사람 洋水의 Adenosine Diphosphoribose Pyrophosphohyrolase

黃炳斗 충남대학교 대학원 1976 論文集 Vol.6 No.-

Adenosine diphosphoribose pyrophosphohydrolase (ADPR-PPase) which catalyzes the hydrolysis at the pyrophosphate linkage of adenosine diphosphoribose (ADP-ribose) has been obtained from human amniotic fluid and partially purified, and it's properties have been investigated. This enzyme shows optimal pH around 9.5 and is heat-labile, being completely inactivated by heat-treatment at 65° for 10 minutes. In contrast to erythrocyte ADPR-PPase of rabbit, the enzyme does not require Mg^(2+), but requires inorganic phosphate for the activity, maximal activity being atained at 20 mM phosphate. The apparent Michaelis constant for ADP-ribose is 1.4 mM. Oxidized and reduced NAD, and pHMB are not inhibitory The enzyme is inhibited markedly by adenine nucleotides which act as a competitive inhibitor, although CMP and GMP are also inhibitory yet lesser in magnitude. These results indicate that the enzyme is controled by related compounds as well as reatcion product.

胎盤 組織의 Protein Methylase II

황병두,이재흔,백문기 충남대학교 의과대학 지역사회의학연구소 1982 충남의대잡지 Vol.9 No.2

Protein methylase Ⅱ has been purified from human term placenta approximately 8,760 folds with a 14.5% yield. The enzyme shows sharp pH optimum around pH 5.8. The enzyme is easily inactivated by heat treatment for 5 minutes at 60℃, and when stored at -20℃ even in the presence of 10% glycerol, approximately 70% of activity has been lost for 8 weeks The enzyme doesn't require any divalent cations and Cu^2+ is a potent inhibitor, the activity being completely inhibited at 2 mM and 86% of the activity is recovered by addition of 4 mM EDTA. Histone IIA and myelin. basic protein arc good substrate for this enzyme. Km for S-adenosyl-L-methicnine and Ki for S-adenosyl-L-homocysteine are 2.03 x 10 exp(-6) M and 5. 8 x 10 exp(-7) M respectively. The methylated membrane protein from human placenta were analyzed by dodecyl sulfate/polyacrylamide gel electrophoresis. Band 1, 2, 4, 15, 17 are identified as 5 major classes of methyl-acceptor protein for protein methylase Ⅱ. The role of placental membrane prctein methylation is discussed with regard to placental function.

胎盤組織의 Adenosine Deaminase

황병두,이재흔,백문기 충남대학교 의과대학 지역사회의학연구소 1981 충남의대잡지 Vol.8 No.2

Adenosine deaminase(ADase) has been partially purified approximately 800-fold from human placenta, and its properties have been investigated. The enzyme exhibits a broad pH optimum from pH 6 to 8 and is heat-labile, being completely inactivated by heat treatment at 70℃ for 10 minutes. In contrast to other tissue ADase, the enzyme is inhibited by 2mM of Cu^2+ and Fe^2+ at pH 7, and by 2 mM of Cu^2+, Fe^2+, Co^2+ and Mn^2+ at pH 8. The inhibitory effect of Cu^2+ on the enzyme is so strong that the enzyme is completely inactivated by 20 μM of Cu^2+. The enzyme activity is easily recovered by EDTA(4 mM) in case of adding Cu^2+ after preincubation of enzyme with substrate, while the recovery is about 50% by treating Cu^2+ prior to addition of substrate. Thus substrate binding to enzyme seems to prohibit the Cu^2+ from binding to the enzyme. The apparant Michaelis constant for adenosine is 77μM, and the kinetic analysis in the presence of 2 μM Cu^2+ shows that Km is independent while Vmax is decreased. The molecular weight of the enzyme is about 800,000 and the apparant activity of the other type of ADase is not detected. From the above results it is suggested that the enzyme is different from other tissue ADase in its nature of protein and possibly biologic role.

