멸균방법에 따른 탈회동종골의 세포독성 여부에 관한 실험적 연구

임창준,김종여,우기선,김세원 大韓顎顔面成形再建外科學會 1997 Maxillofacial Plastic Reconstructive Surgery Vol.19 No.3

Procurement, cutting, cleansing, freezing, freeze-drying, and demineralization of the allogeneic bone must be made under the germ-free stable condition without bacterial and/or viral contamination. Even thought the bone is procured under the germ free condition, we must have confidence on disinfection of all the solutions that come in contact with tissue during the whole procedure. Lots of antibacterial agents have been introduced for chemical sterilization. Recently ethylene oxide gas sterilization or radiation sterilization is frequently selected as a secendary sterilization procedure. The biological and biochemical response of the graft material differs with the type and concentration of the sterilizing agents, and various toxic reactions have been reported due to the graft material itself and the substance released by the chemicals. The authors conducted the Millipore filter test to observe the toxic effect on L929 fibroblasts according to the effect on activity of succinate dehydrogenase, during the secondary sterilization of the demineralized allogeneic bone powder with irradiation or ethylene oxide gas. The result were as follows: 1. Around the copper disk, positive control group, 10mm diameter discoloration was observed. 2. As same as the negative control group, the disk showed no discoloration. 3. The demineralized allogeneic bone which was sterilized with ethylene oxide gas or irradiation showed no cytotoxicity. 4. From this results, it is suggested that treatment with ethylene oxide gas or irradiation should be effective to sterilize the deminineralized allogeneic bone.

폐굴껍질을 산성토양 적용시 토양의 화학적 특성변화에 관한 기초 연구

임진희,문종익,김성우,성낙창,이영형,윤태경 동아대학교 환경문제연구소 1999 硏究報告 Vol.22 No.2

Oyster shell has caused environmental problems in the coast. But it is possible to be used as an acidic soil amendment because it is the alkalic material of pH 9.0. In order to evaluate the utility of the crushed oyster shell as a soil amendment, acidic sandy loam soil was amended with it and then Chinese cabbages were cultivated in pots. The amount of the oyster shell was ⅰ)none, ⅱ)0.5kg/㎥, ⅲ)1.0kg/㎥, ⅳ)1.5kg/㎥, ⅴ)2.0kg/㎥ and ⅵ)2.3kg/㎥. The particle size of oyster shell consists of 34.88% of 40∼100mesh and 14.98% of larger than 100 mesh. It increased pH and the contents of available P₂O_(5) and SiO₂ and exchangeable Ca in used soil The application of oyster shell also increased the height and diameter of Chinese cabbages.

대퇴 골두 골내 결절종 : 증례 보고 A Case Report

임수재,천동일,우승한,임강철,박종석,이병일 대한고관절학회 2004 Hip and Pelvis Vol.16 No.1

갈륨액체금속 이온원과 인듐액체금속 이온원의 빔 특성에 대한 연구

현정우,임연찬,정강원,정원희,박철우,이종향,강승언 한국공작기계학회 2005 한국공작기계학회 춘계학술대회논문집 Vol.2005 No.-

본 연구에서는 인듐 액체금속이온원을 제작하여 빔 특성에 대해 연구를 하였으며, 기존에 연구를 하였던 갈륨 액체금속이온원의 빔특성과 비교 분석 하였다. 빔특성 분석을 위해 빔 안정도, 전류-전압특성곡선, 에너지 퍼짐을 측정하였다. 액체금속이온원에 사용되는 액체금속 저장소 및 바늘전극(tip)은 500μm의 직경을 갖는 텅스텐을 사용하였으며, 국내에서 제작된 제품을 사용하였다. 액체금속 저장소의 구조는 이전에 구상하여 연구가 이루어진 6개의 pre-etching된 텅스텐와이어(wire)가 묶여진 형태를 사용하였다.

