• 한국미생물·생명공학회지
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• 제27권6호
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• pp.427-432
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• 1999

농산물의 증산을 위한 과도한 농약 사용으로 인한 토양 생태계의 파괴를 줄이기 위하여 맹독성 농약을 대신할 수 있는 생물학적 방제제 개발의 일환으로 식물병원성 진균의 생육을 억제하는 siderophore를 생산하는 생물학적 방제균을 선발하여, 이를 동정하고 근부병균억제기작을 연구하여 향후 다기능 생물학적 방제제 개발의 기초자료를 활용하고자 하였다. 이를 위해 저병해 경작지 토양에 장기간 우점화 되어 있는 siderophore 생산 가능성이 높은 토착 길항미생물을 전국 각지의 저병해 경작지에서 분리하였고, 이들 중 siderophore 생산능이 높은 길항미생물 4종의 균주를 chrome azurol S(CAS) agar를 이용하여 선발하였다. 그 중 최고의 길항성을 나타내는 GL7 균주를 최종 선발하였고, API diagnostic test와 지방산 분석, 그리고 각종 생리학적 특성 및 형태학적 특성을 통해 동정한 결과 Pseudomonas fluoresceus의 한 균주임을 확인하였다. 최종 선발된 siderophore 생산균주 P. fluorescens GL7의 길항기작을 병원성 진균인 Fusarium solani를 대상으로 균사 생장과 포자 발아 등의 억제율을 조사한 결과 균사 생장 억제율은 75% 이상이며, 포자 발아억제율은 97% 이상으로 우수한 방제능을 확인할 수 있었다. 선발된 siderophore 생산능이 높은 길항균주 P. fluo-rescens GL7을 대상으로 토양 내에서 실제로 근부병에 방제력이 있는가를 조사하기 위해 강낭콩 종자(Phaseolus vulgaris L.)를 발병 기주식물로 사용하여 방제시험을 한 결과 F. solam만 처리한 경우는 30%의 성장율을, P. fluorescens GL7 균주와 F. solani와 함께 처리한 경우는 95% 정도의 성장율을 나타내어 약 65% 정도의 발병율을 줄일 수 있었다. 또한 F. solani와 P. fluorescens GL7을 처리하지 않은 무처리구에 비해 식물 물게가 147% 정도로 증가하여 식물의 생육도와 뿌리의 발육상태가 양호하였고, 근부병 발생도 거의 볼 수 없었으므로 아주 우수한 siderophore 생산성 생물학적 방제균을 선발할 수 있었다.

• 한국환경성돌연변이발암원학회지
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• 제22권2호
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• pp.112-124
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• 2002

2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) has been reported to exert detrimental toxicities on various organ systems including reproductive, cardiovascular, nervous, or dermal system. Immunomodulatory effects of TCDD is thymic atrophy, downregulation of cytotoxic T or B lymphocyte differentiation and activation, which were demonstrated using experimental animals, whereas immunotoxicity in human has not been investigated well. This study was proceeded to evaluate general immunologic spectrum of the Korean Vietnam War veterans exposed to TCDD during their operation, and compare with that of the non-exposed control subjects with similar age. Regarding composition and quantity, immune cells in peripheral blood collected from the TCDD-exposed was not much different from those of the control except decreased red blood cell, hemoglobin and hematocrit level. Furthermore, plasma IgG2, G3, and G4 isotype distribution was similar between two groups, but IgG1 level was significantly lowered in the TCDD-exposed, indicating a TCDD-mediated functional alteration of B cells. Significantly enhanced level of IgE in plasma, a hallmark of dermal or respiratory allergic response, was also observed in the TCDD-exposed compared with that of the control. Elevated generation of IL-4 and IL-10 was resulted from in vitro stimulation of T cells with PMA plus ionomycin or PHA, respectively, from the TCDD-exposed in comparison to those of the control, suggesting a skewed type-2 response. In addition, the level of IFN${\gamma}$, a multifunctional cytokine for T cell-mediated immunity, was lowered in the TCDD-exposed with upregulation of tumor necrosis factor $\alpha$. The present study suggests that TCDD exposure disturbs immunohomeostasis in humans observed as an aberrant plasma IgE and IgG1 levels and dysregulation of T cell activities.

• 한국축산식품학회:학술대회논문집
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• 한국축산식품학회 2001년도 임시총회 및 제28차 추계학술발표회
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• pp.43-56
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• 2001

