Changes in the Number of Exocrine Granules in Mouse Pancreatic Acinar Cells Induced by Acetylcholine and MNNG in vitro

조응행,최임순,Cho, Eng-Haeng,Choe, Rim-Soon Korean Society of Electron Microscopy 1988 Applied microscopy Vol.18 No.2

췌장비 acinar cell에서 자극-분비 반응 과정을 알아보기 위하여 전자현미경을 이용하였다. 분비 촉진 물질인 아세틸콜린이나 MNNG의 투여에 따른 세포내 반응이 세포의 형태적 변화로서 잘 나타났으며 특히 guanylate cyclase를 활성화시키는 것으로 알려진 MNNG는 투여 후 일정시간 뒤 세포내에 많은 분비물 과립의 형성을 유발하였다. 이러한 결과에서 볼때 췌장의 acinar cell을 아세틸콜린으로 자극할 경우 guanylate cyclase는 지속적 반응 단계의 초기에 분비에 가세하는 것으로 생각되었고 cycloheximide나 dibucaine은 지속적 반응을 억제하는 것으로 나타났다. 또 췌장의 acinar cell에서는 분비물 과립의 형성이 소포체에서 직접 이루어지는 것으로 생각되었다. The stimulation-secretion coupling in the pancreatic acinar cell have been studied by electron microscope. Morphological changes in the cells exhibited the cellular response induced by acetylcholine and MNNG. MNNG, a guanylate cyclase activator, induced the formation of numerous secretory granules in a period after the agent administration. This result suggest that guanylate cyclase potentiated the early sustained response in pancreatic acinar cells stimulated by acetylcholine. Cycloheximide and dibucaine reduced the secretory granules in number during sustained period. In pancreatic acinar cells, the secretion granules were considered to be directly packaged from cisternal space of endoplasmic reticulum.

稻熱病菌(Piricularia oryzae)이 分泌한 β-Glucosidase의 精製 및 그 性質에 關한 硏究

趙應行,金銀壽 연세대학교 자연과학연구소 1981 學術論文集 Vol.8 No.-

稻熱病菌(Piricularia oryzae)의 T-race로부터 Chromatography 方法을 利用하여 部分 純化된 β-glucosidase가 보인 基質 salicin 및 cellobiose에 對한 最適 反應溫度는 55℃이었으며 最適 PH는 4.0∼4.5이였고 이 두 가지 基質에 對한 Km 값은 salicin에 對하여 5.3mM, cellobiose에 4.8mM이었고 Hg^++에 依한 Ki값은 salicin에서 3.6×10^-4M, cellobiose에 8.4×10^-4M이었으며 Sephadex G-200 Column Chromatography를 利用하여 測定한 이 酵素의 分子量은 240,000이였다. 몇 가지 金屬이온과 SDS가 이 酵素의 活性에 影響을 주는 것을 볼 수 있었는데 1mM 濃度에서 Ca^++나 Co^++는 酵素活性을 若干 增加시켰고 反面 Mn^++, Cu^++ 및 Hg^++는 抑制하는 것을 볼 수 있었는데 純化過程에서 同一한 蛋白質로 나타난 이 酵素는 그 Km이나 Ki값에서 볼 수 있듯이 두 가지의 基質 즉 salicin과 cellobiose에 對하여 그 親和性이 다른 것을 알 수 있었다. A tenfold purified β-glucosidase isolated from the T-race of Piricularia oryzae was optimally active at 55℃ on both salicin and cellobiose. The optimum pH of the enzyme activity was at 4.0∼4.5 on both substrates. The Km values of the enzyme estimated using salicin and cellobiose were 5.3mM and 4.8mM respectively. The Ki values of the enzyme estimated for Hg^++ were 3.6×10^-4M on salicin and 8.4×10^-4M on cellobiose. The molecular weight of β-glucosidase measured on the Sephadex G-200 column chromatography was around 240,000. A number of metal ions and SDS were found to influence the enzyme activity at 1mM concentration on both sustrates; ions such as Ca^++ and Co^++ appeared to enhance the enzyme activity, whereas Mn^++, Cu^++ and Hg^++ were found to slightly inhibit its activity. From the data obtained in the present experiment the β-glucosidase isolated was a single protein which works on both salicin and cellobiose with different affinity on both substrates as it was evidenced by such physical data as Km and Ki.