Angiotensin 전환효소 유전자 다형성과 양극성 장애

김경나,김종우,정주호,이기철,정홍경,임성빈 대한생물치료정신의학회 2003 생물치료정신의학 Vol.9 No.2

연구목적 : 감정조절에 Renin-Angiotensin System이 관여하는 것으로 보고되어 왔다. 그 근거로 우울증의 새로운 약물치료로 주목받고 있는 Substance P의 대사에 angiotensin 전환효소(angiotensin-converting enzyme, ACE)가 관여한다는 것과 고혈압 환자에서 angiotensin 전환효소 억제제(ACE inhibitor)를 사용했을 때 다행감이나 우울감을 초래한다는 것 등이 있다. 본 연구에서는 ACE의 유전자 다형성을 분석하여 양극성장애와 ACE와의 연관성을 알아보고자 하였다. 방 법 : 양극성장애로 진단된 환자군 82명과 대조군 135명을 대상으로 16번째 intron의 다형성 부위를 가진 시발체 쌍(primer pair)을 사용하며 중합효소 연쇄반응(Polymerase Chain Reaction, PCR)을 시행하여 490bp 산물(I allele)과 190bp 산물(D allele)을 관찰함으로써 ACE 유전자의 유전자형(genotype)의 발현율과 대립유전자(allele)의 빈도를 비교 분석하였다. 결 과 : 양극성장애 환자군과 대조군 사이에서 유전자형의 발현율과 대립유전자적 빈도는 유의한 차이가 없었다. 결 론 : 양극성장애 환자군과 ACE 유전자 다형성 사이에 유의한 관련성은 없었다. 이 결과는 ACE가 양극성 장애의 원인으로 주요한 역할을 한다는 것을 시사하지 못했다. Objectives : A possible participation of the Renin-Angiotensin Systern(RAS) in regulating of the mood has been suggested by reports as follows : the angiotensrn converting enzyme(AGE1 is involved jn the metabolism of the neuropeptide substance P impficated with novel strategies for the pharmacotherapy of depression and the use of ACE inhibitors in hypertensive patients has been associated with euphoric or depressive stales. The aim of the present study was to analyze the role of the ACE gene I/D polymorphism for bipolar disorder. Methods : We examined the frequency of a polymorphism characterized by the insertion or deletion of a 287-bp Alu repeat sequence in intron 16 of the angiotensin converting enzyme gene(located on chromosome 37q233 in groups of patients with bipolar disorder(n=82) compared to healthy control subjects[n=135). ACE genotype was determined by size-analysis of polymerase chain reaction products. Results : The ACE ID polymorphism did not show any difference in allelic frequencies and genotypic distributions between bipolar disorder patients and control subjects. Conclusions : No significant association was found with bipolar disorder and the polymorphism of ACE gene. This finding does not support that ACE I/D polymorphism is a significant risk factor for bipolar disorder.

한국인 제2형 당뇨병 환자의 골격근에서 인슐린 신호전달체계의 결함

최준혁,이관우,김효정,이동훈,이종우,김정은,엄현채,김경미,최성이,정윤석,김현만 대한내분비학회 2002 Endocrinology and metabolism Vol.17 No.5

