• 제목/요약/키워드: Plaque assay

검색결과 141건 처리시간 0.026초

백작약 조다당분획에 의한 B 세포 증식의 특성 (Characteristics of B cell proliferation by polysaccharide fraction of Paeonia japonica miyabe)

  • 박혜란;함연호;이성태;백상기;조성기
    • IMMUNE NETWORK
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    • 제1권2호
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    • pp.126-134
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    • 2001
  • Background : Paeonia japonica Miyabe is a medicinal plant which has been widely used as a component of blood-building decoctions (Chinese medicinal concept : Bu-Xie). The immunopharmacological characteristics of the extract of Paeonia japonica (PJ) were investigated. Methods : The effects of fractions of PJ extract on lymphocyte proliferation were measured by $H^3$-thymidine incorporation assay. The proliferated lymphocyte subsets were analyzed in flow cytometry. The subset cell populations of spleen cells were separated by magnetic cell separation system, and their proliferation by the extract were investigated. The effect of the extract on antibody production was determined in mice challenged with sheep red blood cells (SRBC) using hemolytic plaque forming cell assay. Results : Spleen cells were proliferated by water extract of PJ. Polysaccharide fraction (PJ-P) of the extract was most active in the proliferation. It was found in flow cytometry that the lymphocyte subset proliferated by PJ-P was B cell population. Among the separated subset cell populations, T cell-depleted cell population and macrophage-depleted cell population were most proliferated by PJ-P. However, positively selected populations of B cells and T cells were not proliferated by PJ-P. These results indicate that B cell proliferation by PJ-P may require the assistance of macrophages or T cells. These results suggest that firstly PJ-P may stimulate macrophages or T cells, and then B cells are activated. The number of antibody-secreting cells was increased by administration of PJ-P in mice immunized with SRBC as a T-dependent antigen. Conclusion : These results suggest that macrophages and accessory cells are directly activated by PJ-P and then helper T cells and B cells are indirectly activated. As the results, immune responses might be coordinately improved. In conclusion, PJ-P, a polysaccharide of P. japonica, may be a characteristic immunostimulator, which is analogous to polysaccharides such as lentinan, PSK and ginsan.

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B 세포의 항체 생산에 대한 게란티 바이오-게르마늄 효모의 영향 (Effect of Geranti Bio-Ge Yeast, a Dried Yeast Containing Biogermanium, on the Production of Antibodies by B Cells)

  • 주성수;원태준;이용진;김민정;박소영;이성희;이도익;황광우
    • IMMUNE NETWORK
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    • 제6권2호
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    • pp.86-92
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    • 2006
  • Background: Germanium compounds are increased to use in nutrient foods and medicines in terms of antibiotics to microbes, anticancer, modulation of immune system and neutralizing heavy metal toxins. Geranti Bio-Ge Yeast, containing stable organic germanium and bound to the yeast protein was developed by Geranti Pharm. LTD. and the modulation effect in the immune system was examined in vivo and in vitro. Methods: The compound, Geranti Bio-Ge Yeast, was fed to female Balb/c mice (each group has 10 mice) for 4 weeks and the yeast powder and steamed red ginseng powder were used as control during the same feeding time points. During 4 weeks there was no symptom to be considered, and after 4 weeks feeding all mice were sacrificed to check the changes of related immune cells and subsidiary responses (i.e. cell counting, FACS, MTT, LDH, PFC assay). Results: In pre-post comparison, B cell population was increased in the group of Geranti Bio-Ge Yeast in a dose dependent manner (100 to 800 mg/kg). However, the population of T cell, dendritic cell and macrophage was not comparably changed in all doses. The ability of cytokine production and proliferation was almost same level as shown in control group. In contrast, PFC assay informed that the compound increase the antibody production ability when fed over 200 mg/kg implying that the increase of PFC number might be due to the increase of B cells. Conclusion: Over the entire study, we concluded that the compound, Geranti Bio-Ge Yeast has better potential in immune response in terms of B cell proliferation than that of positive control, red ginseng, and the compound can be one of the future candidates for a new supplementary source improving immune system activity.

