• Title/Summary/Keyword: ATPase8

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Reproductive cycle and Maturation Induction of Oocytes in Rana rugosa (옴개구리의 생식주기와 난자의 성숙유도)

  • 유명식;나철호
    • The Korean Journal of Zoology
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    • v.38 no.1
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    • pp.96-105
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    • 1995
  • 본 연구는 옴개구리(Rana rugosa)의 생식주기를 파악하고 이들 난자들의 체외 성숙조건을 구하기 위하여 수행하였다 개구리들의 gonadosomatic index(G51)는 4월에서 8월사이에는 비교적 낮았고 9월에서 이듬해 3월까지는 높았다 여포들의 성장은 주로 6월에서 9월 사이에 이루어지는데 난소내에서도 여포들의 성장 속도는 일부 다른 것으로 나타났다. 야외 관찰에서 이 개구리들은 서식지의 온도에 따라 4월에서 7월 사이에 산란을 한다는 것과 10월에서 이듬해 3월까지 동면을 한다는 것을 말았다. 산란기에 취한 여포 난자들은 생체외 배양에서 progesterone에 성숙반응(germinal vesicle breakdown(GVBD))을 일으키지 않았다. 그러나 protein klnase C(PKC)의 촉진제인 12-O-tetradecanoylphorbol 13-acetate(TPA) 혹은 Na+/K+ ATPase의 저해제인 ouabain을 progesterone과 동시에 처리했을 때에는 성숙반응을 일으켰다(각각 86%와 80%). TPA 로 핵붕괴를 일으킨 난자의 세포질을 미성숙 난자에 주입하면 미성숙 난자의 핵붕괴를 유도하였으며 성숙된 다른 종의 난자들의 세포질도 이와 같은 효과를 나타내었다. TPA의 성숙유도 효과는 5분의 노출 기간으로도 충분하였으며 PKC의 저해제인 H-7을 처리하면 그 효과가 없어졌다 이러한 결과들은 옴개구리의 난자는 호르몬에 성숙반응을 일으킨지 않으나 PKC 활성화 이후 단계는 정상이라는 것을 의미한다.

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Characterization of Mitochondrial NADH Dehydrogenase in Lentinus edodes (표고버섯의 미토콘드리아성 NADH 탈수소효소의 특성)

  • Kim, Eun-Mi;Min, Ji-Young;Min, Tae-Jin
    • The Korean Journal of Mycology
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    • v.26 no.1 s.84
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    • pp.119-126
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    • 1998
  • Mitochondria were isolated from Lentinus edodes and properties of the mitochondrial NADH dehydrogenase were studied. Optimal pH, temperature, and thermal stability of the enzyme were estimated to be 7.6, $33^{\circ}C$, and stable for one hour at $50^{\circ}C$. The apparent $K_m$ for the NADH was 0.33 mM. This enzyme catalyzed to transfer electrons from NADH to ferricyanide, decylubiquinone, and 2,6-dichloro-phenol-indophenol. 0.5 mM antimycin A and 0.01 mM dibromothymoquinone strongly inhibited 87.8% and 76.5% of the enzyme activities. 0.01 mM oligomycin known as an inhibitor of ATPase also strongly inhibited 79.2% of activities. 0.5 mM 5,5'-dithiobis-(2-nitrobenzoic acid) and 1.0 mM N-ethylmaleimide known as a modifier of SH group inhibited 50.4% and 36.7% of activities. 1 mM ethyl 2,4-dihydroxy-6-methyl benzoate and 10 mM orcinol, which had been known as an antibiotics isolated from Umbilicaria vellea according to our previous work, stimulated 68.4% and 48.1% of the enzyme activities.

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Histological and histochemical studies of the squirrel epididymis (다람쥐 부고환의 조직학적 및 조직화학적 연구)

