가임녀성(可姙女性)의 월경주기(月經周期)에 따르는 말초혈액상(末稍血液象) 및 L.A.P.의 변동(變動)

권헌영 ( Heun Young Kwon ),한석종 ( Suk Jong Han ) 대한임상검사과학회 1977 대한임상검사과학회지(KJCLS) Vol.9 No.1

Menstruation is the cycle uterine bleeding which occurs at regular intervals during the reproductive period in women``s life. During the active reproductive era menstruation occurs at approximately 28-30 day intervals. The clinical menifestation of menstruation in flow from the uterus, the mucosa of which being shed off. The uterus, however, has no endocrine function of its own, it function is entirely dependent upon physiologically active substances which are carried to the organ by the blood stream. These substances are hormones secreted by two endocrine organs, the ovary and the anterior lobe of the hypophysis, these hormones are the so called sex hormones. These variations in hematological parameters are in good agreement with present knowledge of the hormonal changes during menstrual cycle. In a series of 30 blood samples were tested for Hemoglobin, W.B.C., Lymphocyte, Platelet, L.A.P. and the results plotted against a hormal menstrual cycle.

Purification and Biochemical Characterizations of Calmodulin from Bovine Brain

이상열,권헌영,김정희,이서구,Lee, Sang-Yeol,Kwon, Heun-Young,Kim, Jung-Hye,Rhee, Sue-Goo 생화학분자생물학회 1990 한국생화학회지 Vol.23 No.2

세포내에서 칼슘이 작용하는 반응을 조절하는 calmodulin 단백질을, 소의 뇌로부터 순수 분리하여 여러 가지 생화학적 성질을 연구하였다. 이것은 칼슘의 존재하에서 분자량이 15,000으로 나타나고, 칼슘을 제거한 상태의 분자량은 21,000으로 나타남으로써, 칼슘이 calmodulin에 결합하여 단백질의 3차원적 구조에 상당한 변화를 유발시킨다는 사실을 알 수 있었다. 또한 자외선스팩트럼을 조사해본 결과 259, 265, 279 nm의 세 군데에서 최대 흡수 peak가 나타났는데, 이것은 calmodulin 단백질의 아미노산 조성 중 tryptophan이 존재하지 않으므로 인하여, phenylalanine과 tyrosine의 흡광도에 의한 영향으로 생각된다. 또한 칼슘과 calmodulin 농도를 변화시키면서, Ins(1, 4, 5) $P_3$-Kinase의 효소 활성도를 측정하여 본 결과, $6{\mu}M$ calmodulin이 효소반응용액에 존재할 경우 이들을 첨가하지 않은 때에 비하여 최대 7배까지의 효소 활성도를 증가시켰다. 이러한 실험 결과로부터, 최적 농도의 $Ca^{2+}$/calmodulin 복합체는 기질과 반응하기에 부적합한 구조의 target 효소와 결합하여, 활동적이고 반응하기에 매우 좋은 조건의 구조를 갖는 효소로 바꾸어 줌으로써 생체내의 반응조절에 중요한 역할을 한다는 사실을 알 수 있었다. Calmodulin, a heat stable $Ca^{++}$-binding protein, is a multifunctional modulator, mediating many of the $Ca^{++}$ effects in cellular functions. This protein interacts reversibly with $Ca^{++}$ to form a protein/$Ca^{++}$ complex, whose activity is regulated by cellular flux of $Ca^{++}$. Calmodulin from bovine brain was purified to complete homogeneity by heat, acid treatment and hydrophobic interaction chromatography. The biochemical properties of the purified protein were examined under various criteria. It was estimated to have an apparent molecular weight of 21K and 15K in the absence or presence of $Ca^{++}$ respectively, which would mean the great conformational changes were induced by the binding of $Ca^{++}$ to calmodulin. The calmodulin had distinct ultraviolet absorption peaks at 259, 265, and 279 nm due to phenylalanine and tyrosine residues. Also, this protein activated the Ins(1,4,5) $P_3$-Kinase of which role is very important in cellular signal transduction, about seven folds (at $6{\mu}M$ $Ca^{++}$ and $1.0{\mu}M$ calmodulin) with a $Ca^{++}$-dependent manner.