해바라기씨 추출물의 뇌세포에 대한 사멸 보호 효과 (Protective Effects of Helianthus annuus Seed Extract against Chemical-Induced Neuronal Cell Death)

  • 박자영;우상욱;허진철;이상한
    • 한국식품저장유통학회지
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    • 제14권2호
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    • pp.213-219
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    • 2007
  • 퇴행성 뇌질환의 하나인 알츠하이머는 가벼운 기억력의 장애에서부터 전반적인 인지기능의 장애를 나타내는 질환으로 해마 (hippocampus)를 포함한 신경세포에서 세포사와 관련이 있는 것으로 보고 되어왔다. AP의 자체 독성과 산화 스트레스, $A{\beta}$ plaque에서 나오는 free radical은 세포내 $Ca^{2+}$를 높이고 이로 인해 calpain이 활성화되어 신경 세포사가 촉진되며 microtubule과 같은 cytoskeleton을 파괴시킨다. 이러한 ROS와 $A{\beta}$에 의해 유도되는 세포사멸을 보호하는 물질을 해바라기 씨 추출물을 이용하여 실험을 실시하였다. 우선 추출물이 항산화 효과 (DPPH, FRAP assay), acetylcholinesterase(AChE)의 활성 및 SH-SY5Y cell의 세포사멸에 미치는 영향을 검토하였다. 항산화 활성과 AChE에 대한 억제활성은 해바라기 씨 추출물 처리농도가 높을수록 유의적으로 높게 나타났다. $H_2O_2$$A{\beta}$에 의해 유도된 SH-SY5Y에 대한 세포사멸 억제효과 실험(MTT assay)에서 해바라기 씨 추출물이 모두 높은 활성을 나타내었으므로 이의 성분분리는 뇌세포 보호를 위한 좋은 식품재료가 될 수 있다.

방사선 치료에 내성이 유도된 두경부 편평세포암에 대한 종양살상 헤르페스 바이러스의 유전자 치료 효과 (Therapeutic Effect of Oncolytic Herpes Simplex Virus on Induced Radioresistant Head and Neck Squamous Cell Carcinoma)

  • 김세헌;최은창;이진석;천제영;변형권;송기재;김광문
    • 대한두경부종양학회지
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    • 제22권2호
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    • pp.130-136
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    • 2006
  • Introduction : The sensitivity of tumor cells to radiotherapy is a critical determinant of local control and potential cure in advanced head and neck squamous cell carcinoma(HNSCC). The emergence of radioresistant tumor cells is an obstacle to cancer therapy. Most radioresistant cells have a higher proportion of cells in the Sphase of the cell cycle and a lower apoptotic fraction than radiosensitive cells. HSV replication is increased in cells that have higher S-phase fractions. NV1066 is an oncolytic herpes simplex virus type-1 mutant. We hypothesized that NV1066 replication and cytotoxicity are increased in radioresistant cells. The purpose of this study is to evaluate the antitumor efficacy of NV1066 to treat radioresistant HNSCC. Methods : Radioresistant cells were selected by treating five HNSCC cell lines with repeated conventional fractionated doses of radiation(2Gy/day), using a Cs-137 irradiator, up to a cumulative dose of 70Gy. Clonogenic cell survival and S-phase fractions were compared between radioresistant and parental radiosensitive cells. The two cell populations were then treated with NV1066 to examine viral replication, by the viral plaque assay and viral cytotoxicity. Results : Fractionated irradiation resulted in the selection of radioresistant cells. Radioresistant cells had a higher S-phase fraction(42.9%) compared to parental cells(26.2%). NV1066 replication in radioresistant cells was 7.4 times higher than in parental cells(p<0.01). Treatment with NV1066 resulted in increased cytotoxicity of 24.5% in radioresistant cells compared to parental cells(p<0.05). Conclusion : NV1066 showed increased viral replication and cytotoxicity in radioresistant HNSCC cell lines. These findings suggest a potential clinical application for this oncolytic viral therapy as treatment for radioresistant head and neck cancers.

