Reconstruction of Rabbit Corneal Epithelium on Lyophilized Amniotic Membrane Using the Tilting Dynamic Culture Method

Ahn, Jae-Il,Lee, Doo-Hoon,Ryu, Yang-Hwan,Jang, In-Keun,Yoon, Mun-Young,Shin, Youn Ho,Seo, Young-Kwon,Yoon, Hee-Hoon,Kim, Jae-Chan,Song, Kye-Yong,Yang, Eun-Kyung,Kim, Ki-Ho,Park, Jung-Keug Blackwell Publishing Inc 2007 Artificial Organs Vol.31 No.9

<P>Abstract: </P><P>Rabbit corneal epithelium was reconstructed using tilting dynamic culture with a self-manufactured, amniotic membrane (AM) supporter and a lyophilized amniotic membrane (LAM). Rabbit corneal epithelial (RCE) cells were cultured and cryopreserved after isolation from the limbus. The second- and third-passage RCE cells were plated onto the epithelial side of the LAM of Ahn's AM supporter. Two days later, the air–liquid interface culture was maintained with third-passage RCE cells for 6 days and second-passage corneal epithelial cells for 9 days. The average viability of thawed RCE cells, assessed using trypan blue dye exclusion, was 77.42%. The reconstructed corneal epithelium was characterized by histological (hematoxylin and eosin) and immunohistochemical staining (proliferating cell nuclear antigen) for light microscopy, and by reverse transcriptase-polymerase chain reaction, glucose assay, and transmission electron microscopy. The basal layer of the reconstructed corneal epithelium was well formed, and the epithelium was tightly constructed due to the increase in cell proliferation and differentiation caused by the tilting dynamic culture, as opposed to static culture. Tilting dynamic culture was useful for the reconstruction of the epithelium using easily damaged epithelial cells and resulted in more stratum cell layers. Moreover, cytokeratin (CK3) mRNA expression in tilting dynamic cultured third-passage RCE cells seeded onto AM was greater than in static cultured third-passage RCE cells. The morphology of the reconstructed corneal epithelium on LAM by tilting dynamic culture for 9 days resembled that of the skin epidermis. This was thought to be because the tilting dynamic culture not only accelerated the proliferation and differentiation of cells by physical or mechanical stimulation, but also ensured that the supply of medium was delivered to the basal cells more efficiently. Thus, the reconstruction of the corneal epithelium using LAM and tilting dynamic culture was considered to be a good in vitro model for autologous or allogeneic transplantation of corneal epithelium and skin epidermis in patients with damaged epithelia. </P>

A Comparison of Lyophilized Amniotic Membrane with Cryopreserved Amniotic Membrane for the Reconstruction of Rabbit Corneal Epithelium

Ahn Jae-Il,Jang In-Keun,Lee Doo-Hoon,Seo Young-Kwon,Yoon Hee-Hoon,Shin Youn-Ho,Kim Jae-Chan,Song Kye-Yong,Lee Hee-Gu,Yang Eun-Kyung,Kim Ki-Ho,Park Jung-Keung The Korean Society for Biotechnology and Bioengine 2005 Biotechnology and Bioprocess Engineering Vol.10 No.3

Many researchers have employed cryopreserved amniotic membrane (CAM) in the treatment of a severely damaged cornea, using corneal epithelial cells cultured on an amniotic membrane (AM). In this study, two Teflon rings were made for culturing the cells on the LAM and CAM, and were then used to support the AM, which is referred to in this paper as an Ahn's AM supporter. The primary corneal epithelial cells were obtained from the limbus, using an ex-plantation method. The corneal epithelium could be reconstructed by culturing the third­passage corneal epithelial cells on the AM. A lyophilized amniotic membrane (LAM) has a higher rate of graft take, a longer shelf life, is easier to store, and safer, due to gamma irradiation, than a (AM. The corneal epithelium reconstructed on the LAM and (AM, supported by the two­Teflon rings, was similar to normal corneal epithelium. However, the advantages of the LAM over that of the (AM make the former more useful. The reconstruction model of the corneal epithelium, using AM, is considered as a good in vitro model for transplantation of cornel epithelium into patients with a severely damaged cornea.