소의 뇌로부터 Calmodulin 단백질의 순수분리 및 생화학적 성질 연구

이상열,권헌영,김정희,이서구 ( Sang Yeol Lee,Heun Young Kwon,Jung Hye Kim,Sue Goo Rhee ) 생화학분자생물학회 1990 BMB Reports Vol.23 No.2

Calmodulin, a heat stable Ca^(++)-binding protein, is a multifunctional modulator, mediating many of the Ca^(++) effects in cellular functions. This protein interacts reversibly with Ca^(++) to form a protein/ Ca^(++) complex, whose activity is regulated by cellular flux of Ca^(++). Calmodulin from bovine brain was purified to complete homogeneity by heat, acid treatment and hydrophobic interaction chromatography. The biochemical properties of the purified protein were examined under various criteria. It was estimated to have an apparent molecular weight of 21K and 15K in the absence or presence of Ca^(++) respectively, which would mean the great conformational changes were induced by the binding of Ca^(++) to calmodulin. The calmodulin had distinct ultraviolet absorption peaks at 259, 265, and 279 nm due to phenylalanine and tyrosine residues. Also, this protein activated the Ins(1,4,5) P₃-Kinase of which role is very important in cellular signal transduction, about seven folds (at 6 μM Ca^(++) and 1.0 μM calmodulin) with a Ca^(++)-dependent manner.

Transformed E . coli JM 107 에서 순수분리한 S . marcescens 세포외 Nuclease 의 특징

전영환,김기홍,권헌영,신용철,조무제,이상열 ( Young Hwan Jeon,Kee Hong Kim,Heun Young Kwon,Yong Chul Shin,Moo Je Cho,Sang Yeol Lee ) 생화학분자생물학회 1992 BMB Reports Vol.25 No.5

We have previously described the purification procedures of the extracellular nuclease of Serratia marcescens from a recombinant E. coli JM 107 (Jeon and Lee, 1992). Using the purified nuclease, we examined the various biochemical properties of this enzyme. Kinetic parameters of thermal inactivation followed the first order kinetics. Heating the nuclease to 60℃ or higher temperature reduced enzymatic activity. It required 1 mM Mg^(2+) or Mn^(2+) ions for its maximal activity and no other metal ions can be replaced for the cofactor requirement of this enzyme. It was stable at pH in the range of pH 6 to 10, and its maximal activity was exhibited around pH 8. It was resistant to proteolysis by trypsin and an extracellular protease produced from S. marcescens. However, heat-denatured nuclease and bovine serum albumin were completely degraded when they were treated with the same proteases at the same assay conditions. Its activity was significantly inhibited by the addition of 1 mM ethidium bromide.

Relation of Poly(ADP-ribose) Polymerase Cleavage and Apoptosis Induced by Paclitaxel in HeLa S₃ Uterine Cancer Cells

장정현,김광연,안순철,권헌영,Chang, Jeong-Hyun,Kim, Kwang-Youn,Ahn, Soon-Cheol,Kwon, Heun-Young Korean Society of Life Science 2007 생명과학회지 Vol.17 No.8