생쥐 췌장의 아밀라아제 분비기작에 관한 연구

조응행,최임순 한국통합생물학회 1987 동물학회지 Vol.30 No.2

Patterns of amylase secretion in mouse pancreatic fragments were studied over a period of time after the tissue was stimulated by acetyicholine and MNNG. MNNG is known to activate guanylate cyclase and thus increase the cGMP concentration in the pancreatic acinar cell. These amylase secretion patterns were studied to investigate the role of cGMP in reaction cascade during secretion response of the tissues stimulated by acetyicholine. Cellular response of amylase secretion in the pancreas by acetyicholine was divided into two phases. During the first phase, zymogen granules which had existed in the cells were secreted by the action of $Ca^2$+ and calmodulin immediately after secretagogue administration, this being known as the initial response. When the tissue was stimulated by acetylcholine in a $Ca^2$+-deficient medium or one containing trifluoperazine as a calmodulin antagonist, this initial response was reduced. In the second phase, newly formed zymogen granules were secreted as sustained response after protein synthesis was triggered by secretagogue. This response was provoked by an activation of protein kinase C. When either cycloheximide as a protein synthesis inhibitor or dibucaine as a protein kinase C inhibitor were added to the incubation medium, this sustained response was remarkablely depressed in the pancreatic fragments stimulated with acetylcholine. In the pancreatic acinar cell, phosphatidylinositol turnover plays an important role in the secretion response and hexachlorocyclohexane inhibits this phosphatidylinositol turnover. The pancreatic tissue treated with the hexachlorocyclohexane exhibited inhibition on both initial and sustained responses of amylase secretion by acetylcholine. MNNG also accelerated amylase secretion from the tissue gradually along incubation time. The 22 minutes fraction of the pancratic secretion after administration of both acetylcholine and MNNG showed higher amylase activity than the neighboring fractions. Guanylate cyclase potentiated the sustained response. Even if it is experimented with an indirect method, guanylate cyclase was found responsible for activation of the sustained response of a step prior to the action of protein kinase C. As conclusion, it was considered that amylase secretion in mouse pancreatic fragments stimulated by acetylcholine is a three phasic response.

숙지황이 고과당사료를 식이한 쥐의 혈액상에 미치는 영향

조응행 ( Eng Haeng Cho ),김윤상 ( Yun Sang Kim ) 대한본초학회 2003 大韓本草學會誌 Vol.18 No.1

N/A Objects : To investigate the effects of Rehmanniae Radix on the blood of rats, the effects of Rehmanniae Radix Preparat(RRP) on serum components was examined. Methods : Test group of male Sprague-Dawley rats was fed a fructose-rich(25 %; w/w) diet for 5 weeks. During the last 2 weeks the animals was orally administrated with the decoction prepared form RRP. Control group of rats was also fed a fructose-rich diet for 5 weeks, but administrated with saline during the last 2 weeks. Normal group was fed a normal diet for 5 weeks, and administrated with saline. And total cholesterol(Tc), triglyceride(TG), glucose, AST and ALT in serum were measured at 5 weeks on the fructose-rich diet. Results and Conclusion : The blood levels of Tc, TG, glucose of control group increased, and the activities of AST and ALT of test group decreased compared with those of normal group. The levels of Tc, TG, glucose of teat group decreased, and the activities of AST and ALT of test group increased compared with those of control group. These results suggested that RRP had some regulatory effects on the levels of blood components.

The Inhibitory Effect of Dimethylsulfoxide on the Synthesis of Muscle Specific Protein during Chick Embryonic Myoblast Differentiation

