• Title/Summary/Keyword: atp6

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Development of New Molecular Markers for the Identification of Male Sterile Cytoplasm in Peppers (Capsicum annuum L.)

  • Min, Woong-Ki;Kim, Byung-Dong;Kim, Sung-Gil;Lee, Sang-Hyeob
    • Horticultural Science & Technology
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    • v.29 no.1
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    • pp.53-60
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    • 2011
  • Cytoplasmic male sterility (CMS) induced by mutant mitochondria genome, has been used for commercial seed production of $F_1$ hybrid cultivars in diverse crops. In pepper (Capsicum annuum L.), two sterile cytoplasm specific gene organization, atp6-2 and coxII were identified. An open reading frame, orf456 nearby coxII gene has been speculated to induce male sterility (MS) by mutagenic analysis. Moreover, molecular markers for atp6-2 and coxII of mitochondrial genotype (mitotype) were developed. However, the Cytoplasmic MS specific markers, atp6SCAR and coxIISCAR markers appeared in both N and S cytoplasms when polymerase chain reaction (PCR) cycles prolonged more than 40 cycles. Since the reported molecular markers were dominant markers, the presence of the faint sterile-specific band in normal cytoplasm may lead to the mis-classification of pepper breeding lines. To solve this problem, one common forward primer and two different reverse primers specific to normal coxII and sterile orf456 genes were designed after analyzing their gene organizations. By using these three primers, N and S coxII specific bands were co-amplified in male-sterile lines, but only normal coxII specific band was amplified in maintainer lines. Since the reverse primer for sterile coxII was specifically designed 275 bp downstream of orf456, relatively stable PCR amplification patterns were observed regardless of the number of PCR cycles. These primer sets easily identified different mitotypes among the divergent breeding lines, commercial cultivars and diverse germplasms.

Purification and Characterization of Mitochondrial Malate Dehydrogenase during Ovarian Development in Aedes aegypti L. (Aedes aegypti L. 난성숙과정중 생성되는 Mitochondrial Malate Dehydrogenase의 정제 및 특성)

  • 김인규;이강석;정규회;박영민;성기창
    • Korean journal of applied entomology
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    • v.34 no.3
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    • pp.181-190
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    • 1995
  • Malate dehydrogenase in the mosquito ovary after a blood meal, Aedes aegypti, was purified and characterized. MDH purification steps involved DEAE-Sepharose, S-Sepharose and Cibacron blue affinity chromatography. The purified MDH was 70,000 daltons in molecular weight and was a homodimer consisting of tow identical subunits. Optimal activity of purified MDH was obtained pH 9.0-9.2 in malate-oxaloacetate reaction and pH 9.8-10.2, in oxaloactate-malate reaction. With obtained pH 9.0-92 in malate-oxaloacetate reaction and pH 9.8-10.2, in oxaloactate-malate reaction. With malate as substrate, purified mitochondrial MDH (1.28$\times$${10}^{-4}$ M) had lower Km value than cytoplasmic MDH (8.92x${10}^{-3}$ M). MDH activity was inhibited by citrate, $\alpha$-ketoglutarate, and ATP. Inhibition of MDH activity by ATP and citrate was less in malate-oxaloacetate reaction and in oxaloacetate-malate reaction. MDH activity was completely inhibited by ATP in oxaloacetate-malate reaction and not inhibited by citrate in malate-oxaloacetate reaction. Temporal activity change of MDH is similar to that of isocitrate dehydrogenase in the ovary after blood feeding; their activities in the ovary began to rise at 18 hours after a blood meal, and reached at the maximal level at 48 hours.

