• BMB Reports
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• v.28 no.4
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• pp.359-362
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• 1995

The highly polymorphic variable number of tandem repeat (VNTR) loci in the human genome are informative markers for the genetic characterization of individuals in the paternity test and forensic science as well as for the study of human disease. In this study, VNTR loci D1S80 and D2S123 have been amplified by PCR and the amplified length polymorphic alleles were detected with a discontinuous vertical PAGE system and silver staining. For explicit DNA typing, PCR optimization, in which amplification efficiencies are similar over a wide range of allele sizes, non-specific amplifications are minimal, and new longer alleles have high amplification efficiency, has been performed by changing the PCR reaction buffer composition and thermal cycling conditions. It turned out that adding an appropriate amount of Tween 20 and NP40 to the PCR reaction buffer and raising the annealing temperature to $68^{\circ}C$ in thermal cycling made it possible for optimal VNTR loci amplification. A modified PAGE system for VNTR separation was established. Under these conditions, new longer alleles in the 01580 locus were discovered and 025123 pattern changes in colorectal tumors were observed. These technical tips are valuable for detecting various amplified fragment length polymorphisms.

• BMB Reports
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• v.40 no.6
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• pp.944-951
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• 2007

3-Hydrogenkwadaphnin (3-HK) is a daphnane-type diterpene ester isolated from Dendrostellera lessertii (Thymelaeaceae) with high differentiation and apoptotic potency in leukemic cells without any measurable adverse effects on normal cells (Moosavi et al., 2005b). In this study, we report that 3-HK (12 nM) has the ability to cease proliferation, induce differentiation and apoptosis in chronic myelogenous leukemia (CML) K562 cell line. The treated cells lost erythroid properties and differentiated along the megakaryocytic lineage based on the morphological features apparent after Wright-Giemsa staining, DNA content analysis and the expression of cell surface marker glycoprotein IIb as analyzed by flow cytometry. Moreover, using Hoechst 33258 and Annexin V double staining indicated the occurrence of apoptosis among the treated cells. On the other hand, restoration of the depleted GTP pool size by exogenous addition of guanosine ($50{\mu}M$) reduced the effect of the drug regarding the extent of differentiation while no further enhancement of 3-HK effect was obtained by addition of exogenous hypoxanthine ($100{\mu}M$). These interesting results necessitate further investigation regarding the mechanism of action of this unique anti-leukemic agent.

• Journal of Life Science
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• v.18 no.9
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• pp.1312-1315
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• 2008

High throughput-drug screening plays a pivotal role for early stage of drug discovery process. In the course of assay development for screening of cell death-inducing agents, a protocol that is simple, time-saving, and high throughput-compatible was designed which was confirmed that the protocol can be worked by a HTS-compatible machine. In 96-well format, PC-3 cancer cells (1${\times}10^{4}$ cells/ml) were cultured for 24 hr. After 24 h-incubation with various medicinal plants extracts, the cells were then stained with DAPI for 30 min. The fluorescence intensity of the stained cells was measured semi-automatically with a multilabel counter. To test whether the present assay system effectively works, we screened about 850 medicinal plant extracts, and selected 1 positive crude extracts that contained cell death-inducing activity. These results suggest that the protocol is highly amenable to HTS implementation for a cell death-inducing agent(s).

• Archives of Plastic Surgery
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• v.41 no.1
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• pp.35-39
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• 2014

Background Aurora kinase A (Aurora-A) plays an important role in the regulation of mitosis and cytokinesis. Dysregulated Aurora-A leads to mitotic faults and results in pathological conditions. No studies on Aurora-A expression in human diabetic skin tissue have been reported. In light of this, we explored the expression of Aurora-A in human diabetic skin tissue. Methods Aurora-A protein was evaluated by western blotting in 6 human diabetic skin tissue and 6 normal skin specimens. Results Increased expression of Aurora-A protein was detected in all diabetic skin tissue samples in both western blot analysis and immunohistochemical staining. However, in the case of the normal skin tissue, no bands of Aurora-A protein were detected in either the western blotting analysis or the immunohistochemical staining. Conclusions Thus far, there have been no studies on the expression of Aurora-A in diabetic skin tissue. However, we believe that oxidative DNA damage related to the expression of Aurora-A protein and Aurora-A could be involved inhuman diabetic skin tissue.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.7
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• pp.3073-3076
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• 2012

