• 한국의학물리학회지:의학물리
• /
• 제23권1호
• /
• pp.54-61
• /
• 2012

중합체 겔 선량계를 제작하여 사이버나이프의 광자선의 소조사면에서의 3차원적 선량측정 실험을 수행하였다. 겔 선량계는 아크릴로 제작된 두경부 팬텀내에 설치하여 선량측정에 사용하였다. 두경부 팬텀의 영상은 사이버나이프의 두경부 환자 치료를 위해 사용하는 전산화단층촬영장치(CT)의 프로토콜을 사용하여 획득하였고 치료계획 시스템에 전달되었다. 겔 선량계에 치료계획에서 수립한 방사선을 조사하였으며, 24시간 경과 후에 선량분석을 위하여 3.0T 자기공명영상장치(MRI)로 영상을 획득하였다. 측정된 겔 팬텀의 선량분포는 MATLAB을 이용하여 분석하였고 측정된 결과를 치료계획에서의 계산값과 비교하였다. 선량분포는 축상(axial) 방향의 등선량곡선의 80% 고선량 영역에서 치료계획 선량곡선과 측정된 선량곡선과의 차이가 0.76 mm이었으며, 40% 저선량 영역에서는 차이가 1.29 mm로 저선량 영역보다 고 선량영역에서 잘 일치함을 알 수 있었다. 본 연구에서 중합체 겔 선량계와 자기공명영상을 이용하여 소조사면에 대해 3차원적으로 선량을 분석할 수 있을 뿐 아니라 치료방사선 정도관리에도 활용될 수 있는 가능성을 확인하였다.

• 공업화학
• /
• 제19권2호
• /
• pp.222-227
• /
• 2008

본 연구에서는 광학센서의 응용을 위해 소수성 졸-겔로 개질된 고분자 막의 미생물 비점착 특성과 광학특성을 조사하였다. DiMe-DMOS (Dimethoxy-dimethylsilane)와 TMOS (Tetramethyl-orthosilicate)를 이용하여 제조한 소수성 졸-겔로 코팅된 컨퍼컬디쉬 표면과 유리표면에 E. coli JM109, B. cereus 318 그리고 P. pastoris X-33을 배양하였다. 배양 후, 부유세포를 증류수를 이용하여 제거하고 그람 염색법에 의해서 점착된 미생물을 염색하였다. 점착된 미생물의 수는 SEM을 이용하여 정량적으로 분석하였다. 유리표면에는 $2{\sim}3{\times}10^4$개/$mm^2$의 미생물이 점착되었으나 소수성 졸-겔 표면에는 200~300개/$mm^2$의 미생물이 점착됨으로써 소수성 졸-겔의 비점착 효과를 알 수 있었다. 또한, 소수성 졸-겔과 절광물질인 흑연을 혼합하여 제조한 절광층(Light insulating layer)을 pH나 용존산소 검출막 위에 재코팅한 후, pH나 용존산소의 검출막의 성능이 향상되었음을 알 수 있었다.

• Journal of Photoscience
• /
• 제9권2호
• /
• pp.479-481
• /
• 2002

UV-induced DNA damage causes cell killing and mutations leading to carcinogenesis. In normal human cells, UV damage such as cyclobutane pyrimidine dimers (CPDs) and primidine-prymidone (6-4) photoproducts are mainly repaired by nucleotide excision repair mechanism. The molecular processes have been well characterized recently. To know the influence of mitochondrial genome on the nucleotide excision repair mechanism against CPDs, we comparatively examined the production of CPDs by UVC irradiation and their repair kinetics in human cells completely lacking mitochondrial DNA (mtDNA) and the parental HeLa S cells. Whole DNA extracted from the cells exposed to UVC was treated with T4-endonuclease V to break the phosphodiester bond adjacent to CPDs. The DNA was electrophoresed in a denaturing agarose gel, which was visualized by ethidium bromide staining. The relative amount of CPDs was determined by image analysis using NIH Image software. MtDNA- less (rho-O) cells were apparently more sensitive to UVC than HeLa S cells, while the level of induction of CPDs in rho-O and HeLa cells was comparable. The repair of CPDs was less efficient in rho-O cells compared with HeLa cells. The residual amount of CPDs after 24-h repair was larger in rho-O cells than in HeLa cells where more than 90 % of CPDs were repaired by then. The non-repaired CPDs would lead to apoptosis in rho-O cells. These results suggest that mitochondrial genome may contribute to some ATP-dependent steps in nucletide excision repair by supplying sufficient ATP which is generated through a respiratory chain in mitochondria.

