• 한국동물생명공학회지
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• 제37권3호
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• pp.169-175
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• 2022

Bovine mammary epithelial (MAC-T) cells are commonly used to study mammary gland development and mastitis. Lipopolysaccharide is a major bacterial cell membrane component that can induce inflammation. Autophagy is an important regulatory mechanism participating in the elimination of invading pathogens. In this study, we evaluated the mechanism underlying bacterial mastitis and mammary cell death following lipopolysaccharide treatment. After 24 h of 50 ㎍/mL lipopolysaccharide treatment, a significant decrease in the proliferation rate of MAC-T cells was observed. However, no changes were observed upon treatment of MAC-T cells with 10 ㎍/mL of lipopolysaccharide for up to 48 h. Thus, upon lipopolysaccharide treatment, MAC-T cells exhibit dose-dependent effects of growth inhibition at 10 ㎍/mL and death at 50 ㎍/mL. Treatment of MAC-T cells with 50 ㎍/mL lipopolysaccharide also induced the expression of autophagy-related genes ATG3, ATG5, ATG10, ATG12, MAP1LC3B, GABARAP-L2, and BECN1. The autophagy-related LC3A/B protein was also expressed in a dose-dependent manner upon lipopolysaccharide treatment. Based on these results, we suggest that a high dose of bacterial infection induces mammary epithelial cell death related to autophagy signals.

• Toxicological Research
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• 제24권1호
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• pp.37-44
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• 2008

The in vitro cell transformation assays (CTA) were performed using BALB/3T3 murine fibroblasts and HaCaT human keratinocytes in order to evaluate concordance between both in vitro CTAs and carcinogenicity with compounds differing in their genotoxic and carcinogenic potential. Six test articles were evaluated, two each from three classes of compounds: genotoxic carcinogens (2-amino-5-nitrophenol and 4-nitroquinoline-N-oxide), genotoxic noncarcinogens (8-hydroxyquinoline and benzyl alcohol), and nongenotoxic carcinogens (methyl carbamate and N-nitrosodiphenylamine). Any foci of size $\geq$2 mm regardless of invasiveness and piling was scored as positive in CTA with BALB/3T3. As expected, four carcinogens regardless of their genotoxicity had positive outcomes in two-stage CTA using BALB/3T3 cells. However, of the two genotoxic noncarcinogens, benzyl alcohol was positive CTA finding. We concluded that, of the 6 chemicals tested, the sensitivity for BALB/3T3 system was reasonably high, being 100%. The respective specificity for BALB/3T3 assay was 50%. We also investigated the correlation between results of BALB/3T3 assay and results from HaCaT assay in order to develop a reliable human cell transformation assay. However, evaluation of staining at later time points beyond the confluency stage did not yield further assessable data because most of HaCaT cells were detached after $2{\sim}3$ days of confluency. Thus, after test article treatment, HaCaT cells were split before massive cell death began. In this modified protocol for this HaCaT system, growing attached colonies were counted instead of transformed foci 3 weeks since last subculture. Compared to BALB/3T3 assay, HaCaT assay showed moderate low sensitivity and high specificity. Despite these differences in specificity and sensitivity, both cell systems did exhibit same good concordance between in vitro CTA and rodent carcinogenicity findings (overall 83% concordant results). At present the major weakness of these in vitro CTA is lack of validation for regulatory acceptance and use. Thus, more controlled studies will be needed in order to be better able to assess and quantitatively estimate in vitro CTA data.

• 생명과학회지
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• 제25권1호
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• pp.93-100
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• 2015