Terpinen-4-ol is a terpene found in the rhizome of Plai (Zingiber montanum (Koenig) Link ex Dietr.). In this study apoptogenic activity and mechanisms of cell death induced by terpinen-4-ol were investigated in the human leukemic MOLT-4 cell line. Terpinen-4-ol exhibited cytotoxicity in MOLT-4 cells, with characteristic morphological features of apoptosis by Wright's staining. The mode of cell death was confirmed to be apoptosis by flow cytometric analysis after staining with annexin V-FITC and propidium iodide. A sub-G1 peak in DNA histograms of cell cycle assays was observed. Terpinen-4-ol induced-MOLT-4 cell apoptosis mediated through an intrinsic pathway involving the loss of mitochondrial transmembrane potential (MTP) and release of cytochrome c into the cytosol. In addition, terpinen-4-ol also induced apoptosis via an extrinsic pathway by caspase-8 activation resulting in the cleavage of cytosolic Bid. Truncated-Bid (tBid) translocated to mitochondria and activated the mitochondrial pathway in conjunction with down-regulation of Bcl-2 protein expression. Caspase-3 activity also increased. In conclusion, terpinen-4-ol can induce human leukemic MOLT-4 cell apoptosis via both intrinsic and extrinsic pathways.

• Korean Journal of Microbiology
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• v.30 no.4
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• pp.316-321
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• 1992

Catechol 2, 3-dioxygenase (C230) catalyses the oxidative ring cleavage of catechol to 2-hydroxymuconic semialdehyde. This is one of the key reactions in the metabolism of the widespresd pollutant aniline. We have cloned a gene encoding C230 from cells of the aniline degrading bacteria, Pseudomonas acidovorance KCTC2494 strain and expressed in E. coli, A 11.3-kilobase Sau3A partial digested DNA fragment from KCTC2494 was cloned into phagemid vector pBluescript and designated as pLP201. The C230 gene was mapped to a 2.8-kb region, and the derection of transcription was determined. The cloned C230 gene contains its own promoter which can be recognized and employed by E. coli transcriptional apparatus. C230 activities of subclones were identified by enzyme assay and activity staining. The T7 RNA promoter/polymerase system and maxicell analysis showed that a polypeptide with Mw of 35 kDa is the C230 gene product.

• Korean Journal of Veterinary Research
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• v.39 no.1
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• pp.213-219
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• 1999

Microsatellite allele analysis has been used for individual identification and paternity test. In the present study, the biological father of three puppies was determined by using microsatellite allele amplification analysis. The mother bitch of the litter was a Poongsan dog. The three stud dogs that could have inseminated the bitch, by being in the same residence, were a white Poosan dog, a mixed breed, and a white Jindo dog. DNA was obtained from all the relevant dogs by buccal swabbing. Four loci of tetranucleotide repeat microsatellite were PCR-amplified, and analyzed by polyacrylamide gel electrophoresis and silver staining. The results of genotyping unambigously assigned the Poongsan dog as the biological father. There was no evidence of superfecundation. Therefore, the present study demonstrated the usefulness of microsatellite allele analysis as a simple, efficient method of paternity test in dogs.

• The Korean Journal of Food And Nutrition
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• v.35 no.6
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• pp.464-472
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• 2022

The present study was designed to investigate the antiproliferative activity and molecular mechanisms of Bibimbap in HT-29 human colorectal adenocarcinoma cells. Bibimbap extract inhibited the proliferation of HT-29 cells by 50% at a concentration of 10.1±0.17 mg/mL for 48 h. The population of live cells decreased slightly, and the morphology changed with a reduction in cell volume (pyknosis) with Bibimbap. Treatment with 5 mg/mL of Bibimbap resulted in slight cell shrinkage. Furthermore, as the Bibimbap dose increased to 10 mg/mL, these characteristics were more evident, and HT-29 cells exhibited partial detachment by staining with the DNA-binding dye Hoechst 33342. Flow cytometric analysis by Annexin V and PI double staining showed that Bibimbap increased the levels of apoptosis. Analysis of the mechanism of these events showed that Bibimbap-treated cells exhibited a mitochondria-dependent apoptotic pathway through the modulation of caspase-3, caspase-8, caspase-9, and poly-ADP ribose polymerase, as well as Bax and Bcl-2 expression in dose- and time-dependent manners. Consequently, Bibimbap exerts a significant antiproliferative effect on HT-29 human colorectal adenocarcinoma cells.