설사증 이환견(犬)으로 부터 분리(分離)한 canine coronavirus의 성상(性狀)에 관한 연구(硏究) (Characterization of canine coronaviruses isolated from the dogs with diarrhea in Korea)

  • 이병형;전무형;박종현;황의경;허원
    • 대한수의학회지
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    • 제34권3호
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    • pp.517-527
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    • 1994
  • An attempt was made to isolate a causative viral agents from the fecal specimens of the diseased dogs with the gastroenteritis symptoms. Two coronavirus-like agents were isolated by serial dilution end point method and plaque assay. The isolates were characterized in terms of cytopathology, antigenicity, replication, physicochemical and morphological properties. The results obtained through the experiment were as follows; 1. Among 7 fecal specimens collected from the dogs with enteric disease, 2(28.6%)coronavirus-like agents showing typical cytopathic effects of canine coronavirus were isolated, and designated as CCV D1 and CCV D2, respectively. 2. By the cross-neutralization test and indirect immunofluoresence antibody test, the isolates were antigenically indentified as the standard CCV. The viruses were replicated only in the cytoplasm of A-72 cells. 3. The isolates showed no haemagglutinating activity against the erythrocytes from 11 kinds of animals. 4. The electron microscopic observation for the isolates showed spherical and pleomorphic features, covered with club-shaped projections on the surface. The size of particles was ranged from 70 to 150nm. 5. In one-step growth curve for the isolates in A-72 cells, maximum titers of intracellular vius was $10^{4.6}$ $TCID_{50}/0.1ml$ at 46 hrs postinoculation(pi) of CCV Dl and $10^{4.4}$ $TCID_{50}/0.1ml$ at 34 hrs pi of CCV D2. The maximum titers of extracellular virus was $10^{5.5}$ $TCID_{50}/0.1ml$ at 58 hrs pi of CCV D1 and $10^{5.8}$ $TCID_{50}/0.1ml$ at 46 hrs pi of CCV D2. 6. In physicochemical property test, the isolates were very sensitive to choroform and were found to be RNA virus. The viruses was stable at pH 3.0 for 1 hr and at $22{^{\circ}C}$ for 5 hrs. However, infectivity titers reduced remarkably by treatment with $56{^{\circ}C}$ for 10min.

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서해안 양식장에서 분리한 Edwardsiella tarda의 특이 bacteriophage 특성 (Characterization of Edwardsiella tarda specific phage isolated from fish farms on west coast of Korea)

  • 이찬흔;허용주;백민석;이정은;강자영;한미정;경서봉;최상훈
    • 한국어병학회지
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    • 제24권2호
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    • pp.85-93
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    • 2011
  • Edwardsiella tarda (E. tarda)는 담수 및 해수어에 광범위하게 감염을 일으키는 그람 음성 박테리아이다. 본 연구에서는 서해안 넙치 양식장에서 분리한 E. tarda에 대해 특이 용균을 보이는 파아지를 분리하여 파아지의 활성에 미치는 환경적 요인인 pH와 배양 온도의 영향 등을 조사하였다. 또한 어류모델로서는 담수어종인 Nile tilapia (Oreochromis niloticus)를 이용하였으며 E. tarda와 파아지와의 상호작용을 알아보았다. pH 안정성 실험에서는 담수보다 해수에서 높은 활성을 보였으며 온도 안정성 실험 결과 $50^{\circ}C$까지 일정한 활성을 보였다. Tilapia 감염 실험에서는 파아지와 E. tarda를 접종하였다. 그 결과 실험군은 대조군에 비해 24시간째까지 E. tarda수가 일정하게 감소하는 것을 확인하였으며 파아지는 처음 접종 양보다 약간 증폭됨을 알 수 있었다. 본 연구의 결과는 E. tarda에 의해 발생되는 질병을 특이 파아지를 이용하여 제어할 수 있음을 암시해준다.

오염된 임프란트 표면의 해독 방법에 따른 내독소 제거 효과에 관한 연구 (The Endotoxin Assay of Contaminated Titanium Implants following Various Techniques of Detoxification)

