• Asian-Australasian Journal of Animal Sciences
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• 제15권12호
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• pp.1725-1731
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• 2002

The distribution and activities of hydrolytic enzymes (cellulolyti, hemicellulolytic,pectinolytic and others) in the rumen compartments of Hereford bulls fed 100% alfalfa hay based diets were evaluated. The alfalfa proportion in the diet was gradually increased for two weeks. Whole rumen contents were processed into four fractions: Rumen contents including both the liquid and solid fractions were homogenized and centrifuged, and the supernatant was assayed for enzymes located in whole rumen contents (WRE); rumen contents were centrifuged and the supernatant was assayed for enzymes located in rumen fluids (RFE); feed particles in rumen contents were separated manually, washed with buffer, resuspended in an equal volume of buffer, homogenized and centrifuged and supernatant was assayed for enzymes associated with feed particles (FAE); and rumen microbial cell fraction was separated by centrifugation, suspended in an equal volume of buffer, sonicated and centrifuged, and the supernatant was assayed for enzymes bound with microbial cells (CBE). It was found that polysaccharide-degrading proteins such as $\beta$-1,4-D-endoglucanase, $\beta$-1,4-D-exoglucanase, xylanase and pectinase enzymes were located mainly with the cell bound (CBE) fraction. However, $\beta$-D-glucosidase, $\beta$-D-fucosidase, acetylesterase, and $\alpha$-L-arabinofuranosidase were located in the rumen fluids (RFE) fraction. Protease activity distributions were 37.7, 22.1 and 40.2%, and amylase activity distributions were 51.6, 18.2 and 30.2% for the RFE, FAE and CBE fractions, respectively. These results indicated that protease is located mainly in rumen fluid and with microbial cells, whereas amylase was located mainly in the rumen fluid.

• Asian-Australasian Journal of Animal Sciences
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• 제33권12호
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• pp.1948-1956
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• 2020

Objective: The purpose of this study was to reveal the metabolic shift in the fungus cocultured with the methanogen (Methanobrevibacter thaueri). Methods: Gas chromatography-mass spectrometry was used to investigate the metabolites in anaerobic fungal (Pecoramyces sp. F1) cells and the supernatant. Results: A total of 104 and 102 metabolites were detected in the fungal cells and the supernatant, respectively. The partial least squares-discriminant analysis showed that the metabolite profiles in both the fungal cell and the supernatant were distinctly shifted when co-cultured with methanogen. Statistically, 16 and 30 metabolites were significantly (p<0.05) affected in the fungal cell and the supernatant, respectively by the co-cultured methanogen. Metabolic pathway analysis showed that co-culturing with methanogen reduced the production of lactate from pyruvate in the cytosol and increased metabolism in the hydrogenosomes of the anaerobic fungus. Citrate was accumulated in the cytosol of the fungus co-cultured with the methanogen. Conclusion: The co-culture of the anaerobic fungus and the methanogen is a good model for studying the microbial interaction between H₂-producing and H₂-utilizing microorganisms. However, metabolism in hydrogenosome needs to be further studied to gain better insight in the hydrogen transfer among microorganisms.

• KSBB Journal
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• 제18권5호
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• pp.371-376
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• 2003

프로바이오틱 생균인 B. polyfermenticus SCD의 유용성을 확인하기 위해 항산화 및 콜레스테롤 저하 효과를 검증하였다. DPPH법에 의한 항산화 활성은 48% 정도였으며, TCA법에 의한 활성은 45%로 측정되었다. 그리고 TBARS 법에 의해 측정된 활성은 stimulator가 존재할 때 60%에 달하였으며, SOD 유사활성은 15% 정도였다. 이러한 활성은 기존의 항산화제인 BHT나 $\alpha$-tocopherol의 1% 용액과 비교할 때, 비슷하거나 약간 낮은 수치를 보여 준다. 향후 항산화 물질이 정제되어 이 물질의 1% 용액과 1% 농도의 기존 항산화제의 활성을 비교하여야 한다. 그러나 이 같은 뚜렷한 B. polyfermenticus SCD의 항산화력을 바탕으로 천연 항산화제 개발이 가능하다고 판단된다. 한편 B. polyfermenticus SCD의 콜레스테롤 저하 효과는 0.3% oxgall이 존재할 때, 64% 정도였다. 이 수치는 oxgall이 존재하지 않을 때 수치(67%)와 비교할 때, 큰 변화가 없었다. 그리고 콜레스테롤 저하 기작은 세포내 흡착에 의한 것으로 판단되었다.

