• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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• pp.142.1-142.1
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• 2003

Sensitive and safe method for visualization of DNA in agarose gels using visible dye is described. To improve the sensitivity, we studied a counterion-dye staining method using methyl orange as a counterion-dye which contributes to reduce excessive background staining by indoine blue. Dye concentrations, PH of staining solution, mixing molar ratio of two dyes, and staining times were optimized for the counterion-dye staining. By the staining with a mixed solution of 0.005% indoine blue and 0.00165% methyl orange in 10% ethanol 0.2M sodium acetate, 8 ng of the 3 kb DNA in an agarose gel was detected within 1hr. (omitted)

• 대한임상검사과학회지
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• 제48권4호
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• pp.371-377
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• 2016

탈회방법은 골수조직의 병리학적 진단을 위해서 항상 시행되는 과정이다. HCl 탈회용액과 같이 주로 사용하고 있는 산성용액은 탈회과정 동안에 조직내의 항원성에 손상을 입힌다. 특히, 골수조직 내의 RNA나 DNA에 심하게 손상을 준다. 따라서 조직의 항원성을 보존하기 위한 표준화된 탈회방법이 필요하다. 본 연구는 일반적으로 가장 많이 사용되는 HCl 기반의 상품화된 탈회용액과 직접 제조한 EDTA 탈회용액이 골수조직의 탈회과정에 어떤 영향을 미치는지 분석하였다. 환자로부터 채취된 73예의 골수생검조직을 HCl 탈회와 EDTA 탈회의 두 그룹으로 나누어 탈회과정을 진행하였다. 골수생검조직의 탈회과정 후 결과의 차이는 hematoxylin & eosin 염색과 reticulum 염색, Ki-67, CD20, CD138의 항체를 이용한 면역조직화학염색, DNA 추출 및 분석, in situ hybridization, IGH gene rearrangement 와 같은 분자병리검사를 시행하여 분석하였다. 일반적인 염색과 특수염색에서는 두 탈회용액간의 차이는 없었다. 또한 세포증식 표지자와 같은 세포막 혹은 세포질에서 발현되는 항체는 탈회용액간의 차이 없이 잘 염색되었다. 반면 HCl 탈회 용액에 처리한 후 핵 내 단백질인 Ki-67의 염색상은 현저히 불량한 것으로 관찰되었다. HCl 탈회용액과 비교하여 EDTA 탈회용액에서의 골수생검조직 내의 DNA와 RNA가 잘 보존되었음을 다양한 분자병리검사를 통해 확인할 수 있었다. 특히 HCl 탈회용액에 처리한 28예와 EDTA 탈회용액에 처리한 12예의 DNA의 순도와 농도을 비교한 결과 통계학적으로 유의한 수준으로 차이가 있음을 확인하였다. 이로써 EDTA 탈회용액이 조직 내의 항원성을 잘 유지시키며, 면역조직화학염색과 분자병리검사에 적합한 방법임을 확인 할 수 있었다.

• 치과방사선
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• 제21권2호
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• pp.183-197
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• 1991

After single fraction of 2, 5, 10 Gy irradiation on submandibular gland of 40 male rats, weighing 150gm, respectively, these animal were sacrificed two hours after 0.1㎎/g bromodeoxyuridine (Sigma) peritoneal injection in 1, 3, 7, 15 hours, 1, 3, 7 days after irradiation. And excised submandibular gland were fixed in Carnoy's and Bouin's solution for 2 hours. Paraffin sections were stained with H&E, and PAS for the observation of the change of salivary gland tissue, and with Feulgen for the study of the DNA distribution, and immunohistochemically stained with anti-bromodeoxyuridine (Sanbyo Co.) for detection of DNA synthetic cells in order to study the distribution of DNA synthetic cells of salivary gland tissue and endothelium after irradiation in 5 different sites of 6 slides on X 200 high power field. The results were as followings. 1. In PAS staining 3 days after 5Gy irradiation, decreased mucine secretion of serous cells were found, and 7 days after l0Gy irradiation, decreased mucine secretion of mucous cells were found. 2. In histopathologic features, degeneration of serous cells were found in 3 days after 2 Gy irradiation and there was little change in mucous cells and excretory duct cells. 3. In Feugen staining, 3 days after 2 Gy, 5 Gy irradiation, more high percentage of DNA synthetic cells were found in intercalated duct cells, striated duct cells and excretory duct cells than in BrdU staining. 4. In immunohistochemical features, DNA synethsis of serous cells and granular convoluted tubular cells abruptly decreased in early period after irradiation and showed no recovery in 7 days after irradiation but there was an increase in DNA synthesis of intercalated duct cells, striated duct cells and excretory duct cells, which have less S-phase cells comparatively, in 7 days after 2 Gy, 5 Gy irradiation. 5. In immunohistochemical features, the DNA synthesis of endothelial cells was continuously decreased after irradiation but showed slight increase in 7 days after 2 Gy and S Gy irradiation.

