• Title/Summary/Keyword: nad1/2-3

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Induction of Anticarcinogenic Enzymes by Dichloromethane-soluble Fraction of Physalis alkekengi var. francheti Hort. in Mouse Hepatoma Cells

  • Seo, JiYeon;Kim, Hyo Jung;Kim, Jong-Sang
    • Current Research on Agriculture and Life Sciences
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    • v.32 no.3
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    • pp.119-124
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    • 2014
  • Physalis alkekengi var. francheti Hort. is known as an insecticide and traditional remedy for liver related diseases. Therefore, this study investigated the chemopreventive effects of extracts and several solvent fractions (n-hexane, dichloromethane, n-butanol, water) of Physalis alkekengi var. francheti Hort. First, their cytotoxicity and NQO1 activity were measured using an MTT assay, plus a quinone reductase [NAD(P)H dehydrogenase (quinone); NAD(P)H: (quinone acceptor) oxidoreductase, EC 1.6.99.2]-inducing activity assay was performed using cultured murine hepatoma cells (Hepa1c1c7) and its mutant cells(BpRc1). The reduction of electrophilic quinones by NQO1 is an important detoxification pathway and major mechanism of chemoprevention. When compared with the other solvent soluble fractions with different polarities, the dichloromethane fraction of Physalis alkekengi var. francheti Hort. showed a higher NQO1-inducing activity that was also dose-dependent. Moreover, the dichloromethane fraction of Physalis alkekengi var. francheti Hort. induced ARE-luciferase activities in HepG2-C8 cells that were generated by transfecting the ARE-luciferase gene construct, suggesting the Nrf2-ARE-mediated induction of anti-oxidative enzymes. In conclusion, the dichloromethane-soluble fraction of Physalis alkekengi var. francheti Hort. showed a relatively strong induction of detoxifying enzymes, thereby meriting further study to identify the active components and evaluate their potential as cancer preventive agents.

Practical Algorisms for PCR-RFLP-Based Genotyping of Echinococcus granulosus Sensu Lato

  • Kim, Hye-Jin;Yong, Tae-Soon;Shin, Myeong Heon;Lee, Kyu-Jae;Park, Gab-Man;Suvonkulov, Uktamjon;Kovalenko, Dmitriy;Yu, Hak Sun
    • Parasites, Hosts and Diseases
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    • v.55 no.6
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    • pp.679-684
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    • 2017
  • Echinococcus granulosus sensu lato (s.l.) is a causative agent of cystic echinococcosis or cystic hydatid disease in humans and domestic and wild animals. The disease is a serious health problem in countries associated with poverty and poor hygiene practices, particularly in livestock raising. We introduced a practical algorism for genotyping the parasite, which may be useful to many developing countries. To evaluate the efficiency of the algorism, we genotyped 3 unknown strains isolated from human patients. We found that unknowns 1 and 3 were included in G1, G2, and G3 genotypes group and unknown 2 was included in G4 genotype (Echinococcus equinus) according to the algorisms. We confirmed these results by sequencing the 3 unknown isolates cox1 and nad1 PCR products. In conclusion, these new algorisms are very fast genotype identification tools that are suitable for evaluating E. granulosus s.l. isolated from livestock or livestock holders, particularly in developing countries.

Nitrogen Fixation of Blue Green Algae (Nostoc pruniforme) (남조류(Nostic pruniforme)의 질소고정능에 관한 연구)