Colostrum contains various kinds of cytokines including TGF-${\beta}$ which is known to be multifunctional in immune response and act as an anti-inflammatory agent. First, we measured the amount of TGF-${\beta}$ in bovine and human colostrum. Expression pattern of TGF-${\beta}$ isotypes was dramatically different between human and bovine colostrial samples. Bovine colostrum collected on day 1 post-delivery retained $41.79{\pm}16.96ng/ml$ of TGF-${\beta}$ 1 and $108.4{\pm}78.65ng/ml$ of TGF-${\beta}$ 2 while in human, $284{\pm}124.75ng/ml$ of TGF-${\beta}$ 1 and $29.75{\pm}6.73ng/ml$ of TGF-${\beta}$ 2. Thus, TGF-${\beta}$ is the predominant TGF-${\beta}$ isotype in bovine colostrum and vice versa in human colostrum. Both TGF-${\beta}$ isotypes diminished significantly in human and bovine colostrum with time. Next, biological activity of colostrial samples was examined in vitro. Both human and bovine colostrum increased IgA synthesis by LPS-activated mouse spleen B cells, which is a typical effect of TGF-${\beta}$ on the mouse B cell differentiation. Futhermore, we found that anti-proliferative activity in MV1LU cells by colostrum samples disappeared by addition of anti-TGF-${\beta}$ 1 and anti-TGF-${\beta}$ 2 antibody. In conclusion, there are substantial amounts of biologically active TGF-${\beta}$ 1 and TGF-${\beta}$ 2 in bovine and human colostrum. The results that the colostrum can increase IgA expression has important implications since IgA is the major Ig class produced in the gastrointestinal tract. We have previously shown that the stimulatory effect of Bifidobacteria bifidum on spllen B cells was quite similar to that of LPS which is a well-known polyclonal activator for murine B cells. In the present study, we further asked whether B. bifidum regulate the synthesis of IgA by mucosal lymphoid cells present in Peyers patches (PP) and mesenteric lymph nodes (MLN). B. bifidum alone, but not C. perfringens, significantly induced overall IgA and IgM synthesis by both MLN and PP cells. This observation indicates that B. bifidum possesses a modulatory effect on the mucosal antibody production in vivo. We, therefore, investigated the mucosal antibody prodduction following peroral administration of B. bifidum to mice. Ingested B. bifidum significantly increased the numbers of Ig (IgM, IgG, and IgA) secreting cells in the culture of both MLN and spleen cells, indicating that peroally introduced B. bifidum enhances mucosal and systemic antibody response. Importantly, however, B. bifidum itself does not induce the own specific antibody responses, implying that B. bifidum do not incite any unwanted immune reaction. Subsequently, it was found that excapsulation of B. bifidum further augments the total IgA production by increasing the number of IgA-secreting cells in the culture of both MLN and spleen cells. Finally, we found that the immuno-stimulating activity of B. bifidum is due to its cell wall components but not due to any actively secreting component(s) from bacteria. Thus our data reveal that peroral administration of B. bifidum can enhance intestinal IgA production and that encapsulation of B. bifidum further reinforces the IgA production.

• BMB Reports
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• 제49권5호
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• pp.297-302
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• 2016

Loss of pancreatic β-cells by oxidative stress or cytokines is associated with diabetes mellitus (DM). DJ-1 is known to as a multifunctional protein, which plays an important role in cell survival. We prepared cell permeable wild type (WT) and mutant type (M26I) Tat-DJ-1 proteins to investigate the effects of DJ-1 against combined cytokines (IL-1β, IFN-γ and TNF-α)-induced RINm5F cell death. Both Tat-DJ-1 proteins were transduced into RINm5F cells. WT Tat-DJ-1 proteins significantly protected against cell death from cytokines by reducing intracellular toxicities. Also, WT Tat-DJ-1 proteins markedly regulated cytokines-induced pro- and anti-apoptosis proteins. However, M26I Tat-DJ-1 protein showed relatively low protective effects, as compared to WT Tat-DJ-1 protein. Our experiments demonstrated that WT Tat-DJ-1 protein protects against cytokine-induced RINm5F cell death by suppressing intracellular toxicities and regulating apoptosisrelated protein expression. Thus, WT Tat-DJ-1 protein could potentially serve as a therapeutic agent for DM and cytokine related diseases.

• 폴리머
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• 제24권3호
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• pp.389-398
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• 2000

폴리카보네이트 표면에 spin-coating한 유/무기 복합 코팅막을 자외선 경화와 가열(12$0^{\circ}C$, 12시간)처리함으로써 약 4~13 $\mu\textrm{m}$ 두께의 투명한 내마모성 유/무기 복합코팅막을 제조하였다. 코팅용액은 무기상과 유기상을 각각 0 : 100, 20 : 80, 30 : 70, 50 : 50, 80 : 20 중량비로 혼합한 다음 광개시제, 증감제 및 계면활성제를 첨가하여 제조하였다. 무기상은 TEOS와 실란커플링제 MPTMS을 1 : 2 혹은 2 : 1몰비로 흔합하여 졸ㆍ겔 반응으로 제조하였으며, 유기상은 2관능형 urethane acrylate 올리고머, 다관능형 TMPTA와 HDDA를 4:3:3 wt% 비율로 흔합하여 제조하였다. 유/무기복합상의 화학적 변화는 FT-IR, $^{29}$ Si-NMR로 분석하였으며, 열분석과 코팅층 표면 형상은 TGA/DSC, SEM, AFM으로 분석하였다. 무기물의 함량과 실란커플링제의 첨가량이 증가 할수록 분자수준의 흔화도가 개선되고 표면 평활도와 내마모성도 향상되었다. 즉 약 10 $\AA$ 이하의 표면 거칠기와 약 15$0^{\circ}C$의 T$_{g}$를 나타내는 시료는 500회 마모시험 전후 단지 16%의 광투과도 감소만을 보인 반면에, 무코팅 폴리카보네이트를 46%의 광투과율 감소를 나타내었다..

