• 미생물학회지
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• 제38권2호
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• pp.107-112
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• 2002

남조류 분해세균을 분리하기 위하여 도창, 팔당 저수지와 석촌 호수로부터 저층시료를 채취하였다. Anabaena cylindrica lawn에 각각의 시료를 도말한 후 11 일간 배양하였고 석촌 시료를 접중한 Anabaena cylindrica lawn에서 Anabaena cylindrica에 분해능을 가지는 세균을 분리하였다. 분해능 화인은 남조류와 분해세균을 혼합한 후 chlorophyll a간의 측정과 분리세균의 cell counting방법으로 확인하였고, 세포외 물질이 Anabaena cylindrica를 분해하는 것을 확인하였다. 분리균주는 16S rDNA 염기서열 분석과 형태학적, 생리학적 특징을 기초로 하여 동정하였다. 분리균주의 165 rDNA염기서열을 기초로한 분자계통학적 분석을 통하여 Bacillus속에 포함하는 계통학적 그룹에 속한다는 것을 확인하였고, Bacillus sp. CHSl으로 명하였다.

• 미생물학회지
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• 제25권3호
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• pp.244-248
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• 1987

자연 발효과정중에 GRam 향성 및 음성세균의 수를 신속하고동시에 측정할 t ndlT는 방법을 검토하였다. 본 연구에서는 일반적으로 사용되고 있는 Gram 염색법을 응용하였다. 그 결과로써 Gram 염색법에의 세균수는 분광광도계에 의한 혼탁도와 A610=0.7의 범위내에서 비례관계가 성립하였다. 염색조작중 세척에 의한 균의 제거는 약 8%를 초과하지 않았다. 그리고 Escherichia coli와 Micrococcus luteus의 혼합액에서 이들의 분리 측정에 대한 표준오차는 $5.1\pm2.3$%이었다. 계수가능범위는 $5.5\times 10^{7}-1.0\times 10^{9}$세포 / ml 이었다. 그러므로 일반적인 Gram 염색법은 Gram 양성 및 음성 개체군이 혼합된 배양약에서 이들을 분별 측정하는데도 적용될 수 있다고 본다. 실제적으로, 대마침지와 김치 발효과정에서 균 생장의 동역학적 관계를 검토하였다.

• Applied Microscopy
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• 제51권
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• pp.4.1-4.12
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• 2021

Fluorescence in situ hybridization (FISH) is a technique to visualize specific DNA/RNA sequences within the cell nuclei and provide the presence, location and structural integrity of genes on chromosomes. A confocal Whole Slide Imaging (WSI) scanner technology has superior depth resolution compared to wide-field fluorescence imaging. Confocal WSI has the ability to perform serial optical sections with specimen imaging, which is critical for 3D tissue reconstruction for volumetric spatial analysis. The standard clinical manual scoring for FISH is labor-intensive, time-consuming and subjective. Application of multi-gene FISH analysis alongside 3D imaging, significantly increase the level of complexity required for an accurate 3D analysis. Therefore, the purpose of this study is to establish automated 3D FISH scoring for z-stack images from confocal WSI scanner. The algorithm and the application we developed, SHIMARIS PAFQ, successfully employs 3D calculations for clear individual cell nuclei segmentation, gene signals detection and distribution of break-apart probes signal patterns, including standard break-apart, and variant patterns due to truncation, and deletion, etc. The analysis was accurate and precise when compared with ground truth clinical manual counting and scoring reported in ten lymphoma and solid tumors cases. The algorithm and the application we developed, SHIMARIS PAFQ, is objective and more efficient than the conventional procedure. It enables the automated counting of more nuclei, precisely detecting additional abnormal signal variations in nuclei patterns and analyzes gigabyte multi-layer stacking imaging data of tissue samples from patients. Currently, we are developing a deep learning algorithm for automated tumor area detection to be integrated with SHIMARIS PAFQ.