• Asian-Australasian Journal of Animal Sciences
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• 제18권5호
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• pp.626-631
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• 2005

Nitric oxide (NO) derived from inducible nitric oxide synthase (iNOS) is involved in cell apoptosis, which contributes to luteal regression and luteolysis in some species. In large domestic animals, no direct evidence for the relationship between NO and cell apoptosis in the process of corpus luteum regression is reported. The present study was conducted to investigate the localization of iNOS on porcine corpora lutea (CL) during the oestrus cycle and its relation to cell DNA fragmentation and CL regression. According to morphology, four luteal phases throughout the estrous cycle were defined as CL1, CL2, CL3 and CL4. By isoform-specific antibody against iNOS, the immunochemial staining was determined. Luteal cell DNA fragmentation was determined by flow cytometry. The results showed that no positive staining for iNOS was in CL1 and that iNOS was produced but limited to the periphery of CL2, while in the CL3, the spreading immunochemical staining was found inside the CL. No iNOS positive staining was detected in CL4. Meanwhile, DNA fragmentation increased dramatically when CL developed from CL2 to CL3 (p<0.05). In CL4, higher proportion of luteal cells still had fragmented DNA than that of luteal cells from CL1 or CL2 (p<0.05). These results indicate that iNOS expression is closely related to luteal cell apoptosis and then to luteal regression.

• Asian Pacific Journal of Cancer Prevention
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• 제15권5호
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• pp.2123-2128
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• 2014

A current hurdle in cancer management is the intrinsic or acquired resistance of cancer cells to chemical agents that restricts the efficacy of therapeutic strategies. Accordingly, there is an increasing desire to discover new natural compounds with selective toxicity to combat malignancies. In present study, the cytotoxic and apoptosis-inducing activities of ferutinin, a terpenoid derivative from Ferula ovina, were investigated on human breast (MCF7) and bladder (TCC) cancer cells as well as normal fibroblasts (HFF3).The toxicity and DNA damage inducing effects of ferutinin were studied by MTT and comet assays, DAPI and PI staining and DNA laddering. The $IC_{50}$ values of ferutinin were identified and compared with routine prescribed drugs, doxorubicin and vincristine, by MTT test. Alkaline comet assay and DAPI staining revealed DNA damage due to ferutinin, which was significantly (p<0.001) higher in MCF7 and TCC than HFF3 cells. Apoptosis induction was evidenced by PI staining and DNA laddering. Our results suggest that ferutinin could be considered as an effective anticancer agent for future in vivo and clinical experiments.

• 한국작물학회지
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• 제50권spc1호
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• pp.228-230
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• 2005

형광 염료와 레이저 겔스캐너 장비를 이용하여 변성아크릴 아마이드 겔에서 전기 영동된 DNA를 신속하고 간편한 방법으로 기존의 은염색법과 비슷한 감도로 검출하고자 하였다. 변성아크릴아마이드 겔을 형광 염료인 SYBR Green (Molecular Probes)이나 Vistra Green (Amersham Bioscience) 0.01 X 희석액 (pH 8)으로 염색한 후 480nm 레이져, 520nm filer 옵션으로 스캔하여 DNA를 검출하였으며, 검출감도는 기존의 은염색법과 비슷하면서 염색 단계를 한 단계로 줄일 수 있었다.