연구배경: 제2형 당뇨병에서 나타나는 인슐린 저항성은 간, 지방, 근육 같은 말초조직에서 인슐린의 작용이 떨어지는 것을 말한다. 제2형 당뇨병의 발생기전에 인슐린 저항성과 인슐린 분비능 저하가 같이 관여함은 잘 알려진 사실이다. 인슐린의 세포내에서의 저항성을 규명하기 위한 인슐린의 세포내 신호전달체계에 대한 연구가 활발히 진행되고 있으며 몇몇 연구에서 인슐린 저항성을 가진 골격근육에서 IRS와 관련된 PI3-kinase의 활성감소와 Akt kinase의 활성감소를 보고하고 있으나 아직까지 명확하게 그 기작이 설명되어지지 않고 있다. 본 연구에서는 정상성인, 제2형 당뇨병 환자를 대상으로 경구당부하검사 및 인슐린 클램프검사를 시행하고 인슐린 클램프 검사시 대상인의 근육을 채취하여 인슐린 신호전달 체계(IR-β, IRS, Akt(PKB, Rac) kinase, GSK-3)를 연구하였다. 방법: 연구대상자는 한국인으로서 경구당부하검사상 정상인 및 당뇨병 환자 각각 11명, 9명을 대상으로 하였으며, 정상인은 건강인으로 과거력상 당대사에 영향을 줄 질환이 없고, 현재 당대사에 영향을 줄 약물 복용 및 다른 소견이 없는 경구당부하검사상 정성 내당능을 보이는 대상자로 하였다. 당뇨병환자는 모두 제2형 당뇨병 환자로 이환기간이 만 5년 이내인 경우로 하고, 인슐린으로 치료하는 대상자는 제외하도록 하였다. 대상자의 연령, 체질량지수, 체지방량, 공복시 총콜레스테롤, 중성지방, 고밀도지단백 콜레스테롤 및 HbA1c, 인슐린, C-peptide를 측정하였고, "HOMA model"을 이용하여 베타세포의 기능와 인슐린저항성 정도를 평가하였다. 정상혈당클램프 검사(euhlycemic hyperinsulinemic clamp test)를 시행하고 인슐린 투입 전과 인슐린 투입 30분후, 두차례에 걸쳐 대퇴부 근육생검을 실시한후 western 법으로 IR-β, IRS, Akt kinase, GSK-3의 인산화량을 측정하였다. 결과: 정상인 11명과 당뇨병 환자 9명을 대상으로 하였으며 대상자의 평균 연령, 평균 체질량지수, 체지방량, 허리/엉덩이 둘레의 비(waist hip ratio; WHR)는 두 군간에 차이가 없었다. 평균 공복혈당은 정상군 98.0±0.3㎎/dL, 당뇨병군 208.1±16.5㎎/dL(p<0.05), HbA1c는 정상군 5.4±0.5%, 당뇨병군 9.2±0.6%(p<0.05)였다. "HOMA model"을 이용한 베타세포의 기능과 인슐린저항성은 정상군 56.4±8.5%, 1.4±0.2, 당뇨병군 72.2±52.3%(p<0.01), 10.2±6.3(p<0.01)였으며 정상혈당클램프 검사상 포도당 이용률은 정상군 8.2±0.6㎎/㎏/min, 당뇨병군 3.7±1.1㎎/㎏/min(p<0.01)로 정상군과 당뇨병군간에 유의한 차이를 나타내었다. Western blot법으로 IR-β, IRS, Akt kinase, GSK-3의 인산화량을 인슐린 투입전과 인슐린 투입 30분후 측정하였을 때 IR-β에서는 정상군이 103.9±2.3에서 241.3±18.6, 당뇨군이 108.9±2.2에서 198.7±6.3으로 증가하였다(p=NS). 이는 인슐린 투입후 당뇨병군이 정상군과 비교하여 인산화량의 증가정도가 통계적인 차이가 없었다. IRS에서는 정상군이 111.6±7.3에서 295.6±17.2, 당뇨군이 114.5±6.1에서 222.0±23.2로 증가하였으며(p<0.05), 인슐린 투입후 당뇨병군이 정상군과 비교하여 인산화량의 증가정도가 약 24% 감소하였다. Akt kinase에서는 103.4±6.0에서 416.8±29.5로 증가하였으며(p<0.01), 인슐린 투입후 당뇨병군이 정상군과 비교하여 인산화량의 증가정도가 약 43% 감소하였다. GSK-3에서는 정상군이 107.7±6.7에서 595.7±28.1, 당뇨군이 104.3±4.8에서 443.3±12.9로 증가하였으며(p<0.01), 인슐린 투입후 당뇨병군이 정상군과 비교하여 인산화량의 증가정도가 약 25% 감소하였다. 결론: 제2형 당뇨병 환자에서 정상인과 비교하여 인슐린 자극후 IR-β는 인산화량의 증가정도가 큰 차이가 없었으나, IRS, Akt kinase, GSK-3에서는 인산화량의 증가 정도가 감소되는 것으로 보아 상위 인슐린 신호전단체계부터 결함이 있는 것으로 생각된다. Background: The glucose uptake rate is the limiting step in glucose utilization and storage. The failure of insulin to stimulate glucose uptake in muscle appears to be a primary defect of insulin resistance. This study was undertaken to examine the effect of physiological hyperinsulinemia on the phosphorylation of the insulin receptor (IR-β), insulin receptor substrate (IRS), Akt kinase and GSK-3 in isolated skeletal muscle, in people with type 2 diabetes(n=9) and control subjects(n=11). Methods: 75g OGTT and euglycemic hyperinsulinemic clamp test were done. And vastus lateralis muscle was obtained before and 30 min into the euglycemic clamp. Western blots were performed for tyrosine phosphroylation of insulin receptor substrate (IRS) and phosphorylation of the insulin receptor(IR-β), Akt and GSK-3. Result: There were no statistical differences in the mean age, BMI and body fat between the control subjects and diabetic patients. The fasting blood sugar and HbA_1c in controls and diabetic patients were 98.0±0.3 and 208.1±16.5ng/dl, and 5.4±0.5 and 9.2±0.6%, and 1.4±0.2 in the control subjects, and 72.2±52.3%(p<0.01) and 10.2±6.3(p<0.01) in the diabetic patients, respectively. The insulin resistance from the euglycemic hyperinsulinemic clamp test were 8.2±0.6㎎/㎏/min and 3.7±1.1ng/㎏/min in the control subjects and in the diabetic patients, respectively(p<0.01). Compared with the normal controls, insulin-stimulated IR phosphorylation was no different to that in the diabetic patients. However, insulin-stimulated IRS phosphorylation, insulin-stimulated Akt phosphorylation and insulin-stimulated GSK-3 phosphorylation were reduced in the diabetic patients compared with the normal controls by 24, 43 and 25%, respectively(p<0.05). Conclusion: In Korean type 2 diabetic patients, the insulin resistance may be due to the impairment of the upstream insulin signal molecular network. Further studies will focus on determining whether these signaling defects are the cause of the development of insulin resistance, or secondary to the altered metabolic state, associated with type 2 diabetes mellitus (J Kor Soc Endocrinol 17:685∼697, 2002).

