• 대한수의학회지
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• 제31권3호
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• pp.329-335
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• 1991

The filter elution technique was used to assay $^{60}Co$ $\gamma$ ray-induced DNA strand breaks(SB) in EL4 mouse leukemia cell and mouse spleen lymphocyte. The lymphocytes were stimulated with lipopolysaccharide (LPS, $20{\mu}g/ml$) to label $[^3H]$ thymidine. EL4 cells and lymphocytes in suspension were exposed at $0^{\circ}C$ to 0Gy, 1Gy, 5Gy, 10Gy or l5Gy for DNA single strand breaks(SSB) assay and 0Gy, 25Gy, 50Gy, 75Gy or 100Gy for DNA double strand breaks(DSB) assay of $^{60}Co$ radiation and elution procedure was performed at pH12.1 and 9.6. The number of DNA strand breaks increased with increasing doses of r rays. The strand scission factor(SSF) was estimated in each experiment (eluted volume 21ml). The slope of SSB EL4 cells was $0.01301{\pm}0.00096Gy^{-1}$ (n=5), the slope of SSB for lymphocytes was $0.01097{\pm}0.00091Gy^{-1}$ (n=5) and the slope of DSB for lymphocytes was $0.001707{\pm}0.0000573Gy^{-1}$ (n=5). Thus EL4 cells were more sensitive to induction of DNA SSB by ionizing radiation than lymphocytes (p<0.005). The ratio of slope of dose-response relationship (SSF versus dose) of lymphocytes DNA SSB as compared with the slope of DNA DSB was 6.4.

• 대한수의학회지
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• 제32권1호
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• pp.1-6
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• 1992

The turnover time of the mucosal epithelium in the small intestine(jejunum and ilium) and large interstine(cecum), and the cells in the lamina propria of the small intestine was investigated with the radioautography in mice at various times after single injection of $^3H$-thymidine. Twenty suckling mice were sacrified at each of the following time intervals after injection ; 2 hrs, 1, 3, 5. 7, 14 and 17 days. 1. The labeled index of the epithelial cells in the crypt and the villus of the small intestine averaged 98.7% and 1.3% at 2 hrs, 982% and 1.8% at 1 day, 18.7% and 81.3% at 3 days, 6.3% and 93.7% at 5 days, respectively. The labeled index of the epithelial cells of the crypt-base, the upper-crypt and the mucosal surface in the large intestine averaged 71.8%, 28.2% and 0% at 2 hrs, 45%. 54.2% and 0% at 1 day, 17.2%, 54.5% and 28.2% at 3 days, 10.2%, 32.4% and 57.4% at 5 days, respectively. This result suggested that the turnover time of all the epithelial cells migrating from crypts to villi in the direction of the villus tips was calculated to be less than 5 days, and also the longest turnover time was calculated to be no longer than 7 day. 2. The labeled index of the total cells in the lamina propria of the small intestine averaged 6.2-7% at 2 hrs to 5 days, 4.7% at 7 days 2.6% at 17 days and this index is tend to be decreased moderately at 7 days and severely at 17 days. So this result suggested that the turnover time of the cells with the shorter cycle duration in the lamina propria of the small intestine were less than 5 days and that of the cells with the longer cycle duration more than 17 days.

• 생명과학회지
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• 제18권1호
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• pp.69-74
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• 2008

국내에서 사육중인 말에서 채취한 시료로부터 Stapylococcus aureus를 분리 동정한 결과 39.2%(58/148)의 분리율을 나타내었다. S. aureus의 혈장응고효소 생산능 시험에서는 98.3%가 양성반응을 나타내었고 신선 혈장의 희석농도는 1:2에서 가장 높은 양성반응을 나타내었다. 본 연구에서 최종 동정된 S. aureus의 기본적인 생화학 성상은 일반적으로 알려져 있는 S. aureus의 특성과 유사한 결과를 얻을 수 있었으나, urease test의 경우 전체 균주의 73% 만이 양성 결과를 나타내어 균주별로 다양한 반응 양상을 관찰 할 수 있었다. 또한 34종의 substrate에 대한 분해 양상에 대한 검사 결과 fructose, maltose, ${\alpha}$-hydroxy butyric acid, thymidine-5'-mono phosphate, uridine-5'-mono phosphate 등의 substrate에 대해서는 모든 균주가 이용능력을 가지고 있었지만 ribose, sorbitol, xylose, xylitol 등의 substrate에 대해서는 모든 균주가 기질 이용능력을 가지고 있지 않는 것으로 확인되었다. 그리고 PCR에 의한 S. aureus를 동정한 결과 108bp 크기의 band를 확인하였다. 국내 말에서 분리한 58주의 S. aureus에 대한 약제 내성 양상을 조사한 결과 spectinomycin, sulfonamides, erythromycin, tetracyclin, ciprofloxacin, neomycin, enofloxacin, penicillin 등의 약제에 높은 저항성을 나타내었다.