• 미생물학회지
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• 제44권2호
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• pp.140-146
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• 2008

다양한 염증 질환을 유발하는 사람세포거대바이러스(Human cytomegalovirus: HCMV)는 단핵구 세포주인 THP-1 세포에서 염증반응의 중요한 매개체인 세포간부착분자-1(intercellular adhesion molecule: ICAM-1) 발현을 촉진한다. ICAM-1 발현은 자외선으로 불활화시킨 HCMV (UV-HCMV)에 의해서도 촉진되므로 이 과정에 HCMV 유전자발현은 꼭 필요하지는 않은 것 같다. HCMV에 감염된 THP-1 세포 배양액을 감염되지 않은 THP-1 세포에 처리하거나 공유하게 하였을 시 감염되지 않은 세포에서도 ICAM-1 발현이 증가하였다. 감염된 세포 배양액에 의한 ICAM-1 발현 증가는 $NF-{\kappa}B$ 경로를 거친다. UV-HCMV에 감염된 세포의 배양액은 ICAM-1 발현을 촉진시키지 못하였다. 따라서 HCMV에 의한 THP-1 세포에서 ICAM-1 발현 증가는 바이러스 유전자 발현을 필요로 하지 않지만, 감염된 세포에서 ICAM-1 발현을 촉진하는 물질을 분비하는 과정에는 바이러스 유전자 발현이 필요한 것으로 생각된다.

• 한국습지학회지
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• 제16권3호
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• pp.347-353
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• 2014

여과분리형 생물반응조 운전에서 발생하는 flux 감소 원인을 분석하였고, 상등수 수질과 침강성 변화가 flux에 미치는 영향을 평가하였다. 또한 여과막 모듈과 역세정 장치 일체형 시스템의 특성을 평가하였다. 반응조는 17L 유효 용적의 아크릴 반응조에 $100{\mu}m$ 공경의 여과막 모듈을 통해 배출되는 여과시스템으로, 여과시간이 경과함에 따라 상등수의 수질 특히 탁도와 TOC의 수질이 악화됨에 따라 flux가 점진적으로 감소하였다. MLSS 농도가 18000mg/L 이상에서는 여과저항이 급격히 증가하였다. 상등수의 수질과 flux와의 관계를 규명하기 위해서는 MLSS 농도가 10000mg/L 이상에서는 곤란함에 따라 5000mg/L이 적절한 것으로 판단된다. 활성슬러지 플록을 파괴시켜 SVI를 250으로 조정하여 여과를 시작한 경우, 상등수의 탁도가 증가하여, 침강성이 악화됨에 따라 flux도 점차 감소하였다. 여과막 모듈과 폭기세정 장치를 일체화한 시스템에서 공기 공급은 30~60초 정도로 설정함이 적정한 것으로 나타났다.

• Tuberculosis and Respiratory Diseases
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• 제42권4호
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• pp.447-454
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• 1995

Cytokine release from alveolar macrophages and subsequent interaction of these cytokines with the bronchial epithelium can induce epithelial cells to release inflammatory mediators. Nitric oxide(NO), a highly reactive gas formed from arginine by nitric oxide synthase(NOS), is known to be involved in inflammation and edema formation, and the inducible form of NOS(iNOS) can be increased by cytokines. In this context, we hypothesized that lung epithelial cells could be stimulated by cytokines released by alveolar macrophages to express iNOS. To test this hypothesis, the murine lung epithelial cell line, LA-4, or the human lung epithelial cell line, A549, were stimulated with culture supernatant fluids from alveolar macrophages. NO production was assessed by evaluating the culture supernatant fluids for nitrite and nitrate, the stable end products of NO. Both murine and human cell culture supernatant fluids demonstrated an increase in nitrite and nitrate which were time- and dose-dependent and attenuated by $TNF{\alpha}$ and IL-$1{\beta}$ antibodies(p<0.05, all comparisons). Consistent with these observations, cytomix a combination of $TNF{\alpha}$, IL-$1{\beta}$, and $\gamma$-interferon, stimulated the lung epithelial cell lines as well as primary cultures of human bronchial epithelial cells to increase their NO production as evidenced by an increase in nitrite and nitrate in their culture supernatant fluids, an increase in the iNOS staining by immunocytochemistry, and an increase in iNOS mRNA by Northern blottin(p<0.05, all comparisons). The cytokine effects on iNOS were all attenuated by dexamethasone. To determine if these in vitro observations are reflected in vivo, exhaled NO was measured and found to be increased in asthmatics not receiving corticosteroids. These data demonstrate that alveolar macrophage derived cytokines increase iNOS expression in lung epithelial cells and that these in vitro observations are mirrored by increased exhaled NO levels in asthmatics. Increased NO in the lung may contribute to edema formation and airway narrowing.

