• 한국식품영양과학회지
• /
• 제45권7호
• /
• pp.1041-1048
• /
• 2016

본 연구에서 가금류의 주요 병원성 식중독 균을 인위적으로 오염시킨 훈제오리육을 진공포장 조건에서 10, 15, $24^{\circ}C$에 저장하면서 유통기한 동안 관찰한 미생물의 증식 및 생존 결과 Campylobacter jejuni는 저장기간 이내에 사멸하는 경향을 보였으며, Salmonella Typhimurium과 Listeria monocytogenes는 균주의 성장 속도에는 차이가 있었으나 증식하는 경향을 보였다. 훈제오리의 유통온도는 $10^{\circ}C$이며 유통기한이 약 30일인 것을 고려했을 때, 초기 오염 수준이 Campylobacter 균주에 의한 식중독을 유발하게 되는 균수 500 CFU/g 수준 이하에서는 유통기한 내에 문제가 없을 것으로 생각한다. 그러나 낮은 온도에서 저항성이 증가하며 살아있으나 배양은 불가능한 상태인 VBNC 상태의 C. jejuni의 특성에 따라 적절한 조건에서 회복되어 병원성을 일으킬 가능성이 있으므로 C. jejuni에 대한 지속적인 관리가 필요하다. 또한, S. Typhimurium과 L. monocytogenes의 경우 일반적인 유통/보관 온도인 $10^{\circ}C$에서도 증식이 가능하며, 특히 가공품 및 RTE 식품은 적절한 가열처리 없이 소비할 경우 식중독 발생 가능성이 높다는 점에서 제품 제조 단계에서부터 위생적인 관리가 필요하다. 본 연구에서는 C. jejuni biofilm cells을 인위적으로 오염시킨 훈제오리육을 진공포장 하여 일반 유통/보관 온도인 $10^{\circ}C$와 실온, 그리고 일반적으로 C. jejuni가 증식 가능한 온도인 $36^{\circ}C$에서 저장하였으나, C. jejuni biofilm cell은 훈제오리에서는 모든 온도에서 재증식이 불가능한 것으로 관찰되었다. 또한, $10^{\circ}C$의 저온에서 유도한 VBNC 상태의 C. jejuni를 훈제오리에 인위적으로 오염시키고 혐기적 조건에서 $42^{\circ}C$에 1일간 저장하며 VBNC 상태의 C. jejuni의 재증식 가능성을 분석하였으나, 최적 증식 온도인 $42^{\circ}C$에서도 재증식은 관찰되지 않았다. 이처럼 본 연구에서는 biofilm을 형성한 C. jejuni도 VBNC 상태의 C. jejuni는 살아 있으나 훈제오리에서의 증식은 관찰되지 않았다. 따라서 훈제오리에서의 C. jejuni의 위험성은 매우 적은 것으로 생각한다. 그러나 C. jejuni의 경우 매우 적은 양으로도 식중독을 일으킬 수 있고 C. jejuni biofilm 및 VBNC의 특성에 따라 잠재적인 위험성을 포함하는 동시에 유통/보관 온도인 냉장 온도에서 더 잘 살아남는다는 점에서 식중독 발생의 주요 원인으로 작용할 수 있는 교차오염과 전이를 예방하는 것이 중요하므로 이에 대한 관리가 강조되어야 할 것으로 생각한다.

• 대한세포병리학회지
• /
• 제7권1호
• /
• pp.44-50
• /
• 1996

Small cell carcinoma of the lung is characterized by cells with finely stippled chromatin and scanty cytoplasm as well as a particularly aggressive clinical course and favorable response to the chemotherapy. Recently percutaneous fine needle aspiration (FNA) biopsy has become both widely established and highly respected for the diagnosis of lung cancer. However metastatic small cell carcinoma of lymph node should be cytologically differentiated from the small round cell tumor of particular sites, especially malignant lymphoma, because small ceil carcinoma of classic oat cell type nay simulate small cell non-Hodgkin's lymphoma. We report five cases of metastatic small cell carcinoma of in-termediate cell type diagnosed by FNA of the enlarged lymph nodes of the neck and axilla. The cytologic smears contained diffuse small neoplastic cells larger than lymphocytes with dense, pyknotic nuclei and extremely scanty cytoplasm. Apparently viable large tumor cells have vesicular nuclei with granular, sometimes very coarse chromatin. The characteristic cytologic features of small cell carcinoma as compared to malignant lymphoma were as follows.: 1) small cells with dense pyknotic nuclei are evenly distributed in the background of apparently viable larger tumor cells, admixed with mature lymphocytes and phagocytic macrophages. 2) small loose aggregates of cells with nuclear melding are indicative of small cell carcinoma rather than non-Hodgkin's lymphoma. 3) the cytoplasmic and nuclear fragments of tumor necrosis are more dominant in the smears of small cell carcinoma. 4) nuclear membrane and nucleoli are generally indistinct in small cell carcinoma due to condensation of chromatin.

