• Journal of Animal Science and Technology
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• 제44권5호
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• pp.549-560
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• 2002

본 시험은 패스틴$^{(R)}$ 을 첨가하였을 때, in vitro 상에서 단백질 fraction과 분해율에 미치는 영향과, in vivo 상에서 반추위 성상, 미생물 군집, 암모니아태 질소 농도 및 영양소 소화율에 미치는 영향을 구명하고자 실시하였다. In vitro 실험에서는 1mm로 분쇄된 대두박을 기질로 하여 패스틴$^{(R)}$ ((주)은진인터내셔날)을 첨가하여 borate-phosphate buffer와 중성세제에서의 조단백질 분해율을 측정하였으며, exogenous enzyme (Streptomyces griseus 유래 protease)를 이용하여 39$^{\circ}C$에서 0, 2, 4, 8, 12, 48 시간동안 배양 후 조단백질 분해율을 측정하였다. 반추위 발효성상과 영양소 소화율은 반추위 fistula가 부착된 평균체중 300kg의 홀스타인 수소 4두를 이용하여 무첨가구, 패스틴$^{(R)}$ 첨가구의 두개 처리구에 2마리씩 4마리를 배치하여 측정하였다. Buffer Soluble Protein fraction은 패스틴$^{(R)}$ 첨가 수준별로 차이가 없었으나, 무첨가구에 비해 패스틴$^{(R)}$ 첨가구에서 감소하는 경향을 보였다. 단백질 분해율은 배양 0 시간대에서 4시간대까지는 처리구간 유의성이 없었지만, 12 h과 48 h에서는 패스틴$^{(R)}$ 첨가로 시험구에서 감소되었다. 용해 단백질 분해율 ‘a’는 패스틴$^{(R)}$ 시험구에서 경미하게 높은 수치를 나타내었지만, 소화 가능한 단백질 분해율 ‘a+b’는 패스틴$^{(R)}$ 시험구에서 낮은 경향을 보였다. 패스틴$^{(R)}$ 첨가로 pH와 $NH_3$- N 농도는 증가하는 경향이었으며 휘발성지방산, 미생물 수 및 enzyme activity는 감소하였고 영양소 소화율은 높았으나 유의적인 차이는 없었다.

• Tuberculosis and Respiratory Diseases
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• 제53권4호
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• pp.420-430
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• 2002

연구배경 :임상적으로 만성 기관지염 환자의 호흡기 증상이 급격히 악화되는 것으로 정의되는 만성 기관지염의 급성악화는 기도의 폐쇄를 조장하여 만성 폐쇄성 폐질환으로의 진행을 촉진시키는 것으로 알려져 있다. 방 법 :만성 기관지염의 급성악화 환자 40명을 대상으로 하였으며 대상 환자를 무작위로 moxifloxacin군과 clarithromycin군으로 나누어 항생제를 경구투여 하였다.항생제를 복용 하기 전과 항생제를 7일간 복용한 후 각각 객담을 유도 처리하여 유도객담 상층액 내의 IL-8, SLPI, MMP-1, MMP-9, TIMP-1의 농도를 ELISA 방법으로 측정하였다. 결 과 : 항생제 투여에 의해 유도객담 내 TIMP-l의 농도와 TIMP-1/MMP-1의 분자량 비율이 유의하게 감소하였고 (p<0.05), 유도객담 내 SLPI의 농도는 유의하게 증가하였다(p<0.01) 급성악화 시 유도객담 내의 TIMP-1의 농도(p<0.01, r=0.751)와 TIMP-l/MMP-1의 분자량 비율(p<0.01, r=0.752)은 IL-8과 유의한 상관관계를 보이고 있었으며 항생제 치료로 호전이 된 이후에도 IL-8과의 상관관계는 계속 관찰되었다. 급성악화 시 유도객담 내 SLPI의 농도는 유도객담 내 TIMP-1(p<0.01, r=-0.496)과 TIMP-1/MMP-1(p<0.01, r=-0.456)의 분자량 비율 변화와 유약한 상관관계가 있었다. 그러나 유도객담 내 MMP-1, MMP-9의 농도 그리고 TIMP-l/MMP-9의 분자량 비율은 IL-8이나 SLPI의 농도와 유의한 상관관계가 없었다. 결 론 : 만성 기관지염의 급성 악화에 의해 TIMP와 MMPs의 불균형이 초래되며 TIMP가 상대적으로 많이 증가함으로써 기도 내 ECM이 축적되는데 적절한 항생제를 사용하지 않았을 때에는 이러한 기도벽의 재구성이 반복되어 기도의 폐쇄가 심해질 것으로 생각된다. TIMP와 MMPs의 불균형은 항생제 치료에 의하여 호전이 됨으로 적절한 항생제 치료는 만성 기관지염의 급성악화에 의한 비가역성 기도 폐쇄가 진행되는 것을 어느 정도 예방할 수 있을 것으로 생각된다.

