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      • Helicobacter pylori와 대장균의 Shuttle Vector 개발

        조명제,이우곤,이상룡,김경희,안영숙,김성희,김현주,류복덕,최여정,윤영혜,백승철,전영석,이광호 경상대학교 유전공학연구소 1997 遺傳工學硏究所報 Vol.16 No.-

        In this study, a vehicle vector using cryptic plasmids was constructed for gene transfer in Helicobacter pylori. pHP51(3.9 kb) and pHP489(1.2 kb) were selected for constructing vectors from cryptic plasmid of H. pylori isolates in Korea. The HindⅢ-digestedDNA fragment(1.2kb) of pHP489 and 1.6kb DNA fragment of pHP51 were ligated with a kanamycin resistance gene(aph3'-Ⅲ) from C. jejuni to produce the recombinant plasmids pHP489K and pHP51K, respectively. Transformation frequency of pHP51K by electroporation was low. But pHP489K could be effectively transformed into various H. pylori strains. In order to design an intermdiate vehicle vector for gene transfer into H. pylori, pBlueHP489K was prepared by recloning pHP489K DNA into pBluescript and pTZ19R vector. This vector permitted the DNA fragment containing pHP489 sequence, aph3'-Ⅲ, and cloned DNA to be cut and self-ligated in the SacⅠ site after cloning. ureA and ureB gene were inserted into pBlueHP489K, resulting in pBlueHP489K/AB. The DNA fragment containing pHP489, kanamycin resistance gene(aph3'-Ⅲ), and urease structural gene was cut away from pBlueHP489K/AB and self-ligated to generate pBlueHP489K/AB. pBlueHP489K/AB made urease-negative H. pylori strains restore their urease activity. By this experiment, pBlueHP489K was confirmed to be the vehicle system for transferring H. pylori genes.

      • SCOPUSKCI등재
      • 원예작물의 염류장애 원인과 대책을 위한 생리, 생화학적 연구 2. 토마토 뿌리조직 H+ 펌프 활성 및 Thapsigargin 저해효과

        조광현,사공정,김영기 충북대학교 한국과학재단 지정 첨단원예기술개발 연구센터 1997 연구보고서 Vol.2 No.-

        Thapsigargin은 동물조직 SR/ER-type Ca²+-ATPase에 특이성이 높은 선택적 저해제로서 토마토 뿌리조직으로부터 분리한 마이크로솜에 Ca²+-ATPase의 존재를 확인하기 위하여 사용되었다. 원형질막과 액포막에 위치하는 H+-ATPase들의 활성은 각각의 선택적 저해제인 vanadate와 NO₃를 이용하여 평가하였고, 이들의 활성은 각각 마이크로솜 ATPase 총활성의 ∼26%, ∼36%로 나타났다. 이들 두 가지 저해제 효과는 additive하게 나타났으며, 전체활성의 약 50∼70%를 저해함을 확인하였다. 마이크로솜 ATPase활성에 대한 thapsigargin의 저해효과에서 10 μM thapsigargin은 총활성의 30%를 저해하였으며, 저해효과는 농도 의존적으로 나타났다. Thapsigargin에 의한 활성저해 기작을 조사하기 위하여 H+-ATPase의 선택적 저해제들을 사용하였다. 액포막의 H+-ATPase 활성에 대한 thapsigargin의 효과를 조사하기 위한 NO₃의 dose-response 실험에서, thapsigargin은 NO₃에 의해 저해되는 ATPase활성을 경쟁적으로 감소시켰다. 반면, vanadate의 농도를 10 nM∼2mM로 증가시켜 원형질막의 H+-ATPase 활성을 저해시킨 조건에서는 thapsigargin에 의하여 저해되는 활성이 온전히 나타나 vanadate의 저해효과와는 무관한 결과를 얻었다. 이러한 결과는 액포막 H+-ATPase의 활성이 thapsigargin에 의해 저해된다는 것을 의미한다. 저자들은 액포막 H+-ATPase의 활성이 Ca²+에 의하여 저해됨을 이미 보고하였다 (Cho et al., 1998). Thapsigargin에 의하여 저해되는 마이크로솜 ATPase활성은 반응용액의 Ca²+농도가 증가할수록 감소하였으며, 이것은 thapsigargin이 액포막 H+-ATPase 활성을 저해한다는 가능성을 더욱 지지하였다. 이상의 결과는 토마토 뿌리조직의 마이크로솜 ATPase중 액포막의 H+-ATPase 활성이 thapsigargin에 의해서 저해되는 것을 보여주며, 액포막 H+-ATPase와 ER-type Ca²+-ATPase와의 구조적, 기능적 연관성이 있을 가능성을 제안한다. The effect of thapsigargin, a SR/ER-type Ca²+-ATPase inhibitor, was investigated to determine the presence of thapsigargin-sensitive Ca²+-ATPases in the microsomes of tomato roots. The activities of plasma membrane H+-APTase and vacuolar H+-ATPase were evaluated to ∼26% and ∼36% of total microsomal ATPase activity by using their specific inhibitors, vanadate and NO₃, respectively. The inhibitory effects of vanadate and NO₃- were additive and the simultaneous additions of these two inhibitors decreased the total activity up to 50∼70%. Thapsigargin (10μM) inhibited the microsomal ATPase activity by ∼30% and the thapsigargin-induced inhibition was appeared to be dose-dependent. In order to understand the molecular mechanism of thapsigargin-induced inhibition, the effects of specific H+-ATPase inhibitors were tested and compared to the effect of thapsigargin. The NO₃-induced inhibition was decreased by simultaneous treatment of thapsigargin. In dose-response experiments of vanadate and NO₃-, thapsigargin only blocked the NO₃-induced inhibition, while the effects of vanadate and thapsigargin were independent at the whole range of vanadate concentration (10 nM∼2 mM). Meanwhile, the thapsigargin-induced inhibition was decreased by increasing Ca²+ concentration. Since we have shown that Ca²+ blocks the activity of vacuolar H+-ATPase, these results together suggest that thapsigargin inhibits the NO₃-sensitive vacuolar H+-ATPase but not the vanadate-sensitive plasma membrane H+-ATPase. Our results also suggest that there is structural and functional relations between vacuolar H+-ATPase and SR/ER-type Ca²+-ATPase.