• KSBB Journal
• /
• 제9권1호
• /
• pp.48-54
• /
• 1994

HEK 세포 배양액 중에 존재하는 scu-PA와 tc-PA의 보다 정확한 측정을 위해 fibrin plate법과 기존의 amidolytic 방법을 변형시킨 측정법을 이용했다. 1%의 저혈청 배지를 이용한 T-flask 배양에서 $1.65{\times}10^6$(viable cells/ml)의 최대 세포수에 도달하여 1670(IU/ml)의 scu-PA 생산량을 보였으며 평균 10% 미만이 변환율을 보였다. 또한 Spinner vessel에서의 회분배양시 최대 세포수가 $4.43{\times}10^6(total cells/ml)$ 와 1560(IU/ml)의 scu-PA 생산량을 나타냈으며 평균 11.4%의 변환율을 보였다. 연속배양에서는 0.449(1/day)의 비생육속도와 $3.13{\times}10^{-4}(IU/cell)$의 비생산속도를 보였으며 평균 10.18% 정도의 전환율을 보였다. 이는 회분식 및 유가식 배양 결과와 큰 차이가 없으며 배양공정에 관계없이 약 90% 이상의 회수율이 가능하다는 것을 의미한다.

• 한국어병학회지
• /
• 제31권2호
• /
• pp.93-99
• /
• 2018

Edwardsiella tarda predominantly causes edwardsiellosis in fish at high temperature, but is rarely isolated from water when water temperature is low. However, E. tarda is viable but nonculturable (VBNC) in low water temperature, but it can be revived when water temperature rises and cause disease to fish. Therefore, in order to prevent disease, it is very important to identify pathogens that are in the VBNC state in environmental water. In this study, E. tarda cells in the VBNC state were detected by the ethidium monoazide (EMA)-PCR method using the low-temperature oligotrophic sea water microcosm obtained by inoculation of E. tarda at a concentration of $10^8CFU/ml$. In order to distinguish between live and dead bacteria in E. tarda, each sample was treated with EMA at different concentrations, photoactivated with a 500 W halogen lamp, and PCR was performed with E. tarda specific primer. At the concentration of $10^7CFU/ml$ bacterium, DNA amplification was observed only in the live cells when treated with $60{\mu}g/ml$ of EMA, and smaller amounts of live cells could be distinguished from dead cells by adjusting the EMA concentration. In addition, the VBNC cells of E. tarda in the oligotrophic low temperature seawater microcosm were estimated to be in the range of $10^4{\sim}10^5CFU/ml$ by EMA-PCR. Therefore, it is possible to detect VBNC cells that will act as potential pathogens in environmental water using EMA-PCR method, and quantitative confirmation using concentration change is also possible.

• KSBB Journal
• /
• 제8권5호
• /
• pp.465-472
• /
• 1993

현재 상품화되어 시판되고 있는 4가지의 미립담체를 이용해 각각의 성능을 비교하기 위하여 부착성동물세포배양 실험을 수행한 결과를 요약하면 다음과 같다. Cytodex 3의 경우 세포의 초기 접종농도를 약 $2.0{\times}10^5$ cells/ml로 하였을 때, 3g/l는 약 $1.4{\times}10^6$cells/ml, 5g/l는 약 $2.0{\times}10^6$cells/ml의 최종세포밀도를 얻을 수 있었다. 또한 bead-to-bead trandsfer 실험을 한 결과 3g/l를 간헐적으로 첨가하였을 때는 약 $1.9{\times}10^6$cells/ml, 5g/l 첨가하였을 때는 약 $3.0{\times}10^6$cells/ml까지 최종세포밀도가 증가하였다. Cultispher-G, 3g/l를 이용해 초기 접종농도를 약 $2.0{\times}10^6$cells/ml로 하여 배양하였을 때 약 $1.3{\times}10^6$cells/ml까지 세포농도가 증가했고, 5g/l를 이용해 초기 접종농도를 $4.0{\times}10^5$cells/ml로 접종하였을 때 최종세포농도가 약 $3.2{\times}10^6$cells/ml까지 증가하는 것을 확인할 수 있었다. 그리고 bead-to-bead transfer배양 실험결과에서 3g/l의 미립담체를 간헐적으로 첨가해 약 $1.8{\times}10^6$cells/ml까지 최종세포밀도가 올라갔고 5g/l를 간헐적으로 첨가하였을 때 $2.5{\times}10^6$cells/ml까지 세포밀도를 얻었다. VX-100을 사용하여 세포를 배양하였을 때 초기 접종농도가 약 $2.0{\times}10^5$cells/ml에서 최종세포밀도가 약 $4.4{\times}10^6$cells/ml까지 증가하는 것을 알게 되었다. 따라서 실험에 사용한 다른 종류의 다공성 젤라틴 bead보다 성능이 우수함을 알 수 있었고 Cytodex-3보다는 최종세포농도가 약 2배이상 증가한 결과를 얻었다. Informatrix bead는 초기 접종농도를 약 $3.0{\times}10^5$cells/ml로 하였을 때 최종세포밀도가 약 $2.1{\times}10^6$cells/ml까지 증가하였다. Collagenase효소를 이용하여 젤라틴 bead를 녹인 후 회수한 세포는 대부분 viable하였고 새로 도입된 bead에 성공적으로 부착하여 성장하였다. 따라서 담체 내부에서 자라는 세포도 회수하여 재사용 할 수 있게 되었다.

