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Reunion of the Rabbit Superior Oblique Tendon After Weakening Procedures
강대욱,오지혜,전보영,권정윤 대한안과학회 2009 Korean Journal of Ophthalmology Vol.23 No.3
Purpose: To investigate the degree of reunion in rabbit eyes of the superior oblique tendon after several surgical weakening procedures. Methods: A total of 32 rabbits (64 eyes) were used in this study. The rabbits were randomly assigned to four groups, eight rabbits (16 eyes) in the tenotomy group, eight rabbits (16 eyes) in the tenectomy group, eight rabbits (16 eyes) in the disinsertion group and eight rabbits (16 eyes) in the recession group. The degree of reunion or reattachment of the superior oblique tendon on the globe were examined on four eyes in each group at postoperative weeks two, four, six and eight. Results: At eight weeks, the newly created insertion site remained at the same site in all eyes in the recession group, and the distal end of the superior oblique tendon was reattached at the medial border of the superior rectus muscle in all four eyes in the tenotomy and disinsertion groups, and in three of four eyes in the tenectomy group. Conclusions: From this experimental study, it was speculated that superior oblique recession is more effective than other superior oblique weakening procedures. This result could be helpful in the prediction of time of recurrence for superior oblique overaction after superior oblique weakening procedures.
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항균활성을 보유한 재조합 Pichia pastoris 균주의 개발
강대욱,이준원,허건영,안종석 한국생명과학회 2002 생명과학회지 Vol.12 No.4
곤충에서 유래한 항균 펩티드, defensin을 항균활성이 있는 활성적인 형태로 분비하는 Pichia 균주를 개발하기 위한 일환으로서 MF$\alpha$1 prerpo sequence와 defensin 합성유전자를 pichia 발현 벡터에 재조합하여 형질전환하고 아미노산 histidine을 첨가하지 않은 최소 배지에서 형질전환체를 일차적으로 선별하였다. 선별한 형질전환체를 대상으로 항생제 G-418에 대한 내성과 M. luteus를 시험균으로 사용하여 생육환 저해를 통한 defensin의 세포 외 분비를 조사하여 4 균주를 선택하고 분석하였다. Southern hybridizaion을 통해 숙주의 염색체 DNA에 삽입한 defensin 유전자가 유지됨을 확인하였으며 전체 RNA를 분리하고 RT-PCR을 수행하여 defensin mRNA를 증폭하고 Southern hybridization을 실시한 결과 증폭된 밴드는 probe로 사용한 defensin 유전자에 의해 양성 신호가 나타났다. 배양시간에 따른 4 균주의 세포성장과 항균활성을 비교하기 위해 BMMY 배지에서 96시간 배양하였다. 세포성장은 모두 유사한 양상을 보였다. 세포성장은 48시간 동안 급격하게 증가한 후 그 이후로는 정지기에 도달되었다. 항균활성도 48시간까지 급격히 증가하였으며 항균활성이 가장 높은 형질전환체 균주 3는 72시간 배양 시 550 AU/$m\ell$을 나타내었다. To develop a yeast strain of Pichia pastoris producing an antibacterial peptide, we have attempted the expression and secretion of an insect defensin. The nucleotide sequences corresponding to mature defensin were chemically synthesized by 6 oligomers, assembled in vitro and the synthesized gene was identified by nucleotide sequencing. The prepro sequence of yeast mating factor $\alpha$1 and the defensin gene were recombined into a Pichia expression vector, pPIC9K. The resulting plasmid, pPIDE, was transformed into P. pastoris GSl15 and transformants selected on histidine-deficient minimal plates were tested for antibacterial activity against Micrococcus luteus. Four strains with different antibacterial activity were selected for further analysis. Southern hybridization and RT-PCR verified the defensin gene was maintained and transcribed in a host. Four strains were cultivated in YPD broth for 96 hours to compare cell growth and antibacterial activity, They showed no difference in cell growth, however, each strain showed different antibacterial activity pattern with culture time. The maximal activity was about 550 AU/ $m\ell$.
