• 제목/요약/키워드: Biologic Assay

검색결과 37건 처리시간 0.032초

HeLa.S3(SC) 세포계의 집락형성에 미치는 방사선의 효과 (Radition Effect on Colony Formation of HeLa.S3(SC) Cell Line)

  • 신세원;김성규;김명세
    • Journal of Yeungnam Medical Science
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    • 제10권1호
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    • pp.212-217
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    • 1993
  • 임상적으로 많이 이용되는 범위의 방사선 조사에서는 증식능의 상실을 세포사망으로 정의를 내린다. 본 연구는 실험용 orthovoltage조사기를 이용하여 확립된 세포주인 Hela.53(5C)를 대상으로 각각 100 cGy, 200 cGy, 400 cGy 및 600 cGy 방사선조사 후 2주일간 배양후 crystal violet 염색을 하여 육안적으로 판별가능한 크기의 집락의 수를 산출하여 다음의 결과를 얻었다. HeLa.S3(SC) 세포의 집락 형성능은 0.464이었다. 세포의 생존곡선의 형태는 다표적, 1 hit 형태를 보였다. Do는 150 cGy, Dq는 80 cGy, 외삽치는 1.7이었다. 이 연구결과는 향후 다양한 방사선 생물학 연구에 참고자료로 이용 가능하리라 사료된다.

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Construction and Analysis of Food-Grade Lactobacillus kefiranofaciens β-Galactosidase Overexpression System

  • He, Xi;Luan, MingJian;Han, Ning;Wang, Ting;Zhao, Xiangzhong;Yao, Yanyan
    • Journal of Microbiology and Biotechnology
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    • 제31권4호
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    • pp.550-558
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    • 2021
  • Lactobacillus kefiranofaciens contains two types of β-galactosidase, LacLM and LacZ, belonging to different glycoside hydrolase families. The difference in function between them has been unclear so far for practical application. In this study, LacLM and LacZ from L. kefiranofaciens ATCC51647 were cloned into constitutive lactobacillal expression vector pMG36e, respectively. Furtherly, pMG36n-lacs was constructed from pMG36e-lacs by replacing erythromycin with nisin as selective marker for food-grade expressing systems in Lactobacillus plantarum WCFS1, designated recombinant LacLM and LacZ respectively. The results from hydrolysis of o-nitrophenyl-β-galactopyranoside (ONPG) showed that the β-galactosidases activity of the recombinant LacLM and LacZ was 1460% and 670% higher than that of the original L. kefiranofaciens. Moreover, the lactose hydrolytic activity of recombinant LacLM was higher than that of LacZ in milk. Nevertheless, compare to LacZ, in 25% lactose solution the galacto-oligosaccharides (GOS) production of recombinant LacLM was lower. Therefore, two β-galactopyranosides could play different roles in carbohydrate metabolism of L. kefiranofaciens. In addition, the maximal growth rate of two recombinant strains were evaluated with different temperature level and nisin concentration in fermentation assay for practical purpose. The results displayed that 37℃ and 20-40 U/ml nisin were the optimal fermentation conditions for the growth of recombinant β-galactosidase strains. Altogether the food-grade Expression system of recombinant β-galactosidase was feasible for applications in the food and dairy industry.

조선소 배출수 및 주변 하천수의 생물독성 (Biologic Effect of Effluents from Shipyard and the Adjacent Stream Water on Four Cultured Organisms)

  • 서진영;김기범;안준건
    • 한국해양환경ㆍ에너지학회지
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    • 제9권4호
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    • pp.187-192
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    • 2006
  • 조선소에서 배출되는 처리수, 혼합방류수 및 주변 하천수가 양식 생물에 미치는 생물 독성영향을 알아보기 위해 48시간 급성독성과 DNA 손상을 조사하였다. 조사대상 생물종으로는 조선소 주변에서 양식되고 있는 넙치, 조피볼락, 피조개, 멍게가 사용되었으며, 48 시간 노출 후 치사율을 파악하였고, DNA 손상 정도는 Comet assay을 이용하여 측정되었다. 급성독성 실험 결과, 넙치는 장평천에서 치사가 나타났고(26%), 조피볼락은 혼합방류수 1에서 치사가 나타났다(13%). 멍게는 고현천에서 10%의 치사율을 보였고, 피조개는 어느 시료에서도 치사가 나타나지 않았다. 본 연구에 사용되어진 어떠한 시료에서도 실험생물을 50%까지 치사시키는 독성이 나타나지 않아 $LC_{50}$은 계산될 수 없었다. 넙치는 장평천과 혼합방류수에서 대조구보다 유의하게 높은 DNA 손상을 보여주었고, 조피볼락은 장평천에서 유의하게 높은 DNA손상이 나타났다(p<0.05). 멍게는 세탁폐수에서 유의한 DNA손상이 나타났지만, 피조개에서는 모든 처리구에서 DNA 손상을 보이지 않았다. 치사율과 DNA 손상을 고려하였을 때 조선소의 처리수와 혼합방류수보다는 장평천에서 놀은 생물독성을 보여주는 것으로 나타났다.