Paclitaxel이 암세포에서 세포예정사를 유발할지라도, 아직 정확한 기전은 잘 알려져 있지 않다. 이에 본 연구에서는 HeLa $S_{3}$ 자궁암세포에서의 paclitaxel이 어떠한 영향을 미치는지 알아보고자 한다. 그리하여 방법으로는 세포독성검사, apoptotic cells의 형태학적 변화(DAPI 염색 ), western blot 분석법을 사용하여 수행하였다. 본 연구의 결과로 paclitaxel은 HeLa $S_{3}$ 세포에서 세포독성을 보이며 특히 paclitaxel의 $IC_{50}$ 값은 약 1 ${\mu}M$이며, paclitaxel 처리한 HeLa $S_{3}$ 세포에서 형태학적 변화(분절화)를 관찰하였고, flow cytometric 분석에서는 G2/M기가 차단되어 paclitaxel은 세포주기 특히 Sub-$G_{1}$기를 조절함을 알 수 있다. 그리고 Paclitaxel을 처리한 HeLa $S_{3}$ 세포에서는 PARP cleavage를 유발하였고 Bc1-2의 감소와도 관련되었다. Although paclitaxel induces apoptosis of cancer cells, its exact mechanism of action is not yet known. The present study has been performed to determine whether influence of paclitaxel in HeLa $S_{3}$ uterine cancer cells. Three assays were employed in this study: cell cytotoxicity, morphological assessments of apoptotic cells (DAPI staining assay), and western blot analysis. The results indicated that paclitaxel has cytotoxic effects in HeLa $S_{3}$ cells. Especially, the $IC_{50}$ value of paclitaxel was about 1 ${\mu}M$. And morphological changes (fragmentation) of cells were observed by paclitaxel in HeLa $S_{3}$ cells. The flow cytometric analysis of paclitaxel-treated cells indicated a block of G2/M phase. The results that pacli-taxel regulates the cell cycle, especially Sub-$G_{1}$ phase. Paclitaxel induces apoptosis of HeLa $S_{3}$ cells via PARP-dependent fashion, and this apoptosis is related to disappearance of Bcl-2 proteins.

Effects of Apopotosis on Induction of Resveratrol in Human Lung Cancer A549 Cells

Jeong-Hyun Chang(장정현),Heun-Young Kwon(권헌영) 한국생명과학회 2008 생명과학회지 Vol.18 No.5

Resveratrol (RSVL)은 포도껍질에 많이 함유된 성분으로 다양한 암세포에 대해 antiproliferative activity가 존재한다고 보고된 바 있다. 이에 본 연구에서는 폐암 A549세포에서 RSVL이 세포자멸사를 유도하는지와 관련된 세포자멸사 기작에 대해 알아보고자 한다. 그리하여 본 연구에서는 5가지 방법으로 즉, 세포생존율검사, 세포독성검사, apoptotic cell의 형태학적 변화, 정량적 DNA 분절 분석, western blot 분석법을 사용하여 수행하였다. 본 연구의 결과로 RSVL을 처리한 A549세포에서 형태학적(분절화) 변화가 관찰되었고, flow cytometric 분석에서는 RSVL이 세포에서 세포주기 특히 subG1기를 조절함을 알 수 있었다. 그리고 RSVL을 처리한 A549세포에서는 apoptosis와 관련된 caspase3와 PARP cleavage가 유발되었고 CAD 단백질 증가도 보였다. Resveratrol (3,5,4’-trihydroxystilbene, RSVL) is a supplement found mostly in grape skins. RSVL is known to exhibit potent antiproliferative activity against various cancer cell lines. The aim of this study was to examine the potential of RSVL to induce apoptosis in lung cancer A549 cells and to assess the mechanism of RSVL-induced apoptosis. Five different assays were employed in this study: cell viability and cytotoxicity, morphological assessments of apoptotic cells, quantitative DNA fragmentation analysis, and western blot analysis. The results indicated that RSVL has cytotoxic effects in A549 cells in a concentration- and time-dependent manner. And nuclear morphological changes of cells were observed by RSVL. RSVL regulates the cell cycle, especially Sub-G1 phase. Also, RSVL induces apoptosis of A549 cells via Caspase3, CAD and PARP-dependent fashion.

