• Title/Summary/Keyword: 세포생존

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Development of Acrylic Acid Grafted Polycaprolactone (PCL)/Biphasic Calcium Phosphate (BCP) Nanofibers for Bone Tissue Engineering Using Gamma-Irradiation (감마선을 이용한 아크릴산이 도입된 골조직공학용 PCL/BCP 나노섬유 지지체의 개발)

  • Jeong, Jin-Oh;Jeong, Sung In;Shin, Young Min;Park, Jong-Seok;Gwon, Hui-Jeong;An, Sung-Jun;Huh, Jung-Bo;Shin, Heungsoo;Lim, Youn-Mook
    • Polymer(Korea)
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    • v.39 no.3
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    • pp.418-425
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    • 2015
  • Polycaprolactone (PCL) and biphasic calcium phosphate (BCP) have been considered as useful materials for orthopedic devices and osseous implants because of their biocompatibility and bone-forming activity. However, PCL-based scaffolds have hydrophobic surfaces reducing initial cell adhesion or proliferation. To overcome the limitation, we fabricated surface-modified PCL/BCP nanofibers using gamma-irradiation for bone tissue engineering. PCL/BCP nanofibers were prepared by electrospinning and then we supplemented hydrophilicity by introducing acrylic acid (AAc) through gamma-irradiation. We confirmed the surface of nanofibers by SEM, and then the initial viability of MG63 was significantly increased on the AAc grafted nanofibers, and alkaline phosphatase activity($1.239{\pm}0.226nmole/{\mu}g/min$) improved on the modified nanofibers than that on the non-modified nanofibers($0.590{\pm}0.286nmole/{\mu}g/min$). Therefore, AAc-grafted nanofibers may be a good tool for bone tissue engineering applications.

Effect of Epidermal Growth Factor on Cell Survival of Human Ovarian Teratocarcinoma Cell Line (Epidermal Growth Factor가 난소 기형암종 배아세포주의 생존율에 미치는 효과)

  • Kim, Chung-hui;Kim, Jong-shu;Cuong, Dang Van;Kim, Na-ri;Kim, Eui-yong;Han, Jin
    • Korean Journal of Veterinary Research
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    • v.43 no.2
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    • pp.211-218
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    • 2003
  • Human ovarian cancerous cells survive in a way that they trigger the nucleotide excision repair (NER) or double-strand DNA repair (dsDNA) repair mechanism to show resistance to anticancer drugs and activate many kinds of repair protein, thus removing damaged DNAs. Two experiments on the PA-1 human ovarian teratocareinoma cell line that hardly has any expression of epidermal growth factor receptor (EGFR) were conducted in the study; first, EGF-R was transfected and its receptor was obtained. The receptor was investigated in terms of its mutual relations with many kinds of protein concerning NER or dsDNA repair. Second, it was examined what kind impact cisplatin and adriamycin had on the effects of EGF-R over the PA-1 cell line lacking EGF-R. When being administered with cisplatin and adriamycin, Hey and Hey C2 cell lines showed a high level of resistance while PA-1 cell line a high level of sensitivity. Hey and Hey C2 cell lines that are resistant against anticancer drugs exhibited a high level of EGF-R expression while PA-1 cell line that is sensitive to them did a much lower level of the expression. When PA-1 cell line was transfected for the expression of DNA adduct and EGF-R, it showed a higher level of resistance compared to the control group. There was no difference in the expression of DNA repair proteins (DNA- dependent protein kinase, Ku70, and Ku80) between Hey and the PA-1 cell lines. The results indicate that the Hey cell line that is resistant against cisplatin and adriamycin works along the signaling system responding to the changes of EGF-R while the PA-1 cell line that is sensitive to both of them does to the lack of EGF-R.

Isolation and Characterization of Pyrimidine Auxotrophs from the Hyperthermophilic Archaeon Sulfolobus acidocaldarius DSM 639 (Sulfolobus acidocaldarius 균주로부터 피리미딘 영양요구주의 분리 및 특성 연구)