• 한국환경독성학회:학술대회논문집
• /
• 한국환경독성학회 1997년도 제20회 화학물질의 환경독성과 건강영향
• /
• pp.101-101
• /
• 1997

Comet assay, single cell gel electrophoresis has been known as useful, rapid, simple, visual, and sensitive technique for measuring the DNA breakage in mammalian ce1ls. For evaluation of DNA damage using comet assay, early studies reported a change in comet length and intensity with DNA damage using simple visual technique, such as fluorescence microscopy with eyespiece. In recent, some workers are observing and analyzing nucleotide of comets using quantitative fluorescence image analysis system to estimate 'tail moment', which is defined as the product of the tail length and the fraction of total DNA in tail. Our laboratory also adopted the image analysis software for qualification. In addition, many of the practical features of comet assay render it potentially attractive as useful tool for molecular toxicology and carcinogenesis, because the system is already showing considerable promise as rapid predictor in both in vitro and in vivo experimental designs. Recently, the comet assay becomes a attractive technique to study of apoptosis, because apoptotic fragmentation of nuclear DNA into nucleosomal sizes can be evaluated by the comet assay. So, we attempted to apply the comet assay to studying the effect of various stress on the apoptosis-sensitive cell lines. Particularly, focusing on the hyperthermic apoptosis, we could find that heat shock(44˚C for 60 minutes) was sufficient to induced apoptosis in these cell lines. But using the highly sensitive comet assay, we could not detect DNA breaks immediately after heat shock.

• Asian Pacific Journal of Cancer Prevention
• /
• 제13권4호
• /
• pp.1667-1674
• /
• 2012

Objective: To explore the differential protein expression profile in EC304 gastric cancer cells induced by alphastatin. Methods: Cultured EC304 cells in the exponential phase of growth were randomly divided into alphastatin and control groups. Total proteins were extracted and the two dimensional electrophoresis (2-DE) technique was applied to analyze differences in expression with ImageMaster 2D Platinum 5.0 software. Proteins were identified using the MASCOT database and selected differently expressed proteins were characterised by western blotting and immunofluorescence. Results: $1350{\pm}90$ protein spots were detected by the ImageMaster software in the 2-DE gel images from the control and alphastatin groups. The match rate was about 72-80% for the spectrum profiles, with 29 significantly different protein spots being identified, 10 upregulated, 16 downregulated, two new and one lost. The MASCOT search scores were 64-666 and the peptide matching numbers were 3-27 with sequence coverage of 8-62%. Twenty-three proteins were checked by mass spectrometry, including decrease in Nm23 and profilin-2 isoform b associated with the regulation of actin multimerisation induced by extracellular signals. Conclusion: The proteome in EC304 cells is dramatically altered by alphastatin, which appears to play an important role in modulating cellular activity and anti-angiogenesis by regulating protein expression and signal transduction pathways through Nm23 and profilin-2 isoform b, providing new research directions for anti-angiogenic therapy of gastric cancer.

• 한국환경독성학회:학술대회논문집
• /
• 한국환경독성학회 1996년도 제19회정기학술대회(The 19th Symposium of the Korean Society of Environmental Toxicology)
• /
• pp.61-61
• /
• 1996

Taxol is used as cancer therapeutic agent. It has been known as weak posotive of chromosome aberration assay in vitro in our previous results (Ryu et al., 1996) and potent clastogens in the mouse bone marrow micronucleus (Tinwell and Ashby, 1994). We performed microgel electrophoresis to determine the effect of taxol and it's precursor 10-deacetyl baccatin III(DAB) on DNA. Microgel electrophoresis is useful, rapid, simple, visual, and sensitive technique for measuring DNA breakage and repair mechanisms in mammalian 근ells. The range of concentration used for taxol were 854, 427, 213.5, 106.8, 53.4 Ug/ml, for DAB 910 ,455, 227.5 U9/ml, Cell viability always exceed 85%. We analyzed the results by using the special software of image analyzer for this comet assay (Komet 3.0). By using this image analyzer software , we can get the result as the tail moment ((mean of tail length - mean of head lengh) x tail%DNA/100). A slight increase in DNA migration was observed for taxol at the concentration of 854 Ug/m4 in the absence of S9 mixture. No increased DNA migration was observed after treatment with DAB.

