• Title/Summary/Keyword: C-ELISA

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Investigation of Helicobacter pylori cagE Locus Diversity on Interleukin-8 Induction in AGS Cell-line (Helicobacter pylori cagE 유전자 다형성이 위상피세포주에서 Interleukin-8 유도능에 미치는 영향)

  • Lee Su-Min;Lee Hak-Sung;Lee In-Suk;Co Yu-Kyung;Han Hae-Won;Oh Jung-Hwan;Park Jae-Myung;Choi Myung-Gyu;Chung In-Sik
    • Journal of Life Science
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    • v.14 no.6 s.67
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    • pp.906-912
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    • 2004
  • Helicobacter pylori infection is highly prevalent, as high as 2/3 of whole population infected, in Korea. H. pylori infection initiates inflammation by induction of interleukin-8 through type IV secretion of CagA. It was recently suggested that induction failure of IL-8 is not associated with defect in cag PAI but associated with cagE locus diversity. This study was designed to investigate ability of 11-8 in-duction according to sequence variation within the cagE gene, cagA TP motifs and vacA m-types in vitro study using AGS cell-line, and to evaluate its association with different clinical outcome. Seventy-four H. pylori stains were isolated from 23 patients with gastric cancer (Ca), 24 subjects with gastritis (G) and 27 patients with duodenal ulcer (Du) in Kangnam St. Mary's Hospital, Seoul, Korea. cagE gene diversity was confirmed by the PCR-RFLP methods with MboI/NlaIII and tyrosine phosphate motifs (TPMs) of cagA was determined TPM-A and C by using DdeI/Tsp5091 restriction enzyme and TPM-B was determend by Real time PCR the method of Owen et al. and IL-8 was measured by ELISA assay. IL-8 activity was positively detected in 59 among 74 strains $(79.7\%)$. IL-8 secretion was significantly increased in MboI A and MboI B type compared to MboI C type and in MboI/NlaIII A-C and B-C type than C-C type. 1L-8 activity was not associated with either the number or composition of cagA tyrosine phosphorylation motifs and vacA m-type. There was no significant difference in IL-8 activity among patient groups. cagE gene diversity is thought to be mainly associated with the induction of IL-8 in H. pylori infection.

Cross-reaction of 6B and 19F Specific Antibodies to Serotypes 6A, 6C, and 19A after Immunization with 7-valent Pneumococcal Conjugate Vaccine in Korean Children Aged 12-23 Months (한국 12-23개월 소아에서 7가 폐구균 단백결합 백신 추가접종으로 유도된 6B와 19F 혈청형 특이 방어항체의 교차혈청형 6A, 6C, 19A에 대한 교차 반응)

  • Kim, Kyung-Hyo;Yang, Joo Yun;Park, In Ho;Lim, Soo Young
    • Pediatric Infection and Vaccine
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    • v.20 no.2
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    • pp.53-62
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    • 2013
  • Purpose: The cross-protection of 7-valent pneumococcal conjugate vaccine (PCV7) against vaccine-related serotypes has been controversial. We investigated the serological properties of cross-protective antibodies against vaccine-related serotypes 6A, 6C, and 19A induced in young children aged 12-23 months after booster immunization of PCV7. Methods: IgG and IgM antibody concentrations and opsonic index (OI) against vaccine serotypes 6B and 19F and vaccine-related serotypes 6A, 6C, and 19A were measured by ELISA and opsonophagocytic killing assay (OPA) in 4 selected immunesera. The serological properties and antigenic specificity of protective antibodies were determined by IgM depletion of immunesera, OPA, and competitive OPA against serogroup 6 and 19 pneumococci. Results: Compared to pre-IgM depleted immunesera, OI of IgM-depleted immunesera against 6B and 19F decreased and OI against 6A, 6C, and 19A decreased, too. In competition OPA, free 6B and 19F polysaccharide completely inhibited the immune protection against vaccine-related serotypes 6A, 6C, and 19A as well as vaccine types 6B and 19F. Conclusions: The booster immunization of PCV7 certainly induced cross-protective antibodies against vaccine-related serotypes 6A, 6C, and 19A with both IgG and IgM isotypes. Furthermore, IgM antibodies are more highly contributed to opsonophagocytic activity against vaccine-related serotypes as well as most of vaccine types than do IgG antibodies. Further studies are needed for the more immunized sera in the children as well as adults.