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시험관내 폐암(肺癌), 위암(胃癌) 및 신경교종(神經膠腫) 세포(細胞)에 대한 석웅황(石雄黃)의 항암효과(抗癌效果) (Induction of Apoptosis by Realgar on Lung Cancer Cells(A549), Stomach Center Cells(KATO) and Neuroglioma Cells(SNU-1118, U-87MG, U-373MG))

  • 방대건;김진성;류봉하
    • 대한한방내과학회지
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    • 제28권2호
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    • pp.294-303
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    • 2007
  • Objectives : We are aimed to identify anti-tumor effects of realgar on some kinds of cancer cells through molecular biologic methods. Materials & Methods : We used 5 kinds of cancer cell lines: lung cancer cells(A549). stomach cancer cells(KATO) and neuroglioma cells(SUN-1118. U-87MG, U-373MG). We injected the boiled extracts of realgar $50{\mu}g$. $100{\mu}g$ to cultural media( ml )for 24 hours. We measured the killing effects on 5 kinds of cancer cells through inverted and fluorescence microscope, the suppressive effects on viability of those cells via XTT assay and the effects on the revelation of Bax and Bcl-2 proteins related to apoptosis by western blotting. Results : In the changes of morphology, the extracts of realgar showed more significant killing effects on all cancer cells. especially KATO, SNU-1118, U-87MG, U-373MG, than the control group with dose dependence, which was statistically significant. In XTT assay, the extracts of realgar showed more suppressive effects on viability of all cancer cells, especially KATO and U-373MG, than the control group with dose dependence, which was statistically significant. In the revelation of proteins related to apoptosis, the extracts of realgar increased the level of Bax and decreased that of Bcl-2 in all cancer cells with dose dependence. Conclusions : We identified that realgar had more anti-tumor effects on stomach cancer and neuroglioma than on lung cancer in the experiments above. However, these basic experiments were performed in vitro. We hope the anti-tumor effects of realgar will be practically identified through more progressive research.

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Glycosaminoglycan이 치은섬유아세포의 성장에 미치는 영향 (Effects of Glycosaminoglycan on the Growth of Human Gingival Fibroblast)

  • 이용배;피성희;김탁;이광수;유형근;신형식
    • Journal of Periodontal and Implant Science
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    • 제30권3호
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    • pp.599-610
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    • 2000
  • Gingival fibroblasts are embedded in an extracellular matrix. The matrixs have influence on the development, polarity, and behavior of nearby cells. The major component of periodontal extracellular matrix is a glycosaminoglycan. The glycosaminoglycan are large carbohydrates that are composed of repeating disaccharide units and exist in three main form: dermatan sulfate, chondrotitin sulfate, heparan sulfate. The purpose of present study is to examine the biologic effects of glycosaminoglycan on human gingival fibroblast. Human gingival fibroblasts were supplemented with each glycosaminoglycan, and cellular attachment and proliferation was determined by MTT assay. Dermatan sulfate and chondroitin sulfate did not stimulate the attachment and proliferation of human gingival fibroblasts, but heparan sulfate increased the proliferation and attachment in a time- and dose dependent manner. These results indicated that heparan sulfate seems to have a high potential for gingival regeneration and root surface attachment.

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한국산 P.V.C.의 생물학적 안정도 및 적합성에 대한 실험적 고찰 (An experimental study on the biological safety and compatability of P.V.C. made in Korea)