      • 瓦松의 알콜 抽出物이 心筋 및 腸筋收縮에 미치는 影響

        曺石鉉,金光鎭 충남대학교 의과대학 지역사회의학연구소 1987 충남의대잡지 Vol.14 No.1

        Orostachys malacophyllus F. is a kind of ,Chinese medicine to grow on the tiled roof of the house or on the rock of the mountain. The chemical compositions and actions have not been reported, but, sometimes, it was used as a folk remedy for the treatment of cancerous diseases. This study was attempted to investigate the effects of water fraction separated from alcohol extract of Orostachys malacophyllus F. on the contractilities of cardiac and intestinal muscles comparing with those of the water extract. The muscles used for this study were the ventricular muscle of turtle and the intestinal muscle of mouse, and their contractilities were represented as a change of the muscle tension detected by myograph. The results were summarized as follows: 1. The water fraction decreased the contractility of ventricular muscle, and increased the intestinal contractility. But the water extract was observed the opposite effects on the contractility of each muscle. 2. Sympathetic or parasympathetic blocker and alpha-or beta-blocker had no effect on the decreased or increased contractility of each muscle. 3. Calcium antagonist(verapamil) inhibited significantly the contractility in the case of the water fraction to the intestinal muscle and the water extract to the ventricular muscle. From the above results, it was suggested that the action of water fraction or the water extact affected to calcium transport: release into the sarcoplasmic fluid or active influx into the sarcoplasmic reticulum in the case of the cardiac muscle, and release from ECF or cisternae of reticulum into the muscle cell in the smooth muscle.