• 한국미생물·생명공학회지
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• 제26권2호
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• pp.122-129
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• 1998

최근 생물방제균으로 주목받고 있는 Enterobacter cloacae의 방제기작이 이 균에 의해 토양내에서 생산된 휘발성 ammonia이며 ammonia의 생산에는urease가 관계한다는 보고를 근거로 하여, 항생물질 생산성 균주로 선발된 우수한 길항균주에 암모니아 생성능, 즉 urease 유전자를 유전적으로 부가함으로써 항진균성 길항물질 생산과 암모니아 생산이 동시에 이루어 질 수 있는 새로운 다기능의 생물방제균을 유전적으로 육종하고자 하였다. 저병해 인삼경작지로부터 식물근부균 Fusarium solani의 생육을 강하게 억제하는 길항세균 한 균주 SH14균주를 분리, 선발하였으며, 분리된 균주를 동정한 결과 Bacillus subtilis이거나 그 근연종으로 추정되었다. 억제기작 실험을 통해 길항균주 B. subtilis SH14에 의해 생산되는 항진균성 길항물질은 외막가수분해효소와 같은 고분자 물질이 아니라 열에 안정한 저분자의 항생물질임을 알 수 있었다. 한편 ammonia 생산을 위한 urease의 유전자는 urease 생산력이 강력한 호알칼리성 Bacillus pasteurii의 urease 생산유전자를 E. coli-Bacillus shuttle vector인 pEB203에 subcloning하였고, 이어서 pGU 366으로 명명된 이 recombinant plasmid를 선발된 항진균성 길항균주 B. subtilis SH14에 PEG-induced protoplast transformation 방법으로 도입, 발현시켰으며, 최적조건을 조사하여 90분간의 lysozyme 처리과정 후 1.5 $\mu\textrm{g}$/$m\ell$의 DNA와 40% PEG4000의 첨가로 약 6.5$\times$$10^{-4}$의 형질전환율을 얻을 수 있었다. 아울러 암모니아 생성능이 부가된 생물방제균 B. subtilis SH14(pGU366)에 의해 식물근부균 F. solani에 대한 생육억제력이 증가되는지 여부를 억제거리 측정법과 균체중량법을 통해 확인한 결과 urease 유전자가 도입된 형질전환체 B. subtilis SH14(pGU366)의 근부균 생육억제능이 각각 36.7%, 44.0%정도로 숙주균주인 B. subtilis SH14에 비해 근부균 생육억제능을 보다 강하게 나타내었음을 알 수 있었다. 따라서 항진균성 항생물질 생산성 생물방제균 B. subtilis SH14에 외부의 urease유전자를 도입하여 ammonia 생성능을 부가함으로써 생물방제력의 상승효과를 거둘 수 있었다.

• Journal of Ginseng Research
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• 제47권3호
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• pp.408-419
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• 2023

Background: Ginsenoside compound K (CK), the main active metabolite in Panax ginseng, has shown good safety and bioavailability in clinical trials and exerts neuroprotective effects in cerebral ischemic stroke. However, its potential role in the prevention of cerebral ischemia/reperfusion (I/R) injury remains unclear. Our study aimed to investigate the molecular mechanism of ginsenoside CK against cerebral I/R injury. Methods: We used a combination of in vitro and in vivo models, including oxygen and glucose deprivation/reperfusion induced PC12 cell model and middle cerebral artery occlusion/reperfusion induced rat model, to mimic I/R injury. Intracellular oxygen consumption and extracellular acidification rate were analyzed by Seahorse multifunctional energy metabolism system; ATP production was detected by luciferase method. The number and size of mitochondria were analyzed by transmission electron microscopy and MitoTracker probe combined with confocal laser microscopy. The potential mechanisms of ginsenoside CK on mitochondrial dynamics and bioenergy were evaluated by RNA interference, pharmacological antagonism combined with co-immunoprecipitation analysis and phenotypic analysis. Results: Ginsenoside CK pretreatment could attenuate mitochondrial translocation of DRP1, mitophagy, mitochondrial apoptosis, and neuronal bioenergy imbalance against cerebral I/R injury in both in vitro and in vivo models. Our data also confirmed that ginsenoside CK administration could reduce the binding affinity of Mul1 and Mfn2 to inhibit the ubiquitination and degradation of Mfn2, thereby elevating the protein level of Mfn2 in cerebral I/R injury. Conclusion: These data provide evidence that ginsenoside CK may be a promising therapeutic agent against cerebral I/R injury via Mul1/Mfn2 mediated mitochondrial dynamics and bioenergy.