  • 홍순우;최영길
    • Korean Journal of Microbiology
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    • v.11 no.2
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    • pp.51-58
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    • 1973
  • Three kinds of organic matter such as glucose, oxalic acid, and ethanol were added to the media(N-free or $NO_{3}$-riched) and their effects on the nitrogen fixation of Nostoc pruniforme were measured by manometric technique through the experiments in vivo. 1) The organic matters used in this experiments showed effective results as a role of substrate for the fixation of atmospheric nitrogen. 2) In the nitrogen-free medium treated with the both of flucose nad ethanol, the highest nitrogen uptakes were detected in the treated of low concentrations (glucose ; 0.1%, 0.5%, ethanol : 0.1%, 0.5%). On the contrary, the highest nitrogen uptakes in $NO_{3}$-riched medium were measured at the treated of high concentrations (glucose ; 2%, 1%, ethanol ; 1.5%, 1.0%). 3) The highest nitrogen uptakes in N-free medium treated with oxalic acid were measured at the concentration of 2% and 1%, respectively. In the medium of $NO_{3}$-riched, the nitrogen uptakes were in the opposite directions.

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Molecular and Morphologic Identification of Spirometra ranarum Found in the Stool of African Lion, Panthera leo in the Serengeti Plain of Tanzania

  • Eom, Keeseon S.;Park, Hansol;Lee, Dongmin;Choe, Seongjun;Kang, Yeseul;Bia, Mohammed Mebarek;Lee, Sang-Hwa;Keyyu, Julius;Fyumagwa, Robert;Jeon, Hyeong-Kyu
    • Parasites, Hosts and Diseases
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    • v.56 no.4
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    • pp.379-383
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    • 2018
  • The present study was performed with morphological and molecular analysis (cox1 and nad1 mitochondrial genes) to identify the proglottids of spirometrid tapeworm found in the stool of an African lion, Panthera leo, in the Serengeti plain of Tanzania. A strand of tapeworm strobila, about 75 cm in length, was obtained in the stool of a male African lion in the Serengeti National Park ($34^{\circ}$ 50' E, $02^{\circ}$ 30' S), Tanzania, in February 2012. The morphological features of the adult worm examined exhibited 3 uterine coils with a bow tie appearance and adopted a diagonal direction in the second turn. The posterior uterine coils are larger than terminal uterine ball and the feature of uteri are swirling rather than spirally coiling. The sequence difference between the Spirometra species (Tanzania origin) and S. erinaceieuropaei (GenBank no. KJ599680) was 9.4% while those of S. decipiens (GenBank no. KJ599679) differed by 2.1% in the cox1 and nad1 genes. Phylogenetic tree topologies generated using the 2 analytic methods were identical and presented high level of confidence values for the 3 major branches of the 3 Spirometra species in the cox1 gene. The morphological and molecular findings obtained in this study were nearly coincided with those of S. ranarum. Therefore, we can know for the first time that the African lion, Panthera leo, is to the definitive host of this tapeworm.

Mitochondrial Genome of Spirometra theileri Compared with Other Spirometra Species

  • Ndosi, Barakaeli Abdieli;Park, Hansol;Lee, Dongmin;Choe, Seongjun;Kang, Yeseul;Nath, Tilak Chandra;Bia, Mohammed Mebarek;Eamudomkarn, Chatanun;Jeon, Hyeong-Kyu;Eom, Keeseon S.
    • Parasites, Hosts and Diseases
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    • v.59 no.2
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    • pp.139-148
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    • 2021
  • This study was carried out to provide information on the taxonomic classification and analysis of mitochondrial genomes of Spirometra theileri. One strobila of S. theileri was collected from the intestine of an African leopard (Panthera pardus) in the Maswa Game Reserve, Tanzania. The complete mtDNA sequence of S. theileri was 13,685 bp encoding 36 genes including 12 protein genes, 22 tRNAs and 2 rRNAs with absence of atp8. Divergences of 12 protein-coding genes were as follow: 14.9% between S. theileri and S. erinaceieuropaei, 14.7% between S. theileri and S. decipiens, and 14.5% between S. theileri with S. ranarum. Divergences of 12 proteins of S. theileri and S. erinaceieuropaei ranged from 2.3% in cox1 to 15.7% in nad5, while S. theileri varied from S. decipiens and S. ranarum by 1.3% in cox1 to 15.7% in nad3. Phylogenetic relationship of S. theileri with eucestodes inferred using the maximum likelihood and Bayesian inferences exhibited identical tree topologies. A clade composed of S. decipiens and S. ranarum formed a sister species to S. erinaceieuropaei, and S. theileri formed a sister species to all species in this clade. Within the diphyllobothridean clade, Dibothriocephalus, Diphyllobothrium and Spirometra formed a monophyletic group, and sister genera were well supported.