• 미생물학회지
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• 제27권1호
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• pp.48-55
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• 1989

HA 비드에 부착한 S. sanguis 세포는 방사능 측정시 칵테일 속에서 비드에 여전히 부착되어 있음으로 비드에 의한 방사능의 sel-absorpiton이 일어났으며 그 정도는 염산용액으로 비드를 용해시킬 때 측정한 반사능값의 34.5% 이었다. 현탁액으로 준비한 세포의 방사능 측정과는 달리 HA 또는 SHA에 부착한 세포의 경우에는 소량의 scintillation 용액(2.5ml) 사용에서 더 좋은 측정값을 얻었다 . HA 비드에 의한 칭은 약 18% 이었다. 부착 assay 에 사용할 HA 비드 준비에 보통 이용되는 3가지의 dqkdqjqdm 비교에서는 차이가 없었다. 부착실험용의 세균세포는 초소리파로 세균사슬을 끊었는데 30초간의 파절로 세포하나 또는 두개로 주로 구성된 현탁액을 얻을 수 있었다. 본 연구에서는 HA 비드에 부착한 S. sanguis 세포를 측정할 때 self-absorpiton이 일어남을 발견하였고 부착한 세균을 비드에서 탈락하거나 또는 비드를 용해시킨 후 측정함으로써 self-absorption을 제거할 수 있었다. 그리고 부착한 세균의 방사능 측정에서는, 적정의 칵테일량을 발견하여 사용하므로 측정값을 유의하게 증가시킬 수 있었다. 수 있었다.

• 대한한의학회지
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• 제23권1호
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• pp.24-36
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• 2002

Objectives: The chemotherapeutic potential of Gagamgilgyung-tang for the treatment of human lung cancer, the antitumorigenic effects of Gagamgilgyung-tang on the proliferation and apoptosis of human lung cancer cell line A427 were investigated using molecular biological approaches, Methods: To determine Gagamgilgyung-tang concentrations which do not evoke cytotoxic damage to the cell line, cell viability was examined by MTT assay. To prove Gagamgilgyung-tang's antitumorigenic potential to human lung cancer, [3H]thymidine incorporation assay, trypan blue exclusion and Cpp32 protease activity assays and quantitative RT-PCR analysis were examined. Results: While A427 cells treated with $0.1-2.0{\mu\textrm{g}}/ml$ of Gagamgilgyung-tang showed no recognizable effect, marked reductions of cell viability were detected at concentrations over $5.0{\;}\mu\textrm{g}/ml$. DNA replication of A427 cells was inhibited by Gagamgilgyung-tang in a dose-dependent manner and Gagamgilgyung-tang induced the G1 cell cycle arrest through inhibition of DNA replication. Gagamgilgyung-tang triggered apoptotic cell death of A427 and enhanced the apoptotic sensitivity of the cells that were injured by a DNA damage-inducing chemotherapeutic drug etoposide. Gagamgilgyung-tang induces expression of growth-inhibiting genes such as p53 and p21/Wafl whereas it inhibited expression of growth-promoting genes such as c-Myc and Cyclin D1. Expression of a representative apoptosis-inducing gene Bax was also found to be induced by Gagamgilgyung-tang while apoptosis-suppressing Bcl-2 expression was not changed. Conclusions: Gagamgilgyung-tang could suppress the abnormal growth of tumor cells by suppressing the survival of genetically altered cells via induction of apoptosis. This study suggests that Gagamgilgyung-tang might have an antitumorigenic potential to human lung cancer cells, which might be associated with its growth-inhibiting and apoptosis-inducing properties.

• The Korean Journal of Physiology and Pharmacology
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• 제3권3호
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• pp.283-291
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• 1999

We have synthesized novel platinum (II) coordination complex containing cis-1,2-diaminocyclohexane (DACH) as a carrier ligand and 1,2-bis(diphenylphosphino)ethane (DPPE) as leaving group. Furthermore, nitrate was added to improve the water-solubility. A new series of [Pt(cis-DACH)(DPPE)] $2NO_3(PC)$ was evaluated its antitumor activity on various MKN-45 human gastric adenocarcinoma cell-lines and normal primary cultured kidney cells. The new platinum complex demonstrated high efficacy in the cytotoxicity on MKN-45 cell-lines as well as adriamycin-resistant (MKN-45/ADR) and cisplatin-resistant (MKN-45/CDDP) cells. The cytotoxicities of PC were found quite less than those of cisplatin in rabbit proximal renal tubular cells, human renal cortical cells and human renal cortical tissues using MTT assay, $[^3H]-thymidine$ uptake and glucose consumption tests. Based on these results, this novel platinum (II) coordination complex, was considered as better a valuable lead for improving antitumor activities with low nephrotoxicities in the development of a new clinically available anticancer chemotherapeutic agents.