Pachymic acid는 복령에서 분리된 lanostane-type인 triterpenoid의 일종이다. 최근 pachymic acid가 항암 및 항염증 효능과 산화적 스트레스에 대한 항산화 효능 등과 같은 약리적인 효능이 있는 것으로 밝혀지고 있으나, 그에 대한 구체적인 분자생물학적 기전 연구는 매우 미비한 실정이다. 본 연구에서는 pachymic acid의 항암활성 및 관련 기전 조사의 일환으로 T24 인체 방광암세포 모델을 이용하여 pachymic acid에 의한 apoptosis 유발 여부를 검증하였다. 본 연구의 결과에 의하면 pachymic acid는 T24 세포의 증식을 유의적으로 억제시켰으며, 이는 apoptosis 유발과 연관성이 있음을 다양한 방법으로 확인하였다. Pachymic acid에 의한 apoptosis 유도에는 pro-apoptotic 인자들의 발현 증가와 anti-apoptotic 유전자 산물들의 발현 감소가 동반되었으며, MMP의 소실과 tBid의 발현 증가가 관찰되었다. 아울러 pachymic acid는 extrinsic 및 intrinsic apoptosis 경로의 개시에 관여하는 caspase-8 및 -9의 활성뿐 만 아니라, caspase-3의 활성도 증가시킴으로서 PARP와 같은 기질 단백질의 단편화를 초래하였다. 따라서 pachymic acid는 항암활성을 지니는 천연생리활성 물질로서의 잠재력이 매우 높음을 알 수 있었다.

• Journal of Periodontal and Implant Science
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• 제38권sup2호
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• pp.299-308
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• 2008

Purpose: The aim of this study was to evaluate adhesion and gene expression of the MC3T3-E1 cells cultured on machined titanium surface (MS) and anodized titanium surface (AS) using MTT test, Scanning electron micrograph and cDNA microarray. Materials and Methods: The MTT test assay was used for examining the proliferation of MC3T3-E1 cells, osteoblast like cells from Rat calvaria, on MS and AS for 24 hours and 48 hours. Cell cultures were incubated for 24 hours to evaluate the influence of the substrate geometry on both surfaces using a Scanning Electron Micrograph (SEM). The cDNA microarray Agilent Rat 22K chip was used to monitor expressions of genes. Results: After 24 hours of adhesion, the cell density on AS was higher than MS (p < 0.05). After 48 hours the cell density on both titanium surfaces were similar (p > 0.05). AS had the irregular, rough and porous surface texture. After 48 hours incubation of the MC3T3-E1 cells, connective tissue growth factor (CTGF) was up-regulated on AS than MS (more than 2 fold) and the insulin-like growth factor 1 receptor was down-regulated (more than 2 fold) on AS than MS. Conclusion: Microarray assay at 48 hours after culturing the cells on both surfaces revealed that osteoinductive molecules appeared more prominent on AS, whereas the adhesion molecules on the biomaterial were higher on MS than AS, which will affect the phenotype of the plated cells depending on the surface morphology.

• 한국멀티미디어학회논문지
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• 제24권10호
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• pp.1326-1335
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• 2021

As part of the cell division method, we proposed a method for segmenting images generated by topography microscopes through deep learning-based feature generation and graph segmentation. Hybrid vector shapes preserve the overall shape and boundary information of cells, so most cell shapes can be captured without any post-processing burden. NIH-3T3 and Hela-S3 cells have satisfactory results in cell description preservation. Compared to other deep learning methods, the proposed cell image segmentation method does not require postprocessing. It is also effective in preserving the overall morphology of cells and has shown better results in terms of cell boundary preservation.

• 한국환경보건학회지
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• 제21권4호
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• pp.44-48
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• 1995

26 bacterial strains capable of growing on Terephthalic acid (TPA) in minimal medium were isolated from soil and wastewater by selective enrichment culture, and among them, one isolate which was the best in the cell growth and TPA degradation was selected and identified as Pseudomonas sp. T-1 by its characteristics. Cell growth almost revealed a stationary phase at 24 hrs after cultivation. Cell growth dramatically increased in a minimal medium containing 0.1% of TPA as a sole carbon source and TPA was not detected any more at 80 hrs after cultivation. Therefore, it is suggested that Pseudomonas sp. T-1 could be effectively used for the biological treatment of wastewater containing TPA.