  • Ryu, Si-yun;Cho, Sung-whan;Kim, Moo-kang;Kim, Sung-ho;Lee, Cha-soo
    • Korean Journal of Veterinary Research
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    • v.29 no.3
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    • pp.215-222
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    • 1989
  • In order to investigate the morphological characteristics of epididymal duct of the squirrel, the histological and histochemical studies were carried out. The results obtained were summarized as follows: The epididymal duct can be divided into 9 segments by histological and histochemical features. Segments 1 to 5 were located in the head, segments 6 and 7 in the body, and segments 8 and 9 in the tail of the epididymis. The apical cells were numerous in the segment 1. Clear cells which has a compact, deeply staining nucleus and a characteristically clear cytoplasm were scattered in the epithelium throughout the duct. Interepithelial clear cells which had PAS-positive granules tended to increase in number caudally. Strong PAS-positive reaction was detected at the intralumen of the segments 3,8 and 9. Acid phosphatase activity was relatively high in the basal cytoplasm of the segment 7, and then in the supranuclear region of the segments 8 and 9. Alkaline phosphatase activity was weakly positive or negative except the segments 3 and 4. ATPase activity was strong in the free surface of the epithelium in the head and the entire cytoplasm in the body and tail, a,nd SDH activity was generally weak except for the body where it was more intense.

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Effect of Ethanol on $Na^+-P_i$ Uptake in Opossum Kidney Cells: Role of Membrane Fluidization and Reactive Oxygen Species

  • Park, In-Ho;Hwang, Moon-Young;Woo, Jae-Suk;Jung, Jin-Sup;Kim, Yong-Keun
    • The Korean Journal of Physiology and Pharmacology
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    • v.3 no.5
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    • pp.529-538
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    • 1999
  • This study was undertaken to examine the effect of ethanol on $Na^+ -dependent$ phosphate $(Na^+-P_i)$ uptake in opossum kidney (OK) cells, an established renal proximal tubular cell line. Ethanol inhibited ^Na^+-dependent$ component of phosphate uptake in a dose-dependent manner with $I_{50}$ of 8.4%, but it did not affect $Na^+-independent$ component. Similarly, ethanol inhibited $Na^+-dependent$ uptakes of glucose and amino acids (AIB, glycine, alanine, and leucine). Microsomal $Na^+-K^+-ATPase$ activity was not significantly altered when cells were treated with 8% ethanol. Kinetic analysis showed that ethanol increased $K_m$ without a change in $V_{max}$ of $Na^+-P_i$ uptake. Inhibitory effect of n-alcohols on $Na^+-P_i$ uptake was dependent on the length of the hydrocarbon chain, and it resulted from the binding of one molecule of alcohol, as indicated by the Hill coefficient (n) of 0.8-1.04. Catalase significantly prevented the inhibition, but superoxide dismutase and hydroxyl radical scavengers did not alter the ethanol effect. A potent antioxidant DPPD and iron chelators did not prevent the inhibition. Pyrazole, an inhibitor of alcohol dehydrogenase, did not attenuate ethanol-induced inhibition of $Na^+-P_i$ uptake, but it prevented ethanol-induced cell death. These results suggest that ethanol may inhibit $Na^+-P_i$ uptake through a direct action on the carrier protein, although the transport system is affected by alterations in the lipid environment of the membrane.

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Effect of Salviae Radix on Impairment of Membrane Transport Function in Rabbits with Myoglobinuric Acute Renal Failure (마이오글로빈뇨성 급성 신부전 토끼에서 신장 세포막 수송 기능 장애에 대한 단삼의 효과)

  • Ji-Cheon, Jeong;Hyun-Soo, Kim
    • The Journal of Korean Medicine
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    • v.21 no.3
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    • pp.119-128
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    • 2000
  • This study was carried out to determine if Salviae Radix extract (SRE) exerts protective effect against alterations in membrane transport function in rabbits with rhabdomyo lysis-induced acute renal failure. Acute renal failure was induced by intramuscular administration of glycerol (50%, 10 ml/kg). GFR in the glycerol-injected animals was reduced to 11% of the basal value and the fractional $Na^{+}$ excretion was increased to 7.8-fold, indicating generation of acute renal failure. When animals received SRE pretreatment for 7 days prior to glycerol injection, such changes were significantly attenuated. The fractional excretion of glucose and phosphate was increased more than 43-fold and 27-fold, respectively, in rabbits treated with glycerol alone. However, they were increased to 17-and 4.3-fold, respectively, in SRE-pretreated rabbits, and these values were significantly lower than those in rabbits treated with glycerol alone. Uptakes of glucose and phosphate in purified isolated brush-border membrane, the $Na^{+}-K^{+}-ATPase$ activity in microsomal fraction, and cellular ATP levels all were reduced in rabbits treated with glycerol alone. Such changes were prevented by SRE pretreatment. Uptakes of organic ions, PAH and TEA, in renal cortical slices were inhibited by the administration of glycerol, which was prevented by SRE pretreatment. Pretreatment of an antioxidant DPPD significantly attenuated the increase in the fractional excretion of glucose and phosphate induced by rhabdomyolysis. These results indicate that rhabdomyolysis causesimpairment inreabsorption of solutes in the proximal tubule via the generation of reactive oxygen species, and SRE pretreatment may provide the protection against the rhabdomyolysis-induced impairment by its antioxidant action.