• Applied Biological Chemistry
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• 제33권1호
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• pp.93-100
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• 1990

Petunia, 당근 및 대두의 Protoplast를 전기자극법으로 융합시킬 때의 적정 전기자극 조건과, petunia protoplast의 융합율 그리고 또는 viability에 대한 calcium, magnesium, pretense, trypsin, triton X-100, concanavalin A, dimethyl sulfoxide(DMSO), glycerol monooleate 그리고 spermine의 영향을 조사하였다. Protoplast pearl chain 형성을 위한 교류의 주파수와 전압은, Petunia의 경우 10kHz-20 V/cm와 1 MHz-60 V/cm, 당근의 경우에 100 kHz-40 V/cm와 $1\;MHz-40{\sim}80\;V/cm$, 그리고 대두의 경우에 $1\;MHz-40{\sim}80\;V/cm$가 양호하였다. Protoplast융합을 위한 최적 직류 처리조건은, petunia의 경우에 교류를 1MHz, 60 V/cm (15 sec) 처리한 후 직류를 2.5 kV/cm, $40\;{\mu}sec$ 처리했을 때였는데, 이 때 Protoplast의 생존율 과 융합율은 각각 45 %와 10 %였다. 당근과 대두의 경우는 처리시간에 관계없이 직류 2.5 V/cm 이하에서는 융합이 일어나지 않았고, 2.5 V/cm 이상에서는 protoplast가 파괴되었다. Calcium은 전기자극융합을 촉진시켰는데, 최적 농도는 140 mM이었고 이 때 protoplast의 생존율과 융합율은 43 %와 11 %였다. Magnesium은 융합촉진효과가 없었으나 calcium이 존재할 때는 융합을 촉진하였는데, calcium 140 mM일 때 최적 magnesium농도는 140mM이었고, 이때의 생존을과 융합율은 각각 45%와 13%였다. 융합배지에 protease, trypsin, triton X-100, concanavalin A, DMSO, glycerol monooleate, spermine을 처리했을 때 융합율은 대조구(11.2 %)에 비해 각각 2.4, 2.1, 1.6, 1.4, 1.8, 1.5, 2.2배로 증가하였고, 생존율은 각각 37 %, 35 %, 38 %, 41 %, 31 %, 37 %, 42 %로 대조구(45.2 %)에 비해 모두 저하되었다.

• The Plant Pathology Journal
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• 제31권3호
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• pp.278-289
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• 2015