  • Choi, Kyoung-Hwa;Cha, Jae-Ho
    • Journal of Life Science
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    • v.21 no.10
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    • pp.1370-1376
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    • 2011
  • To study the functional genomic analysis of a crenachaeon Sulfolobus acidocaldarius, we have constructed an auxotrophic mutant based on pyrEF, which encodes the pyrimidine biosynthetic enzymes orotate phosphoribosyltransferase and orotidine-5'-monophosphate decarboxylase. S. acidocaldarius was shown to be sensitive to 5-fluoroorotic acid (5-FOA), which can be selected for mutations in pyrEF genes within a pyrimidine biosynthesis cluster. Spontaneous 5-FOA-resistant mutants by ultraviolet, KH1U and KH2U, were found to contain two point mutations and a frame shift mutation in pyrE, respectively. Mutations at these sites from KH1U and KH2U decreased the activity of orotate phosphoribosyltransferase encoded by the pyrE gene and blocked the degradation of 5-FOA into toxic 5-FOMP and 5-FUMP that kill the cells. Therefore, KH1U and KH2U were uracil auxotrophs. Transformation of Sulfolobus-Escherichia coli shuttle vector pC bearing pyrEF genes from S. solfataricus P2 into S. acidocaldarius mutant KH2U restored 5-FOA sensitivity and overcame the uracil auxotrophy. This study establishes an efficient genetic strategy towards the systematic knockout of genes in S. acidocaldarius.

Isolation of Temperature-sensitive Mutant Escherichia coli YrdC Involved in Universal t6 A tRNA Synthesis (대장균에서 t6 A tRNA의 생합성에 관여하는 필수 단백질 YrdC의 온도 민감형 돌연변이 분리)

  • Hwang, Jihwan
    • Journal of Life Science
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    • v.28 no.2
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    • pp.257-264
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    • 2018
  • The YrdC superfamily is a group of proteins that are highly conserved in almost all organisms sequenced so far. YrdC in Escherichia coli was suggested to be involved in ribosome biogenesis, translation termination, cold adaptation, and threonylcarbamoyl adenosine formation in tRNA. In this study, to unambiguously demonstrate that yrdC is essential in E. coli, we constructed two yrdC mutant strains of E. coli and examined their phenotypes. In the temperature-sensitive yrdC mutant strain, cell growth stopped almost immediately under nonpermissive conditions and it appeared to accumulate 16S ribosomal RNA precursors without significant accumulation of 30S ribosomal subunits. We also cloned yeast and human homologs and demonstrated that they complement the E. coli yrdC-deletion strain. By mutational study, we demonstrated that the concave surface in the middle of the YrdC protein plays an important role in E. coli, yeast, and human versions. By comparison of two yrdC-deletion strains, we also unambiguously demonstrated that yrdC is essential for viability in E. coli and that the functions of its yeast and human homologs overlap with that of E. coli YrdC.

Improvement of Chromosome Doubling Efficiency in Cymbidium Hybrids by Colchicine and Oryzalin Treatment (Colchicine과 oryzalin 처리에 의한 Cymbidium hybrids의 기내 배수화 효율 증진)

  • Hwang, Sook-Hyun;Kim, Mi-Seon;Park, So-Young
    • Horticultural Science & Technology
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    • v.33 no.6
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    • pp.900-910
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    • 2015
  • This study aimed to investigate the efficiency of colchicine and oryzalin in inducing polyploidy in two Cymbidium hybrids [Showgirl 'Silky' and Mystery Island 'Silk Road' (Silk Road-4)]. Colchicine was used at concentrations ranging from 50 to $500mg{\cdot}L^{-1}$, with treatments lasting 1 to 3 weeks. Oryzalin was used at concentrations ranging from 3 to $20mg{\cdot}L^{-1}$, with treatments lasting 3 to 6 days or 1 to 3 weeks. The survival rate of PLBs was better in colchicine than in oryzalin solutions. The ploidy levels were screened using flow cytometry. In C. Showgirl 'Silky', the highest chromosome doubling efficiencies were obtained with the 1-week treatment in $50mg{\cdot}L^{-1}$ colchicine (60%) and the 2-week treatment in $5mg{\cdot}L^{-1}$ oryzalin (46.7%). In C. Mystery Island 'Silk Road' (Silk Road-4), the highest chromosome doubling efficiencies were obtained with the 1-week treatment in $50mg{\cdot}L^{-1}$ colchicine (16.7%) and the 3-day treatment in $10mg{\cdot}L^{-1}$ oryzalin (6.7%). Colchicine was more efficient than oryzalin in terms of polyploidy induction. Furthermore, pre-treatment, which entailed poking 10 times with forceps, improved the efficiency of chromosome doubling.