  • 선경
    • Journal of Chest Surgery
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    • 제17권1호
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    • pp.157-166
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    • 1984
  • These biologic test procedures are designed to test the suitability of P.V.C. made in Korea intended for parenteral preparation, which were based on the U.S. Pharmacopeia XIX "Biologic Test-Plastic Container", Official from July 1, 1975. Healthy adult human blood and rabbits weighing 2\ulcorner.2Kg were used for test materials. Sample P.V.C. were sampled from the medical equipments made in Korea randomly and Control P.V.C. were sampled from the standardized Cobe and Polystan P.V.C. tubes. P.V.C. extract was prepared from a homogeneous P.V.C. samples by incubating 60 square centimeters of the sample per 20 millimeters of sterile pyrogen-free saline at 70\ulcorner for 72 hours or autoclaving at 120\ulcorner for 1 hour. The Implantation Test was designed to evaluate the reaction of living tissue to the plastic by the method of the implantation of the Sample itself into animal tissue. The Systemic Injection Test, the Intracutaneous Test, and the remainders were designed to determine the biological response of animals to plastics by the single-dose injection of specific extracts prepared from a Sample. The results are as follows; 1.Implantation Test - No significant difference for reactions was noted between the Sample treated animal and the Control after 72 hours of implantation. 2.Systemic Toxicity Injection Test - No sign of toxicity and/or death immediately after injection and at 4, 24, 48 hours respectfully after injection. 3.Intracutaneous Test - None of the animals treated with the Sample showed a significantly greater reaction than the observed in the animals treated with Blank. 4.Pyrogen Assay-Only one animal treated with the Sample showed the maximal rise of rectal temperature about 0.2\ulcorner after 3 hours of injection, but remainders showed no change. 5.Hemolytic Index - The positive Control tube of distilled water exhibited complete hemolysis while the negative Control tube and P.V.C. extract were negative demonstrating no hemolysis. 6.Cell Morphology of Erythrocytes and Leukocytes on Stored, Heparinized Human Blood -- There was no significant difference in the morphology of either the Control or Sample extract. 7.Clotting Mechanism of Human Blood in vitro - After allowing to the P.V.C. extract at room temperature for 5 Hours and at 10\ulcorner for 24 hours, there was no appreciable difference in Prothrombin Time under these conditions. 8.Clotting Mechanism of Rabbit in vivo - At the termination of 5 days after intraperitoneal injection of the P.V.C. extract, no significant changes in Clotting Time were observed. According to the above results, it could be concluded that the P.V.C. made in Korea was acceptable for parenteral preparation, especially treated with physiologic saline and/or human blood.man blood.

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석웅황의 시험관내 위암, 신경교종 및 전립선암 세포에 대한 항암 연구 (Anti-tumor effects of Realgar on Stomach Cancer Cells (AGS), Glioma Cells (T98G, A172, SNU-489) and Prostate Cancer Cells (LNCaP))

  • 김선량;윤성우;류봉하
    • 대한한방내과학회지
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    • 제28권3호
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    • pp.409-420
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    • 2007
  • Objectives : The purpose of this study was to identify the anti-tumor effects of realgar on various cancer cells through molecular biologic and cellular biologic methods. Materials & Methods : We used 5 kinds of cancer cell lines:stomach cancer cell (AGS), glioma cells (T98G, A172, SNU-489) and prostate cancer cells (LNCaP). We injected the boiled extract of realgar. $50{\mu}$g/ml and $100{\mu}$g/ml to culture media (ml) for 24 hours. We examined the morphological changes under an inverted microscope and a fluorescence microscope. We measured the suppressive effect on viability of 5 kinds of cancer cells via XTT assay. We examined the effect on the revelation of PARP cleavage, Bcl-2 protein and Bax protein by western blot analysis. Results : The extract of realgar caused markedly morphological changes on AGS, T98G, SNU-489, and LNCaP. All of them showed withdrawn and floating appearance. The suppressive effect on viability of AGS, T98G, A172, SNU-489, and LNCaP showed that each test group had more suppressive effect on viability of AGS, T98G, A172, SNU-489, and LNCaP than the control group, which was statistically significantly (p<0.01). The extract of realgar did not induce PARP cleavage in AGS, T98G, A 172, SNU-489, or LNCaP. In the revelation of protein related to apoptosis, the protein levels of Bcl-2 decreased and the protein levels of Bax increased in AGS, T98G, SNU-489, and LNCaP treated with realgar. The protein levels of Bcl-2 decreased and the protein levels of Bax did not change in A172 treated with realgar. Conclusions : This experiment showed that realgar has anti-tumor effect on stomach cancer cells (AGS), glioma cells (T98G, SNU-489L and prostate cancer cells (LNCaP)

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편도암의 발암 원인으로 Human Papilloma Virus를 통한 발암 기전과의 상관 관계 (Correaltion of Human Papilloma Virus Infection Status with Tonsillar Squamous Cell Carcinoma)