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Occurrence of Leaf Spot Caused by Stemphylium lycopersici on Cirsium setidens in Korea (Stemphylium lycopersici에 의한 고려엉겅퀴 점무늬병의 발생)

  • Choi, Hyo-Won;Kim, Seok Gu;Hong, Sung Kee;Lee, Young Kee;Lee, Jae Guem;Kim, Hyo Won;Lee, Eun Hyeong
    • The Korean Journal of Mycology
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    • v.44 no.3
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    • pp.201-205
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    • 2016
  • In August 2015, leaf spot symptoms were observed on Korean gondre thistle (Cirsium setidens) in Youngwol, Korea. During the early stage, the symptoms appeared as one or more small gray-brown to brown spots on plant leaves. The spots showed extensive enlargement over time and eventually became large dark brown to black lesions on the whole leaf. Stemphylium species were consistently isolated from affected leaves. All isolates were identified as S. lycopersici, S. solani, or S. xanthosomatis based on morphological and cultural characteristics. The isolates were confirmed as S. lycopersici based on a multilocus sequence analysis using the ribosomal internal transcribed spacer (ITS) region, elongation factor 1, GAPDH (glyceraldehyde-3-phosphate dehydrogenase), and the noncoding region between the vacuolar membrane ATPase catalytic subunit A gene and a gene involved in vacuolar biogenesis. Pathogenicity was tested by spore suspension inoculation on wounded or unwounded gondre leaves. The lesions were observed on inoculated leaves within 3 days after inoculation, regardless of wound. To our knowledge, this is the first report of the leaf spot on gondre thistle caused by S. lycopersici in Korea or elsewhere.

Effects of Cholesterol Diet and Exercise on Plasma and Liver Lipids, Platelet Aggregation and Erythrocyte Na Efflux in Rats (콜레스테롤 식이와 운동이 흰쥐의 체내 지질수준, 혈소판 응집 및 적혈구막 Na 유출에 미치는 영향)

  • Kang, Min-Sook;Kim, Young-Pyo;Kang, Young-Hee;Kang, Jung-Sook
    • Journal of Nutrition and Health
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    • v.41 no.5
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    • pp.381-390
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    • 2008
  • Present study was conducted to investigate the effects of exercise and cholesterol diet on plasma and liver lipids, platelet aggregation, erythrocyte Na efflux and liver index such as GOT and GPT using Sprague Dawley rats. Forty rats were divided into four groups and fed control or 0.5% cholesterol diet with and without treadmill for six weeks. The final body weight of group fed cholesterol diet with exercise was somewhat decreased compared with group fed cholesterol diet without exercise. L.W/B.W ratio was significantly increased in groups fed cholesterol diet (p < 0.01), but exercise tended to decrease this ratio. Plasma total cholesterol was significantly increased and HDL-cholesterol was decreased in groups fed cholesterol diet (p < 0.01). Plasma triglyceride was significantly decreased in groups fed cholesterol diet compared with groups fed control diet (p < 0.01). Plasma triglyceride of groups with exercise was significantly decreased compared with their non exercising counterparts regardless diet (p < 0.05). Liver total cholesterol and triglyceride was significantly increased in groups fed cholesterol diet (p < 0.01), but exercise did not affect on these levels. Na-K ATPase was somewhat decreased in groups fed cholesterol diet, and exercise tended to recover the reduced Na-K ATPase. Na passive transport was significantly decreased in group fed control diet without exercise and significantly increased in group fed cholesterol diet with exercise, there were significant differences between groups (p < 0.05). There were no differences in total Na efflux and intracellular Na among groups, and total Na efflux was not correlated with intracellular Na. Hematocrit was significantly lower (p < 0.05) in group fed cholesterol diet without exercise compared with other groups. Platelet aggregation in the initial slope and the maximum was increased in groups fed cholesterol diet, but not statistically significant. Exercise especially increased the initial slope of aggregation. Plasma GOT and GPT was significantly increased in groups fed cholesterol diet (p < 0.01), and exercise in group fed cholesterol diet significantly decreased both GOT and GPT compared with the non exercising counterpart (p < 0.01). This study showed that cholesterol diet increases plasma and liver lipids and GOT and GPT, and exercise improves plasma and liver lipid profile and liver index of GOT and GPT preventing fatty liver.

