• Clinical and Experimental Reproductive Medicine
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• 제30권1호
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• pp.5-14
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• 2003

Objective: Present study was aimed to verify the effect of granulocyte macrophage-colony stimulating factor (GM-CSF) in the preimplantation development of mouse embryos and the involvement of the mitogen activated protein kiase (MAPK) in the GM-CSF signaling. Methods: Two-cell embryos were cultured for 96 h in the presence or absence of GM-CSF (0, 0.4, 2, 10 ng/ml) and PD98059, a MEK inhibitor (10 ${\mu}M$). Morphological development, cell number per blastocyst, and apoptotic nuclei, were eamined. MAPK activity of embryonic immunoprecipitate by MAPK (ERK1/2) antibody was measured by in vitro phosphorylation of myelin basic protein. Results: At post hCG 122 h the embryonic development among the experimental groups was significantly different (p=0.018). The rate of blastocyst development and cell number per embryo were the highest in 2 ng/ml GM-CSF treatment group. The percent of apoptotic cells of the GM-CSF-treated embryos was the lowest among the group. In blastocysts, GM-CSF treatment transiently increased MAPK activity. PD098059 attenuated the effect of GM-CSF on the morphological development, increase in cell number per blastocyst, down regulation of apoptosis, and upregulation of MAPK activity, suggesting that activation of MAPK cascade possibly mediated the embryotropic effect of GM-CSF. Conclusion: This result suggested that GM-CSF potentiated the development of preimplantation mouse embryos by activation of MAPK.

• 한국발생생물학회지:발생과생식
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• 제13권3호
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• pp.191-198
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• 2009

In an attempt to simultaneously produce two human proteins, hGH and hG-CSF, in the milk of transgenic mice, we constructed goat $\beta$-casein-directed hGH and hG-CSF expression cassettes individually and generated transgenic mice by co-injecting them into mouse zygotes. Out of 33 transgenic mice, 29 were identified as double transgenic harboring both transgenes on their genome. All analyzed double transgenic females secreted both hGH and hG-CSF in their milks. Concentrations ranged from 2.1 to $12.4\;mg/m{\ell}$ for hGH and from 0.04 to $0.13\;mg/m{\ell}$ for hG-CSF. hG-CSF level was much lower than hGH level but very similar to that of single hG-CSF mice, which were introduced with hG-CSF cassette alone. In order to address the causes of concentration difference between hGH and hG-CSF in milk, we examined mRNA level of hGH and hG-CSF in the mammary glands of double transgenic mice and tissue specificity of hG-CSF mRNA expression in both double and single transgenic mice. Likewise protein levels in milk, hGH mRNA level was much higher than hG-CSF mRNA, and hG-CSF mRNA expression was definitely specific to the mammary glands of both double and single transgenic mice. These results demonstrated that two transgenes have distinct transcriptional potentials without interaction each other in double transgenic mice although two transgenes co-integrated into same genomic sites and their expressions were directed by the same goat $\beta$-casein promoter. Therefore goat $\beta$-casein promoter is very useful for the multiple production of human proteins in the milk of transgenic animals.

• 한국구조물진단유지관리공학회 논문집
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• 제17권5호
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• pp.1-12
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• 2013

본 연구는 보강재를 사용하지 않은 기존 RC기둥과 CSF (고연성재로 보강한 RC기둥) 등 2가지 형태의 기둥에 대한 내진성능특성과 변위연성도 특성 분석을 연구목적으로 한다. 이러한 특성들을 유한요소법에 의한 해석과 실험으로 분석한 결과, CSF의 균열양상과 하중-변위 곡선에 대한 실험치와 해석치는 유사함을 보였다. 보강하지 않은 기둥 (CNF)은 전단균열이 지배적이나 보강기둥 (CSF)은 휨균열이 지배적이다. 보강기둥의 최대변위 크기와 변위연성도는 CNF와 비교하여 큰 증가를 나타낸다. 그러므로 기존기둥의 내진성능과 변위연성도 향상시킬 때 CSF는 CNF의 대체구조로 사용할 수 있다.

• Pediatric Infection and Vaccine
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• 제3권2호
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• pp.163-167
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• 1996

Aseptic meningitis, the most common infection of the central nervous system, is an acute illness mostly caused by enteroviruses. Cerebrospinal fluid(CSF) has been used for the detection of enteroviral RNA but the detection has been mostly performed in a single CSF specimen obtained during the illness. A major objective was to evaluate the relation of sampling time to the recovery of enteroviral RNA in CSF. Thirty seven CSF specimens were obtained from 24 patients between May and August 1993, when an outbreak of asceptic meningitis by echovirus type 9 occurred. Enteroviral RNA in CSF was detected by polymerase chain reaction(PCR). Data about onset of symptom development were obtained by review of medical records. Enteroviral RNA was detected by PCR in 29 of 37 CSF specimens. PCR yielded positive results in 4 of 5 CSF specimens obtained on day 1 to 3, 10 of 11 on day 4 to 6, 8 of 10 on day 7 to 9, 6 of 8 on day 10 to 12, 1 of 3 on day 13 to 15 postonset. Of 11 patients from each of whom more than one CSF were obtained on different day postonset, PCR yielded positive resutls in 2 of 3 cases in whom enteroviral RNA detection was negative in the first CSF. These results indicate that two or more CSF specimens obtained within 12 days postonset are required for improving the accuracy of the diagnosis of enteroviral meningitis.

