• 대한의용생체공학회:의공학회지
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• 제41권2호
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• pp.67-74
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• 2020

Brain tumors or gliomas are fatal cancer species with high recurrence rates due to their strong invasiveness. Therefore, the goal of surgery is complete tumor resection. However, the surgery is difficult to distinguish the border because tumors and blood vessels have the same color tone and shape. The fluorescein sodium is used as a fluorescence contrast agent for boundary separation. When the external light source is irradiated, yellow fluorescence is expressed in the tumor, which helps distinguish between blood vessels and tumor boundaries. But, the fluorescence expression of fluorescence sodium depends on the concentration of fluorescein sodium and such analytical data is insufficient. The unclear fluorescence can obscure the boundaries between blood vessels and tumors. In addition, reduce the efficiency of fluorescence sodium use. This paper proposes a protocol of concentration range for fluorescence expression conditions. Fluorescent expression was observed using a near-infrared (NIR) color camera with corresponding dilution using normal saline in 1 ml microtube. The flunoresence emission density range is 1.00 mM to 0.15 mM. The fluorescence emission begin to 1.00 mM and the 0.15 mM discolor. The discolor is difficult to fluorescence emission condition obserbation. Thus, the maximum density range of the bright fluoresecein is 0.15 mM to 0.30 mM. When the concentration range of fluorescein sodium is analyzed based on the gradient of fluorescence expression and the power measurement, the brightest fluorescence is expected to facilitate the complete resection of the tumor. For the concentration range protocol, setting concentration ranges and analyzing fluorescence expression image according to saturation and brightness to find optimal fluorescence concentration are important. Concentration range protocols for fluorescence expression conditions can be used to find optimal concentrations of substances whose expression pattern varies with concentration ranges. This study is expected to be helpful in the boundary classification and resection of brain tumors and glioma.

• 한국안광학회지
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• 제5권1호
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• pp.83-87
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• 2000

본 연구는 각막상피가 저절로 탈락해 나가는 과정을 이해하기 위해 사람의 각막상피 세포를 배양하여 배지의 영양이 고갈될 때까지 약 7일 정도 배지를 교환하지 않고 계속 배양하여 세포고사를 조사하였다. 영양이 고갈된 배지인 Exhausted Medium을 여과하여 새로운 각막 상피세포에 넣어 2일 정도 배양을 계속하였다. Exhausted Medium에서 배양된 세포를 수확하여 세포고사를 검정하기 위해 Agarose gel electrophoresis로 DNA 단편을 확인하고 세포고사를 초기에 감지할 수 있는 방법인 M30 $CytoDEATH^*$. Fluorescein으로 확인하였다. 또한 Exhausted Medium에 의한 세포고사의 유발 경로를 조사하기 위해 사이토카인에 의한 유도인자 중 가장 많이 알려진 FAS나 FAS Ligand에 의한 유발여부를 soluble FAS and FAS Ligand에 대한 sandwich ELISA로 조사하였다. 그 결과 전형적인 DNA Ladder패턴을 보였으며 M30 $CytoDEATH^*$. Fluorescein에도 선명하게 염색되어 세포고사를 확인할 수 있었다. 또한 soluble FAS and FAS Ligand ELISA Kit로 Exhausted Medium이 FAS와 관련이 있는지를 조사한 결과 거의 무관한 것으로 나타났다. 본 연구의 결과는 영양 부족상태의 배지는 각막상피 세포가 세포고사를 거쳐 소모되는데 FAS and FAS Ligand system과는 무관한 것으로 나타났다.

• The Plant Pathology Journal
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• 제32권4호
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• pp.340-346
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• 2016

To understand how Bursaphelenchus xylophilus kills pine trees, the differences between the effects of B. xylophilus and B. mucronatus on pine trees are usually compared. In this study, the migration and attacking ability of a non-pathogenic B. mucronatus in Pinus thunbergii were investigated. The distribution of B. mucronatus and the number of dead epithelial cells resulting from inoculation were compared with those of the pathogenic B. xylophilus. Although B. mucronatus is non-pathogenic in pines, its distribution pattern in P. thunbergii was the same as that of B. xylophilus. We therefore concluded that the non-pathogenicity of B. mucronatus could not be attributed to its migration ability. The sparse and sporadic attacking pattern of B. mucronatus was also the same as that of B. xylophilus. However, the number and area of the dead epithelial cells in pine cuttings inoculated with B. mucronatus were smaller than in those cuttings inoculated with B. xylophilus, meaning that the attacking ability of B. mucronatus is weaker than that of B. xylophilus. Therefore, we concluded that the weaker attacking ability of B. mucronatus might be the factor responsible for the non-pathogenicity.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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• pp.695-698
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• 2000