Effects of Dietary n-3 Highly Unsaturated Fatty Acids on Growth and Biochemical Changes in the Korean Rockfish Sebastes schlegeli II. Changes of Blood Chemistry and Properties of Liver Cells (사료의 n-3계 고도불포화지방산 함량에 따른 조피볼락 Sebastes schlegeli의 성장 및 생화학적 변화 II. 혈액성분 변화 및 간세포 성상)

  • LEE Sang-Min;LEE Jong Yun;KANG Young Jin;HUR Sung Bum
    • Journal of Aquaculture
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    • v.6 no.2
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    • pp.107-123
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    • 1993
  • To define the effects of various levels $(0\~1.5\%)$ of dietary n-3HUFA on the physiological changes in the Korean rockfish, variations in blood variables and hepatocytes were studied. Biochemical serum analyses, lactate dehydrogenase (LDH) activity of the liver cytosol and ATPase activity of the liver microsomal membrane were also studied. The haematological values (red blood cell, hemoglobin, hematocrit, MCHC, MCV and MCH) were not significantly different in the experimental groups $(P\geq0.05)$. The total protein and glucose levels in the serum were affected by dietary n-3HUFA levels. These levels in groups fed n-3HUFA insufficient diets were significantly lower than those of n-3HUFA sufficient groups (P<0.05). Serum levels of total cholesterol, free cholesterol, glutamic pyruvic transaminase (GPT) and glutamic oxaloacetatic transaminase (GOT) showed significantly higher values in the fish fed n-3HUFA deficient diets (P<0.05). The LDH in the serum was dropped with increasing dietary n-3HUFA levels, but the LDH activity of the liver cytosol was elevated. Histologically, the hepatic cell in the fish fed n-3HUFA free diet was abnormal and showed a necrotic condition. $Ca^{2+}-ATPase$ activities of the liver microsomal membrane were significantly lower in the fish fed n-3HUFA deficient diets than in those fed n-3HUFA sufficient diets (p<0.005). These results suggested that the liver cell membrane was affected by dietary fatty acid compositions and cell membrane of the fish fed n-3HUFA insufficient diets showed abnormalities.

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생쥐 초기 배아에서 Aquaporin 8과 9의 발현에 관한 연구