      • Fallot 四徵症의 根治手術

        曺洸鉉 인제대학교 1985 仁濟醫學 Vol.6 No.1

        One of the most common of the more serious congenital malformations of the heart is the tetralogy of Fallet, which is said as the watershed between the simple and complex cardiac anomalies. And, therefore, all cardiac surgeons have been much interested in surgical approach of this anomaly. Although Stensen deserves credit for the first description in 1972 of what is new termed the tetralogy of Fallot, nevertheless it is Etienne-Louis Arthur Fallot (1888) of Marseille, France, whose name is characteristically attached to this congenital cardiac disorder. In the original description of this, Fallot stated, "This malformation consists of a true anatomopathological type represented by the following tetralogy: (1) stenosis of the pulmonary artery; (2) interventricular communication; (3) deviation of the origin of the aorta to the right; (4) hypertrophy, almost always concentric, of the right ventricle. Failure of obliteration of the foramen ovale mar ocassionally be added in a wholly accessory manner." Despite the fact that accurate clinical diagnosis could often be established after these contributions by fallot, nevertheless, many years passed before definitive treatment of this was initiated by Blalock and Taussig (1945) with the establishment of the subclavian artery to pulmonary artery anastomosis. Thereafter, several types of this kind of palliative operation were established by Potts (1946), Waterston (1962), Sellors (1948), Brock (1948) and Glenn (1954). In an imaginative and daring effort, in 1954, Lillehei using controlled crass-circulation, carried out the first intracardiac repair of this anomaly by closing the VSD and relieving the pulmonary stenosis under direct vision. The results of surgery for this have steadily improved over the years, thanks to important contributions of many surgeons. Nevertheless because of its protean physiologic and anatomic presentations, tetralogy of Fallot continues to offer challenges to cardiologists and cardiac surgeons. Auther wanted to study and summarize about the radical operation of this anomaly with literature review.

      • 원예작물의 염류장애 원인과 대책을 위한 생리, 생화학적 연구 3.토마토 뿌리조직에서 마이크로솜 Ca²+ uptake 특성연구

        조광현,김영기 충북대학교 한국과학재단 지정 첨단원예기술개발 연구센터 1998 연구보고서 Vol.3 No.-

        식물세포의 세포질 Ca²+ 이동과 관련된 Ca²+ transport 특성을 조사하기 위하여, 토마토 뿌리조직으로부터 마이크로솜을 분리하고, 45Ca²+ uptake 실험을 수행하였다. 반응용액에 Ca²+-ATPase의 선택적 저해제인 1 mM vanadate와 액포막 H+-ATPase의 선택적 저해제인 50 mM NO₃를 각각 첨가하였을 때, 45Ca²+ uptake는 각각 20%와 30% 저해되었고, 두 가지 저해제를 동시에 첨가하였을 때 50% 저해되었다. 또한 마이크로솜 45Ca²+ uptake는 vanadate 또는 NO₃-의 농도를 증가시킴에 따라 저해되었다. 이러한 저해효과는 토마토 뿌리조직 마이크로솜에 Ca²+-ATPase와 Ca²+/H+ antiport가 동시에 존재할 가능성을 시사한다. Protonophore인 gramicidin의 처리에 의해 45Ca²+ uptake는 30% 가량 저해되어, 토마토 뿌리조직 마이크로솜에 Ca²+/H+ antiport가 존재하고 있음을 확인하였다. NO₃-를 처리하여 45Ca²+ uptake를 저해시킨 후 gramicidin을 처리하였을 때, gramicidin에 의한 추가저해는 거의 관측되지 않았으나, vanadate의 경우 gramicidin에 의한 추가저해가 현저히 나타났다. 이러한 결과는 토마토 뿌리조직 마이크로솜 45Ca²+ uptake가 vanadate에 의하여 저해되는 Ca²+-ATPase와 NO₃-에 의하여 저해되는 Ca²+/H+ antiport에 의해 이루어지고 있음을 의미한다. 한편, 동물조직 ER/SR-type Ca²+-ATPase의 특이성 높은 선택적 저해제인 thapsigargin은 마이크로솜 45Ca²+ uptake를 농도 의존적으로 저해하였으며, 10μM 농도에서 최대저해효과를 나타냈다. Thapsigargin에 의한 45Ca²+ uptake의 저해효과는 NO₃를 사용하여 액포막 H+-ATPase 활성을 저해하였을 때 현저하게 감소하였다. 반면, vanadate를 처리하여 원형질막 H+-ATPase의 활성을 저해한 후에도 thapsigargin에 의한 추가적 저해효과가 관측되어 thapsigargin이 액포막 H+-ATPase의 활성을 저해하여 간접적으로 Ca²+/H+ antiport를 저해함을 확인하였다. 이상의 결과들은 토마토 뿌리조직 마이크로솜에 Ca²+-ATPase와 액포막 Ca²+/H+ antiport가 존재하며, thapsigargin이 액포막 Ca²+/H+ antiport를 선택적으로 저해함을 보여준다. In order to characterize the property of Ca²+ transport in plant tissue, microsomes were prepared from the roots of tomato and microsomal 45Ca²+ uptake was measured. When 1mM vanadate, a selective inhibitor of P-type ATPases, 50 mM NO₃, a specific inhibitor of vacuolar H+-ATPase, and both of these inhibitors were treated, the microsomal 45Ca²+ uptakes were inhibited by 20%, 33%, and 47%, respectively. The inhibitory effects of these two inhibitors were investigated by using a protonophore, gramicidin. When the chemical gradient of H+ was relieved by gramicidin, the uptake was decreased by 30%, implying the presence of Ca²+/H+ antiporter in the microsomal membrane. In the 45Ca²+ uptake experiment, the effect of gramicidin was independent of vanadate-induced inhibition. However, when the activity of vacuolar H+-ATPase was inhibited by NO₃, the effect of gramicidin was severely decreased. Meanwhile, thapsigargin, a specific antagonist of ER/SR-type Ca²+-ATPase, inhibited the microsomal 45Ca²+ uptake and the maximum inhibitory effect was obtained at 10μM. The effect of thapsigargin was blocked by NO₃and gramicidin, but not by vanadate. These results imply that vanadate directly inhibits the activity of Ca²+-ATPase; however, NO₃- and thapsigargin block the activity of Ca²+/H+ antiporter by inhibiting the vacuolar H+-ATPase. In conclusion, the microsomal 45Ca²+ uptakes are mediated by two major enzymes, Ca²+-ATPase and Ca²+/H+ antiporter in tomato root tissue.

