• Title/Summary/Keyword: 항원

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Determination of Antigenicity and Characterization of Proteinase from Tissue Invading Nematode Larvae (조직기생 선충류 유충에서 분리한 단백 분해 효소의 특성 및 항원성 검토)

  • Rim, Han-Jong;Joo, Kyeong-Hwan;Choi, Sung-A;Lee, Hye-Jeong;Joo, Chong-Yoon;Chung, Myung-Sook
    • Journal of agricultural medicine and community health
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    • v.22 no.1
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    • pp.61-74
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    • 1997
  • In case of tissue invading nematode, proteolytic enzyme was required at their parasitic life. Proteinases obtained from these parasites(Toxocara canis, Ansakis spp. and Trichinella spiralis) were extracted, isolated and further purified. And then the analysis for activity and inhibitory effect of proteinases were performed by appropriate substrate. Determination of protein as a circulating antigen was done in use of infected animal serum with above parasites, respectively. For above experimental objects, following procedures were performed. First, enzymatic activity was measured in use of azocasein and inhibitory effect of porteinase were studied by various inhibitors. Second, partially purified proteins containing enzymatic activity were obtained by ion exchange chromatography, ultrafiltration and electrophoretic elution. Third, role of the partially purified protein as a circulating antigen. The results obtained were as follows : 1. Enzymatic activity of each nematode proteinase was varied according to pH. Optimal pH of Toxocara canis, Ansakis spp. and Trichinella spiralis were pH 6.0, pH 5.5 and pH 6.5, respectively. The optimal molarity of buffer was 0.1M phosphate buffer. Although little difference between these proteinases was observed, temperature stability was at least maintained at $4^{\circ}C$ until 5 days. 2. In case of Ansakis spp. and Toxocara canis, enzymatic activity of these proteinases was considerably inhibited by Leupeptin and EDTA. For maximum enzymatic activity of above proteinases, it was required that cysteine residue of enzyme should be protected. And it was suggested that metallo type was contained in enzyme active site. Proteinase of Trichinella spiralis contained metallo type also. 3. Although partial purification was performed in Ansakis spp. and Toxocara canis, proteins maintaining enzymatic activity were identified as a circulating antigen. From SDS-PAGE and immunoblot, 25 kDa was presented in Ansakis spp.. Specific antigen of Toxocara cains was 110 kDa protein fraction. 55 and 42 kDa proteins were reacted with normal serum. Trichinella spiralis 60 kDa protein fraction was successfully purified from excretory materials in culture. As a result of immune-reaction with Trichinella spiralis infected serum, highly purified 60 kDa protein was maintained antigenicity until final purification step.

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Effect of High-fat Diet on Peritoneal Macrophage Immunocompetence in C57/BL6 Mice (고지방 식이가 C57/BL6생쥐의 복강대식세포 면역능력에 미치는 영향)

  • Jeong, Jun-Hyun;Park, Hee-Geun;Lee, Wang-Lok
    • Journal of Life Science
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    • v.23 no.6
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    • pp.779-788
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    • 2013
  • This study was to investigate the effect of high-fat diet on macrophage immunocompetence in C57BL/6 mice. C57BL/6 male mice (4 weeks aged, n=16) were divided into two groups. HD groups fed high-fat diet (45% of fat) and ND groups fed chow diet (10% of fat). Peritoneal macrophages were obtained from each mouse intra-peritoneal by sterile lavage method. Macrophage were stimulated with $1{\mu}g/ml$ of lipopolysaccharide (LPS) for 24 hr. Body weight was significantly increased by high-fat diet. Macrophage phagocytosis of HD was significantly lower than that of ND. After 24 hr of LPS stimulation, NO, IL-$1{\beta}$ and IFN-${\gamma}$ production of HD were significantly lower than those of ND. There were no significant differences in the production of TNF-${\alpha}$ and IL-12 between HD and ND. These findings suggest that high fat diet-induced obesity is associated with decreased Immunocompetence and antigen-stimulated sensitivity of peritoneal macrophage, and lower production of NO, IL-$1{\beta}$ and IFN-${\gamma}$ may contribute to these changes.