정신과 환자들의 의료서비스 이용에 관한 연구 : 경기도 연천 지역에서 In Yonchon Area of Kyunggi Province

황성혜,홍진표,권준수,우종인,김중술,조두영,이부영,조맹제,양병국,배재남 大韓神經精神醫學會 1996 신경정신의학 Vol.35 No.4

본 연구는 농촌지역 정신과 환자들의 의료서비스 이용 형태를 알아보고자 하여 행하여졌다. 경기도 연천군 주민중 1993년 1년간 지역 의료보험과 의료보호자료에서 정신과적 진단을 받은 사람들 1295명 중 209명을 대상으로 하였다. 연구자들이 제작한 반구조화된 면담도구를 사용하여 13명의 훈련된 면담자에 의해 증상을 처음 느낀 후 이용한 의료서비스를 차례대로 조사하였다. 결과는 다음과 같다. 정신증적 장애 환자들은 정신과 치료만을 받은 경우가 28.8%이며 처음 방문한 의료서비스는 정신과 45%, 일반의 28%, 약국 6%, 한방치료 5%, 요양원 6%, 기타 10%였다. 비정신증적 장애 환자들은 정신과를 한번이라도 이용한 경우는 32.1%였고 처음 방문한 의료서비스는 정신과 15%, 일반의59%,약국 17%, 한방치료8%, 기타1%였다. 진단에 따라서는, 정신분열병의 경우 64.5%가 처음에 정신과를 방문하였고 정동형 정신병의 경우는 20.8%에 불과했고 64.1%가 일반이나 약국을 처음에 방문하였다. 신경증적 장애(ICD-9 300)는 40.0%가 일반의만을 방문했다. 이상의 결과로 정신분열병 환자들은 대다수가 정신과 치료를 받는 것으로 나타났으나 정동형 정신병과 신경증적 장애는 다수가 일반의를 이용하는 것으로 나타났다. 지역사회 정신보건에서 일반의의 역할이 매우 중요하며 일반의에 대한 정신과 교육이 절실하고, 보다 전문적인 치료환경과 기술을 요하는 경우에는 정신과와의 연계 체계의 필요성도 요구된다. The purpose of this study was to investigate the pathways of health care seeking by the psychiatric patients at a well - defined rural area in Korea. Among 1,295 patients diagnosed to have mental disorders by ICD-9 in a year, all the psychotic patients(184 in number) and randomly selected 15% of nonpsychotic patients(162 in number) were selected as subjects of this study. Thirteen well - trained interviewers collected the data using semi-structurd inter-view instrument developed by the authors. Two hundred nine patients(60.4%) fully responded to the interviews. As for psychotic patients, 28.8% used only psychiatric services, and the first visits for help were psychiatrists, general practitioners, pharmacists, asylums and oriental herb medicine in the order of decreasing frequency. Only 32.1% of nonpsychotic patients used mental health specialties during the course of their illness, and their most common first visits for care were general practitioners(59%) followed by pharmacists(17%) and psychiatrists(15%). Majority(64%) of schizophrenic patient visited psychiatrist first, while 64.1% of affective psychosis patients visited general practitioners and pharmacists first. The forty percent of neurotic patients(ICD-9 300) visited general practitioners only for their health care.

사람 胎盤組織 膜 結合 Adenosine Deaminase의 性狀에 關한 硏究

황병두,홍승원 충남대학교 의과대학 지역사회의학연구소 1982 충남의대잡지 Vol.9 No.1

The properties of plasma membrane bound adenosine deaminase(ADase) from human placenta have been investigated. The enzyme exhibits a broad pH optimum from pH 6 to 8 and is heat labile, being completely inactivated by heat treatment at 70℃ for 10 minutes. The enzyme is not effected at pH 7 by 2 mM. Li^+, Mg^2+, Ca^2+, Mn^2+, Co^2+, Fe^2+ and Zn^2+, while the enzyme is activated by 2 mM Ca^2+ and Co^2+ at pH 8 and inhibited by 2 mM Cu^2+ independent of pH. The enzyme activity is completely recovered by EDTA (4 mM) only in case of adding Cue+ after preincubation of enzyme with substrate. The apparent Michaelis constant of the enzyme for adenosine is 58 μM. From the above result, it is suggested that the enzyme is different from ADase of soluble fraction of human placenta.