• Molecules and Cells
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• 제24권3호
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• pp.424-430
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• 2007

The biological effects of low-dose radiation have been investigated and debated for more than a century, but its cellular effects and regulatory mechanisms remain poorly understood. This study shows the human cellular responses to low-dose radiation in CCD-18 Lu cells, which are derived from normal human lung fibroblasts. We examined a colony-forming assay for cell survival by ionizing radiation. Live cell counting and cell cycle analysis were measured for cell proliferation and cell cycle progression following low-dose irradiation. We examined Raf and Akt phosphorylation to determine the proliferation mechanism resulting from low-dose radiation. We also observed that p53 and p21 were related to cell cycle response. We found that 0.05 Gy of ionizing radiation enhanced cell proliferation and did not change the progression of the cell cycle. In addition, 0.05 Gy of ionizing radiation transiently activated Raf and Akt, but did not change phospho-p53, p53 and p21 in CCD-18 Lu cells. However, 2 Gy of ionizing radiation induced cell cycle arrest, phosphorylation of p53, and expression of p53 and p21. The phosphorylation of Raf and Akt proteins induced by 0.05 Gy of ionizing radiation was abolished by pre-treatment with an EGFR inhibitor, AG1478, or a PI3k inhibitor, LY294002. Cell proliferation stimulated by 0.05 Gy of ionizing radiation was blocked by the suppression of Raf and Akt phosphorylation with these inhibitors. These results suggest that 0.05 Gy of ionizing radiation stimulates cell proliferation through the transient activation of Raf and Akt in CCD-18 Lu cells.

• Restorative Dentistry and Endodontics
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• 제20권1호
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• pp.71-95
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• 1995

Bacteria have been regarded as a one of major etiologic factors in root canal infections. In endodontic treatment the effective removal of pathogenic microorganisms in the root canal is the key to successful outcome. Bacterial cell wall components may play an important role in the development of pulpal and periapical disease. The purpose of this study was to evaluate the effect of sonic extracts of Streptococcus spp. on cultured L929 cells and to characterize cell wall protein profiles of Streptococcus spp. Streptococcus spp. were isolated from infected root canals and identified with Vitek Systems(Biomeriux, USA). Five streptococci, namely S. sanguis, S. mitis, S uberis, S. mutans (ATCC 10449) and S. faecalis (ATCC 19433) weere enriched in brain heart infusion broth. Cell pellets were sonicated and cell wall extracts were dialyzed and membrane filtered. Prepared cell wall proteins were applied to cultured L929 cell. The cell reaction were evaluated by monitoring DNA synthesis, cell numbers and the change of cell morphology. The total cell wall protein profiles of microorganisms were characterized by sodium dodecyl sulfate polyacrylamide-gel eledruphoresis(SDS-PAGE). DNA synthesis of L929 cells were reduced by the increasing concentration of sonic extracts. DNA synthesis was significantly suppressed in more than $50{\mu}g$/ml of sonic extract conentration in five streptococci. S. nutans (ATCC 10449) showed stronger suppression on DNA synthesis than remaining four streptococci, which had the similar effect on DNA synthesis. Analysis of DNA synthesis measured by [$^3H$]-thymidine uptake was more sensitvie method than cell counting. Sonic extracts affected the microscopic findings of L929 cells. The protein profiles indicated that all five strains shared two major proteins with molecular masses of 70.8 and 57.5 kD respectively. S. uberis and S. mutans shared common minor proteins of which molecular weights were 147.9 and 112.2 kD respectively. However some minor proteins were unique for S. mitis, S. uberis and S. faecalis.

• Journal of Gastric Cancer
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• 제10권3호
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• pp.99-105
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• 2010

Purpose: Currently, the two most influential gastric stem cell marker candidates are CD44 and CD133. The aim of this study was to make a comparison and determine the appropriate marker for use in gastric cancer stem cell research. Materials and Methods: We analyzed the expressions of CD44, CD133, and CD24 from the gastric cancer cell lines MKN45, MKN74, KATO-III, NCI-N87, SNU-1, SNU-216, SNU-601, SNU-638, and SNU-688 using flow cytometry. In addition, we measured the change in viability after applying 5 fluorouracil (5-FU) to the MKN45, MKN74, KATO-III, and NCI-N87 cell lines using a Cell Counting Kit 8. Results: CD133 expression was above moderate in the KATO-III, SNU-216, SNU-601 cell lines, whereas it was below 1% in the remaining cell lines. CD44 was expressed at levels above 5% in all gastric cancer cell lines. The effect of 5-FU on viability and CD133 or CD44 expression in the cell lines were not related. Conclusions: Expression of CD133 positive cells was insufficient in the gastric cancer cell lines. Therefore, of the cell lines tested, CD44 was the most appropriate tumor maker for research on gastric cancer stem cells.