Effects of Arbuscular Mycorrhiza Inoculation and Phosphorus Application on Early Growth of Hot Pepper(Capsicum annum L.) (Arbuscular mycorrhiza의 접종방법 및 인산시용량이 고추(Capsicum annum L.)의 초기생장에 미치는 영향)

  • Park, Hyang-Mee;Kang, Hang-Won;Kang, Ui-Gum;Park, Kyeong-Bae;Lee, Sang-Sun;Song, Sung-Dahl
    • Korean Journal of Soil Science and Fertilizer
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    • v.32 no.1
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    • pp.68-75
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    • 1999
  • This study was conducted to examine the effects of Arbuscular Mycorrhizae inoculation and phosphorus application on early growth of hot pepper. Gigaspora margarita and Acaulospora spinosa were chosen for this investigation and inoculated into soils of different P levels by varying inoculation time and density. After treatment, some relevant growth responses of hot pepper were measured. Regardless of soil P levels, hot peppers treated with arbuscular mycorrhizal fungi had 5~34% more fresh weight than those untreated, but the effect of inoculation time and density was not different between two species. With decreased P levels, the infection rate and dependency of hot peppers increased. The content of P and K of AMF-inoculated hot peppers increased with increasing P levels, but the shoot to root ratio of those elements decreased. The results of this study showed that inoculation of AMF would be effective in promoting growth of hot pepper seedlings and increase transplant adaptation due in part to the resulted higher root development.

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Strain Recognition and Classification of Korean Native Rhizobium japonicum by Seroimmunological Method (토착대두근류균(土着大豆根瘤菌)의 균주(菌株) 인식(認識)과 분류(分類)를 위한 혈청면역적(血淸免疫學的) 검정(檢定))

  • Lim, Sun-Uk;Kim, Min-Kyun
    • Korean Journal of Soil Science and Fertilizer
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    • v.21 no.2
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    • pp.141-148
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    • 1988
  • Though the cultivation history of soybean in Korea is relatively long, taxonomical study on symbiont nodule bacteria, Rhizobium japonicum is not carried out yet systematically. This work was for the taxonomical study on Korean native R. japonicum by recognizing isolates seroimmunologically as well as for the elucidation of its affinity with host soybean variety. Twenty seven isolates from 13 soybean cultivars cultivated at Seoul National University's experiment field and 6 strains of R. japonicum preserved in our laboratory have been tested. Tube agglutination test, agglutinin adsorption test, and gel immune diffusion test were used. The results obtained are as follows: 1. Twenty five isolates and strains of R. japonicum among 33 were classified into 4 serogroups and identified as indivisual serotype. 2. Two isolates isolated from Hill and Milyang cultivars, 2 isolates from Bangsa and Jangbaek, and 4 isolates from Paldal, Sae-al, and Jangbaek were identified as the same serotype respectively. 3. Seroimmunological tests may be adapted for the elucidation of the affinities between the strains and soybean cultivars as well as strain recognition and systematic classification of Korean native R. japonicum.

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Growth and Yield Responses of Soybean to Planting Density in Late Planting (남부지방 콩 만파 재배 시 재식밀도에 따른 생육 및 수량변이)