• The Korean Journal of Physiology and Pharmacology
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• 제19권5호
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• pp.421-426
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• 2015

The increased potential for vascular smooth muscle cell (VSMC) growth is a key abnormality in the development of atherosclerosis and post-angioplasty restenosis. Abnormally high activity of platelet-derived growth factor (PDGF) is believed to play a central role in the etiology of these pathophysiological situations. Here, we investigated the anti-proliferative effects and possible mechanism(s) of murrayafoline A, a carbazole alkaloid isolated from Glycosmis stenocarpa Guillamin (Rutaceae), on PDGF-BB-stimulated VSMCs. Murrayafoline A inhibited the PDGF-BB-stimulated proliferation of VSMCs in a concentration-dependent manner, as measured using a non-radioactive colorimetric WST-1 assay and direct cell counting. Furthermore, murrayafoline A suppressed the PDGF-BB-stimulated progression through $G_0/G_1$ to S phase of the cell cycle, as measured by [$^3H$]-thymidine incorporation assay and cell cycle progression analysis. This anti-proliferative action of murrayafoline A, arresting cell cycle progression at $G_0/G_1$ phase in PDGF-BB-stimulated VSMCs, was mediated via down-regulation of the expression of cyclin D1, cyclin E, cyclin-dependent kinase (CDK)2, CDK4, and proliferating cell nuclear antigen (PCNA), and the phosphorylation of retinoblastoma protein (pRb). These results indicate that murrayafoline A may be useful in preventing the progression of vascular complications such as restenosis after percutaneous transluminal coronary angioplasty and atherosclerosis.

• Journal of Microbiology and Biotechnology
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• 제6권6호
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• pp.445-450
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• 1996

Gelatinase A is a member of the matrix metalloproteinases that play an important role in cancer invasion and metastasis. In the course of screening gelatinase A inhibitors from microbial sources, a fungal strain PT-262 showed a strong inhibitory activity. The strain was identified as Chaunopycnis alba on the basis of its morphological characteristics. The inhibitor was isolated from acetone extract of mycelial cake by sequential chromatographies on MCI-gel, Sephadex LH-20, and a reverse-phase HPLC column. The purified inhibitor was identified as pyridoxatin by its physico-chemical properties and spectroscopic analysis. Pyridoxatin is not a peptide analog and has cyclic hydroxamic acid moiety. It inhibited activated gelatinase A with an $IC_{50}$ value of 15.2 ${\mu}M$ using fluorescent synthetic peptide. It also had a strong cytotoxicity against human cancer cell lines in vitro. Furthermore, this compound inhibited DNA synthesis with an $IC_{50}$ value of 2.92 ${\mu}M$ in PC-3 prostate cancer cells by [$^3H$]thymidine incorporation assay.

• Journal of Periodontal and Implant Science
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• 제23권3호
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• pp.454-460
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• 1993

Abnormalities of the T cell subsets have been detected in the immunologically mediated disease sites such as periodontal lesions which are attributable to the regulatory effect of cell differentiation and specific chemokinetic effect of various cytokines. Macrophage Inflammatory protein$(MIP)-1{\alpha}$ and gammain terferon$({\gamma}-IFN)$ serve as important immunoregulatory molecules through which growth and differentiation of specific T cell subsets are known to be negatively regulated. Murine cytolytic T cell line CTLL-2 were used to perform the [$^3H$]-thymidine incorporation test, by which we obtained more comprehensive view in regulatory actions of cytokines on the T cell subset proliferation. 1. $rMIP-{\alpha}$(200ng/ml) and $r{\gamma}-IFN$(100U/ml) appreared to suppress the proliferation rate to CTLL-2 by 74 and 86% respectively, and the suppressive action of two cytokines were synergisic. 2. Culture supernatant of anti-CD3 mAb-stimulated mouse splenocyte enhanced the proliferation rate of CTLL-2 up to 10-fold with dose-dependent manner. However, culture supernatant of unstimulated splenocyte showed only 2-fold increase in the proliferation rate. 3. CTLL-2 cell proliferation was strictly IL-2 dependent.

• Advances in pediatric surgery
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• 제8권2호
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• pp.101-106
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• 2002

The antiangiogenic effects of novel agent KJ3, Betulinic acid, and Fumagillin on the neovascularization were studied by examining ultrastructural alterations in the vasculature of synthetic gelform and mouse neuroblastoma C1300. Small pieces of gelform with 0.4% agar were introduced subcutaneously (s.c.) in 7 week old male CH3/HeJ mice. After the $LD_{50}s$ were determined by FACS analysis, a third of $LD_{50}$ of three drugs were injected either locally or intraperitoneally every other day for 14 days. A/J mice were inoculated s.c. with the C1300 neuroblastoma cell line, then either saline or three drugs were injected in the same manner. The antiangiogenic effects of three drugs were studied by measuring the histologic changes in tumors, and immunostaining for CD34, VIII/vWF, CD105, and thymidine phosphorylase. In the drug treated groups, the number of vessels in gelform experiments and C1300 neuroblastoma experiments were lower than the corresponding values in the control. The histologic findings were significantly different in drug treated groups on day 7, but these were not significant on day 14. These results imply that antiangiogenic agents were effective when the tumor burden is minimal.