Effect of Disease Resistance on Oral Administration of Lightyellow Sophora Extract in Olive Flounder (생약제 고삼 뿌리 열수추출물의 넙치 투여시 질병 저항성에 미치는 영향)

  • SEO, Jung-Soo;JEON, Eun-Ji;KWON, Mun-Gyeong;HWANG, Jee-Youn;KIM, Jin-Do;JUNG, Sung-Hee;KIM, Na-Young;JEE, Bo-Young;PARK, Myoung-Ae
    • Journal of Fisheries and Marine Sciences Education
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    • v.27 no.6
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    • pp.1656-1664
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    • 2015
  • The effects of lightyellow sophora, Sophora flavescens extract were tested on non-specific immune response and a disease resistance of olive flounder, Paralichthys olivaceus. For feeding trial, weight gain of fish fed different treated groups were not significant differences. Serum analysis of treated and control group did not demonstrated any differences. Lysozyme activity in the kidney and spleen of the 0.05% treated group on $4^{th}$ and $8^{th}$ week showed significant increase, respectively. In addition, phagocytic activity of the 0.05% treated group on $4^{th}$ and 8th week showed significant increase compared to the control group. Histopathology of the liver and kidney tissue of the administrated group for $4^{th}$, $8^{th}$ and $12^{th}$ week showed no particular signs of tissue degeneration. The treated group was higher than control group by analyzing the relative percent survival (RPS) of the experimental challenge of Edwardsiella tarda. Therefore, the lightyellow sophora showed effective in preventing fish-pathogenic edwardsiellosis for a certain period of time.

Incidence and Control of Coliform Bacteria in the Manufacturing of Commercial Kimchi (상업용 김치 생산과정에서 대장균유사세균의 발생과 억제)

  • Lee, Su-Jin;Han, Tae-Won;Kim, Ma-Rie;Seul, Keyung-Jo;Park, Yu-Mi;Jin, Ing-Nyol;Ghim, Sa-Youl
    • Microbiology and Biotechnology Letters
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    • v.37 no.2
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    • pp.91-98
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    • 2009
  • As consumption of kimchi has increased, factories have begun to produce this traditional Korean fermented vegetable dish on a large scale. Following the rise in manufacturing, the hygienic conditions under which commercial kimchi is being made have become an issue. We isolated 17 coliform bacteria from commercial kimchi that had not been fully fermented. These bacteria were partially identified as one of seven different species from three genera by 16S rDNA sequence analysis as follows: Enterobacter intermedius, Ent. cloacae, Ent. amnigenus, Klebsiella terrigena, K. ornithinolytica, K. oxytoca, and Hafnia alvei. Lactobacillus paraplantarum KNUC25 has been isolated from over-fermented Chinese cabbage kimchi and its antimicrobial activity reported in the literature. In our study, the KNUC25 strain showed antibacterial activity against isolated coliform bacteria and some pathogenic coliform bacteria through spot-on-the-lawn tests and viable cell tests. Through development and use of a cell-free supernatant of L. paraplantarum KNUC25, we effectively controlled coliform bacteria in commercial kimchi.

THE EFFECT OF DIFFERENTIAL MODULATION OF N-METHYL-D-ASPART ATE RECEPTOR ON THE VIABILITY OF PRIMARY CULTURED NORMAL HUMAN ORAL KERATINOCYTES (N-methyl-D-aspartate 수용기의 다양한 조절이 일차 배양된 정상사람구강각화세포의 생존에 미치는 영향)

  • Kim, In-Soo;Lee, Won;Kim, Seong-Hun;Choi, Bohm
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.28 no.4
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    • pp.277-286
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    • 2006
  • In the present study, I investigated the effects of N-methyl-D-aspartate (NMDA), arachidonic acid (AA), and Nitric Oxide Synthase Inhibitor (NOS-I), alone or in combination, on the viability of cultured primary normal human oral keratinocytes (NHOK). Specifically, we examined whether AA and NOS-I could protect primary NHOK from glutamate cytotoxicity. The purpose of this study was therefore the preliminary study for the examination of the interaction between these agents and NHOK in order to elucidate the mechanisms by which epithelial growth and regeneration are regulated. NHOK were obtained from gingival tissue of 20 individuals aged 20 to 29, and third passage (P3) cells were used for this study. Cell viability was measured by the MTT assay. NMDA and NNA, a calcium dependent NOS inhibitor, induced an initial increase in cell number, which subsequently decreased by the $7^{th}$ day. Low concentration of AA ($0.5\;{\mu}M$ & $1\;{\mu}M$) induced an increase in cell number while high concentrations of AA ($5\;{\mu}M$ & $10\;{\mu}M$) induced a decrease in cell number. The decrease in cell number induced by NMDA at the $7^{th}$ day was abolished by the addition of low concentrations of AA ($0.5\;{\mu}M$ & $1\;{\mu}M$) or NOS inhibitors. Low concentrations of AA ($0.5\;{\mu}M$ & $1\;{\mu}M$) or NOS inhibitors may protect the NHOK from NMDA induced cytotoxicity. These reactions might be related to the NMDA receptor in the cell and the change of the intracellular calcium ion concentration.