  • 김세헌;변형권;천제영;박영민;정진세;이소윤
    • 대한두경부종양학회지
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    • 제23권1호
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    • pp.21-25
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    • 2007
  • Background:Squamous cell carcinoma(SCC)of the palatine tonsils represents approximately 15-23% of all intraoral SCC. The most frequently reported risk factors for oropharyngeal cancer are smoking and alcohol. In a recent overview of HPV and tonsillar squamous cell carcinoma(TC), 51% contained HPV DNA, and HPV-16 being the most frequent type. We aimed to clarify whether HPV directly effects on the oncogenesis and biologic behavior of TC by comparison with infection prevalence, and physical status of virus. Material and Method:We used HPV genotyping DNA chip(Biocore, Korea, Seoul) arrayed by multiple oligonucleotide probes of L1 sequence of 26 types of HPV and HPV genotypes are identified by fluorescence scanner. The copy numbers of HPV E2 and E6 open reading frames(ORF) were assessed using a TaqMan-based 5'-exonuclease quantitative real-time PCR assay. The ratio of E2 to E6 copy numbers was calculated to determine the physical status of HPV-16 viral gene. Results:We observed a significant difference in HPV prevalence between 52 TCs and 69 CFTs(73.1% vs. 11.6%), and most of the HPVs were type 16(87.2%)and non-episomal(94.1%) state. Conclusions:This study regarding HPV infection prevalence and mechanism in the largest population of palatine tonsillar squamous cell carcinoma with chronic follicular tonsillitis revealed significant difference pf HPV prevalence between TC and CFT. Most of HPV were 16 type and integrated or mixed, HPV-16 integration could be directly related to tonsillar carcinogenesis.

소각장 근로자에서 GSTM1의 유전자 다형성이 glycophorin A변이 발현율과 소변내 PAH 대사산물 농도와의 관계에 미치는 영향 (Association of glycophorin A mutant frequency and urinary PAH metabolites influenced by genetic polymorphisms of GSTM1 in incineration workers)

  • 이경호;하미나;최재욱;조수헌;박정규;황응수;강대희
    • 한국환경성돌연변이발암원학회지
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    • 제21권2호
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    • pp.149-155
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    • 2001
  • Eighty-one workers including 38 employees directly incinerating industry wastes were recruited from a company located in South Korea. To evaluate the association between urinary 1-hydroxypyrene glucuronide (1-OHPG) levels, as internal dose of polycyclic aromatic hydrocarbon (PAH) exposure, and glycophorin A (GPA) mutation frequency, as an early biologic effect indicator. Urinary 1-OHPG levels were measured by synchronous fluorescence spectroscopy after immunoaffinity purification using monoclonal antibody 8E11. Erythrocyte GPA variant frequency (NN or NO) was assessed in MN heterozygotes with a flow cytometic assay. The GSTM1 and GSTT1 genotypes were assessed by a multiplex PCR method. The GPA NN phenotype frequency was higher in occupationally exposed group (n=14, mean$\pm$S.D. 6.6$\pm$12.0 in 10/SUP 6/ erythrocyte cells) than in non-exposed group (n=22, 2.1$\pm$3.5). Similarly, the GPA(NO or NN) phenotype frequency was higher in exposed group (n=14, 9.7$\pm$17.3) than non-exposed group (n=22, 4.2$\pm$6.3). The above differences failed to reach statistical significance, but a significant increase was seen in GPA variant frequency levels with increase in urinary 1-OHPG levels (Spearman's correlation: p=0.06 (NO), p=0.07 (NO or NN)). When this association was evaluated by GSTM1 genotype status, the association between GPA mutation and urinary 1-OHPG levels was stronger in individuals with GSTM1 present genotype (Spearmans correlation; r=0.50, p=0.02). These results suggest that the association between urinary 1-OHPG and GPA mutation is be modulated by the GSTM1 genotype.

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Benzochloroporphyrin Derivative Induced Cytotoxicity and Inhibition of Tumor Recurrence During Photodynamic Therapy for Osteosarcoma

  • Gong, Hai-Yang;Sun, Meng-Xiong;Hu, Shuo;Tao, Ying-Ying;Gao, Bo;Li, Guo-Dong;Cai, Zheng-Dong;Yao, Jian-Zhong
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권5호
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    • pp.3351-3355
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    • 2013
  • Photodynamic therapy (PDT) is a promising cancer treatment modality that uses dye-sensitized photooxidation of biologic matter in target tissue. This study explored effects of the photosensitizer BCPD-17 during PDT for osteosarcoma. LM-8 osteosarcoma cells were treated with BCPD-17 and cell viability after laser irradiation was assessed in vitro with the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay. The effects of BCPD-17 during PDT recurrence were then examined on tumor-bearing mice in vivo. BCPD-17 had dosedependent cytotoxic effects on LM-8 osteosarcoma cells after laser irradiation which also had energy-dependent effects on the cells. The rate of local recurrence was reduced when marginal resection of mice tumors was followed by BCPD-17-mediated PDT. Our results indicated BCPD-17-mediated PDT in combination with marginal resection of tumors is a potentially new effective treatment for osteosarcoma.