Mechanism of Alcoholic Liver Disease

  • 문전옥
    • Journal of Life Science
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    • v.4 no.3
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    • pp.102-112
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    • 1994
  • 알콜성 간장해의 발생, 진전에는 많은 인자가 관여하고 있으며 극히 복잡한 병태를 형성하는데 그 기전으로는 1)간내 [NADH]/[NAD] 비의 상승, 2)에탄올의 주 대사산물인 아세트알데히드에 의한 간장해, 3)면역기구에 의한 간장해, 4)과산화지질, 활성산소 및 free radical 에 의한 장해와 5) 중심정맥역의 hypoxia에 의한 간세포장해 기전을 들 수 있다. 본 총설에서는 알콜을 장기 섭취할 경우 간에서의 대사경로와 현재 고려되고 있는 몇 가지 알콜성 간장해 발생기전에 대한 최근의 연구들을 정리하였다.

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Rapid Purification of Glucose-6-Phosphate Dehydrogenase by Affinity Chromatography (Affinity Chromatography를 이용한 Glucose-6-Phosphate Dehydrogenase의 신속한 정제방법 개발)

  • 이한수;임정빈
    • Korean Journal of Microbiology
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    • v.21 no.4
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    • pp.221-228
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    • 1983
  • An improved procedure for the rapid purification of glucose-6-phosphate dehydrogenase from extracts of Saccharomyces cerevisiae was developed by using affinity chromatography. Among six affinty media tested, $NADP^+ -agarose$ and Affi-gel Blue were more effective than others (i.e., Affi-gel Red, AMP-agarose, ATP-agarose, and $NAD^+ -agarose$). Conditions to desorb the enzyme bound to the affinity media were examined to increase the purity as well as yield. The best result was obtained when the column was developed with a linear gradient of KCl (0-1.0M). In case of Affi-gel Blue, introduction of $NAD^+$ (15mM) washing step prior to the salt gradient was most effective to remove $NAD^+ -binding$ proteins. For a large scale preparation of G-6-P dehydrogenase higher recovery was obtained by Affi-gel Blue than $NADP^+ -agarose$, however, the purity of the enzyme was decreased by 10 times if the former was used as the affinity medium. The capacity of Affi-gel Blue for G-6-P dehydrogenase was found to be 5 times higher than that of $NADP^+ -agarose$. Furthermore Affi-gel Blue could be reused repeatedly and its preparation is relatively easier and less expensive than $NADP^+ -agarose$.

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Purification and Properties of Homoseine Dehydrogenases in Canavalia Iineata Cotyledons (해녀콩(Canavalia lineata) 자엽에서 Homoserine Dehydrogenase의 정제 및 특성)

  • 최영명
    • Journal of Plant Biology
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    • v.39 no.1
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    • pp.41-48
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    • 1996
  • Two forms of homo serine dehydrogenase have been isolated from 8-day-old cotyledons of Canavalin lineata by a heat denaturation, ammonium sulfate fractionation, DEAE-8ephacel ion exchange and Sephacryl 8-300 gel filtration chromatographies, and Pro cion red dye, Cibacron blue dye and Resource Q column chromatographies. The molecular weights of T -form (threonine-sensitive) and K-form(threonine- insensitive) were estimated to 230 kD and 135 kD, respectively. In the presence of 10 mM threonine, the activity of T-form was inhibited with almost 70%, but that of K-form was not at all. The Km values tor homo serine of T- and Kform were 1.6 mM and 0.3 mM, respectively. The Km values for NAD of T- and K-form were 2.34 mM and 0.03 mM, respectively. And Km values for NADP of two isozymes were the same as 0.01 mM. The activities of T- and K-form were markedly stimulated up to 4.9and 2.8-fold, respectively, by 400 mM KCI. The partial purified(gel filtration) enzymes(Tform and K-form) can be reversibly converted.verted.