AN EXPERIMENTAL STUDY ON FAT CELL VIABLITY ACCORDING TO DIFFERENT HARVESTING TECHNIQUES (지방 채취 방법에 따른 지방 세포의 생존성에 대한 연구)

  • Lee, Won-Deok;Choi, Jin-Young
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.30 no.1
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    • pp.22-29
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    • 2008
  • Purpose: The purpose of this study is to test the efficacy of various methods of fat harvesting in animal model by viability comparison with assay including cell counting, MTT assay, and histologic evaluation. Materials and methods: New Zealand white rabbits experiments were used. Groin fat pads were subjected to different harvest method varying ingredients of solution(Experiment 1: T1 solution= lidocaine 1000mg/L, epinephrine 1mg/L, sodium bicarbonate 10mgEq/L, Triamcinolone 10mgEq/L; T2 solution=lidocaine 1000mg/L, epinephrine 1mg/L, sodium bicarbonate 0mgEq/L, Triamcinolone 0mgEq/L) and pressure exerted on harvesting with Luer-Lock syringe connected to suction cannula.(Experiment 2: P1 group=3cc intermittent pressure; P2 group=10cc sustained pressure) Fat cell viability was assessed with cell counting with a hemocytometer, MTT assay, and histologic evaluation. Results: Experiment 1 Cell count: T1=2.4/3.4/4.2, T2=9.6/8.4/7.2($\times10^5$ per mL); MTT assay: T1=0.516/0.41/0.453/0.412/0.421, T2=0.925/0.765/0.54/0.634/0.614 in 21 days(absorbance); Histology: T1 showed elongated and, different in size and shape, and ruptured adipocytes with only a few normal adipocytes whereas T2 showed central core of fat with almost intact fat cells Experiment 2 Cell count: P1=1.2/3.2/4.2, P2=1.2/2.4/3.8($\times10^5$ per mL); MTT assay:P1=0.256/0.245/0.258/0.21/0.264, P2=0.12/0.231/0.245/0.313/0.281 in 21 days(absorbance); Histology: P1 showed somewhat evenly distributed normal-looking fat cells and P2 showed relatively irregular shape of fat cells with small blood vessel amongst adiopocytes. Conclusion: Viability was higher in ‘modified tumescent solution’without sodium bicarbonate and triamcinolone and we also found no significantly different viability between using intermittent pressure and using sustained pressure. But in terms of initial viability of fat cell, we can assume that lower intermittent pressure would make better clinical results.

THE EFFECT OF PHOTODYNAMIC THERAPY ON THE VIABILITY OF STREPTOCOCCUS MUTANS ISOLATED FROM ORAL CAVITY (광역동 치료가 구강 내에서 분리한 수종의 Streptococcus mutans의 생존력에 미치는 영향)

  • Jung, Ji-Sook;Park, Ho-Won;Lee, Ju-Hyun;Seo, Hyun-Woo;Lee, Si-Young
    • Journal of the korean academy of Pediatric Dentistry
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    • v.39 no.3
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    • pp.233-241
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    • 2012
  • Photodynamic therapy (PDT) is a technique that involves the activation of photosensitizer by light in the presence of tissue oxygen, resulting in the production of reactive radicals capable of inducing cell death. The aim of this study was to evaluate the effect of PDT on Streptococcus mutans in planktonic conditions, previously treated with different photosensitive concentrations of erythrosine, using halogen and LED curing unit as a light source. And we compared the effects of PDT on six strains of S. mutans isolated from oral cavity and reference strain. As a result, S. mutans was susceptible to the combination of hand held photopolymerizer (HHP) and erythrosine. The higher concentration of erythrosine in the presence of light irradiation induced greater effects in reduction of viability of S. mutans. Isolated S. mutans showed a significant reduction in bacterial counts of the groups submitted to PDT compared to the control groups. And they appeared to be similar or slightly lower antimicrobial effect compared with reference strain. However, the difference was not significant (p < 0.05). In conclusion, PDT using erythrosine as a photosensitizing agent and HHP as a light source could be an efficient option for diseases caused by S. mutans.