Regulation of Ethylene Emission in Tomato (Lycopersicon esculentum Mill.) and Red Pepper (Capsicum annuum L.) Inoculated with ACC Deaminase Producing Methylobacterium spp.

Yim, Woo-Jong,Woo, Sung-Man,Kim, Ki-Yoon,Sa, Tong-Min Korean Society of Soil Science and Fertilizer 2012 한국토양비료학회지 Vol.45 No.1

Improvement of plant growth by Methylotrophic bacteria can be influenced through alterations in growth modulating enzymes or hormones, especially by decreasing ethylene levels enzymatically by 1-aminocyclopropane-1-carboxylate (ACC) deaminase or by production of indole-3-acetic acid (IAA). In this study, the effect of seven strains of Methylobacterium on seedling ethylene emission of tomato and red pepper plants was evaluated under greenhouse condition. Ethylene emission was lowest in Methylobacterium oryzae CBMB20 inoculated tomato plants and CBMB110 inoculated red pepper plants at 47 days after sowing (DAS). However, at 58 DAS all inoculated plants showed almost similar pattern of ethylene emission. Methylobacterium inoculated tomato and red pepper plants showed significantly less ethylene emission compared to control. Our results demonstrated that Methylobacterium spp. inoculation promotes plant growth due to the reduction of ethylene emission and therefore can be potentially used in sustainable agriculture production systems.

Ubiquitous Presence and Activity of Thiosulfate Oxidizing Bacteria in Rhizosphere of Economically Important Crop Plants of Korea

Woo-Jong Yim,R. Anandham,P. Indira gandhi,In-Soo Hong(서평자),M.R. Islam,P. Trivedi,M. Madhaiyan,Gwang-Hyun Han,Tong-Min Sa 한국토양비료학회 2008 한국토양비료학회지 Vol.41 No.1

The presence of thiosulfate oxidizing bacteria was examined in rhizosphere soils of 19 economically important plant species belonging to 10 different families. The results showed that the thiosulfate oxidizing bacteria were present in all the tested rhizosphere soils, and the total 32 thiosulfate oxidizing bacteria were recovered. Furthermore, the biochemical characterization revealed that 56% and 44% of the isolates belonged to the obligate chemolithoautotrophs and facultative heterotrophs, respectively. The isolates ATSR15P utilized 19.17 mM of thiosulfate and accumulated 11.65 mM of sulfate in the medium. Concurrently, the decrease in pH of the medium was observed. This study comprehensively demonstrates that the active sulfur oxidation is a ubiquitous phenomenon in the rhizosphere of crop plants in Korea.