  • Park, Hyeon-Jin;Han, Won-Young;Oh, Ki-Won;Ko, Jong-Min;Bae, Jin Woo;Jang, Yun Woo;Baek, In Youl;Kang, Hang-Won
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.60 no.3
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    • pp.343-348
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    • 2015
  • Soybean is one of the important food crop around the world. Especially in East Asia, it is the main ingredient for traditional food like soy sauce and soy paste. The double cropping system including soybean following onion, Chinese cabbage, and potato is widely adopted in Southern region of Korea. In this system, sowing date of second crop (soybean) can be delayed depending on first crops' growth period and weather condition. When planting date is delayed it is known that soybean yield is declined because of shorter vegetative growth period and earlier flowering induced by warm temperature and changes in photoperiod. The objective of this study was to determine soybean growth and yield responses as plant populations at late planting date. Field experiment was conducted at Department of Functional Crop, National Institute of Crop Science, RDA located in Miryang, Gyeongsangnam-Do for two years ('13-'14) in upland field with mid-late maturity cultivar Daewon. A split-plot block design was used with three replications. Main plots were three sowing dates from June 20 to July 20 with 15 days intervals, and subplots were 4 levels of planting densities. Data of maturity (R8) was recorded, yield components and yield were examined after harvesting. Experimental data were analyzed by using PROC GLM, and DMRT were used for mean comparison. Optimum planting population for maximizing soybean yield in late planting which compared with standard population. In mid-June planting, higher planting density causes increased plant height and decreased diameter which lead to higher risk of lodging, however, reduced growth period due to late planting alleviated this problem. Therefore higher seeding rates can provide protection against low seedling emergence caused by late planting in this region.

Protective Efficacy of Recombinant Proteins Adenylate Kinase, Nucleoside Diphosphate Kinase, and Heat-Shock Protein 70 against Mycobacterium tuberculosis Infection in Mice (유전자 재조합 단백질 Adenylate Kinase, Nucleoside Diphosphate Kinase와 Heat-Shock Protein 70의 결핵균에 대한 방어면역효능 분석)

  • Lee, Seung-Heon;Lee, Eun-Gae;Kim, Su-Yeon;Cho, Sang-Nae;Park, Young-Kil;Bai, Gill-Han
    • Tuberculosis and Respiratory Diseases
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    • v.58 no.2
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    • pp.142-152
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    • 2005
  • Background : Priming and boosting vaccination strategy has been widely explored for new vaccine development against tuberculosis. As an effort to identify other vaccine candidates, this study was initiated to evaluate protective efficacy of adenylate kinase (AK), nucleoside diphosphate kinase (NdK), and heat shock protein 70 (Hsp70) of Mycobacterium tuberculosis. Method : M. tuberculosis genes encoding AK, NdK, and Hsp70 proteins were amplified by PCR and cloned into E. coli expression vector, pQE30. Recombinant AK, NdK, and Hsp70 was purified through Ni-NTA resin. To evaluate immune responses, we performed enzyme-linked immunosorbent assay (ELISA) for IgG isotype and $IFN-{\gamma}$ after mice were immunized subcutaneously with recombinant proteins delivered in dimethyl dioctadecylammonium bromide (DDA). Immunized- and control groups were challenged by aerosol with M. tuberculosis. The spleens and lungs of mice were removed aseptically and cultured for CFU of M. tuberculosis. Result : Vaccination with recombinant proteins AK, NdK, and Hsp70 delivered in DDA elicited significant level of antibody and $IFN-{\gamma}$ responses to corresponding antigens but no protective immunity comparable to that achieved with Mycobacterium bovis BCG. Conclusion : Recombinant proteins AK, NdK, and Hsp70 do not effectively control growth of M. tuberculosis in mice when immunized with DDA as an adjuvant.

Comparison of Grain Filling Characteristics by Source-Sink Size Control in Glutinous and Non-glutinous Near Isogenic Line of Rice (근동질유전자 계통인 찰벼와 메벼의 전엽과 절영처리에 따른 등숙특성 비교)