• 대한소아치과학회지
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• 제27권2호
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• pp.185-191
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• 2000

Streptococcus mutans가 치태를 형성할 때, 유산구균 1370 (Streptococcus lactis 1370)이 생산하는 수용성 글루캔이 영향을 미친다. 본 연구에서는 여러 인자에 의한 수용성 글루캔 형성 정도를 유산구균 1370배양 상청액의 흡광도로 측정하여 다음과 같은 결과를 얻었다. 1. 5% 자당이 첨가된 M17 broth에서 유산구균 1370 배양 상청액의 흡광도는 높고 Streptococcus mutans 배양 상청액의 흡광도는 낮았으나, 통계학적인 유의성은 없었다. 2. M17 broth에 10% 자당을 첨가하여 유산구균 1370을 배양할시 배양 상청액의 흡광도는 첨가하지 않을 때보다 높았다. (p<0.05), 배지 pH가 5에서보다 7에서 배양 상청액의 흡광도가 더 높았다(p<0.05). 3. $32^{\circ}C,\;37^{\circ}C,\;42^{\circ}C$중 $37^{\circ}C$에서 배양시 배양 상청액의 흡광도가 가장 높았으나 통계학적인 유의성이 없었고, 호기성 배양시보다 혐기성 배양시 배양 상청액의 흡광도가 더 높았다(p<0.05). 4. 배지의 $CaCl_2$ 농도가 1.0mM에서 (p<0.05), KCl 농도가 2.5mM에서 (p<0.05) 배양 상청 액의 흡광도가 높았다. 5. M17 broth에 5% 자당을 첨가한 배지에 유산구균 1370을 접종하여 배양한 배양 상청액과 배지를 1:3으로 가한 비커와이어 검사에서 Streptococcus mutans에 의하여 형성된 인공치태 무게는 5.6mg으로 5% 자당을 첨가하지 않을 때의 103.0mg에 비교하여 현저히 감소하였다 (p<0.05). 이상의 결과를 요약하면 유산구균 1370에 의한 수용성 글루캔의 형성은 자당이 함유된 배지나 세균 증식이 잘 되는 조건에서 증가하였으며, 수용성 글루캔이 Streptococcus mutans에 의한 인공치태 형성을 억제하였다.

• 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.

• Toxicological Research
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• 제14권3호
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• pp.301-306
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• 1998

This experiment was carrid out to know the effects of heating and serum on hydroxyl radicals in embryonic mouse forebrain (cerebrum) culture. The heating to mouse embryonic cerebrum cells in culture was done in a water bath at 43${\circ}C$ for 60min. After that, two supernatants were prepared at 20 hrs and 48 hrs respectively after heat treatment to the brain cells. To find out the heating effects on neuron cells, mouse cerebrum cells (13 embryonic day) were cultured in hydroxyl radical generation system composed of 20mU/ml glucose oxidase (GO system), using condition of normal culture media (MEM, 5% serum, 5% $CO_2$or supernatant prepared after heating at 43${\circ}C$ for 60 min in a water bath. Supernatant prepared at 20 hrs after heat treatment had a greater protective effects against hydroxyl radical than supernatant prepared at 48 hrs after heat treatment . Otherwise, the protective effect of serum against hydroxyl radicals in the cultured brain cells is higher than that in the heat treatment. These results indicated that serum in culture media reduced cytotoxicity of hydroxyl radicals in mouse forebrain culture, also that heat treatment showed the protective effects against hydroxyl radicals generated with 20mU/ml GO system in mouse forebrain culture.

• 한국임상수의학회지
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• 제21권3호
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• pp.236-242
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• 2004

1,2-benzopyrone can stimulate macrophages to increase the ability of phagocytosis. Peripheral blood polymorphonuclear cells (PMN) and macrophages destroy microbial organisms with reactive oxygen species (ROS), called oxidative burst activity (OBA). This study was undertaken to determine whether 1,2-benzopyrone affects the OBA on the phagocytic response of canine peripheral blood phagocytes. The OBA of phagocytes in the addition or absence of latex beads was analyzed by flow cytometry system using dihydrorhodamine 123 (DHR). The direct treatments of 1,2-benzopyrone have no effect on the OBA of peripheral blood mononuclear cells (PBMC), PMN and monocyte-rich cells regardless of addition of latex beads. When latex beads are added to PMN, the OBA of PMN was remarkably enhanced by culture supernatant from PBMC but not PMN treated with 1,2-benzopyrone. Similary, it was also enhanced by human recombinant (hr) $TNF-\alpha.$ However, when latex beads were not added to PMN, its OBA was not enhanced by culture supernatant from either PBMC or PMN treated with 1,2-benzopyrone. The OBA of latex beads-phagocytized PBMC and monocyte-rich cells was not enhanced by culture supernatant from either PBMC or PMN treated with 1,2-benzopyrone. These results strongly suggested that 1,2-benzopyrone has an immunoenhancing effect on the OBA of PMN when phagocytic response occurred only. This enhanced OBA may be mediated through active humoral substance(s), such as $TNF-\alpha,$ produced by PBMC stimulated with 1,2-benzopyrone.