Immunomodulatory and antitumor effects in vivo by the cytoplasmic fraction of Lactobacillus casei and Bifidobacterium longum

Hee Jong Woo,Jung-Woo Lee,Jung-Gul Shin,Eun Hee Kim,Hae Eun Kang,In Been Yim,주홍구,Ji Yeon Kim 대한수의학회 2004 Journal of Veterinary Science Vol.5 No.1

The immunomodulatory and antitumor effects of lactic acid bacteria (LABs) were investigated. Cytoplasmic fraction of Lactobacillus acidophilus, Lactobacillus casei and Bifidobacterium longum were tested for the antiproliferative activity in vitro to SNUC2A, SNU1, NIH/ 3T3 and Jurkat cell lines by crystal violet assay. All cytoplasmic fraction suppressed proliferation of tumor cells, though L. casei and B. longum were more effective. From these results, cytoplasmic fraction of L. casei and B. longum with Y400 as a control were administered as dietary supplements to Balb/c mice for 2, and 4 consecutive wks. Administration for 4 wks enhanced the number of total T cells, NK cells and MHC class II+ cells, and CD4−CD8+ T cells in flow cytometry analysis. To determine of antitumor activity of LABs preparation in vivo, F9 teratocarcinoma cells were inoculated on mice at 14th day. Body weight was decreased with increased survival rate in all groups with the cytoplasm of LABs. Our results showed that cytoplasmic fraction of LABs had direct antiproliferative effects on tumor cell lines in vitro, effects on immune cells in vivo, and antitumor effects on tumor-bearing mice with prolonged survival periods.

Ralstonia Solanacearum에 감염된 토마토에서 ACC Deaminase를 생산하는 Methylobacterium spp.의 스트레스 에틸렌 수준 조절

임우종 ( Woo Jong Yim ),김기윤 ( Ki Yoon Kim ),우성만 ( Sung Man Woo ),이선미 ( Seon Mi Lee ),홍보희 ( Bo Hui Hong ),사동민 ( Tong Min Sa ) 한국환경농학회 2012 한국환경농학회 학술대회집 Vol.2012 No.-

Biosynthesis of ethylene is usually enhanced with severity of pathogenic infection. Some ethylene synthesis inhibitors are known to significantly decrease the severity of pathogen infections in plants. Bacteria of genus Methylobacterium have been found to stimulate plant growth and regulate the level of ethylene in crop plants. A selective decrease of ethylene can be achieved by plant growth promoting bacteria, for instance Methylobacterium, containing ACC (1-aminocyclopropane-1- carboxylate) deaminase, which cleaves ACC into -ketobutyrate and ammonia in higher plants which leads to reduced ethylene level. This work aimed to test the induction of defense responses in tomato against bacterial wilt by stress ethylene level reduction mediated by the ACC deaminase activity of Methylobacterium strains. In greenhouse experiments, the disease index value observed in Methylobacterium strains treated tomato and red pepper plants were lower than that of uninoculated treatment. Plants treated with Methylobacterium strains challenge inoculated with Ralstonia solanacearum (RS) showed significantly reduced disease symptom and lowered ethylene emission under greenhouse condition. The ACC and ACO (1-aminocyclopropane-1- carboxylate oxidase) accumulated in the tomato leaves were significantly reduced with Methylobacterium strains inoculation compared to treatment with pathogen alone. Similar to ACC oxidase activity, ACC oxidase related gene expression also increased in tomato treated with Ralstonia solanacearum alone compared to Methylobacterium strains treatment. In addition pathogenesis-related (PR) proteins related to ISR (Induced systemic resistance) including -1,3-glucanase, phenylalanine ammonia-lyase (PAL), peroxidase (PO) and polyphenol oxidase (PPO) were increased in Methylobacterium strain inoculated tomato plants. A significant increase in -1,3-glucanase and PAL related genes expression was found in all of the Methylobacterium spp. treatments compared to the Ralstonia solanacearum alone treatment. This study supports that the Methylobacterium strains might increase the activity of defense enzymes by modulating the ethylene biosynthesis pathway, suggesting the possibility of using Methylotrophic bacteria as potential bio-control agents.