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Studies on the Development of Photoreceptor in the Nonchromatophore Organisms (III) -Light- Induced Mitochondrial ATP Synthase in the Lentinus edodes (Berk.) Sing.- (무흡광 색소 생물의 감광수용체 개발 연구(III) -표고버섯 중의 광감응성 Mitochondrial ATP Synthase-)

  • Min, Tae-Jin;Lee, Wan-Gie;Park, Sang-Shin
    • The Korean Journal of Mycology
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    • v.17 no.2
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    • pp.91-98
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    • 1989
  • Mitochondria in the L. edodes was separated and purified by stepped sucrose density gradient centrifugation. The activity of mitochondrial ATP synthase has been investigated during various illumination times at each wavelength within the range of 400 nm to 700 nm. The stimulation of above activity increased by two times compared with nonilluminated control group when the illumination was given for 15 seconds at 470 nm wavelength. The optimal pH and temperature of this light-induced mitochondrial ATP synthase were 7.5 and $54^{\circ}C$, respectively. The activity of this enzyme increased by 26%, 25% and 14%, respectively, when there were 1 mmole $Fe^{3+}$, 0.5 mmole $Fe^{2+}$, and 5 mmole ${SO_4}^{2-}$ ion, and was inhibited by 5 mmole $Co^{2+}$, 5 mmole $Mn^{2+}$, 1 mmole $Ca^{2+}$, 0.1 mmole $Na^+$, 5 mmole $CN^-$, and 0.1 mmole ${CO_3}^{2-}$ ion. But $Na^+$ and $K^+$ ion did not affect the activity of enzyme.

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A Study on the Protective Effect and Its Mechanism of Zinc against Immuno-cytotoxicity of Methylmercury (유기수은의 세포면역독성과 이에 대한 아연의 방어효과 및 기전)

  • 고대하;염정호;오경재
    • Journal of Environmental Health Sciences
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    • v.27 no.2
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    • pp.82-91
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    • 2001
  • This study was carried out to elucidate the protective effect of zinc chloride(ZnCl$_2$) and its mechanism against the immuno-cytotoxicity of methylmercury chloide($CH_3$HgCl). This study was observed in the culture of EMT-6 cells which are originated from mammary adenocarcinoma of Balb/c mouse. Cytotoxicity of metals was measured by cell viability and NO$_2$$^{[-10]}$ , and mitochondrial function was evaluated by adenosine triphosohate (ATP) production. $CH_3$HgCl significantly decreased the sythesis of nitric oxide(NO), ATP and glutathione(GSH) in a dose-dependent manner. ZnCl$_2$ significantly increased the synthesis of GSH in a dose-dependent manner, but synthesis of NO and ATP were not changed. The immuno-cytotoxicity of $CH_3$HgCl was not fully protected when combined addition of ZnCl$_2$, whereas ZnCl$_2$ prior to addition of $CH_3$HgCl completly protected the Hg-induced immuno-cytotoxicity. Similarly, intracellular accumulation of mercury significantly decreased by ZnCl$_2$. Degree of diminution of intracellular mercury was larger in ZnCl$_2$ prior to addition of $CH_3$HgCl than in combined addition of ZnCl$_2$ and $CH_3$HgCl.. Dithiothreitol(DTT) or buthionine sulfoximine(BSO) addition at 50$\mu$M or less, which was not toxic to the cells, did not affect synthesis of NO and ATP. DTT increased intracellular GSH level and DTT pretreatment protected toxicity induced by $CH_3$HgCl as shown complete recover in the NO and ATP values. BSO decreased intracellular GSH level and BSO pretreatment exaggerated toxicity induced by $CH_3$HgCl as shown synergistic reduction in the NO and ATP values. These results indicated that the protective effects of zinc against immuno-cytotoxicity of methylmercury associated with increasing cellular level of GSH. Increased intracellular GSH transports methylmercury to out of cells. In accordance with intracellular level of mercury decreased, immuno-cytotoxicity of methylmercury decreased. These result also suggest that the protective mechanism of zinc against the mercury toxicity would be exerted in the immune system in vivo.