실험적 지하철 소음에서의 지적활동이 혈액생리에 미치는 영향

최석철,현경예,박재현,권헌영,Choi, Seok-Cheol,Hyun, Kyung-Yae,Park, Jae-Hyun,Kwon, Heun-Young 한국생명과학회 2006 생명과학회지 Vol.16 No.7

이 연구는 지하철 소음 하에서 지적활동이 혈액생리에 어떤 영향을 미치는지 규명하기 위해 계획되었다. 56명의 건강한 지원자를 대상으로 지하철 소음노출 하에서 지적활동(암산)을 50분간 수행한 후 60분간의 휴식을 취하게 하였다. 스트레스 노출 전(기준치), 노출 50분, 노출 종료 후 60분의 휴식 등의 세 시기 때 정맥혈액을 채취하여 CBC, prothrombin time (PT), activated partial thromboplastin time (aPTT), 적혈구침강률, 섬유소원 농도, D-dimer 농도, 고감도 C-반응단백(H-CRP) 농도를 각각 분석하였다. 총 백혈구 수, 호중구 수, 림프구 수는 스트레스 종료 후 60분때는 모두 기준치보다 유의하게 상승하였다. 적혈구 수는 스트레스 노출 50분 때 기준치에 비해 유의한 증가를 보인 반면, 단구 수는 스트레스 노출 50분과 종료 후 60분 때 모두 기준치보다 유의하게 저하되었다. 혈소판 수와 적혈구침강률은 스트레스 종료 후 60분 때 기준치보다 낮았다. aPTT는 스트레스 노출 50분과 종료 후 60분 때 기준치보다 유의하게 단축되었는데 비해, PT는 스트레스 종료 후 60분때만 유의하게 단축되었다 D-dimer와 H-CRP 농도 둘 다 스트레스 50분과 종료 후 60분 때 유의한 상승을 보였다. 이 연구의 결과들은 지하철 소음과 병행된 지적활동이 백혈구수의 증가, 혈액농축, PT 및 aPTT의 단축, ESR의 감소, D-dimer 및 H-CRP 생성의 증가를 일으키며 이는 좋지 않은 지하철 환경이 잠재적으로 염증반응 및 혈전형성 반응의 발생 가능성을 유도할 수도 있음을 시사하는 것이다. The present study was sought to clarify whether the combination of mental activity with subway noise affects hematological variables. Fifty-six healthy volunteers participated in this experiment and underwent a stress task consisting of combination_of mental activity (mental arithmetic) with subway noise for 50 min and 60min of recovery after the end of the stress task. Venous blood samples were collected for measuring CBC, prothrombin time (PT), activated partial thromboplastin time (aPTT), erythrocyte sedimentation rate (ESR), fibrinogen concentration, D-dimer and high sensitive C-reactive protein (H-CRP) levels before (baseline), 50min of stress task (S-50m), and 60 min of recovery (R-60m). Total leukocyte, neutrophil and lymphocyte counts significantly increased at R-60m compared with baselines. RBC count at S-50m was higher, while monocyte counts at S-50m and R-60m were lower than those of baselines. aPTTs shortened at S-50m and R-60m, but PT reduced at R-60m as compared with baselines. D-dimer and H-CRP levels at S-50m and R-60m were significantly higher than those of baselines. These findings imply that a combination of mental activity with subway noise nay cause leukocytosis, homo-concentration, shortened PT and aPTT, decreased ESR, and raised D-dimer and H-CRP levels, suggesting possible development of inflammation and prothrombogenic reaction attributable to a subway environment.

Alterations in Blood Electrolyte of Rabbits with Experimental Injection of Escherichia coli Endotoxin

Seok-Cheol Choi(최석철),Jai-Young Kim(김재영),Heun-Young Kwon(권헌영),Tae-Un Kim(김태운),Soo-Myung Hwang(황수명),Won-Jae Lee(이원재) 대한의생명과학회 2000 Biomedical Science Letters Vol.6 No.2