We studied the viability and function of islet with monomethoxy polyethylene glycol (mPEG) grafted onto its membrane. Islets were isolated from rat and were repeatedly reacted with activated mPEG (mw 5000) in order to increase grafting density. The density of grafted PEG on the islet membrane was confirmed by Fluorescein-PEG-NHS. An assessment of islet viability using AO / PI staining method showed that multiple PEGylation did not reduce islet viability. The function of PEG grafted islets was evaluated by measuring released insulin from islets. Insulin secreted from the PEGylated islets for 1 h did not show any significant difference compared to control (non-PEGylated) islets. In addition, PEGylated islets responded in the same pattern as control islets in the perifusion test.

• 한국안광학회지
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• 제17권4호
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• pp.389-394
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• 2012

목적: 본 연구는 역방향(inside out or everted)으로 착용한 실리콘 하이드로겔렌즈가 각막 형상변화에 미치는 영향을 알아보기 위해 실시하였다. 방법: 역방향 실리콘 하이드로겔 렌즈의 형상과 플루오레신 패턴을 알아보고 이를 수면 중 8시간 이상 착용하여 타각적 굴절이상도와 각막지형도의 변화를 관찰하였다. 검사는 렌즈 착용 전, 렌즈 착용 후 1일, 2일, 3일, 5일, 7일 후에 시행하였으며 렌즈의 착용을 중지한 후 1일, 2일, 3일, 4일 후에 동일하게 시행하였다. 결과: 역방향 실리콘 하이드로겔 렌즈 착용 후 피팅패턴을 검사한 결과 역기하렌즈와 유사한 눌림이 있었다. 타각적 굴절이상도의 근시도와 각막 굴절력이 감소하였으며 각막지형도의 변화를 보였고 이는 렌즈 착용 중지 후 4일 동안 대부분 회복되었다. 렌즈를 뒤집어 착용하는 것에 대한 불편함은 크게 없는 것으로 나타났다. 결론: 역방향으로 착용한 실리콘 하이드로겔 렌즈의 수면 중 착용(Overnight wear)을 통하여 각막지형도상의 유의한 차이를 야기하였다. 추후 각막의 형상변화(reshaping)를 위해 실리콘 하이드로겔 렌즈의 사용이 가능할 것으로 보고, 교정효과에 대해 예측이 가능(predictable)하고 효과적인(effective) 방법을 개발하기 위해 여러 변수를 확인해야 할 것으로 생각된다.

• 한국안광학회지
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• 제17권1호
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• pp.37-45
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• 2012

목적: 구면 RGP 렌즈 및 비구면 RGP 렌즈를 양호하게 피팅할 때 중심부와 주변부의 플루레신 염색 면적이 난시도 및 각막형상에 따라 어떻게 달라지는 지를 비교하여 렌즈와 각막과의 정렬상태가 렌즈의 디자인에 따라 어떠한 차이가 있는지를 알아보고자 하였다. 방법: 직난시를 가진 19~30세($25.12{\pm}3.52$세)의 90안을 대상으로 구면 및 비구면 RGP 렌즈를 착용시키고 플루레신 염색 패턴을 분석하였으며, 이의 정량적인 평가를 위하여 중심부와 주변부의 면적을 계산하여 비교하였다. 결과: 양호한 피팅이 되는 렌즈의 베이스커브가 구면 RGP 렌즈와 비구면 RGP 렌즈가 동일한 경우는 72%에 달하였으나, 대칭나비형 각막과 높은 난시도를 가진 경우에는 동일한 베이스커브에서 구면 RGP 렌즈와 비구면 RGP 렌즈가 양호한 피팅이 될 가능성이 적어졌다. 양호하게 피팅된 비구면 RGP 렌즈의 주변부 플루레신 염색 면적은 구면 RGP 렌즈보다 더 작았으며, 두 디자인의 렌즈 모두 난시가 심해질수록 주변부 면적은 감소하였고 원형 각막보다도 대칭 나비형 각막에서의 주변부 면적이 더 작았다. 동일한 난시도의 각막에서는 두 각막 형상 모두에서 양호하게 피팅된 비구면 RGP 렌즈의 주변부 플루레신 염색 면적이 더 작았다. 결론: 본 연구를 통하여 RGP 렌즈의 각막과의 정렬상태의 달라지는 정도가 렌즈 디자인, 각막의 난시도, 각막 형상에 의해 차이가 있음을 알 수 있었다. 따라서 렌즈 디자인과 각막의 정렬상태의 정량적인 고찰을 통한 본 연구결과는 RGP 렌즈의 피팅 가이드라인 설정, 적합한 디자인 개발, 부분별 누액양에 대한 기초자료로 사용될 수 있을 것이다.