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Metabolic Flux Analysis of a Poly-${\beta}$-hydroxybutyrate Producing Cyanobacterium, Synechococcus sp. MA19, Grown under Photoautotrophic Conditions

  • Nishioka, Motomu;Nishiuma, Hajime;Miyake, Masato;Asada, Yasuo;Shimizu, Kazuyuki;Taya, Masahito
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.7 no.5
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    • pp.295-302
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    • 2002
  • To understand the utilization property of light energy, Synechococcus sp. MA19, a poly-${\beta}$-hydroxybutyrate (PHB) producer, was cultivated at the different incident light intensities of 15.3, 50.0 and 78.2 W/$m^2$ using media with and without phosphate. From the results of metabolic flux analysis, it was found that the cell yield based on ATP synthesis was estimated as $3.5{\times}10^{-3}$ kg-biomass/mol-ATP in these cultures. Under the examined conditions, there were no significant differences in the efficiency of light energy conversion to chemical energies estimated as ATP synthesis and reducing potential (NADH + NADPH) formation whether the PHB synthesis took place or not. The energy converted from light to ATP was kept relatively high around the energy absorbed by the cells of $2.5-3.0{\times}10^{6} J\;h^{-1}\;kg^{-1}$, whereas the energy of reducing potential was hardly changed in the examined range of the energy absorbed by the cells.

Changes of Nucleotides in the Raw Fishes during the Aquarium Storage (수족관 저장기간에 따른 횟감생선의 핵산관련성분 변화)

  • Hong, Cheul-Hee;Lee, Jeong-Min;Kim, Kyong-Su
    • Korean Journal of Food Science and Technology
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    • v.36 no.3
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    • pp.379-384
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    • 2004
  • Changes in nucleotide concentrations of aquarium-stored flounder, sea bass, and sea bream were studied. ATP, ADP, and AMP slowly decreased, whereas IMP, HxR and Hx slightly increased with increasing storage ported. ATP was converted into IMP at initial storage stage. Changes in concentrations of nucleotides differed depending on fish type and season. Freshness indicators, $K,\;K_{p}\;G,\;P,\;H,\;and\;F_{r}$ values during 14 days storage showed no significant differences. Changes in nucleotide concentrations during 14 days storage had no significant effect on taste of raw fishes.

Degradation of Nucleotides and Their Related compounds in Sea Foods during processing and Storage -3 . Degradation of Nucleotides and Their Related compounds in Conger-eel Astroconger myriaster Muscle during Drying- (수산식품(水産食品)의 가공(加工) 및 보관중(保藏中)의 핵산(核酸) 관련물질(關聯物質)의 변화(變化)에 관한 연구(硏究) -3. 붕장어 천일건조중(天日乾燥中)의 핵산관련물질(核酸關聯物質)의 변화(變化)-)

  • Lee, Eung-Ho;Han, Bong-Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.1 no.1
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    • pp.17-24
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    • 1972
  • In this paper, the degradation of nucleotides and their related compounds in conger eel muscle during sun drying was studied. The results showed that IMP was dominant in fresh conger eel, showing 75.5% of total nucleotides while the contents of ATP, ADP, AMP, inosine and hypoxanthine were low. The nucleotides tended to degrade rapidly during drying, only 6.2% of IMP remained after seven day's sun drying and ATP, ADP and AMP were also entirely disappeared. In consideration of flavor quality, it was consumed that sun drying is not an effective processing method of conger eel, as far as IMP is concerned.

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Characteristics of Purinergic Receptor Expressed in 3T3-L1 Preadipocytes

  • Lee, Hyung-Joo;Baik, Joon-Heum;Kim, Min-Jeong;Kim, Na-Hyun;Kong, In-Deok
    • Biomedical Science Letters
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    • v.15 no.4
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    • pp.319-326
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    • 2009
  • Extracellular ATP elicits diverse physiological effects by binding to the G-protein-coupled P2Y receptors on the plasma membrane. In addition to the short-term effects of extracellular nucleotides on cell functions, there is evidence that such purinergic signalling can have long-term effects on cell proliferation, differentiation and death. The 3T3-L1 cell line derived from mouse embryo is a well-established and commonly utilized in vitro model for adipocytes differentiation and function. However, the distributions and roles of P2Y subtypes are still unknown in the preadipocyte. In this study, we identified the distributions and roles of P2Y subtypes in preadipocyte using $Ca^{2+}$ imaging and realtime PCR. ATP increased the $[Ca^{2+}]_i$ in a concentration-dependent manner. ATP increased $Ca^{2+}$ in absence and/or presence of extracellular $Ca^{2+}$. Suramin, non-selective P2Y blocker, largely blocked the ATP-induced $Ca^{2+}$ response. U73122, a PLC inhibitor, completely inhibited $Ca^{2+}$ mobilization in 3T3-L1 cells. The mRNA expression by realtime PCR of P2Y subtypes was $P2Y_2:P2Y_5:P2Y_6=1.0:12.5:0.3$. In conclusion, we showed that $P2Y_5$ receptor is a dominant purinergic receptor in preadipocytes, and multiple P2Y receptors could involve in differentiation and migration via regulating of intracellular calcium concentration.

