• 한국미생물생명공학회:학술대회논문집
• /
• 한국미생물생명공학회 2000년도 추계 학술대회
• /
• pp.181-188
• /
• 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.

• 한국응용약물학회:학술대회논문집
• /
• 한국응용약물학회 1994년도 제2회 추계심포지움
• /
• pp.147-172
• /
• 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.

• Applied Microscopy
• /
• 제45권3호
• /
• pp.135-143
• /
• 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.

• Asian-Australasian Journal of Animal Sciences
• /
• 제8권4호
• /
• pp.353-356
• /
• 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.

• 멤브레인
• /
• 제8권1호
• /
• pp.22-28
• /
• 1998

혈중 글루코우즈의 농도를 측정하기 위한 진단막의 제조를 위해 다층 젤라틴 필름이 사용되었다. 혈당치를 얻기위해 glucose dehydrogenase와 diaphorase를 이용하여 다층 젤라틴 필름을 통과하는 글루코우즈의 확산속도를 측정하였다. 글루코우즈의 최대 확산속도에 미치는 코팅의 간격거리, 효소의 양, 필름의 두께, 그리고 외부 온도의 영향에 대해 조사하였다.

• 대한약침학회지
• /
• 제4권1호
• /
• pp.125-139
• /
• 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.

• Journal of Acupuncture Research
• /
• 제25권3호
• /
• pp.179-187
• /
• 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.

• 생물정신의학
• /
• 제3권2호
• /
• pp.191-202
• /
• 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.

• 한국산림과학회지
• /
• 제80권1호
• /
• pp.97-102
• /
• 1991

포플러류의 식물체(植物體)와 그 식물체(植物體)에서 분리(分離)한 미토콘드라아를 동위효소(同位酵素) 분석(分析)하여, 각 동위효소(同位酵素)의 세포내(細胞內) 위치(位置)를 규명(糾明)하였다. 분석(分析)한 동위효소(同位酵素)는 ADH, 6-PGD, MDH, PGI와 Diaphorase였다. ADH는 세포질(細胞質)에 있으며, 핵내(核內)에 있는 한개의 유전자좌(遺傳子座)에 의(依)해 지배(支配)되는 것으로 나타났고, 6-PGD는 mitochondria에 있는 것으로 나타났다. MDH는 세포질(細胞質)에 3개의 band, mitochondria에 4개의 band가 있는 것으로 나타났다. PGI는 세포질(細胞質)에 3개의 band가 있는 듯하며, Diaphorase는 mitochondria에 1개의 band, 세포질(細胞質)에 2개의 band가 있는 것으로 나타났다. 세포질(細胞質)에 있는 두개의 band는 핵내(核內)에 있는 1개의 유전자좌(遺傳子座)에 있는 유전자(遺傳子)에 의(依)해 지배(支配)되는 것으로 판단되었다.

• Journal of Acupuncture Research
• /
• 제18권2호
• /
• pp.79-90
• /
• 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.