• Nuclear Engineering and Technology
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• 제30권3호
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• pp.245-261
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• 1998

In order to validate the variable aperture channel model that can deal with the non-uniform How rate in flow domain, migration experiments of conservative tracer were performed in two artificial fractures, a parallel and a wedge-shaped fracture. These different fracture shapes were designed to give different flow pattern. The fractures were made from a transparent acrylic plastic plate and a granite slab with dimensions of 10 $\times$ 61 $\times$ 61 cm. Uranine (Fluorescein sodium salt) was used as a conservative tracer. The volumetric flow rates of uranine feed solution were 30 mL/ hr, giving a mean residence time in the fracture of approximately 24 hours for the parallel fracture and 34 hours for the wedge-shaped fracture. The migration plumes of uranine were photographed to obtain profiles in space and time for movement of a tracer in fractures. The photographed migration plume was greatly affected by the geometric shape of fractures. The variable aperture channel model could have predicted the experimental results for the parallel fracture with a large accuracy. It is expected that the variable aperture channel model would be effective to predict the transport of the contaminant, especially, with the flow rate variation in a fracture.

• 한국안광학회지
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• 제17권3호
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• pp.255-264
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• 2012

목적: 본 연구에서는 형상 및 난시도가 상이한 각막에 구면 및 비구면 RGP 렌즈를 피팅 상태를 달리하여 착용시켰을 때 각막과 렌즈와의 간극에 미치는 영향을 알아보아 적절한 RGP 렌즈의 선택에 도움이 되고자 하였다. 방법: 직난시를 가진 $(25.12{\pm}3.52)$세의 90안에 구면과 비구면 RGP 렌즈를 각기 다른 피팅 상태로 착용시키고 플루레신 염색의 정량적인 분석을 위하여 중심부, 중간주변부 및 주변부로 나눠 면적을 계산하여 각막과 콘택트렌즈의 간극을 비교하였다. 결과: 양호한 피팅 시 중심부와 주변부 면적은 렌즈 디자인에 따른 유의한 차이가 있었으나 스팁 혹은 플랫하게 피팅되었을 때의 중심부 면적은 각막형상과 렌즈 디자인에 따른 차이는 없었다. 각막 형상에 따라 분석하였을 때 양호한 피팅과 플랫 피팅에서는 두 렌즈 디자인 모두에서 중심부와 주변부 면적이 각막 형상에 따라 통계 적으로 유의하게 달라졌으나, 스팁 피팅에서는 차이가 없었다. 각막 난시도에 따라 분석하였을 때 각막형상에 상관없이 각막난시가 증가함에 따라 일정하게 중심부 및 주변부의 면적이 변하는 양호한 피팅의 경우와는 달리 스팁 피팅과 플랫한 피팅의 경우는 원형 각막과 대칭나비형 각막에서의 면적 변화 양상이 상이하였다. 결론: 본 연구에서 는 RGP 렌즈를 스팁이나 플랫하게 피팅 되었을 경우 양호한 피팅시 보다 각막난시나 각막 형상에 따른 렌즈와 각막과의 간극 변화가 더 커지며 이러한 결과는 구면 RGP 렌즈와 비구면 RGP 렌즈 모두에서 공통적으로 나타나는 것임을 알 수 있었다.