• Journal of Microbiology and Biotechnology
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• 제9권4호
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• pp.504-508
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• 1999

This report compares the selective cytotoxic effects of Doenjang fermented by various Bacillus strains (Bacillus sp. SS9, SSA3, and PM3) on human liver cell lines with that of conventional Doenjang (DTY, DTG, and DTK) and commercial Doenjang (DCM, DCD, and DCS). To investigate selective cytotoxic effects of Doenjang extracts, the cell density of HepG2 (Hepatocellular carcinoma) and CCL-13 (cells derived from human normal liver) was estimated after addition of the extracts by using a viable cell counting method. The maximum selectivity ratio ($IC_{50}$value against CCL-13/$IC_{50}$ value aganist HepG2) was observed by PM3 (extracts of Doenjang fermented with Bacillus sp. PM3). As for morphological changes shown by the addition of PM3 into HepG2 and CCL-13 cultures, HepG2 was significantly disrupted, however, CCL-13 was not affected. Also, the growth rate of HepG2 was decreased significantly by the addition of PM3. Consequently, PM3 showed a more detrimental effect on HepG2 than that on CCL-13.

• 동의생리병리학회지
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• 제27권5호
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• pp.617-624
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• 2013

The purpose of this study is to investigate the protective effects of 25 herbal formulas on acetaminophen (APAP) or D-galactosamine (D-GalN)-induced hepatotoxicity in primary rat hepatocytes. Cell viability was measured using by Cell Counting Kit-8. 15 kinds of herbal formulas significantly reversed the cell viabilities of D-GalN-treated rat hepatocytes compared with D-GalN alone (p<0.05). In particular, 9 herbal formulas (Bangpungtongseong-san, Bojungikgi-tang, Galgeun-tang, Gumiganghwal-tang, Guibi-tang, Sagunja-tang, Samsoeum, Pyeongwi-san and Yijin-tang) showed the potent protective effects. However, 8 herbal formula exerted weak protective effects and 2 herbal formula did not exert effects on hepatotoxicity by D-GalN. On APAP-induced hepatotoxicity, 7 kinds of herbal formulas increased the viabilities of hepatocytes compare with APAP alone (p<0.05). These results could be provide a valuable information for the future in vivo or clinical studies to predict the hepatoprotective effects of herbal formulas.

• Nutrition Research and Practice
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• 제14권5호
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• pp.453-462
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• 2020

BACKGROUND/OBJECTIVES: Platycodon grandiflorum (PG), an oriental herbal medicine, has been known to improve liver function, and has both anti-inflammatory and antimicrobial properties. However, little is known about the immune-enhancing effects of PG and its mechanism. In this study, we aimed to investigate whether fermented PG extract (FPGE), which has increased platycodin D content, activates the immune response in a murine macrophage cell line, RAW 264.7. MATERIALS/METHODS: Cell viability was determined by Cell Counting Kit-8 assay and the nitric oxide (NO) levels were measured using Griess reagent. Cytokine messenger RNA levels of were monitored by quantitative reverse transcription polymerase chain reaction. To investigate the molecular mechanisms underlying immunomodulatory actions of FPGE in RAW 264.7 cells, we have conducted luciferase reporter gene assay and western blotting. RESULTS: We found that FPGE treatment induced macrophage cell proliferation in a dose-dependent manner. FPGE also modulated the expression of NO and pro-inflammatory cytokines, such as tumor necrosis factor-α, interleukin (IL)-1β, and IL-6. The activation and phosphorylation levels of nuclear factor kappa B (NF-κB) were increased by FPGE treatment. Moreover, 5-aminoimidazole-4-carboxamide ribonucleotide, an activator of AMP-activated kinase (AMPK), significantly reduced both lipopolysaccharides- and FPGE-induced NF-κB reporter gene activity. CONCLUSIONS: Taken together, our findings suggest that FPGE may be a novel immune-enhancing agent acting via AMPK-NF-κB signaling pathway.

• KSBB Journal
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• 제11권1호
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• pp.52-57
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• 1996

평활근 세포에 대한 shear stress의 영향을 알아보기 위해 슬라이드 글라스위에 배양된 牛 대통맥의 평활근 세포를 120시간 동안 0~26 dyn/$cm^2$의 각기 다른 일정한 shear stress의 유체에 노출하였다. 실험 장치를 순환하는 배지의 lactate dehydrogen­a ase의 농도측정 결과, 본 연구에서 적용된 shear stress 범위에서는 유체의 흐름으로 인하여 세포가 슬라이드 글라스 표면으로부터 이탈되는 현상은 없었다. 표면 $cm^2$ 당 존재하는 세포의 수를 측정한 결과, 평활근 세포는 정체배양시 가장 빨리 성장하였다. 표변에서의 shear stress가 증가할수록 i영활근 세포의 성장은 늦었으며, shear stress가 17 dyn/$cm^2$ 이상에서는 세포의 성장이 관찰되지 않았다.