뇌 자기공명영상상 뇌량 병변의 호전을 보인 Marchiafava-Bignami병 1예

정재훈,성기범,안무영,신현길,박형국 대한신경과학회 1999 대한신경과학회지 Vol.17 No.5

상업스포츠시설 이용자의 스포츠 소비행동에 관한 연구

안영필,정종훈,조재기,김종백 東亞大學校 附設 스포츠科學硏究所 1995 스포츠科學硏究論文集 Vol.13 No.-

With 509 regular members of commercial sports centers as the subjects, such variables as their consumption behavior, expectation & satisfaction were analyzed. For the purpose of setting up a strategic theory of marketing segmentation, this sports market's consumer characteristics and responses were segmented. The following sums up the conclusion : THE Segmentation Matrix of the Commercial Sports Center Market -------------------------------------------------------------------------------- Division Large Users (27.0%) Small Users (24.2%) -------------------------------------------------------------------------------- Class by skill(beginning, Expectation intermediate, advanced, sex, age) Parking lot. Shuttle buses Friendship matches with some invited athletes. ------------------------------------------------------------------------------- Satisfaction Convenient facilities and Price of using the center VIP or discount tickets. -------------------------------------------------------------------------------- Unsatisfactory in convenient Unsatisfactory in divers facilities, clean weter, programs,health counselling, periods and contens of leaders' advise and programs, technicques, class of by skill, number of price of using,shuttle buses, members a class,various and general environment. Satisfaction programs,health counselling, A little satisfactory in the Against leaders' advise,coaching items of parking lot and Expectation echnicques,price of using, special events for shuttle buses,parking lot, member's friendship and environment. Rather satisfactory in the items of convenient membership and attitude to customers. --------------------------------------------------------------------------------

沙上地域 産業體 勞動者의 職場體育 參與擴大 方案

安永弼,鄭琮勳,鄭三鉉,金海禮,趙在基,朴俊東,金垠利,張禎允,金福善,孔庚鎬 東亞大學校附設스포츠科學硏究所 1989 스포츠科學硏究論文集 Vol.7 No.-

톨루엔 디이소시아네이트 폭로 근로자들의 생물학적 모니터링

안연순,노재훈,김치년,박윤정,정상혁 大韓産業醫學會 1995 대한직업환경의학회지 Vol.7 No.2

Toluene diisocyanate(TDI) is widely used in the production of flexible polyurethane foams, as well as in the formulation of polyurethane paints and coatings. The commercial material is generally a mixture of 2,4- and 2, 6-TDI, the predominant mix being 80% 2,4 and 20% 2,6-TDI. The 2,4-isomer is considerably more reactive than the 2,6-TDI at ambient temperatures due to steric factors involving the positions of the isocyanate groups relative to the ring methyl group. Because of this difference in the reactivities of the isomers, it seemed probable that there might be an increase in the amount of 2,6-TDI offgased relative to the 2,4-isomer. Therefore a relative enrichment of the 2,6-TDI has been found in industrial atmospheres. Toluene diamines, which are metabolites of TDI, in urine have a linear relation with exposure to TDI, so that urianry TDA could be used as a biological index of the exposure to TDI. This study was conducted to investigate the distribution of TDI isomer in industrial atmospheres and to propose proper biological monitoring methods by identifying the relationships between the environmental TDI exposure and concentration of TDA in urine. Concentrations of 2, 4-TDI and 2, 6-TDI in air were 4.38㎍/㎥ and 25.43㎍/㎥, respectively. The Threshold Limited Value of 40㎍/㎥ was exceeded for the 2, 6-TDI in about 46.8%(22 samples) of the samples, while the 2, 4-TDI was not at all exceeded. The ratio between 2, 4-TDI and 2, 6-TDI varied in air samples in the range of 2.4%:97.6%-51.0%:49.0%. There was an enrichment of 2, 6-TDI in air relative to the 2, 4-TDI. Concentrations of 2, 4-TDA and 2, 6-TDA in urine were 1.31㎍/g creatinine and 4.16㎍/g creatinine, respectively. The ratio between 2, 4-TDA and 2, 6-TDA varied in urine samples in the range of 1.4%:98.6%-99.9%:0.1%. There was an enrichment of 2, 6-TDA in urine relative to the 2, 4-TDA. No relation between the concerations of TDA isomer in urine and concerations of TDI isomer in air was found. Above results of this study, workers were more exposed to the 2, 6-TDI relative to the 2, 4-TDI in industrial atmospheres. Therefore, the establishment of TLV for 2, 6-TDI should be considered. Also, the further studies on biological monitorigs of workers exposed to TDI should be continued.