Cho, Eng Haeng,Ha, Doo Bong,Choe, Rim Soon 연세대학교 자연과학연구소 1988 學術論文集 Vol.21 No.-

계배 근세포를 분리하여 배양하면서 근세포 분화기간 동안 몇 가지 근특이 단백질들의 합성에 대한 dimethyl sulfoxide (DMSO)의 억제 양상을 검토하였다. 전기영동겔상의 각 단백질 성분들을 densitometer로 scanning한 결과 DMSO를 처리한 구와 무처리구간에 비슷한 분포를 보였으나 근특이단백질인 actin과 myosin의 상대적 양은 현저한 차이를 보였는데 이것은 근세포 분화기간 동안 총단백질의 합성이 DMSO를 처리한 세포에서 감소한 때문이라고 보여지며 그 결과 근세포 융합이 억제된 것이라고 생각된다. 그외에 아세틸콜린 수용체와 creatine phosphokinase도 또한 DMSO에 의하여 그 합성이 억제된 것을 볼 수 있었으며 배양액에서 DMSO를 제거 하였을 때 근세포 융합이나 총단백질 합성량 및 아세틸콜린 수용체의 합성은 다시 증가하였고 무처리 상태 가까이 회복되었다. The pattern of inhibition by DMSO on several muscle specific proteins synthesis during myogenesis was studied. The profiles of protein in densitometric scans on SDS-polyacrylamide electrophoresis gel showed similar pattern in DMSO treatment and untreatment. But the amount of actin and myosin in the densitometric peak was revealed significant difference. Therefore, it seemed to be that total protein synthesis during muscle cell differentiation was decreased in DMSO-arrested cells, and this resulted in inhibition of cell fusion phenotypically. The synthesis of other proteins, acetylcholicne receptor and creatine phosphokinase, was also decreased by DMSO. The inhibition of total protein and acetylcholine receptor by DMSO was reversed and cell fusion was restored nearly to control in 2-days after DMSO removal.

인공적혈구의 제조 및 이용

하종식,조응행,김구자 이화여자대학교 생명과학연구소 1990 생명과학연구논문집 Vol.1 No.-

Hemoglobin was purified from the outdated human red blood cells. Phospholipids were purified from egg yolk and stored in chloroform. The artificial red blood cells (hemosome) were prepared by encapsulation of hemoglobin wht phospholipid mutilayer using rotary vacuum evaporator. The shape and size of hemosomes were measured by phase contrast microscope and image analyzer. The function of hemosomes was tested by measuring oxygen dissociation curve using blood gas analyzer. In order to test whether hemosomes are useful as blood substitute they were infused into rats of which one third of total blood were drawn. The results obtained are summarized as followings. 1) Hemosomes were spherical shape and their mean diameter was 0.7um. 2) Oxygen dissociation curve of hemosomes showed the same figure as that of normal red blood cells. 3) All rats given 1/3transfusion with hemosomes survived until sacrificed whereas three of four rats given 1/3 transfusion with saline died within 1 hour and the rest of them died within 24 hours.

담배나방 의각태별(各態別) 발생소장(發生消長)과 란기생봉(卵寄生蜂)의 기생률(寄生率)에 관(關)한 조사(調査)

최궤문,조응행,소재선,황창연 한국식물병리학회 1975 한국식물보호학회지 Vol.14 No.3

인공적혈구의 제조 및 이용

하종식,조응행,김구자,Hah, Jong-Sik,Cho, Eng-Haeng,Kim, Ku-Ja 대한생리학회 1990 대한생리학회지 Vol.24 No.1

Hemoglobin was purified from the outdated human red blood cells. Phospholipids were purified from egg yolk and stored in chloroform. The artificial red blood cells (hemosome) were prepared by encapsulation of hemoglobin with phospholipid mutilayer using rotary vacuum evaporator. The shape and size of hemosomes were measured by phase contrast microscope and image analyzer. The function of hemosomes was tested by measuring oxygen dissociation curve using blood gas analyzer. In order to test whether hemosomes are useful as blood substitute they were infused into rats of which one third of total blood were drawn. The results obtained are summarized at followings. 1) Hemosomes were spherical shape and their mean diameter was 0.7 um. 2) Oxygen dissociation curve of hemosomes showed the same figure as that of normal red blood cells. 3) All rats given 1/3 transfusion with hemosomes survived until sacrificed whereas three of four rats given 1/3 transfusion with saline died within 1 hour and the rest of them died within 24 hours.

멸강나방(Mythimna separata Walker)의 發生消長에 관한 調査

崔貴文,趙應行 한국응용곤충학회 1975 한국응용곤충학회지 Vol.14 No.1

명강나방(Mythimna separata Walker)의 발생소장에 관한 조사

최귀문,조응행,Choi K. M.,Cho E. H. 한국응용곤충학회 1975 한국식물보호학회지 Vol.14 No.1