• IMMUNE NETWORK
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• 제24권2호
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• pp.14.1-14.26
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• 2024

The inflammatory response during cutaneous leishmaniasis (CL) involves immune and non-immune cell cooperation to contain and eliminate Leishmania parasites. The orchestration of these responses is coordinated primarily by CD4⁺ T cells; however, the disease outcome depends on the Th cell predominant phenotype. Although Th1 and Th2 phenotypes are the most addressed as steers for the resolution or perpetuation of the disease, Th17 cell activities, especially IL-17 release, are recognized to be vital during CL development. Th17 cells perform vital functions during both acute and chronic phases of CL. Overall, Th17 cells induce the migration of phagocytes (neutrophils, macrophages) to the infection site and CD8⁺ T cells and NK cell activation. They also provoke granzyme and perforin secretion from CD8⁺ T cells, macrophage differentiation towards an M2 phenotype, and expansion of B and Treg cells. Likewise, immune cells from the inflammatory infiltrate have modulatory activities over Th17 cells involving their differentiation from naive CD4⁺ T cells and further expansion by generating a microenvironment rich in optimal cytokines such as IL-1β, TGF-β, IL-6, and IL-21. Th17 cell activities and synergies are crucial for the resistance of the infection during the early and acute stages; however, if unchecked, Th17 cells might lead to a chronic stage. This review discusses the synergies between Th17 cells and the inflammatory infiltrate and how these interactions might destine the course of CL.

• Journal of electromagnetic engineering and science
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• 제15권3호
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• pp.142-150
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• 2015

Several reports supported that continuous exposure to 60 Hz magnetic field (MF) induces testicular germ cell apoptosis in vivo. We recently evaluated duration- and dose-dependent effects of continuous exposure to a 60 Hz MF on the testes in mice. BALB/c male mice were exposed to a 60 Hz MF at $100{\mu}T$ for 24 hours a day for 2, 4, 6, or 8 weeks, and at 2, 20 or $200{\mu}T$ for 24 hours a day for 8 weeks. To induce the apoptosis of testicular germ cell in mice, the minimum dose is $20{\mu}T$ at continuous exposure to a 60 Hz MF for 8 weeks, and the minimum duration is 6 weeks at continuous exposure of $100{\mu}T$. Continuous exposure to a 60 Hz MF might affect duration- and dose-dependent biological processes including apoptotic cell death and spermatogenesis in the male reproductive system of mice. The safety guideline of the International Commission on Non-Ionizing Radiation Protection (ICNIRP) indicates that the permissible maximum magnetic flux density for general public exposure is $200{\mu}T$ at 60 Hz EMF (ICNIRP Guidelines, 2010). In the present study, we aimed to examine the expression of pro- and anti-apoptotic genes regulated by the continuous exposure to 60 Hz at $200{\mu}T$ in Sprague-Dawley rats for 20 weeks. The continuous exposure to 60 Hz at $200{\mu}T$ does not affect the body and testicular weight in rats. However, exposure to 60 Hz MF significantly affects testicular germ cell apoptosis and sperm count. Further, the apoptosis-related gene was scrutinized after exposure to 60 Hz at $200{\mu}T$ for 20 weeks. We found that the message level of endonuclease G (EndoG) was greatly increased following the exposure to 60 Hz at $200{\mu}T$ compared with sham control. These data suggested that 60 Hz magnetic field induced testicular germ cell apoptosis through mitochondrial protein Endo G.

• 대한한의학회지
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• 제24권3호
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• pp.1-10
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• 2003