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Changes in Contents of Some Taste Compounds of Dried Mussel and Baby Clam during Storage (건조 홍합 및 바지락의 저장 중 핵산관련물질 및 유기염류의 변화)

  • Joo, Ok-Soo;Seo, Kwon-Il;Lee, Young-Soo;Lee, Jong-Ho;Choi, Sang-Do;Shim, Ki-Hwan
    • Korean Journal of Food Science and Technology
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    • v.28 no.5
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    • pp.882-887
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    • 1996
  • Some taste compounds such as nucleotide and their related compounds, trimethylamine oxide (TMAO), trimethylamine (TMA) and total creatinine of sea mussel and baby clam during drying at 40, 50 and 60^{\circ}C$ and storage at low temperature$(4^{\circ}C)$ and room temperature$(20^{\circ}C)$ were investigated. Six kinds of nucleotide and their related compounds such as adenine triphosphate (ATP), adenine diphosphate (ADP), adenine monophosphate (AMP), inosine, adenosine and hypoxanthine were analyzed. The contents od adenosine in raw sample was high in sea mussel and baby clam. The contents of ATP, ADP and AMP decreased, while those of inosine and hypoxanthine increased during drying and storage periods. The contents of TMAO, TMA and total creatinine were low in sea mussel and baby clam. TMAO and total creatinine decreased but TMA increased during drying and storage periods. The rate of change was high in room temperature storage and for long storage periods than that of low temperature storage and for short storage periods.

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Knowledge and Information Need for Prenatal Genetic Screening and Diagnosis and Attitude toward Terminating Pregnancy among Pregnant Women in South Korea (임산부의 산전 기형아 검사에 관한 지식과 정보 요구 및 낙태에 대한 태도)

  • Jun, Myunghee;Shin, Gyeyoung;Kim, Hye-Kyung
    • The Journal of Korean Academic Society of Nursing Education
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    • v.24 no.4
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    • pp.463-477
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    • 2018
  • Purpose: This study identifies correlations among information needs and knowledge about prenatal genetic screening and diagnosis (I-PGSD & K-PGSD), and attitude toward terminating pregnancy (ATP) among pregnant women in South Korea. Methods: A descriptive survey was conducted from January 2013 to April 2014 in South Korea. 222 pregnant women responded to three questionnaires developed by the authors. The questionnaire for I-PGSD consisted of 19 questions; 18 questions for K-PGSD; and 10 questions for ATP. Results: Mean scores were $80.46{\pm}11.73$ for I-PGSD; $14.86{\pm}3.74$ for K-PGSD; and $33.71{\pm}6.13$ for ATP. The ATP score was positively correlated with the I-PGSD and K-PGSD scores, but statistically significant with only I-PGSD (p=.006). I-PGSD scores were higher than average on three genetic syndromes (Down, Patau, and Edwards syndrome), on management after the diagnosis of positive fetal aneuploidy, and on test result interpretation after the amniocentesis and level II fetal ultrasonogram. Conclusions: In light of current legal and moral controversy regarding terminating pregnancy and rapidly advancing prenatal genetic testing technology, more prenatal genetic education for nurses and nursing students who teach pregnant women is needed. In addition, more professional counseling services provided by trained nurses are also required.