  • 신현상;계명찬;강수만;이성은;이지원;강한승;김문규
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.68-68
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    • 2003
  • Aquaporin은 막관통 통로 단백질(transmembrane channel protein)로서, 삼투압의 농도구배에 따라 세포막을 가로질러 물분자를 이동시키는 기능을 하고 있다. 포유류 초기배아에서 포배강 형성은 영양외배엽세포에서 $Na^+ / K^+$ATPase에 의한 이온 농도 구배가 형성되면 auqaporin에 의해 물이 포배강으로 유입되면서 이루어진다. 본 연구에서는 생쥐 초기배아에서 반정량적인 역전사 중합효소 연쇄반응 방법(semi-quantitative RT-PCR)과 실시간 역전사 중합효소 연쇄반응 방법(real-time RT-PCR)을 통하여 AQP8과 9의 mRNA발현을 조사하고 다중 면역형광현미경 방법(confocal immunofluorescence microscopy)을 통해 단백질 발현양상을 분석하였다. AQP8 mRNA는 상실기까지 발현되지 않다가 포배기에 이르러 발현되었고 AQP9 mRNA는 수정란에서부터 발현되어 포배기에는 유의할 정도로 증가하였다. 따라서 AQP8 mRNA는 배아유전자가 활성화되어 나타나는 것이고 AQP9 mRNA는 모계유전자 기원임을 알 수 있었다. AQP8 단백질은 상실배 단계까지 발현되지 않다가 포배시기에 영양외배엽세포사이의 접합면에 발현되었고 AQP9 단백질은 상실배 시기에 할구 사이의 인접 부위에서 강하게 발현되었다가 포배시기에는 세포간의 접합면에 약하게 발현하는 경향을 나타내었다. 실시간 역전사 중합효소 연쇄반응 방법으로 조사한 결과 포배에서 물과 글리세롤을 통과시키는 AQP9는 mRNA의 발현양이 AQP8보다 약 4배 정도 많았다. 또한 포배기에 이르러서야 물만을 통과시키는 AQP8의 발현이 나타나는 것을 보아 포배강 형성시 외부에서 영양외배엽을 통해 포배강으로 유입되는 물의 이동(trans- trophectodermal water movements)에 AQP9보다 AQP8이 더 중요하게 관여할 것으로 사료된다., K, Pb, Cd, Cr, Co, Cu, Ni)을 측정하였다. 실험 조건1의 결과로서 각 국의 유아용 일회용 기저귀의 중금속 함량은 거의 유사한 경향을 나타내었으며 Cr, Zn, Pb, Ni, Mn, Mg, Li, K는 detection limit(2 ppm) 이하였고, Cd, Fe, Co, Cu, Ca, Al, Sr는 검출되었지만 기준치 이하였다. 실험 조건2의 결과로서 측정 항목(Cr, Sb, Cd, Pb, Ni, Co, Cu)중 Cr, Cd, Ni, Cu는 detection limit(0.1 ppm) 이하였고, Sb, Pb, Co는 검출되었지만 기준치 이하였다.았다. 4%의 경우에는 8$0^{\circ}C$이하로 온도를 낮추는 것이 좋은 상태를 나타내었다. 이와 같은 결과는 일반적으로 화학적 레팅을 4%, 7%에서한 선행결과와 상당히 다른 결과이다.염 농도가 증가할수록 감소 현상을 보였다.X>, 75BG30은 8.6$\mu\textrm{m}$, 75BG40은 7.02$\mu\textrm{m}$로 나타났다. 따라서 경화제 양에 관계없이 10$\mu\textrm{m}$ 이하로 나타나, 경화제 10$m\ell$만으로 미세한 크기를 얻을 수 있음을 알 수 있다. 젤리 강도 변화에 따른 차이는 300BF는 78.09$\mu\textrm{m}$ 300BG는 56.32$\mu\textrm{m}$로, 75BF나 75BG에 비하여 현저히 증가하여, 젤라틴의 젤리 강도는 캡슐 제조 조건의 주요한 변수임을 알 수 있다.추출물 투여시 혈당강하 및 혈중콜레스테롤 강하가 나타났으며, 상엽복합추출물 투여와 운동을 병행시 이러한 감소 효과가 더 뚜렷하게 나타났다.교육의 적임자로 보는 시각이 비교적 높았고 약 1/2정도는 영양교육에 참여하겠다는 의지를 가지고 있을 뿐만 아니라 실제로 영양지도를

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Characterization of proteases of Toxoplasma gondii (Toxoplasma gondii에서 단백질 분해 효소의 특징)

  • Choe, Won-Yeong;Nam, Ho-U;Yun, Ji-Hye
    • Parasites, Hosts and Diseases
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    • v.27 no.3
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    • pp.161-170
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    • 1989
  • The proteases of Toxoplasma gcndii were purified partially and characterisrd for some biochemical properties including various chromatographic patterns, major catalytic classes, and conditions to promote the activity of these enzymes. When Toxoplasma extract was incubated with 3H-casein at various pH, peak hydrolysis of casein was observed at pH 6.0 and at pH 8.5. Proteasfs working at pH 6.0 and at pH 8.5 were purified partially by conventional methods of chromatographies of DE52 anion rxchange, Sephadex G-200 gel permeation, and hydroxylapatite chromatography. Partially purified enzymes were tested by site-specific inhibitors and promotorf. The protease working at pH 6.0 was inactivated by iodoacetamide with LDso of 10-5 M and promoted by dithiothreitol, while the protease working at pH 8.5 was inhibited by phenylmethylsulfonyl fluoride with LD50 of 10-5 M and was Promoted by ATP (excess ATP beyond 2 mM inhibited the activity reversely). The protease of pH 8.5 had the activity of ATPase which might exert the energy to its action. Therefore the former was referred to as a cysteinyl acid protease and the latter, ATP-dependent neutral serine protease.

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