Background : Asthma is a chronic inflammatory disorder under immunological influence. Gamichunggumgangwha-tang (CG, Jiaweiqingjinjianghuo-tang) and Gamiyukmigiwhang-tang (YM, Jiaweiliuweidihuang-tang) are herbal tonics for asthma from traditional herbal medicine. Objective : To evaluate the effect of CG and YM on immune cell & serum OA-specific IgE in broncho-alveolar lavage fluid (BALF) in a rat asthma model. Materials and Methods : Rats were sensitized with OA; at day I sensitized group and CG and YM groups were systemically immunized by subcutaneous injection of 1mg OA and 300mg of Al(OH)3 in a total volume of 2ml. At the same time, 1 ml of 0.9% saline containing $6{\times}10^9$ B. pertussis bacilli was injected by Lp. 14 days after the systemic immunization, rats received local immunization by inhaling 0.9% saline aerosol containing 2% (wt/vol) OA. A day after local immunization, BALF was collected from the rats. Rats were orally administered with each of CG and YM extract for 14 days since the day after local immunization. Lymphocyte, CD4＋ T cell and CD8＋ T cell counts, CD4＋/CD8＋ ratio in BALF, change of serum OA-specific IgE level, CD4＋ T cell and CD8＋ T cell percentages in the peripheral blood were measured and evaluated. Results : CG and YM showed an alleviating effect on asthmatic responses of rats. CG decreased total cell, lymphocyte, CD4＋ T cells in BALF, and serum OA-specific IgE level as compared with the control group. YM decreased lymphocytes as compared with the control group. CD4＋/CD8＋ ratio in BALF from the CG and YM groups and serum OA-specific IgE level from the YM group didn't show any significant variation from the control group. Conclusion : CG alleviated asthmatic hyperreactivity of the immune system through CD4＋ T cells and serum IgE. Further the study of this immune system modulating mechanism is expected.

• Parasites, Hosts and Diseases
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• 제31권3호
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• pp.249-258
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• 1993

원발성 아메바성 수막뇌염을 야기하는 Naegleria fowleri를 사망율에 따라 $1{\;}{\times}{\;}10^4$개 아메바 영양형 접종군과. $1{\;}{\times}{\;}10^5$개 아메바 영양형 접종군으로 나누어 비장세포를 비특이 mitogen인 Phytohemagglutinin(PHA)과 특이항원인 N. fowleri Iysates로 자극하여 T 세포의 interleukin-2 생성정도를 측정하고 T 림프구의 아형 및 아세포화정도를 측정한 결과 N.fowleri 아메바 영양형 $1{\;}{\times}{\;}10^5$개 접종군에서의 마우스 사망율은 72.2%였으며 $1{\;}{\times}{\;}10^4$개 접종군에서의 마우스 사망율은 14.3%였다. 또한 IL릭 생성정도를 접종후 7, 14, 24일째 측정한 결과. 접종 후 14일째에는 두 실험군 모두에서 대조군과 비교하여 IL닉의 생성이 유의하게 감소하였으며 접종 후 24일째에는 두 실험군 모두에서 접종 후 14일째 보다는 증가하였으나 대조군과 비교하여 유의하게 억제되었다 비장세포내 T 림프구 아형의 변동은 전체 비장 림프구에 대한 $Thy-1.2^{+}{\;}T.{\;}L3T4^{+}{\;}T,{\;}Ly^{2+}{\;}T$ 세포는 N.fowleri아메바 영양형 $1{\;}{\times}{\;}10^5$개 접종군에서 접종 후 7일째 대조군과 비교하여 유의하게 증가하였고 접종 후 14일째와 24일째에는 대조군과 비교하여 유의한 차이가 없었다. 아메바 영양형 $1{\;}{\times}{\;}10^4$개를 접종시킨 실험군은 경과 일수에 관계없이 대조군과 비교하여 유의한 차이가 없었다. 비장세포내 T 세포의 DNA의 분획을 관찰한 결과 두 실험군 모두에서 접종 후 7일째에 S phase 분획이 가장 높이 증가하였으며 접종 후 14일과 24일째에도 대조군에 비하여는 유의하게 증가하여 있었다. 이상의 결과를 종합하여 볼 때, N.fowleri를 사망율을 기준으로 접종량을 달리하여 아메바 영양형 $1{\;}{\times}{\;}10^4$개 접종군과 $1{\;}{\times}{\;}10^5$개 접종군으로 나누어 접종하였을 때, 접종 후 7일을 전후하여 IL-2를 매개로 활성화되는 세포매개성 면역이 N.fowleri감염의 방어기작으로 작용하는 것으로 생각되며 아메바 접종 후 14일째에는 치명적인 수막뇌염으로 진행되어 비장세포의 IL-2의 생성능력이 매우 억제된 것으로 생각된다. 또한 IL-2 생성능력과 T 세포의 아세포화의 증가 및 T세포 아령의 수의 변동과는 잘 일치되지 않는 것으로 나타났다.