Indigenous strains of Trichoderma species isolated from rhizosphere soils of Tea gardens of Assam, north eastern state of India were assessed for in vitro antagonism against two important tea fungal pathogens namely Pestalotia theae and Fusarium solani. A potent antagonist against both tea pathogenic fungi, designated as SDRLIN1, was selected and identified as Trichoderma viride. The strain also showed substantial antifungal activity against five standard phytopathogenic fungi. Culture filtrate collected from stationary growth phase of the antagonist demonstrated a significantly higher degree of inhibitory activity against all the test fungi, demonstrating the presence of an optimal blend of extracellular antifungal metabolites. Moreover, quantitative enzyme assay of exponential and stationary culture filtrates revealed that the activity of cellulase, ${\beta}$-1,3-glucanase, pectinase, and amylase was highest in the exponential phase, whereas the activity of proteases and chitinase was noted highest in the stationary phase. Morphological changes such as hyphal swelling and distortion were also observed in the fungal pathogen grown on potato dextrose agar containing stationary phase culture filtrate. Moreover, the antifungal activity of the filtrate was significantly reduced but not entirely after heat or proteinase K treatment, demonstrating substantial role of certain unknown thermostable antifungal compound(s) in the inhibitory activity.

• KSBB Journal
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• 제23권3호
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• pp.187-192
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• 2008

Lysozyme-producing microorganisms were isolated to obtain bacteria which can efficiently solubilize microbial cells. Cells of normal and chloroform-treated Escherichia coli and Micrococcus Iysodeikticus were used as model substrates to isolate lysozyme-producing microorganisms and investigate the efficiency of cell lysis. The culture supernatant of the isolate New1 (98% similarity of 16S rDNA sequence with Thermomonas haemolytica) showed different lytic characteristics for different substrates. Thermal treatment (autoclave) of substrate cells showed a significant effect on cell solubilization by culture supernatant of the New1. For autoclaved substrate cells, E. coli, M. Iysodeikticus and chloroform-treated E. coli were solubilized by 58.7%, 49.4% and 79.1%, respectively, in the culture supernatant of New1. The lytic activity of New1 was mainly caused by lysozyme produced by the isolate. It was also showed that New1 exhibited high protease activity and a little cellulase activity.

• Nutrition Research and Practice
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• 제9권6호
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• pp.586-591
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• 2015

BACKGROUND/OBJECTIVES: Reactive oxygen species (ROS) formation is closely related to miconazole-induced heart dysfunction. Although rhamnetin has antioxidant effects, it remained unknown whether it can protect against miconazole-induced cardiomyocyte apoptosis. Thus, we investigated the effects of rhamnetin on miconazole-stimulated H9c2 cell apoptosis. MATERIALS/METHODS: Cell morphology was observed by inverted microscope and cell viability was determined using a WelCount$^{TM}$ cell proliferation assay kit. Miconazole-induced ROS production was evaluated by fluorescence-activated cell sorting with 6-carboxy-2',7'-dichlorofluoroscein diacetate ($H_2DCF$-DA) stain. Immunoblot analysis was used to determine apurinic/apyrimidinic endonuclease 1 (APE/Ref-1) and cleaved cysteine-aspartic protease (caspase) 3 expression. NADPH oxidase levels were measured using real-time polymerase chain reaction. RESULTS: Miconazole (3 and $10{\mu}M$) induced abnormal morphological changes and cell death in H9c2 cells. Rhamnetin enhanced the viability of miconazole ($3{\mu}M$)-treated cells in a dose-dependent manner. Rhamnetin (1 and $3{\mu}M$) treatment downregulated cleaved caspase 3 and upregulated APE/Ref-1 expression in miconazole-stimulated cells. Additionally, rhamnetin significantly reduced ROS generation. CONCLUSIONS: Our data suggest that rhamnetin may have cytoprotective effects in miconazole-stimulated H9c2 cardiomyocytes via ROS inhibition. This effect most likely occurs through the upregulation of APE/Ref-1 and attenuation of hydrogen peroxide levels.