혈액 전해질 성분들에 대한 대장균 내독소의 영향을 조사하기 위해 토끼를 대상으로 한 동물실험을 실시하였다. 대장균 내독소 (혈청형 O55 : B5)를 토끼의 귀정맥을 통해 0.10 ㎎/㎏ 혹은 0.50 ㎎/㎏ 농도로 주입한 후 3, 6, 12, 24시간대에 채혈하여서 Ca??, Mg??, Na?, K?, Cl? 농도를 측정하였다. 대조군에 비해, 내독소투여 토끼의 Ca?? 농도는 6시간대에 증가하였고, Mg?? 농도는 3, 6, 12시간대에, Na?과 K?는 모든 채혈시간대에, 그리고 Cl? 농도는 3, 12, 24시간대에 각각 유의하게 높았다 (p<0.05). 흥미롭게도, 고 Mg? 혈증 (약 4.0 ㎎/dL)을 보인 내독소투여 토끼들은 심각한 임상징후들로 인식되는 분비불의 증가, 쇼크, 빈호흡, 경련, 혹은 설사와 같은 증세를 보였다. 본 연구의 결과들은 대장균 내독소가 혈액 전해질 농도의 항상성 혼란을 유도하며 이러한 생리적 불균형은 치명적 상황과 그로 인한 죽음을 야기할 수도 있음을 시사하고 있다. We studied the effects of Escherichia coli (E. coli) endotoxin on blood electrolytes levels in rabbits. Endotoxin (E. coli serotype O55 : B5) was injected via rabbits' ear vein : 0.10 ㎎/㎏ (Group A) or 0.50 ㎎/㎏ (Group B). Blood samples were taken at postendotoxemic 3, 6, 12 and 24 hrs and were analyzed for detections of the levels of blood electrolytes such as Ca??, Mg??, Na?, K? and Cl?. As compared to control group, in endotoxin-treated rabbits Ca?? levels elevated at 6 hrs but decreased at 24 hrs, Mg?? levels were high at 3, 6 and 12 hrs, Na? and K? levels increased at all sampling times and Cl? levels rose at 3, 12 and 24 hrs (p<0.05). Interestingly, endotoxic rabbits having hypermagnesemia (about 4.0 ㎎/dL) showed severe syndromes such as increased secretion, shock, tachypnea, seizure and/or diarrhea, suggesting that these may be clinical signs of imminent death in rabbits. These observations testify that bacterial endotoxin leads to dyshomeostasis of blood electrolytes and the physiological imbalances may cause fatal disorders and subsequent death.

Biochemical Characterization of a Protease with Fibrinolytic Activity from Maggots of Protaetia brevitarsis

장정현,조지영,김영진,이선이,조효진,유선녕,김광연,박병근,안순철,권헌영,Chang, Jeong-Hyun,Jo, Ji-Young,Kim, Yeong-Jin,Lee, Sun-Yi,Cho, Hyo-Jin,You, Sun-Nyoung,Kim, Kwang-Youn,Park, Byoung-Keun,Ahn, Soon-Cheol,Kwon, Heun-Young Korean Society of Life Science 2007 생명과학회지 Vol.17 No.5