• 대한의생명과학회지
• /
• 제15권1호
• /
• pp.73-80
• /
• 2009

Bloodstream infections (BSI) are caused by planktonic microorganisms, sometimes leading to serious infections such as bacteremia and sepsis. BSI occurs more frequently to the patients wearing the central venous catheter (CVC). The ciprofloxacin-incorporated CVC (CFX-CVC) has been reported previously to possess antimicrobial activity. In this study, the antibacterial activity of CFX-CVC and its mechanism against planktonic BSI cells were explored by using the shake flask test and by examining the release rate of 260 nm-absorbing substances from the bacterial cells indicative of the membrane damage of the bacterial cells. CFX-CVC reduced more than 99.9% of the viable planktonic BSI cells demonstrating its potent antibacterial activity. It provoked bacteriolysis causing leakage of a large amount of 260 nm-absorbing materials from the planktonic bacterial cells like S. aureus and E. coli. These results provide evidence that the antibacterial activity of CFX-CVC came from the inhibition of the stability of the planktonic bacterial cells.

• 한국근적외분광분석학회:학술대회논문집
• /
• 한국근적외분광분석학회 2001년도 NIR-2001
• /
• pp.4105-4105
• /
• 2001

Near infrared spectroscopy (NIRS) was employed to qualify and quantify on survival, the injury rate and apoptosis of the human breast cancer cell line MCF-7 cells. MCF-7 cells were cultured in RPMI medium supplemented with 10% FCS in a 95% air and 5% CO2 atmosphere at 37$^{\circ}C$. For the viable cells preparation, cells were de-touched by 0.1% of trypsin treatment and washed with RPMI supplemented with 10% FCS medium by centrifugation at 1000 rpm for 3min. For the dead cells preparation, cells were de-touched by a cell scraper. The cells were counted by a hemacytometer, and the viability was estimated by the exclusion method with frypan blue dye. Each viable and dead cells were suspended in PBS (phosphate bufferred saline) or milk at the cell density desired. For the quantitative determination of cell death by measuring the LDH (lactate dehydrogenase) activity liberated from cells with cell membrane injuries, LDH-Cytotoxic Test Wako (Wako, Pure Pharmaceutical Co. Ltd., Japan) was used. We found that NIRS measurement of MCF-7 cells at the density range could evaluate and monitor the different characteristics of living cells and dead cells. The spectral analysis was performed in two wavelength ranges and with 1,4, 10 mm pathlength. Different spectral data pretreatment and chemometrics methods were used. We applied SIMCA classificator on spectral data of living and dead cells and obtained good accuracy when identifying each class. Bigger variation in the spectra of living cells with different concentrations was observed when compared to the same concentrations of dead cells. PLS was used to measure the number of cells in PBS. The best model for measurement of dead cells, as well as living cells, was developed when raw spectra in the 600-1098 nm region and 4 mm pathlength were used. Smoothing and second derivative spectral data pretreatment gave worst results. The analysis of PLS loading explained this result with the scatter effect found in the raw spectra and increased with the number of cells. Calibration for cell count in the 1100-2500 nm region showed to be very inaccurate.