      • KCI등재

        CAD/CAM 으로 제작된 Ceramic Inlay의 변연누출에 관한 비교연구

        조병훈,이명종,임미경,이광원,손호현 大韓齒科保存學會 1997 Restorative Dentistry & Endodontics Vol.22 No.2

        To compare the marginal leakage of CAD/CAM-fabricated ceramic inlay, gold inlay and amalgam, forty extracted caries-free premolars were prepared with Class Ⅱ MO cavity design. The teeth were divided into four groups of ten samples each. Group 1 was restored with CAD/CAM-fabricated ceramic inlays cemented with Scotchbond Resin Cement / Scotchbond Multi-purpose plus. Group 2 was restored with gold inlays cemented with Scotchbond Resin Cement / Scotchbond Multi-purpose plus. Group 3 was restored with gold inlays cemented with zinc phosphate cement. And, Group 4 was restored with amalgam. All samples were thermocycled, and stored in 1% methylene blue. Marginal leakage was examined at four margins, that is, occlusal distal, priximal gingival, occlsal facial and occlusal lingual margins from sectioned samples under steeomicroscope(x15). The results were as follows: 1. Group 1 and 2 showed no statistically significant difference among marginal leakage at all four examined margins(p>0.05). 2. Group 3 and 4 showed significant marginal leakage at proximal gingival margin compared with other margins(p<0.05). 3. Significantly increased marginal leakage at priximal gingival, occlusal facial and occlusal lingual margins in group 3 were observed compared with other groups(p<0.05).

      • 심실중격결손증-치험 1예-

        조광현,우종수,황윤호,이양행,박철호,김종성,김철호,이경순,조영일,박태인 인제대학교 1985 仁濟醫學 Vol.6 No.4

        A successful treatment of infants and children with congenital heart defects depends upon several factors: diagnosis must be accurate, the operative technique, including techniques of perfusion and myocardial preservation, must be suited to the needs of patients and intensive cares, including accurate anesthesia, must be applied through all stages of investigation and treatment, i.e. before, during and after the operation. Recently we performed a open heart surgery successfully for a six-years old girl who had suffered from frequent upper respiratory tract infection and mild to moderate exertional dyspnea with congenital heart defects (ventricular septal defect, Kirklin type II, with patent foramen ovale). The operation and postoperative course were very smooth. And now we report this with review of literatures.

      • HGCMP기반 서버 시스템상의 전달지연 연구

        조현주,김한수,이광휘 昌原大學校 基礎科學硏究所 1995 基礎科學硏究所論文集 Vol.7 No.-

        In this thesis, the message transmission delay on a server system using HGCMP for group communication protocol was analyzed. The queue model of a server system was modeled by queueing theoretic approach. As the feedback structure was imbedded in the queue model, it is difficult to analyze the delay. To solve this problem, the simplification of the queue model was performed. After the simplification process, the queue model was modeled M/M/1. The transmission delay of a server system is calculated using average waiting time and service time in a server system.

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