• Toxicological Research
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• 제7권2호
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• pp.157-163
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• 1991

The effect of patulin, a mycotoxin, on the growth of Escherichia coli cell was investigated. E. coli cell elongation usually shown in SOS-response for DNA repair was induced by 20 mg of patulin per ml. After staining the E. coli chromosome with fluorescence dye(DAPI, 4', 6-diamino-2-phenyl-indole), chromosomal DNA partitioning was not affected by patulin. The observation indicateds that patulin acts as a DNA damaging agent which is effective for E. coli cell elongation introduced by the inhibition of septum formation.

• Biomolecules & Therapeutics
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• 제20권5호
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• pp.487-491
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• 2012

The aim of this study was to investigate the inhibitory effects of rutaecarpine on DNA strand breaks and apoptosis induced by hydrogen peroxide ($H_2O_2$) in murine Hepa-1c1c7 cells. Oxidative DNA damage was estimated by nuclear condensation assessment, fluorescence-activated cell sorting analysis, and Comet assay. Rutaecarpine inhibited cell death induced by $500{\mu}M$ $H_2O_2$, as assessed by 4',6-diamidino-2-phenylindole (DAPI) staining. Treatment with rutaecarpine reduced the number of DNA strand breaks induced by $H_2O_2$, as assessed by DAPI staining and Comet assay, and increased quinone reductase, phosphatidylinositol 3-kinase, and pAkt protein levels, as assessed by western blotting.

• Asian Pacific Journal of Cancer Prevention
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• 제14권3호
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• pp.1643-1647
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• 2013

Background: To identify the risk factors and assess the role of survivin in predicting progessivity precancerous cervical lesions. Materials and Methods: This case-control study was conducted from October 2009 until May 2010. We obtained 74 samples, classified according to the degree of cervical intraepithelial neoplasia (CIN): 19 samples for CIN 1, 18 samples for CIN 2, 18 samples for CIN 3, and 19 samples as controls. Demographic profiles and risk factors assesment, histopathologic examination, HPV DNA tests, immunocytochemistry (ICC) and immunohistochemistry (IHC) staining for survivin expression were performed on all samples. Data was analyzed with bivariate and multivariate analysis. Results: Multivariate analysis revealed significant risk factors for developing precancerous cervical lesions are age <41 years, women with ${\geq}2$ sexual partners, course of education ${\geq}13$ years, use of oral contraceptives, positive high-risk HPV DNA, and high survivin expression by ICC or IHC staining. These factors were fit to a prediction model and we obtained a scoring system to predict the progressivity of CIN lesions. Conclusions: Determination of survivin expression by immunocytochemistry staining, along with other significant risk factors, can be used in a scoring system to predict the progressivity of CIN lesions. Application of this scoring system may be beneficial in determining the action of therapy towards the patient.

• Journal of Korean Neurosurgical Society
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• 제39권6호
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• pp.432-437
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• 2006

Objective : In order to establish the index of degeneration, the authors performed a histochemical study with Safranin-O staining and investigated the occurrence of apoptosis in the human intervertebral disc. Methods : Eighteen intervertebral disc specimens surgically extracted from the patients and two additional specimens from the autopsied cases were stained with Safranin-O for proteoglycan according to a standard protocol. Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate- biotin nick end labeling[TUNEL] was used to detect the fragmented DNA known to be associated with apoptotic cell death and classification scheme was formulated for categorization of the degree of Safranin-O staining [normal, moderate reduction, faint] by modification of Makin's histological-histochemical grading. The Kruskal-Wallis H test and Chi-square test were used for statistical analysis. Results : The statistical results showed a significant difference in the mean age between "normal" Safranin-O staining group and the others [19.3 versus 55, 43.4, p=0021]. However, there was no statistically significant correlation between Safranin-O staining and MR grading of disc degeneration. Only six of eighteen surgical specimens and none in autopsies showed positive apoptotic cells in TUNEL staining. Conclusion : The determination of the degree of degeneration in surgically obtained disc tissue per se by histochemical staining or by the degree of apoptosis that corresponds to its morphologic change was not feasible.