十全大補湯이 癌轉移 抑制에 미치는 影響

윤재호,최승훈,안규석 대한한방종양학회 1998 대한한방종양학회지 Vol.4 No.1

To examine the effect of Shiquandabutang on the metastasis of cancer, the following experiments were made. Before the main experiments, the cytotoxieity was measured by putting Shiquandabutang sample in HT1080, Then zymography was made to examine the change of gelatinolytic activity. And western blotting was carried out to examine the changes of Fos, Jun, Ets, the transcription factors of MMP-2. MMP-9, and Erk. JNK on signal transduction pathway to AP-1, Third, in vitro invasion assay with tranwells coated by collagen and matrigel was carried out. From the results of the above the following conclusions were obtained. 1. The experimental result about cytotoxicity of Shiquandabutang against HT1080 was as below. The stained cell count after being treated by Shiquandabutang sample 400ug/ml for 24 hours was 0.9% of total cells. and the stained cell count by Shiquandabutang sample 100ug/ml was 1.5% of total cells. Both were near the level ot control groupwhich showed 0.6% stained. 2. The restlt of collagenase assay was as bclow. In shiquandanbutang sample 400ug/ml, MMP-2 was reduced as compared with TPA control group. and the band of MMP-9 induced by TPA disappeared. In Shiquandabutang sample 800ug/ml. both bands of MMP-2 and MMP-9 disappeared. 3. The reasults of western blots for Jun, Fos, Ets, Erk, JNK were as below. In Shiwaandabutang sample 200ug/ml, Ets was reduced, and Jun, Fos were increased. 4. The result of invasion assay was as below. The number of cells which migrated across transwell membrane in shiquandabutang-lteated group was less than that of +TPA control group. From the above results, it was concluded that Shiquandabutang might control the appearting and acting of collagenase not by the MMP-2, -9 promoter but by other way.

니켈담지촉매의 제조과정에서 액상산화의 효과

최재석,임태훈,홍성안,이호인 한국공업화학회 2004 응용화학 Vol.8 No.1

The deactivation of reforming catalyst in the DIR-MCFC was reported to be affected by dispersion, loading amounts, kind of support and so on. In this research, we checked the resistivities of catalysts prepared by the different methods, and examined the effect of liquid phase oxidation. The oxidized catalyst in a liquid phase showed the increased activity and dispersion even though the nickel species was supported by deposition-precipitation method. In order to investigate the role of the oxidation, TGA, TPR and XRD analyses were conducted. The increase in activity was ascribed to the strong metal-support interaction which was induced by liquidphase oxidation.

지역사회획득 폐렴의 치료지침 권고안

송재훈,정기석,강문원,김도진,배현주,서지영,심태선,안중현,안철민,우준희,이남용,이동건,이미숙,이상무,이영선,이혁민,정두련,지역사회획득 폐렴 치료지침 제정위원회 대한감염학회 2009 감염과 화학요법 Vol.41 No.3

A successful therapy of community-acquired pneumonia requires appropriate empirical antimicrobial therapy. Etiology and antimicrobial susceptibility of major pathogens of pneumonia can differ by country. Therefore, an ideal treatment guideline of community-acquired pneumonia should be based on the studies performed in each country. We developed a treatment guideline for community-acquired pneumonia in immunocompetent adults in Korea. This guideline was developed by the joint committee of the Korean Society for Chemotherapy, the Korean Society of Infectious Diseases, and the Korean Academy of Tuberculosis and Respiratory diseases.

벤지딘계 염료제조 사업장 근로자의 벤지딘 폭로

노재훈,안연순,김규상,김치년,김현수 大韓産業醫學會 1995 대한직업환경의학회지 Vol.7 No.2

To evaluate the differences of benzidine exposure patterns of the workers in two benzidine-based dye manufacturing factories, the concentration of benzidine in air, blood and urine were measured. The air levels of benzidine dihydrochloride and benzidine-based dye were measured by high performance liquid chromatography with ultraviolet detector. Blood samples were collected at 3 hours after exposure and urine samples were collected at the end of shift. Blood and urine samples were analyzed by high performance liquid chromatography with electrochemical detector. The level of benzidine in reaction process (input, diazotization, and coupling) was 0.381±7950g/㎥. The blood benzidine was deteced in 25 workers among 38 in reaction process and their mean levels were 0.0153±.0376 ng/mg Hb. The urinary benzidine was detected for 11 workers among 38 workers in the reaction process. The level of benzidine-based dye in drying and packing process was 52.1748±14.4111g/㎥. The blood benzidine was deteced in 6 workers among 38 in drying and packing process and their mean levels was 0.0062±.0274 ng/mg Hb. The urinary benzidine was detected for 1 worker among 38 workers exposed to benzidine-based dye. The blood and urinary benzidine were detected in workers exposed to benzidine-based dye. Such results suggested that some part of benzidine-based dye was metaboized to benzidine. Therefore, some regulations for manufacturing and use of the benzidine-based dye are needed to prevent its hazards in industries.