  • 김춘송;안종국;정일민;강항원;이재생;고지연;박성태
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.49 no.3
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    • pp.243-250
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    • 2004
  • Two cultivars which are glutinous (Hawcheongchalbyeo) and non-glutinous (Hawcheongbyeo) near isogenic line of rice, were used for this study. The objective of this experiment was to gain the basic information for increasing grain yield of waxy rice by means of source and sink size control. In both Hwacheongbyeo and Hawcheongchalbyeo, the trend of decrease in total and average spikelet weight was ranked as follows; removal of penultimate leaf< removal of flag leaf< removal of flag leaf and 3ya leaf from the top < removal of flag leaf and penultimate leaf < removal of flag leaf, penultimate leaf, and 3rd leaf from the top. The reduction yale of total and average spikelet weight per panicle of Hwacheongbyeo was higher than those of Hwacheongchalbyeo according to the removal of flag leaf, penultimate leaf, and 3rd leaf from the top. In both cultivars, high-density grain ratio and grain filling ratio of the primary branches were higher Hun those of the secondary branches by leaf clipping treatment. The spikelet number and total spikelet weight per pinicle in both Hwacheongbyeo and Hwacheongchalbyeo were decreased by removal of spikelets on branches compared with control, whereas average spikelet weight and grain filling ratio were increased. The increase rate of average spikelet weight of Hwacheongchalbyeo was much higher than that of Hwacheongbyeo by sink size control. High-density grain ratio by removal of spikelets on branches was higher in Hwacheongchalbyeo, but filled grain ratio was higher in Hwacheongbyeo.

Development of Tumor Screening ELISA kit by Using Novel Tumor Antigen, CM1 (새로운 종양항원인 CM1을 이용한 종양 진단 ELISA kit의 개발)

  • Kang, Jae-Seung;Kim, Dae-Jin;Kim, Young-In;Chang, Ka-Young;Lee, Wang-Jae
    • IMMUNE NETWORK
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    • v.5 no.2
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    • pp.124-129
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    • 2005
  • Background: CM1 (centrocyte/-blast marker 1) is originally defined as a germinal center B cell marker. It is known that CM1 plays a critical role on B cell development in germinal center. In addition, we have found that CM1 is expressed on lymphoma cell lines, such as Raji, Ramos and IM-9. This means that CM1 might be served as a tumor marker as well. In the present study, we examined the expression of CM1 on the surface of the other tumors and the possibility of the development of tumor screening ELISA kit by using CM1. Methods: First, we have examined the expression of CM1 on stomach cancer and hepatoma, which are predominantly (discovered) occurred in Korean, by flow cytometry analysis. After purifying of CM1 antigen from Raji and Ramos, the optimal ELISA condition was determined. And then we compared the level of CM1 between normal individuals and cancer patients by ELISA. To decrease the non-specific binding of anti-CM1 mAb with serum components except CM1 and to enhance the diagnostic accuracy, albumin depletion spin column was used. Results: CM1 was highly expressed on stomach cancer and hepatoma cell lines. In addition, we have also confirmed the increased CM1 expression on cancer patients. The difference of CM1 expression between normal individuals and cancer patients were more clearly observed, after deletion of serum albumin by using albumin depletion spin column. Conclusion: Based on the results from this study, CM1 might be a useful molecule for the early diagnosis of cancer. In addition, further studies for the increase of ELISA sensitivity and appropriate albumin depletion methods should be needed.

IL-12 and TNF-α productions from human peripheral blood mononuclear cells in untreated patients with active pulmonary tuberculosis stimulated with 30-kDa or TSP antigen of Mycobacterium tuberculosis H37Rv (결핵균 PPD, 30-kDa 및 TSP 항원에 의한 치료전 폐결핵환자 말초혈액 단핵구의 IL-12 및 TNF-α 생성능)