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Generation and characterization of calmodulin-DHFR sandwich fusion protein

  • Han, Chang Hoon
    • Korean Journal of Veterinary Research
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    • v.48 no.3
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    • pp.243-250
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    • 2008
  • A calmodulin-dihydrofolate reductase (DHFR) sandwich fusion protein was generated by insertion of calmodulin into the $\beta$-bulge region of DHFR to observe the effects of structurally constraining the calmodulin structure. The calcium binding properties of the sandwich protein were almost identical to calmodulin. Similar to calmodulin ($10.7 {\mu}M$), the sandwich protein bound four equivalents of calcium, with half saturation ($K_{0.5}$) observed at a [$Ca^{2+}$] of $8{\mu}M$. However, nicotinamide adenine dinucleotide (NAD) kinase activation property of the sandwich protein was lower than that of calmodulin. The sandwich protein activated NAD kinase, but to only half of the level obtained with calmodulin. The K 0.5 for both calmodulin and the sandwich protein were approximately the same (1-2 nM). Methylation analyses of the sandwich protein show that insertion of calmodulin into DHFR results in a large decrease in methylation. The $V_{max}$ observed with the sandwich protein (95 nmole/min/ml) was only 22% of the value observed with calmodulin (436 nmol/min/ml) in the presence of calcium. Addition of trimethoprim to the reaction significantly inhibited the observed methylation rate. Overall, the data suggest that the insertion of calmodulin into the DHFR structure has little effect on calcium binding by the individual lobes of calmodulin, but may constrain the lobes in a manner that results in altered interaction with the calmodulin-dependent proteins, and severely perturbed the methyltransferase recognition site.

Effect of Pyruvate and Aspartate Enriched University of Wisconsin Solution on Myocardial Protection (피루브산염과 아스파라진산염을 첨가한 위스콘신대학 용액의 심근보호 효과)

  • 이정렬;김준석;한재진;강문철
    • Journal of Chest Surgery
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    • v.35 no.1
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    • pp.11-19
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    • 2002
  • Background: Ischemia-reperfusion myocardial injury is an important factor to determine the early and the late mortality of transplanted patients. Recently, modulation of the cytosolic NADH/NAD+ ratio by Pyruvate and aspartate was tested to Protect the heart from ischemia-reperfusion injury. Material and Method: We added pyruvate and aspartate to the University of Wisconsin solution, and evaluated their effect on myocardial protection. We used 16 piglet(age 1 to 3 days) hearts. Eight hearts were arrested with and stored in the University of Wisconsin solution(UW solution) for 24 hours(control group), and the other eight hearts were arrested with and stored in the modified UW solution added pyruvate(3mmol/L) and aspartate(2 mmol/L)(test group). All hearts underwent modified reperfusion with blood cardioplegic solution followed by conversion to a left-sided working model with perfusion from a support pig. And then, we measured stroke work index(SWI), high-energy phosphate stores, and myocardial water content of the hearts. SWI was calculated at left ventricular end-diastolic pressures of 3, 6, 9, and 12 mmHg after 60 and 120 minutes reperfusion, respectively, Result: At 60 minutes and 120 minutes after reperfusion, SWI was higher in the test group than in the control group significantly. The levels of AMP, ADP, ATP of the test group were also higher. But, the creatine phosphate level and myocardial water content were similar in the two groups. Conclusion: From these results, we could Prove that pyruvate and aspartate enhance cardiac contractility and high-energy phosphate stores after ischemia.