cortex of rat with adjuvant induced rheumatoid arthritis. Experimental groups were divided into 6 groups ; Normal, Control, $ST_{36}$, $SP_9$, $ST_{36}+SP_9$ and Non-Acupoint. Normal group, non-arthritic group, was injected normal saline, and the other groups were injected FCA. Each acupoint groups were treated by 2Hz electroacupuncture at each acupoints and NA group was treated by 2Hz electroacupuncture at non-acupoint. Each groups were evaluated by the number of NADPH-d positive neurons in primary somatosensory area(S1), secondary somatosensory area(S2), motor area and caudate putamen by using an image analyzer and a microscope. Results : 1. In S1, the number of NADPH-d positive neuron cells in the $ST_{36}$ group were significantly(p<0.05) increased compared with the control group. 2. In S2, the number of NADPH-d positive neuron cells in all electroacupuncture groups were not significantly changed compared with the control group. 3. In motor area, the number of NADPH-d positive neuron cells in $ST_{36}$ group, $SP_9$ group, NA group were significantly(p<0.05) increased compared with the control group. 4. In Caudate putamen, the number NADPH-d positive neuron cells in all electroacupuncture groups were significantly(p<0.05) decreased compared with the control group. Conclusions : Our result demonstrated that low frequency electroacupuncture on $ST_{36}$ & $SP_9$ normalized expression of NADPH-d positive neurons in the brain cortex of the rheumatoid arthritis model in rats.

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A Study on NADPH-Diaphorase and Neuropeptide Y-Positive Neurons in The Brain Stem of Aged Rats (노화된 흰쥐의 뇌줄기에 분포하는 NADPH-Diaphorase 및 Neuropeptide Y양성 신경세포에 관한 연구)

  • Kim, Soon-Hee;Chang, Hwan-Il;Huh, Young-Buhm;Ahn, Hee-Kyung
    • Korean Journal of Biological Psychiatry
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    • v.3 no.2
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    • pp.191-202
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    • 1996
  • This study was aimed to charily the change of NADPH-diaphorase(NADPH-d) and neuropeptide Y(NPY) associated with aging of the rots. To verily the effect of aging of NPY and NADPH-d-positive neurons in the brain stem, the neurons were stained by the immunohistochemical and histochemical method. In the aged group, the number of NADPH-d-positive neurons was significantly decreased in substantia nigra lateralis, supragenual nucleus, raphe magnus nucleus and raphe obscurus nucleus as compared with control group. The number of NADPH-d-positive neurons was not significantly decreased in superior colliculus, interpeduncular nucleus, central gray. dorsal raphe nucleus, retrorubral nucleus, pedunculopontine tegmental nucleus, laterodorsal tegmental nucleus, pontine reticular nucleus, prepositus hypoglossal nucleus and nucleus solitarius of the aged rats. The NADPH-d and NPY-positive neurons were found in the interpeduncular nucleus, central gray, substantia nigra lateralis, laterodorsal tegmental nucleus, nucleus solitarius, raphe magnus nucleus, raphe obscurus nucleus of the control and aged groups. The coexistence of NADPH-d and NPY in the some cell was not found in the brain stem of both groups.

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Subcellular Location of Five Isozymes in Populus Species by Isozyme Analysis with Whole Plant and Mitochondria (포플러류(類)의 식물체(植物體)와 미토콘드리아의 동위효소(同位酵素) 분석(分析)에 의(依)한 몇 동위효소(同位酵素)의 세포소기관적(細胞小器管的) 위치(位置))

  • Ryu, Jang Bal;Noh, Eun Woon;Son, Doo Sik
    • Journal of Korean Society of Forest Science
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    • v.80 no.1
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    • pp.97-102
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    • 1991
  • Subcellular location of five isozymes in Populus species was studied by isozyme analysis with plant and mitochondria. Isozymes analyzed were ADH, 6-PGD, MDH, PGI, and Diaptrorase. ADH seems to be cytosolic isozyme encoded with nuclear genes at one locus, while 6-PGD seems to be mitochondrial one. MDH showed three hands in cytosol and four other bands in mitochondria. Three cytosolic hands were showed for PGI. Diaphorase showed one mitochodrial band and two cytosofic bands which seemed to the encoded by nuclear genes at one locus.

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Effects of Auricular Acupuncture on NADPH-diaphorase Neurons in Brain Cortex of Fasted Rats (이침자극(耳鍼刺戟)이 절식(絶食) Stress로 인(因)한 흰쥐 대뇌피질(大腦皮質)의 NADPH-diaphorase 신경세포(神經細胞)에 미치는 영향(影響))

  • Lee, Jung-hyun;Kim, Ee-Hwa;Lee, Eun-yong
    • Journal of Acupuncture Research
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    • v.18 no.2
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    • pp.79-90
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    • 2001
  • Objective : The aim of the present study is to investigate whether stimulating a specific auricular point(胃點) is effect on suppression of appetite. Methods : This study evaluated the changes of Reduced-Nicotinamide-Adenine-inucieotide-Phosphate-diaphorase(NADPH-d)-positive neurons using a histoehemical method. Staining intensities of NADPH-d-positive neurons were assessed in a quantitative fashion using a microdensitometrical method based on optical density by means of an image analyzer. Results : The results show as follows ; 1. In the RSG/RSA, FRI/FR2, HL, PAR1 area, The optical density of NADPH-d-positive neurons of fasted group was significantly decreased in comparison to fed group. 2. In most cortical area, It was not significant statistically, But the optical density of NADPH-d-positive neurons of fed with auricular acupuncture group was decreased in comparison to fed group. 3. In most cortical area, It was not significant statistically, But fasted with auricular acupuncture group increased the optical density of NADPH-d-positive neurons comparison to fasted group. Conclusions : We conclude that stimulating the specific auricular points is able to effect on NO(Nitric Oxide)-expression in the brain cortex of sprague dawley rats, The results suggest that auricular acupuncture may be useful for controlling obesity.

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