  • Song, Chang-Hwa;Jo, Eun-Kyeong;Lee, Ji-Suk;Kim, Dae-Su;Lim, Jae-Hyun;Kim, Un-Ok;Nam, Hyeon-Hui;Kim, Hwa-Jung;Paik, Tae-Hyun;Park, Jeong-Kyu
    • IMMUNE NETWORK
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    • v.1 no.3
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    • pp.250-259
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    • 2001
  • To determine if initial infection with Mycobacterium tuberculosis changes the balance of cytokines between T cells and macrophages, we evaluated interferon (IFN)-${\gamma}$), interleukin-12 (IL)-12, and tumor necrosis factor (TNF)-${\alpha}$ productions by peripheral blood mononuclear cells (PBMC) from 15 untreated active pulmonary tuberculosis (TB) patients and 12 healthy tuberculin reactors (HTR). Freshly isolated PBMC were stimulated with Triton X-100 solubilized protein (TSP), 30-kDa or purified protein derivatives (PPD) antigen for 6, 18 and 96 hours. IL-12 p40 production by antigen-stimulated PBMC from TB patients was significantly decreased compared with that in HTR. In addition, IFN-${\gamma}$ production was significantly depressed in TB patients than that in HTR at a 96-hr stimulation. However, TNF-${\alpha}$ production was significantly higher in antigen-stimulated PBMC from TB than that of HTR. A pronounced increase in IFN-${\gamma}$ protein followed neutralization of IL-10 in early TB patients. However, neutralization of TNF-${\alpha}$ did not significantly alter IFN-${\gamma}$ induction in PBMC from TB patients. There were no significantly differences in the cytokine productions among three proteins, TSP, 30-kDa or PPD antigen. These results indicate that development of TB may be strongly associated with dysregulated productions of IL-12, IFN-${\gamma}$ and TNF-${\alpha}$, during the initial immune responses to M. tuberculosis. Further understanding of operative cytokine networks during human immune cell responses to protein antigens of M. tuberculosis may improve strategies for vaccine development.

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Effect of Conditioning Regimen Intensity on Cytomegalovirus Infection and Related Risk Factors Analysis in Allogeneic Hematopoietic Stem Cell Transplantation (동종조혈모세포이식에서 거대세포바이러스 감염에 미치는 전처치요법 강도의 영향과 관련위험인자 분석)

  • Lee, Hye-Min;Heo, Yoon-Jeong;Im, Hyun-Jeong;Chung, Seon-Young;In, Yong-Won;Jung, Chul-Won;Lee, Young-Mee;Sohn, Kie-Ho
    • Korean Journal of Clinical Pharmacy
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    • v.22 no.2
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    • pp.123-130
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    • 2012
  • 거대세포바이러스(Cytomegalovirus; CMV) 감염은 동종조혈모세포이식 환자의 주요 사망원인 중 하나이다. 용량감소전처치(Reduced-intensity conditioning; RIC)를 이용한 조혈모세포이식은 골수억제전처치(Myeloablative conditioning; MAC)에 비해 골수억제 및 면역억제가 적으므로 CMV 감염 발생율을 감소시킬 것이라 예상되었으나 예방적 면역억제요법, T세포 제거 약제의 사용 등으로 서로 상이한 결과가 보고되고 있다. 2007년 1월부터 2009년 12월까지 총 141명의 환자(MAC 113명, RIC 28명)가 동종조혈모세포이식을 받았으며, CMV 감염은 MAC 62.8%, RIC 57.1% (p = 0.310), CMV 질환은 각각 12.4%, 14.3% (p = 0.785)에서 발생하였다. CMV 감염/질환 발생빈도와 CMV 항원 혈증검사 지속기간, 초기/최고치, 생존율은 두 군간 유의한 차이가 없었다. CMV 감염 위험인자에 대한 다변량분석 결과, 환자가 고령일수록(HR 1.024, 95% CI 1.002-1.045; p = 0.031) 또는 grade 2 이상의 급성 이식편대숙주병이 발생한 경우에(HR 1.849, 95% CI 1.031-3.315; p=0.039) CMV 감염 발생 위험율이 유의하게 높았다. 결론적으로, 전처치요법 강도에 따른 CMV 감염의 발생빈도와 발현양상의 차이는 없었으나, 고령이거나 grade 2 이상의 급성 이식편대숙주병이 발생한 환자의 경우 CMV 감염 발생과 유의한 연관성을 보였다. 이상과 같은 결과에 비춰 봐서 CMV 질환이 대부분 이식 100일 이후에 발생한 점을 고려할 때, 이식 후 CMV 감염 발생 시 ganciclovir 선제요법과 함께 이들 환자들에게 지속적인 모니터링을 실시하는 것이 필요할 것으로 사료된다.