• Journal of Ginseng Research
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• 제44권2호
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• pp.274-281
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• 2020

Background: Ultraviolet (UV) goes through the epidermis and promotes release of inflammatory cytokines in keratinocytes. Granulocyte-macrophage colony-stimulating factor (GM-CSF), one of the keratinocyte-derived cytokines, regulates proliferation and differentiation of melanocytes. Extracellular signal-regulated kinase (ERK1/2) and protein kinase C (PKC) signaling pathways regulate expression of GM-CSF. Based on these results, we found that ginsenoside Rh3 prevented GM-CSF production and release in UV-B-exposed SP-1 keratinocytes and that this inhibitory effect resulted from the reduction of PKCδ and ERK phosphorylation. Methods: We investigated the mechanism by which ginsenoside Rh3 from Panax ginseng inhibited GM-CSF release from UV-B-irradiated keratinocytes. Results: Treatment with 12-O-tetradecanoylphorbol-13-acetate (TPA) or UV-B induced release of GM-CSF in the SP-1 keratinocytes. To elucidate whether the change in GM-CSF expression could be related to PKC signaling, the cells were pretreated with H7, an inhibitor of PKC, and irradiated with UV-B. GM-CSF was decreased by H7 in a dose-dependent manner. When we analyzed which ginsenosides repressed GM-CSF expression among 15 ginsenosides, ginsenoside Rh3 showed the largest decline to 40% of GM-CSF expression in enzyme-linked immunosorbent assay. Western blot analysis showed that TPA enhanced the phosphorylation of PKCδ and ERK in the keratinocytes. When we examined the effect of ginsenoside Rh3, we identified that ginsenoside Rh3 inhibited the TPA-induced phosphorylation levels of PKCδ and ERK. Conclusion: In summary, we found that ginsenoside Rh3 impeded UV-B-induced GM-CSF production through repression of PKCδ and ERK phosphorylation in SP-1 keratinocytes.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XIII)
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• pp.351-355
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• 2003

Screening 실험을 통하여 결정된 주요인자인 sucrose, nitrogen, 배양온도로 인자의 최적수준을 결정하는 표면반응법중 하나인 Box-Behnken design을 수행하였다. 각 인자의 상관관계를 통하여 sucrose는 90 g/L, nitrogen은 41 mM, 온도는 $22^{\circ}C$가 최적수준으로 결정이 되었으며 확인실험을 통하여 대조구보다 세포생장이 증가하였을 뿐만 아니라 hGM-CSF의 생산도 약 2배 정도 증가되었음을 확인하였다. 이것은 고농도의 sucrose로 인해 배지내로의 hGM-CSF의 분비가 촉진되었기 때문이며 또한 저온으로 인해 분비된 hGM-CSF를 분해하는 protease 활성이 감소되었기 때문인 것으로 사료된다.

• 대한치과교정학회지
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• 제39권4호
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• pp.248-256
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• 2009

이 연구의 목적은 배양된 사람 치주인대 세포에서 파골세포의 형성에 관련된 물질을 합성, 분비할 수 있는지를 알아보고 압박력이 M-CSF, IL-$1{\beta}$, RANKL 및 OPG mRNA의 발현에 미치는 영향을 알아보고자 하였다. 교정치료를 목적으로 발치된 소구치에서 얻은 치주인대세포를 배양한 후 다양한 크기(0.5, 1.0, 2.0, 3.0, $4.0\;g/cm^2$)의 기계적 자극을 다양한 기간(0.5, 1.5, 6, 24, 48 hours) 동안 적용하고, M-CSF, IL-$1{\beta}$, RANKL, OPG mRNA 발현양의 변화를 검사하였다. 각각의 실험군에서 얻어진 mRNA에 대해 역전사 중합효소 연쇄반응검사를 시행하였다. 검사 결과 압박력은 사람 치주인대 세포에서 M-CSF mRNA를 발현시켰으며 M-CSF, IL-$1{\beta}$, RANKL mRNA의 발현양은 자극의 크기와 기간에 따라 증가하였다. 그러나 압박력은 사람 치주인대 세포에서 OPG mRNA의 발현양에 영향을 미치지 않는 것으로 나타났다. 이상의 결과는 기계적 자극이 치주인대 세포에서 M-CSF, IL-$1{\beta}$, RANKL mRNA의 발현양을 조절함으로 파골세포의 분화에 영향을 미칠 수 있음을 시사한다.