胎盤組織 Cytidine Deaminase에 關한 硏究 (Ⅰ) : Purification & Characterization

황병두,곽상태,임규,이재흔,백문기 충남대학교 의과대학 지역사회의학연구소 1984 충남의대잡지 Vol.11 No.2

Cytidine deaminase has been partially purified from human term placenta approximately 26-fold by combination of ammoninm sulfate saturation, sephacryl S-300, DEAE-cellulose and hydroxyapatite chromatography, and then its properties have been investigated. The enzyme shows a broad optimum pH from 5 to 9 and is heat-stable being, almost remained its activity by heat treatment at 70℃ for 8 minutes. The enzyme activity is inhibited to 23% and 77% by 2 mM of Cu^2+ and Co^2+, respectively, but, other cations do not affect the enzyme activity. The apparant Michaelis constant for cytidine and deoxycytidine are 74 uM and 69 uM, respectively. Kinetic analysis of cytidine deminase in presence of 0.25 mM Cu^2+ shows that the nature of the inhibition to cytidine deaminase is competitive, considering that Vmax is independent while km value is increased. The enzyme activity is completely inhibited by 0. 5 mM of p-hydroxymecuribenzoicacid(PHMB)but all of the activity is recovered in adding mercaptoethanol. The enzyme activity is decreased about 50% in prescence of 5 mM of polyamines. The enzyme activity is mainly located in the nuclei (80%) and cytosol(16%) fractions. he molecular weight of the enzyme is about 770, 000.

胎盤組織 S-Adenosyl-L-Methionine : Protein-Carboxyl-O-Methyltransferase에 關한 硏究 Protein-Carboxyl-O-Methyltransferase from Human Placenta

黃炳斗,白汶基 충남대학교 의과대학 지역사회의학연구소 1983 충남의대잡지 Vol.10 No.2

S-Adenosy-L-methionine: protein-carboxyl O-methyltransferase (protein methylase Ⅱ) has been purified from human term placenta approximately 8,700-fold with a 14% yield. the enzyme shows a sharp pH optimum around pH 5.8. The enzyme is easily inactivated by heat treatment for 5 minutes at 60℃, and when stored at -20℃ even in the presence of 10% glycerol, approximately 70% of activity has been lost in 8 weeks. Copper ion (Cu^2+) is a potent inhibitor, the activity being completely inhibited at 2 mM and 86% of the activity is recovered by addition of 4mM EDTA. The other divalent cations are almost without effect. Human chorionic gonadotropin, immunoglobulin A, and histone ⅡA are good substrates for this enzyme with preference for HCG. The Km value for S-adenosyl-L-methionine and Ki value for S-adenosyl-L-homocysteine are 2.08 × 10 exp (-6) M and 5.8 × 10 exp (-7) M, respectively. The methylated membrane protein from human placenta are analyzed by dodecyl sulfatepolyacrylamide gel electrophoresis. The bands 9 and 17 of the membrane protein are identified as two major classes of methyl acceptors, but in the presence of epinephrine, methylation of the band 9 is decreased and the bands 13, 14, 16 and 17 become major methyl-acceptors. Especially methylation of the band 17 is increased about 3-fold compared with that of the band 17 in the absence of epinephrine. The nuclear membrane proteins are found to be a good methyl-acceptor protein. The enzyme activity with nuclear fraction as substrate is increased about 17% when 39,000 x g supernatant and prednisolone are added together to reaction mixture. Methylated histone prefers to bind methylated nucleus rather than nonmethylated nucleus. The enzyme activity is changed with gestational age, the activity at first and second trimester being about 2 times higher than that of term placenta. From the above results, the possible roles of this enzyme, such as a regulation of hormone action, transport of histone and steroid hormone into the nucleus and signal transmission, are discussed.

初期 胎盤組織의 Adenosine Deaminase

황병두,변해공,임규,이재흔,백문기 충남대학교 의과대학 지역사회의학연구소 1983 충남의대잡지 Vol.10 No.2

Two forms of adenosine deaminase (ADase) having molecular weights of 1,500,000 & 800,000 in the soluble fraction of the early placenta and plasma membrane bound ADase of the placenta were demonstrated respectively. Their properties and activity changes with gestational age have been investigated. ADases exhibit a broad pH optimum from 6 to 9 and heat labile, being completely inactivated by heat treatment at 70℃ for 10 min. Soluble fraction ADases and plasma membrane bound ADase are inhibited about 100% and 80% by s mM Cu^2+. The apparent Michaelis constant of pADase-cytotrophoblast(pADase-C) for adenosine is 110μM, and for deoxyadenosine 21μM, that of pADase-syncytiotrophoblast(pADase-S) is 80μM and 35μM, and that of plasma membrane bound ADase is 31μM and 26μM, respectively. Kinetic analysis of pADase-C and pADase-S in the presence of 10μM Cu^2+ show that the nature of the inhibition to pADase-C and pADase-S are noncompetitive and uncompetitive, respectively. The pADases (pADase-C + pADase-S) activity decreased with gestational age, the activity at term being about one-tenth that of the early placenta, while plasma membrane bound ADase increased with gestational age by about 2-fold. From above results it is discussed that pADase-C is related to cytotrophoblast, pADase-S to syncytiotrophoblast and plasma membrane bound ADase to cellular differentiation of placenta, and suggested that soluble fraction ADases of human placenta is different from plasma membrane bound ADase in their biological roles.