• Parasites, Hosts and Diseases
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• 제41권3호
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• pp.165-169
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• 2003

This study describes the characterization of 80 kDa pretense showing gelationlytic property among three pretenses in the excretory/secretory proteins (ESP) from Toxoplasma gondii. The pretense activity was detected in the ESP but not in the somatic extract of RH tachyzoites. This pretense was active only in the presence of calcium ion but not other divalent cationic ions such as $Cu^{2+},{\;}Zn^{2+},{\;}Mg^{2+},{\;}and{\;}$Mn^{2+}$, implying that $Ca^{2+}$ is critical factor for the activation of the protease. The 80 kDa pretense was optimally active at pH 7.5. Its gelatinolytic activity was maximal at $37^{\circ}C$, and significant level of enzyme activity of the pretense remained after heat treatment at $56^{\circ}C$ for 30 min or $100^{\circ}C$ for 10 min, This thermostable enzyme was strongly inhibited by metal chelators, i.e., EDTA, EGTA, and 11 10-phenanthroline. Thus, the 80 kDa pretense in the ESP secreted by T. gondii was classified as a calcium dependent neutral metalloprotease.

• The Plant Pathology Journal
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• 제32권3호
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• pp.251-259
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• 2016

The present study investigated the suppression of the disease development of anthracnose caused by Colletotrichum gloeosporioides and C. acutatum in harvested apples using an antagonistic rhizobacterium Paenibacillus polymyxa APEC128 (APEC128). Out of 30 bacterial isolates from apple rhizosphere screened for antagonistic activity, the most effective strain was APEC128 as inferred from the size of the inhibition zone. This strain showed a greater growth in brain-heart infusion (BHI) broth compared to other growth media. There was a reduction in anthracnose symptoms caused by the two fungal pathogens in harvested apples after their treatment with APEC128 in comparison with non-treated control. This effect is explained by the increased production of protease and amylase by APEC128, which might have inhibited mycelial growth. In apples treated with different APEC128 suspensions, the disease caused by C. gloeosporioides and C. acutatum was greatly suppressed (by 83.6% and 79%, respectively) in treatments with the concentration of $1{\times}10^8$ colony forming units (cfu)/ml compared to other lower dosages, suggesting that the suppression of anthracnose development on harvested apples is dose-dependent. These results indicated that APEC128 is one of the promising agents in the biocontrol of apple anthracnose, which might help to increase the shelf-life of apple fruit during the post-harvest period.

• Bulletin of the Korean Chemical Society
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• 제35권12호
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• pp.3571-3575
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• 2014

Proteasome, a multicatalytic protease complex, has been validated as a promising therapeutic target in oncology. Carfilzomib (Kyprolis$^{(R)}$), a tetrapeptide epoxyketone, irreversibly inhibits the chymotrypsin-like (CT-L) activity of the proteasome and has been recently approved for multiple myeloma treatment by FDA. A chemistry effort was initiated to discover the compounds that are reversibly inhibit the proteasome by replacing the epoxyketone moiety of carfilzomib with a variety of ketones as reversible and covalent warheads at the C-terminus. The newly synthesized compounds exhibited significant inhibitory activity against CT-L activity of the human 20S proteasome. When the compounds were tested for cancer cell viability, 14-8 was found to be most potent in inhibiting Molt-4 acute lymphoblastic leukemia cell line with a $GI_{50}$ of $4.4{\mu}M$. Cytotoxic effects of 14-8 were further evaluated by cell cycle analysis and Western blotting, demonstrating activation of apoptotic pathways.

• Journal of Dairy Science and Biotechnology
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• 제38권1호
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• pp.53-57
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• 2020

장관 흡착능 등을 비롯한 프로바이오틱 활성이 우수한 Lactobacillus johnsonii PF01의 배양상등액을 황산암모늄으로 70% 농축한 후에 항균능을 조사한 결과, 황색포도상구균을 억제하는 활성을 나타내었다. PF01 균주가 분비하는 항균물질은 단백질분해효소 proteinase K에 의해 항균활성이 소실되지 않는 특성을 나타냄으로써, 항균물질이 박테리오신일 가능성을 시사하였다. L. johnsonii PF01 유전체에서 박테리오신을 코딩하는 유전자를 검색한 결과, helveticin과 상동성이 높은 유전자가 존재함을 확인하였다.