• Reproductive and Developmental Biology
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• 제31권3호
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• pp.153-159
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• 2007

본 연구에서: hG-CSF의 발현을 유도적으로 조절하기 위한 FIV-Tet-On lentivirus vector system을 구축하고자 하였다. hG-CSF는 호중성구 계열 세포의 증식과 분화, 생존에 영향을 미치는 물질로서, 이 유전자의 발현을 증가시키기 위하여 FIV-Tet-On vector 상의 hG-CSF나 $rtTA2^SM2$ 서열의 3' 위치에 WPRE 서열을 도입하였다. 구축된 각각의 vector는 293FT 세포에 일시적으로 transfection하여 virus를 생산하였으며, 이 virus를 일차 배양 세포인 CEF와 PFF에 감염시켰다. 각 세포에 전이되 hG-CSF의 발현 양상을 관찰하기 위하여 doxycycline을 첨가하거나 첨가하지 않은 배지에서 배양한 후 quantitative real-time PCR, Western blot과 ELISA를 이용하여 hG-CSF 유전자의 발현 정도를 비교 측정한 결과, CEF에서는 WPRE가 hG-CSF의 3' 위치에 도입된 경우에 발현량과 유도율이 가장 높은 것으로 나타났고, PFF에서는 rtTA 서열의 3'위치에 도입된 경우에 발현량과 유도율이 가장 큰 것으로 확인되었다. 이 FIV-Tet-On vector system은 형질 전환 동물의 생산이나 유전자 치료에서 문제시되는 외래 유전자의 지속적인 과다 발현에 의한 개체의 생리적인 부작용을 최소화하기 위한 해결 방법으로 제시될 수 있을 것이다.

• 한국정밀공학회:학술대회논문집
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• 한국정밀공학회 2000년도 춘계학술대회 논문집
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• pp.1051-1054
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• 2000

We developed the micro CSF (celebrospinal fluid) shunt valve with surface and bulk micromachining technology in polymer MEMS. This micro CSF shunt valve was formed with four micro check valves to have a membrane connected to the anchor with the four bridges. The up-down movement of the membrane made the CSF on & off and the valve characteristic such as open pressure was controlled by the thickness and shape of the bridge and the membrane. The membrane, anchor and bridge layer were made of the $O_2$ RIE (reactive ion etching) patterned Parylene thin film to be about 5~10 microns in thickness on the silicon wafer. The dimension of the rectangular nozzle is 0.2＊0.2 $\textrm{mm}^2$ and the membrane 0.45 mm in diameter. The bridge width is designed variously from 0.04 mm to 0.12 mm to control the valve characteristics. To protect the membrane and bridge in the CSF flow, we developed the packaging system for the CSF micro shunt valve with the deep RIE of the silicon wafer. Using this package, we can control the gap size between the membrane and the nozzle, and protect the bridge not to be broken in the flow. The total dimension of the assembled system is 2.5＊2.5 $\textrm{mm}^2$ in square, 0.8 mm in height. We could precisely control the burst pressure and low rate of the valve varing the design parameters, and develop the whole CSF shunt system using this polymer MEMS fabricated CSF shunt valve.

• IMMUNE NETWORK
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• 제5권2호
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• pp.110-116
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• 2005

Background: As an attempt to develop a strategy to improve the protective immune response to GM-CSF-secreting CT26 (GM-CSF/CT26) tumor vaccine, we have investigated whether the apoptogenic treatment of GM-CSF/CT26 prior to vaccination enhances the induction of anti-tumor immune response in mouse model. Methods: A carcinogeninduced mouse colorectal tumor, CT26 was transfected with GM-CSF gene using a retroviral vector to generate GM-CSF-secreting CT26 (CT26/GM-CSF). The CT26/GM-CSF was treated with ${\gamma}$-irradiation or mitomycin C to induce apoptosis and vaccinated into BALB/c mice. After 7 days, the mice were injected with a lethal dose of challenge live CT26 cells to examine the protective effect of tumor vaccination in vivo. Results: Although both apoptotic and necrotic CT26/GM-CSF vaccines were able to enhance anti-tumor immune response, apoptotic CT26/GM-CSF induced by pretreatment with ${\gamma}$-irradiation (50,000 rads) was the most potent in generating the anti-tumor immunity, and thus 100% of mice vaccinated with the apoptotic cells remained tumor free for more than 60 days after tumor challenge. Conclusion: Apoptogenic pretreatment of GM-CSF-secreting CT26 tumor vaccine by ${\gamma}$-irradiation (50,000 rads) resulted in a significant enhancement in inducing the protective anti-tumor immunity. A rapid induction of apoptosis of CT26/GM-CSF tumor vaccine at the vaccine site might be critical for the enhancement in anti-tumor immune response to tumor vaccine.