• Journal of Microbiology and Biotechnology
• /
• 제12권3호
• /
• pp.457-462
• /
• 2002

Low Electromagnetic Field (EMF) intensity in the range of $1{\mu}T\;to\;10{\mu}T$(Tesla) was found to enhance the growth of CHO cells and the production of tPA in batch and perfusion cultivations. At $1{\mu}T\;intensity,\;1.3{\times}10^7$ viable cells/ml of maximum cell density and 80 mg/l of maximum tPA production were obtained in batch cultivation, compared to $2.8{\times}10^6$ viable cells/ml and 59 mg tPA/1 in unexposed case (control). A similar trend was observed in the perfusion process, where it was possible to obtain $1.2{\times}10^7$ viable cells/ml of maximum cell density and 81 mg tPA/l of maximum tPA production by more than 80 days of cultivation. However, there was not much difference between $1{\mu}T\;and\;10{\mu}T$ in perfusion cultivation, possibly due to better environmental growth conditions being maintained by continuous feeding of fresh medium into the reactor. On the contrary, both cell growth and tPA production were severely inhibited at higher than 1 mT intensity, showing no growth at 10 mT exposure. Specific growth rate was linearly correlated to specific tPA production rate at $1{\mu}T$EMF intensity, which represents a partially growth-related relationship. It was also found that a large amount of $Ca^2+$ was released at low EMF intensity, even though the cell growth was not much affected. Low EMF intensity significantly improved both cell growth and tPA production, and tPA production seemed to be more affected than the cell growth, possibly due to the changes of cell membrane characteristics. It can be concluded that the elaboration of EMF intensity less than $10{\mu}T$ could improve cell growth and tPA production, but mainly tPA secretion through batch or perfusion process in a bioreactor.

• Journal of Periodontal and Implant Science
• /
• 제30권3호
• /
• pp.687-700
• /
• 2000

Antigen-specific T cell clones were obtained from mice immunized with Fusobacterium nucleatum ATCC 10953(F .nucleatum) and/or Porphyromonas gingi valis 381(P. gingivalis). 10 Balb/c mice per group were immunized with F. nucleatum followed by P. gingivalis, or with P. gingivalis alone by intraperitoneal injection of viable microorganisms. Spleen T cells were isolated and stimulated in vitro with viable P. gingivalis cells to establish P. gingivalisspecific T cell clones. T cell phenotypes and cytokine profiles were determined along with T cell responsiveness to F .nucleatum or P. gingivalis. Serum IgG antibody titers to F. nucleatum or P. gingivalis were also determined by ELISA. All the T cell clones derived from mice immunized with F. nucleatum followed by P. gingivalis demonstrated Th2 subsets, while those from mice immunized with P. gingivalis alone demonstrated Th1 subsets based on the flow cytometric analysis and cytokine profiles, All T cells clones from both groups were cross-reactive to both P. gingivalis and F. nucleatum antigens. Phenotypes of T cell clones were all positive for CD4. Mean post-immune serum IgG antibody levels to F. nucleatum or P . gingivalis were significantly higher than the pre-immune levels(p <0.01, respectively). There were no significant differences in the antibody titers between the two groups. It was concluded that P. gingivalis-specific T cells initially primed by cross-reactive F. nucleatum antigens were polarized to Th2 subsets, while T cells stimulated with P. gingivalis alone maintained the profile of Th1 subset.

• Journal of Korean Neurosurgical Society
• /
• 제49권1호
• /
• pp.13-19
• /
• 2011

Objective: The purpose of this study is to investigate the combined effects of ginkgo biloba extract, ginkgolide A and B and aspirin on SK-N-MC, human neuroblastoma cell viability and mRNA expression of growth associated protein43 (GAP43), Microtubule-associated protein 2 (MAP2), B-cell lymphoma2 (Bcl2) and protein53 (p53) gene in hypoxia and reperfusion condition. Methods: SK-N-MC cells were cultured with Dulbecco's Modified Eagle's Medium (DMEM) media in $37^{\circ}C$, 5% $CO_2$ incubator. The cells were cultured for 8 hours in non-glucose media and hypoxic condition and for 12 hours in normal media and $O_2$ concentration. Cell survival rate was measured with Cell Counting Kit-8 (CCK-8) reagent assay. Reverse transcriptase polymerase chain reaction (RT-PCR) was used to estimate mRNA levels of GAP43, MAP2, Bcl2, and p53 genes. Results: The ginkgolide A and B increased viable cell number decreased in hypoxic and reperfused condition. The co-treatment of ginkgolide B with aspirin also increased the number of viable cells, however, there was no additive effect. Although there was no increase of mRNA expression of GAP43, MAP2, and Bcl2 in SK-N-MC cells with individual treatment of ginkgolide A, B or aspirin in hypoxic and reperfused condition, the co-treatment of ginkgolide A or B with aspirin significantly increased GAP43 and Bcl2 mRNA levels. In MAP2, only the co-treatment of ginkgolide A and aspirin showed increasing effect. The mRNA expression of p53 had no change in all treating conditions. Conclusion: This study suggests that the combined treatments of Ginkgo biloba extracts and aspirin increase the regeneration of neuroblastoma cells injured by hypoxia and reperfusion.