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효모에서 활성형의 곤충유래 항균펩티드 defensin의 발현
강대욱,안순철,김민수,안종석 한국생명과학회 2002 생명과학회지 Vol.12 No.4
효모 glucoamylase의 promoter와 분비신호서열 그리고 MF$\alpha$1의 prosequence를 이용하여 곤충 defensin을 S. cerevisiae 2805에서 항균활성을 보유한 형태로 발현 및 분비하는데 성공하였다. 발현된 defensin의 대부분이 세포 외로 분비되어 거의 모든 항균활성이 배양 상등액에 존재하였다. 이것은 S. cerevisiae에서 발현된 defensin이 glucoamylase의 분비신호서열과 MF$\alpha$1의 prosequenre에 의해 효율적으로 processing되어 분비됨을 시사한다. Defensin의 다른 미생물에 대한 항균활성을 조사한 결과 병원균인 St. aureus와 L. monocytogenes에 대해서도 항균활성이 존재하였다. As a biological model system for the production of an active antibacterial peptide, we have attempted the expression and secretion of insect defensin in Saccharomyces cerevisiae. Nucleotide sequences encoding mature defensin composed of 40 amino acids were fused in frame with promoter and signal sequence of Saccharomyces diastaticus glucoamylase, and mating factor $\alpha$ l[MF $\alpha$1] prosequence. The host strain, S. cerevisiae 2805 was transformed with the resulting plasmid, pSMFll The secretion of functional defensin was confirmed by growth inhibition zone assay using Micrococcus luteus as a test organism. Insect defensin was secreted to the culture supernatant in biologically active form by glucoamylase signal sequence and mating factor $\alpha$1 prosequence. Most of antibacterial activity was detected in the culture supernatant. Defensin was also active against Staphylococcus aureus and Listeria monocytogenes.
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Saccharomyces cerevisiae에서 발현한 곤충 항균펩티드, defensin의 정제 및 특성 조사
강대욱,이준원,김보연,안종석 한국생명과학회 2002 생명과학회지 Vol.12 No.4
S. cerevisiae에서 glucoamylase 유전자의 promoter와 signal sequence 그리고 MF$\alpha$1의 prosequence를 이용하여 합성 곤충 defensin를 발현하고 항균활성을 보유한 형태로 분비하는데 성공하였다. Defensin의 여러 생화학적인 특성을 조사한 결과 열 안정성이 높아 10$0^{\circ}C$에서 30분간 가열하여도 항균활성을 온전히 유지하였으며 조사한 pH 영역, 2.0-12.0에서 항균활성의 변화가 없었다. 또한 여러 단백질 분해효소를 처리하면 항균활성이 완전히 사라졌으나 전분질 분해효소, 섬유소분해효소 및 지질분해효소의 처리는 항균 활성에 전혀 영향이 없었다. 황산암모늄침전, SP-Sepharose column cormatography, RP-HPLC 등의 조작을 통해 defensin을 순수한 형태로 정제하였으며 Tricine-SDS-PAGE를 통해 분자량이 약 4.0 kDa임을 확인하였고 정제한 defensin은 항균활성을 보유하였다. We investigated the biochemical properties of insect defensin expressed and secreted from Saccharomyces corevisiae. The defensin showed extremely high resistance to boiling for up to 30 min and to pH values tested from 2.0 to 12.0. The treatment of defensin with various proteases abolished antibacterial activity. However, amylases, cellulase, lipase and catalase had no effect on the activity. The defensin was purified to homogeneity through ammonium sulfate concentration of culture supernatant, SP-Sepharose column chromatography and RP-HPLC. Tricin-SDS-PAGE analysis revealed that the molecular weight of the defensin was about 4.0 kDa. The antibacterial activity of the purified defensin was verified by renaturation of stained gel and gel pouring assay using Micrococcus luteus as a test organism.
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大單位 住居團地의 規模計劃的 特性에 관한 硏究 : 釜山市 宅地開發事業地를 中心으로 Focus on the Housing Lot Development Projects in Pusan
姜大旭 釜山大學校 都市問題硏究所 1997 都市硏究報 Vol.5 No.-
The purpose of this paper is to find out the characteristics of pattern in large scale residential districts. For the above purpose, literature and field survey was carried out with the large scale residential district projects which were executed in Pusan. The main results obtained from the study were as follows. 1. The pattern of development project is affected by the conditions of location and easiness of execution. 2. The developing circumstance of peripheral district is good prepared, the ratio of housing lot is shown upward tendency. 3. The density of residential district is increased by the change of times.
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