• 한국정보과학회논문지:소프트웨어및응용
• /
• 제30권5_6호
• /
• pp.444-453
• /
• 2003

최근 생명공학(BT)에 대한 관심이 집중되면서, 새로운 생리활성 물질을 찾거나 유전자 정보를 분석하기 위한 목적으로 전기영동 젤의 영상 분석 기술에 대한 요구가 급증하고 있다. 이를 위해서는 젤 영상의 레인에서 각 밴드의 위치와 양을 정확히 측정해야 한다. 기존 연구에서는 주로 레인의 프로파일에서 피크를 탐색하는 접근방법을 사용하는데, 이 피크의 위치는 밴드에 있는 최대 자기 화소의 위치도 아니고 더욱이 밴드 무게중심의 위치도 아니기 때문에 밴드의 대표 위치로 인정하기 어렵다. 또한, 피크 추출을 쉽게 하기 위해 다양한 영상 향상 처리를 적용하기 때문에 밴드의 양을 측정하기에는 부적절한 경우가 많다. 본 논문에서는 영상의 상대적인 밝기를 변화시키지 않으면서 먼저 밴드의 영역을 추출한 후, 밴드 영역의 밝기 합으로 양을 구하고 이의 무게중심을 밴드 위치로 정하는 방식을 채택한다. 실제로, 먼저 젤 영상 히스토그램에 엔트로피기반 임계치를 설정하여 레인을 추출한 후, 밴드 영역 추출을 위해 서로 다른 세 가지 방법을 시도한다. 첫째, 추출된 레인을 이등분하는 중심선을 탐색하여 피크와 밸리를 찾고, 피크의 상하 밸리를 각 밴드의 최소 포함 박스영역으로 지정하는 방법(MER), 둘째, 앞의 방법에서와 같이 구한 피크를 영역 성장의 시드로 사용하여 이웃하는 밴드와의 중첩을 해결하면서 밴드 영역을 추출하는 방법(RG-1), 셋째, 이와 달리 레인을 삼등분하는 두 탐색선에서 피크를 찾고 동일한 밴드에 속하는 피크 쌍을 결정한 후 영역을 성장하는 방법(RG-2)을 제안한다. 이상의 세 방법을 비교하기 위해 밴드의 위치 및 양을 측정한 결과, 밴드 위치의 평균 오차는 레인의 길이를 단위 크기로 정규화 할 때, MER 방법이 6%, RG-1 방법이 3%, RG-2 방법이 1%로 나타났다. 또한, 밴드 양의 평균 오차는 레인 내 밴드들의 양의 합을 단위 크기로 정규화 할 때, MER 방법이 8%, RG-1 방법이 5%, RG-2 방법이 2%로 나타났다. 결과적으로, RG-2 방법이 밴드의 위치 및 양 추출에 있어서 정확도가 가장 높은 것으로 판명되었다.

• Journal of Plant Biotechnology
• /
• 제39권1호
• /
• pp.81-92
• /
• 2012

단백질체학에서 이차원 전기영동 (2-DGE)은 고해상도 단백질 분리가 가능한 중요 기술 중 하나이다. 본 논문에서는 2-DGE 이미지 분석 일치도를 향상시킴으로써 작물 단백질 검출과 정량분석이 가능하도록 하는 전반적인 주요 장비, 시약 등을 자세히 기술한 프로토콜을 제시하고 있다. 이 프로토콜은 식물의 발달, 생물학적, 비생물학적 스트레스 반응 등과 관련된 작물 단백질 바이오마커를 개발하기 위한 목적에서 2-DGE를 처음 시도하는 연구자들에게 유용할 것으로 기대한다.

• 한국한의학연구원논문집
• /
• 제13권1호통권19호
• /
• pp.153-160
• /
• 2007

In the post-genome era, analysis of the cellular transcriptome using microarray or the cellular proteome using a 2-D gel electrophoresis and MALDI-TOF mass spectrometry are most widely used. Stroke is one of the most important causes of death along with cancer and cardiac disease. When pathological change of cells in developed from cerebral ischemia accompanied by stroke administration of neuroprotective drugs before stroke can decreases the degeneration of neuronal cells. The purpose of the present study was to assess the neuroprotective effect and protein expression after administration of P004, middle cerebral artery model of cerebral ischemia in rats. SD rats were subjected to middle cerebral artery occlusion. P004 (1,000 mg/kg) was administered 2 times at 0, 90 minutes after middle cerebral artery occlusion (MCAo). Rats were killed at 48 hours, and infarct area and volume were determined by histology and computerized image analysis. We investigated the protein expression profile on the global ischemia induced by MCAo. This proteomic analysis enable us to identify several proteins differently expressed in infarct brain tissue. The aims of this study were to do investigation comparing the neuroprotection activities of P004 and to understand the mechanism of acted as neuroprotective drug.

• 대한의생명과학회지
• /
• 제13권4호
• /
• pp.341-347
• /
• 2007

Streptococcus mutans, one of a major causal agents of dental caries, is component of the dental plaque and produces various organic acids such as lactic acid as the end-product of glycolysis. In this study, we isolated S. mutans from Korean children with caries and also investigated the expression of protein under acid stress. S. mutans was identified at the species level using a 16S ribosomal DNA sequencing comparison method. The primer specificity was tested on eleven S. mutans strains isolated from Korean children with caries. The data showed that eleven strains are S. mutans. At treatment of concentration of 20 mM lactic acid in the mid-log phage, K-7 exhibited the highest maximum culture OD compared with those of other groups. As a consequence, we examined the expression of protein under 20 mM lactic acid stress using S. mutans K-7. The results of 2D gel electrophoresis by image analysis showed that thirteen proteins are up-regulated under the stress. Further study is being focused on amino acid analysis by mass spectrometry in order to analyze those spots.