• The Korean Journal of Nuclear Medicine
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• v.34 no.2
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• pp.144-153
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• 2000

Purpose: The purpose of this study was to evaluate the relationship between radiation-induced activation of DNA repair genes and radiation induced apoptosis in A431 cell line. Materials and Methods: Five and 25 Gys of gamma radiation were given to A431 cells by a Cs-137 cell irradiator. Apoptosis was evaluated by flow cytometry using annexin V-fluorescein isothiocyanate and propidium iodide staining. The expression of DNA repair genes was evaluated by both Northern and Western blot analyses. Results: The number of apoptotic cells increased with the increased radiation dose. It increased most significantly at 12 hours after irradiation. Expression of p53, p21, and hRAD50 reached the highest level at 12 hours after 5 Gy irradiation. In response to 25 Gy irradiation, hRAD50 and p21 were expressed maximally at 12 hours, but p53 and GADD45 genes showed the highest expression level after 12 hours. Conclusion: Induction of apoptosis and DNA repair by ionizing radiation were closely correlated. The peak time of inducing apoptosis and DNA repair was 12 hours in this study model. hRAD50, a recently discovered DNA repair gene, was also associated with radiation-induced apoptosis.

• The Korean Journal of Zoology
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• v.15 no.4
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• pp.214-224
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• 1972

작년 9월 2$\\sim$4일에 프랑스, 파리에서 사람의 染色體에 관한 國際會議(The Standardization Conference on Human Cytogenetics)가 열렸는데, 이것은 Denver, Londan 및 Chicago 會議(1960, 1963, 1966)에 계속해서 개최된 4번째 會議가 된다. 계속해서 이틀후인 9월 6일부터 1주일간 같은 장소인 파리에서 4차 國際人類遺傳學會가 개최되었다. 이들 양 會議에서 사람의 染色體에서 밴드 구조를 얻을 수 있는 特殊染色法과 그에 따른 染色體의 새로운 同定과 命名法이 화제의 중심이 되었던 것이다. 이 사람은 다행히 이들 회의에 참석할 기회를 얻어 이 문제를 토의하는 회합에 참석했기에 여기 간단히 그 문제를 해설해볼까 한다. 1956년 Tjio 및 Leven이 사람의 染色體의 정확한 수를 확인했고, 그 뒤 Denver 및 London 國際會議를 거쳐서 사람 染色體의 同定과 命名에 관한 國際的인 規約이 정해지기는 했지만, 23쌍의 사람의 染色體 하나 하나를 정확히 구분하기란 어려운 일로되었다. 종래의 방법으로 얻어진 사람의 染色體를 顯微鏡 밑에 色體 1쌍(X-Y, X-X)으로 구분을 하지만 실제로 확신을 갖일 수 있는 것은 常染色體에서 6쌍과 Y染色體만이였다. 다음 1960년대에 染色體 연구에 널리 이용되어온 오토래디오그리피(Autoradiography)는 DNA 複製의 시간적인 차이를 통해 染色體 固定에 큰 도움을 주었지만 실제로 정확한 식별이 가능케 한 것은 常染色體쌍 5과 여성의 X染色體 1개만 이라고 하겠다. 이렇게 생각한다면 남어지 11쌍의 常染色體와 1개의 X染色體는 1970년에 이르기까지 정확한 同定은 불가능했다고 말할 수 있다. 그러나 1970년을 전후해서 이문제에 관한 정세는 일변했으며, 여러 가지 特殊染色法이 개발되어 사람의 染色體 하나 하나를 정확히 식별 할 수 있는 단계에 접어들게 되었다. 그중에서도 식별 할 수 있는 단계에 접어들게 되었다. 그중에서도 대표적인 것이 키나크린 螢光染色法(quinacrine fluorescence staining method)와 김자分染法(heat-giemsa staining method)이며 이들 방법을 통하면 染色體의 縱軸에 따라 특유한 明暗의 밴드 패턴(banding patterns)이 나타나게 되어, 사람만이 아니라 고등한 동물의 모든 染色體가 쉽게 同定이 된다는 것이다.