  • 박중희;임성빈
    • Journal of Periodontal and Implant Science
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    • 제34권1호
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    • pp.71-81
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    • 2004
  • Peri-implantitis could be the result of biomechanical and occlusal overload as well as microbiologic invasion. The dental implant may be more susceptible to dental plaque than the natural tooth, as the predictability of a stable soft tissue attachment complex has not yet been confirmed. With the development of peri-implantitis, the implant surface would be exposed to the oral environment and becomes coated with bacteria. The objective of therapy for this condition is to regain integration of the implant with bone. Since fibroblast adherence to surfaces is impeded by endotoxin, it would seem that decontamination would be desirable to obtain maximum osseointegration. The purpose of this study was to determine whether various chemotherapeutic and mechanical treatments(distilled water, air-powder abrasive, hypersaturated citric acid, tetracycline) can detoxify contaminated titanium implant surface by means of kinetic LAL test. Experimental rough surface titanium disks were fabricated. All of them were divided into two groups(A.a group and P.g group) and each contaminated by A. actinomycetemcomitans and P. gingivalis suspension. Contaminated disks were treated with distilled water, air-powder abrasive, citric acid and tetracycline, and then all disks were placed into LPS-free water for elution. The results were as follows : 1. In A.a group, LPS elute level of all test groups were significantly lower than control group(p<0.05). 2. In A.a group, LPS elute level of test 2, test 3 and test4 groups were significantly lower than that of control group(p<0.05). But, among the test 2, test 3, test4 groups, the significant differences were not detected. 3. In P.g group, LPS elute level of test 2, test 3 and test 4 groups were lower than that of control group(p<0.05). But, among the test groups, the significant differences were not detected. From the result of this study, it would be concluded that air-powder abrasive, hypersaturated citric acid and tetracycline treatments may be effective at reducing endotoxin level on rough titanium implant surfaces, and can be clinically used. But the treatments in peri-implantitis differentially impact osseointegration making one method clinically superior. To gain this knowledges, further molecular biologic and histopathologic studies should be developed.

Kimchi attenuates fatty streak formation in the aorta of low-density lipoprotein receptor knockout mice via inhibition of endoplasmic reticulum stress and apoptosis

  • Woo, Minji;Kim, Mijeong;Noh, Jeong Sook;Park, Chan Hum;Song, Yeong Ok
    • Nutrition Research and Practice
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    • 제11권6호
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    • pp.445-451
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    • 2017
  • BACKGROUND/OBJECTIVES: Endoplasmic reticulum (ER) stress is positively associated with atherosclerosis via elevating macrophage cell death and plaque formation, in which oxidative stress plays a pivotal role. Antioxidative, lipid-lowering, and anti-atherogenic effects of kimchi, a Korean fermented vegetable, have been established, wherein capsaicin, ascorbic acid, quercetin, 3-(4'-hydroxyl-3',5'-dimethoxyphenyl)propionic acid, and lactic acids were identified. In this study, mechanisms of action of kimchi methanol extracts (KME) on fatty streak formation via suppression of ER stress and apoptosis in aorta were examined in low-density lipoprotein receptor knockout mice. MATERIALS AND METHODS: Mice fed a high cholesterol diet with an oral administration of KME (KME group, $200 mg{\cdot}kg-bw^{-1}{\cdot}day^{-1}$) or distilled water (control group) for 8 weeks (n = 20 for group). Plasma lipid and oxidative stress levels were evaluated. Protein expression was measured by western blot assay. Fatty streak lesion size and the degree of apoptosis were examined in the aorta. RESULTS: Compared to the control group, in the KME group, plasma lipids levels were decreased and oxidative stress was alleviated (P < 0.05). Protein expression levels of nuclear factor (erythroid-derived 2)-like 2-mediated antioxidants in aorta were increased whereas those for ER stress markers, glucose regulated protein 78, phospho-protein kinase RNA-like ER kinase, phospho-eukaryotic initiation factor 2 subunit ${\alpha}$, X-box binding protein 1, and C/EBP homologous protein were decreased in the KME group (P < 0.05). Moreover, apoptosis was suppressed via downregulation of phospho-c-Jun N-terminal kinase, bcl-2-associated X protein, caspases-9, and -3 with a concomitant upregulation of anti-apoptotic protein, B-cell lymphoma 2 (P < 0.05). Fatty streak lesion size was reduced and the degree of apoptosis was less severe in the KME group (P < 0.05). CONCLUSIONS: In conclusion, antioxidant activity of KME might prevent fatty streak formation through, in part, inhibition of ER stress and apoptosis in aortic sinus where macrophages are harbored.