혈전(fibrin clot)은 심혈관계 질환을 일으키는 주요 인자로서 전신의 미세동맥이나 모세혈관 내에서 형성되어 주의 조직이나 장기에 혈류의 공급방해가 생겨 허혈, 경색, 나아가 괴사까지도 발생시킨다. 혈전이 원인이 되어 발생한 심혈관계 질환을 예방 혹은 치료할 목적으로 기존으로 사용되고 있는 항혈전제(antithrombolytic drug)나 혈전용해제(thrombolytic drug)의 개발을 위해 많은 연구들이 진행되고 있다. 본 연구에서는 치료목적으로 사용할 혈전용해제를 분리하고자 Protaetia brevitarsis의 maggot로부터 ammonium sulfate 분획과 desalting column을 이용하여 fibrinolytic protease를 분리하고 생화학적 특성을 조사하였다. 활성의 최적 pH는 9.0였고 최적온도는 $50^{\circ}C$였다. pH 7.0-9.0 사이와 온도 $60^{\circ}C$이하에서는 비교적 활성이 안정성을 나타냈다. 효소활성이 phenylmethanesulfonyl fluoride에 의해 강하게 저해되고 있는 것으로 보아 serine protease로 추정되며 금속이온에 의한 영향을 조사해 본 결과 $Ca^{2+}$과 $Zn^{2+}$에 의해서는 저해되지만 $Mg^{2+}$와 $Fe^{2+}$에 의해서는 저해를 받지 않았다. Fibrin clots remained in blood vessels can be one of the serious factor caused cardiovascular disease, such as ischemia, infarction and necrosis The development of an antithrombotic and thrombolysis solvent is necessary to prevent and treat these diseases. In this study, the fibirinolytic protease was prepared from the maggots of Protaetia brevitarsis using ammonium sulfate fractionation and desalting column. The optimum pH and temperature for the enzyme activity were pH 9.0 and $50^{\circ}C$, respectively. The enzyme activity was relatively stable at pH 7.0-9.0 and temperature below $60^{\circ}C$. The activity of the enzyme was strongly inhibited by phenylmethanesulfonyl fluoride. And the activity of the enzyme was inhibited by $Ca^{2+}\;and\;Zn^{2+}$, but it was not by $Mg^{2+}\;and\;Fe^{2+}$ ions. In these experimental results, we have speculated that the enzyme derived from maggots of Protaetia hrevitarsis is a serine protease with a strong fibrinolytic activity.

Gamma Irradiation Induces a Caspase-dependent Apoptotic Mechanism in Human Prostate Cancer PC-3 Cells

Jeong Hyun Chang(장정현),Dong Hyun Kim(김동현),Gye Rok Jeon(전계록),Heun Young Kwon(권헌영) 한국생명과학회 2008 생명과학회지 Vol.18 No.8

전립선 암은 현대 남성에게 걸리기 쉬운 질환으로 매년 점점 증가하는 암 사망률 중 하나이다. 하지만 남성호르몬 비의존형 전립선 암 치료에 대한 효과가 거의 없어 이에 본 연구에서는 남성호르몬 비의존형 전립선 암세포인 PC-3 세포에서 감마선이 세포 성장 억제와 세포자멸사 기작에 대해 알아보고자 한다. 그리하여 본 연구에서는 5가지 방법으로 즉, 세포증식 억제, apoptotic cell의 형태학적 변화, DNA 분절 분석, AV/PI 염색, western blot 분석법을 사용하여 수행하였다. 본 연구의 결과로 감마선을 처리한 PC-3세포에서 형태학적(분절화) 변화와 DNA ladder가 관찰되었다. 또한 감마선을 처리한PC-3세포에서는 apoptosis와 관련된 Caspase3와 PARP cleavage가 유발되었고 Bax 단백질 증가도 보였다. Prostate cancer is the most predominant cancer in men and related death rate increases every year. Till date, there is no effective therapy for androgen independent prostate cancer. To investigate the mechanism for cell growth inhibition or apoptosis in human androgen independent prostate cancer PC-3 cells after gamma irradiation. The aim of this study was to examine the potential of gamma irradiation to induce apoptosis in PC-3 cells and to assess the mechanism of gamma irradiation-induced apoptosis. Five different assays were employed in this study: cell proliferation assay, morphological assessments of apoptotic cells, DNA fragmentation analysis, quantification of apoptosis by annexin V (AV) and propidium iodide (PI) staning, and western blot analysis. Cell viability was inversely related to radiation dose. DAPI-positive cells were detected 48 hr after 40 Gy radiation exposure. And nuclear morphological changes of cells were observed by gamma irradiation. DNA ladder patterns in the cells exposed to gamma-radiation were appeared at 24 hr. Also, gamma irradiation induces apoptosis of PC-3 cells via Caspase3, Bax and PARP-dependent fashion.