• Journal of Pharmaceutical Investigation
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• 제31권4호
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• pp.241-256
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• 2001

Alginate microspheres, containing fluorescein isothiocyanate-bovine serum albumin (FITC-BSA) or green fluorescent protein (GFP) were prepared and used as a model drug to develop the oral vaccine delivery system. The alginate microspheres were coated with poly-L-lysine or chitosan. Two methods, w/o-emulsion and spray, were used to prepare alginate microspheres. To optimize preparation conditions, effects of several factors on the particle size and particle morphology of microsphere, and loading efficiency of model antigen were investigated. In both preparation methods, the particle size and the loading efficiency were enhanced when the concentration of sodium alginate increased. In the w/o-emulsion preparation method, as the concentration of Span 80 was increased from 0.5% to 2%, the particle size was decreased, but the loading efficiency was increased. The higher the emulsification speed was, the smaller the particle size and loading efficiency were. The concentration of calcium chloride did not show any effect on the particle size and loading efficiency. In the spray preparation method, the particle size was increased as the nozzle pressure $(from\;1\;kgf/m^2\;to\;3\;kgf/m^2)$ and spray rate was raised. Increasing calcium chloride concentration (<7%) decreased the particle size, in contrast to no effect of calcium chloride concentration on the w/o-emulsion preparation method. Alginate microspheres prepared by two methods were different in the particle size and loading efficiency, the particle size of microspheres prepared by the spray method was about $2-6\;{\mu}m$, larger than that prepared by the w/o emulsion method $(about\;2{\mu}m)$, and the loading efficiency was also higher with spray method. Furthermore, drying process for the microspheres prepared by the spray was simpler and easier, compared with the w/o emulsion preparation. Therefore, the spray method was chosen to prepare alginate microspheres for further experiments. Release pattern of FITC-BSA in alginate microspheres was evaluated in simulated intestinal fluid and PBS (phosphate buffered saline). Dissolution rate of FITC-BSA from alginate/chitosan microsphere was lower than that from alginate microsphere and alginate/poly-L-lysine microsphere. By confocal laser scanning microscope, it was revealed that alginate/FITC-poly-L-lysine microspheres were present in close apposition epithelium of the Peyer's patches of rabbits following inoculation into lumen of intestine, which proved that microspheres could be taken up by Peyer's patch. In conclusion, it is suggested that alginate microsphere prepared by spray method, showing a particle size of & $10\;{\mu}m$ and a high loading efficiency, can be used as a model drug for the development of oral vaccine delivery system.

• Clinical and Experimental Reproductive Medicine
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• 제26권3호
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• pp.419-432
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• 1999

Objective: To establish the evaluation system of the quality of oocytes on the basis of the incidence of cumulus cells apoptosis, to investigate the relationships beween the incidence of cumulus cells and the outcomes of IVF-ET. Method: Thirth-four cycles undergoing controlled ovarian hyperstimulation for IVF-ET with tubal infertility (23 cycles) or unexplained infertility (11 cycles) were included in this study. Cumulus cell masses surrounding mature oocyte and co-culture of embryos with autologous cumulus cells during IVF-ET process. The incidence of apoptosis in cumulus cells was assessed by apoptosis detection kit fluorescein. The effect of co-culture using cumulus cells and the incidence of cumulus cells apoptosis. Results: The results were as follows: 1. The incidence of apoptosis in cumulus cells markedly increased in patients aged 40 or over, while the fertilization rate was greatly decreased in those age group. 2. Apoptosis in cumulus cells was found in both the fertilized oocytes and unfertilized oocytes, but the incidence of apoptosis was higher in unfertilized oocytes. 3. There is no clear correlation between apoptosis in cumulus cells and the number of oocytes retrieved. However, the incidence of apoptosis was increased when the number of oocytes retrieved was 5 and fewer in comparison with $6{\sim}10$. 4. Embryo grade was significantly affected by the incidence of apoptosis in cumulus cells. 5. Pregnancy rate of IVF-ET per cycle was 29.4%, and the pregnant group had the higher fertilization rate and a significantly lower incidence of apoptosis in cumulus cells compared with the nonpregnant group. 6. When cumulus cells were used as helper cells in the co-culture of the embryo, in vitro activity of cumulus cells based on morphological change and proliferation did not influence the quality of embryo, but was closely associated with the implantation rate and pregnancy rate, which was enhanced when morphological changes and proliferation of cumulus cells was more active. 7. This difference in the outcome of IVF-ET according to in vitro activity of cumulus cells used for co-cultue was not associated with the incidence of apoptosis in cumulus cells; but rather had likely relations with the different secretion pattern of protein, which may be an embryo trophic factor by cumulus cells. Conclusion: These results suggest that the incidence of apoptosis in cumulus cells can be used in predicting oocyte qualities and the outcomes of IVF-ET. And the effect of co-culture largely depends on the in vitro activity of cumulus cells as well.