A Point Mutation at the C-Terminal Half of the Repressor of Temperate Mycobacteriophage L1 Affects Its Binding to the Operator DNA

  • Ganguly, Tridib;Chattoraj, Partho;Das, Malabika;Chanda, Palas K.;Mandal, Nitai.C.;Lee, Chia Y.;Sau, Subrata
    • BMB Reports
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    • 제37권6호
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    • pp.709-714
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    • 2004
  • The wild-type repressor CI of temperate mycobacteriophage L1 and the temperature-sensitive (ts) repressor CIts391 of a mutant L1 phage, L1cIts391, have been separately overexpressed in E. coli. Both these repressors were observed to specifically bind with the same cognate operator DNA. The operator-binding activity of CIts391 was shown to differ significantly than that of the CI at 32 to $42^{\circ}C$. While 40-95% operator-binding activity was shown to be retained at 35 to $42^{\circ}C$ in CI, more than 75% operator-binding activity was lost in CIts391 at 35 to $38^{\circ}C$, although the latter showed only 10% less binding compared to that of the former at $32^{\circ}C$. The CIts391 showed almost no binding at $42^{\circ}C$. An in vivo study showed that the CI repressor inhibited the growth of a clear plaque former mutant of the L1 phage more strongly than that of the CIts391 repressor at both 32 and $42^{\circ}C$. The half-life of the CIts391-operator complex was found to be about 8 times less than that of the CI-operator complex at $32^{\circ}C$. Interestingly, the repressor-operator complexes preformed at $0^{\circ}C$ have shown varying degrees of resistance to dissociation at the temperatures which inhibit the formation of these complexes are inhibited. The CI repressor, but not that of CIts391, regains most of the DNA-binding activity on cooling to $32^{\circ}C$ after preincubation at 42 to $52^{\circ}C$. All these data suggest that the 131st proline residue at the C-terminal half of CI, which changed to leucine in the CIts391, plays a crucial role in binding the L1 repressor to the cognate operator DNA, although the helix-turn-helix DNA-binding motif of the L1 repressor is located at its N-terminal end.

흰털오가피와 더덕 추출물을 첨가한 발효유 급여가 마우스의 면역기능에 미치는 영향 (Effects of Fermented Milk Containing Herb Extract from Acanthopanax divaricatus var. albeofructus and Codonopsis Ianceolata on the Immune Status of Mouse)

  • 임상동;성기승;김기성;한동운
    • 한국축산식품학회지
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    • 제27권1호
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    • pp.95-101
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    • 2007
  • 흰털오가피의 면역활성 증진 효과를 알아보기 위하여 선발된 수컷 마우스를 대상으로 하여 발효유에 흰털오가피 (뿌리 : 잎 : 줄기)와 더덕 열수추출건조물을 (5 : 2 : 1.5) : 1.5 비율로 혼합하여 투여한 그룹 1 mg/mL(A), 3 mg/mL(B), 9 mg/mL(C) 3그룹으로 나누어 임상적용 경로인 경구투여를 선택하여 발효유를 각각 3 mL/kg씩 위내로 직접 투여하였다. 대조군은 발효유만 먹인 그룹(D)과 식염수만 먹인 그룹(E)을 두었다. 흰털오가피의 함량이 ICR계 수컷 마우스의 면역기능에 미치는 영향에 대해 알아본 결과 모든 그룹에서 마우스의 증체량과 체중 증가는 유의성 있는 차이를 보이지 않았고, 7주와 10주령에 안락사 시킨 마우스의 각 장기 무게에서도 간, 신장, 심장, 폐, 고환의 무게에서는 유의성 있는 차이를 보이지 않았다. 비장계수는 7주령 C군과 10주령 B군에서 유의성(p<0.05)있는 증가가 있었으며, 식이 지속기간이 길어질수록 증가된 반면, 흉선계수는 모든 군에서 유의성 있는 증가가 관찰되지 않았다. 항체생산 세포수는 흰털오가피 투여군이 대조군에 비해 증가하였으며, 7주령에 비해 10주령이 감소하는 경향을 보였다. 항원에 대한 항체 생성량을 알아보기 위하여 혈구응집반응을 실시한 결과 흰털오가피 투여군이 대조군에 비하여 증가하는 경향을 보였고 10주령 C군에서 유의성 있는 차이를 보였다. 면역글로블린 농도는 흰털오가피 투여군에서 증가하였고, 7 및 10주령 C군에서 유의성이 있었다.