精神分裂症의 生物學的 硏究 : 液體 크로마토그라프와 放財酵素法에 依한 Catecholamines 微量定量 A Rapid Microassay of Catecholamines by High Performance Liquid Chromatography-Radioenzymatic Method

고병학,송옥헌,임규,황병두,백문기 中央醫學社 1983 中央醫學 Vol.45 No.4

A direct rapid radioenzymatic assay in combination with high performance liquid chromatography (HPLC) for measuring simultaneously picogram quantities of nore pinephrine, epinephrine and dopamine has been developed, and the relationship of changes in cerebrospinal fluid (CSF) norepinephrine concentration during haloperidol treatment to changes in psychosis rating of twelve schizophrenic patients has been investigated. The three catecholamines in 20?l of CSF are converted to their 0-methylated ana-logues by catechol-0-methyltransferase in the presence of S-adenosyl-L-1{methyl-3H} methionine. After the addition of tetraphenylborate, these derivatives are extracted. with diethylether at pH 8.2 followed by re-extraction with 0.1N HCI and then isol-ated by HPLC using, ?Bondapak C18column and 0.17 M acetic acid-60% methanol- 0.02 M Na2SO4 as solvent. Manual collection of each fraction is started two seconds after in appeared mark. The HPLC fraction of norepinephrine and epinephrine deri-vatives are oxidized to vanillin followed by extracting the vanillin with toluene, and dopamine derivatives fraction is extracted with toluene/isoamylalcohol and then re-extracted with 0.1N HCl. Internal standard curve shows a good linearity from 5 to 30 picogram per samples, and incorporated radioactivity of norepinephrine, epinephrine and dopamine derivatives are 163±11.4, 125±10.5 and 112±3.9cpm per picogram and calculated sensitivities are 0.4, 0.3 and 1 picogram, respectively. 3H-Normetanephrine, 3H-metanephrine and 3H-methoxytytamine are well separated by HPLC, and each retention times are 3.52, 4.08 and 5.62 minutes and yields are 75.6?0.9, 76.1?0.8 and 70.1?0.4%. Separation of the methylated derivatives by HLPC are performed within 8 minutes, and thus enabled us to complete the differential analysis of 15 samples (30 tubes) in a day. CSF norepinephrine concentration in schizophrenic patients show rostral-caudal gradient and haloperidol induced decreases of norepinephrine and the decreases correlated significantly with psychosis rating, From the above rsults, it is suggested that this method is more excellent than any others in sensitivity, accuracy, reproducibility and specificity of the differential assay, and CSF norepinephrine can be used for evaluation of neuroleptics treatment of schizophrenia by this method.

Tubulin Polymerization에 대한 Colchicine의 영향

최병훈,권기량,임규,황병두 충남대학교 의과대학 지역사회의학연구소 1990 충남의대잡지 Vol.17 No.2

Although the inhibition of microtubule assembly by colchicine is well established, the paradoxical effect of colchicine in the centriole, cilia and basal body is not clear. The microtubule(MADBP^+) was prepared from rabbit brain through 2 cycles of glycerol-and GTP-mediated polymerization and the tubulin without DNA binding protein(MADBP^-) was prepared from MADBP^+ using double strand DNA column chromatography and the properties of these tubulin preparation were compared with those of MADBP^+. The time course of tubulin polymerization was different, with MADBP^+ being sigmoidal and MADBP^- being hyperbolic. MADBP^+ polymerization could be inhibited by colchicine, whereas MADBP^- polymerization could not. There was little difference in colchicine binding activity between MADBP^+ and MADBP^- and the initial binding velocity of MADBP^- was faster than MADBP^-. From the above results, the possible existence of colchicine sensitive protein among microtubule associated DNA binding proteins was discussed.