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The Effect of Antioxidant Vitamins on Aflatoxin $B_{1}-DNA$ Adduct the Formation in Aflatoxin $B_1$ Administered Mice Liver (항산화비타민이 Aflatoxin $B_1$ 투여 마우스의 간세포에서 Aflatoxin $B_{1}-DNA$ Adduct 형성에 미치는 영향)

  • Park, Seon-Ja;Kang, Sung-Jo;Park, Jung-Hyun;Oh, Sang-Suk;Chung, Duck-Hwa
    • Korean Journal of Food Science and Technology
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    • v.33 no.6
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    • pp.669-675
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    • 2001
  • The objective of this study was to examine the effects of antioxidant vitamins on the formation of $AFB_{1}-DNA$ adduct and $AFB_{1}-inducing$ cellular oxidative damage. Intraperitoneal(i.p.)injections of 10 mg/kg vitamin C(VC) and 63.8 mg/kg vitamin E(VE) were repeatedly administrated 4 times with 2 days interval to 6 week old male ICR mice. After one hour of vitamin treatments, 0.4 mg/kg $AFB_1$ was injected in $AFB_1$ plus vitamin treated groups by same way. On the other hands, $AFB_1$ treated group was only injected with $AFB_1$ by the same method described above without vitamins. According to quantitative analysis of the $AFB_1$ in mice serum by indirect competitive ELISA, 12.28 and 18.78 ng/mL were detected in $AFB_1-treated$ groups, but 7.60 and 4.85 ng/mL in $AFB_1$ plus VC and VE treated groups, respectively. 23.78, 25.48 ng/mL of $AFB_1-DNA$ adduct were detected in mice liver of $AFB_1$treated groups, while 5.26, 7.81 ng/mL in $AFB_1$ plus VC and VE treated groups, respectively. Consequently, the differences in the concentrations of $AFB_1$ related materials between vitamin treated and non-treated groups were significant. Immunohistochemistry revealed brownish infiltration of $AFB_1$ around central vein and sinusoid in $AFB_1-treated$ group. This manifestation was distinctly reduced in $AFB_1$ plus VC and VE treated groups.

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Seroepidemiology of helminthic diseases in Yanbian of the People's Republic of China (중국 연변지역 주요 기생충질환에 대한 혈청역학적 조사)

  • Joo, Kyoung-Hwan;Quan, Fu Shi;Kwon, Hye-Soo;Cho, Sung-Weon;Li, Shun Yu
    • Journal of agricultural medicine and community health
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    • v.23 no.2
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    • pp.275-285
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    • 1998
  • Antibody responses(IgG) to Paragonimus westermani. Clonorchis sinensis, Cysticercus cellulosae, Sparganum Anisakis simplex, Toxocara canis and Trichinella spiralis were studied. The ELISA technique was performed to determine the prevalence of above helminthic diseases. 975 cases obtained from Yanbian of China during October, 1995 to July, 1997 were examined with a positive antibody titer of 5.74% in clonorchiasis, 4.92% in paragonimiasis, 1.54% in cysticercosis. 8.51% in sparganosis, 1.85% in anisakiosis, 12.51% in toxocariasis, and 7.08% in trichinosis respectively. And 23.87% in showed positive antibody titer at least one of the seven helminths. The differences of the age and sex in the positive sera were analysed by the Chi-squared test and the level of significance accepted was p<0.05. The significant differences in positive antibody production were P.W.(p<0.01). C.S.(p<0.01), A.S.(p<0.05). T.C.(p<0.001), and T.S.(p<0.01) respectively in age groups. sparganosis(p<0.05) in sex groups. Other parasites showed that there were no significant differences among age groups and sex groups(males and females). Higher positive antibody rate of C.S. and P.W. occured in the 50-59 years old and those of T.C. and T.S. happened in the 20-29 years old. Patients of internal disease showed more positive antibody titer, that is to say, there was significant difference between positive rate of internal diseases and that of control (p<0.01. p<0.05) in 6 helminths except cysticercosis. The result showed that some cross reactions existed among nematodes, and the developed techniques(EITB) should be done for a correct diagnosis. Also the prevalence of some important helminths may be evaluated from the result and it would be a basic data for controlling parasitic diseases in Yanbian.

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Antioxidant Status and its Relationship to Plasma Cytokine Levels in Korean Elderly Women Living in Seoul

  • Kim, Mi-Joung;Kim, Jung-Hee
    • Journal of Community Nutrition
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    • v.6 no.2
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    • pp.103-109
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    • 2004
  • Body antioxidant status is an important factor for the prevention of many chronic diseases in the elderly. This study was done to investigate antioxidant status and its relationship to immune response by measuring plasma cytokine (IL-2 and IL-6) levels in elderly women. Subjects were 76 elderly women aged over 60 years, visiting Jangwhi Social Welfare Center of Seongbook-Gu in Seoul. Subjects were divided into 3 groups according to age (< 65, 65 - 74, > 75). Dietary intakes were assessed by semi-quantitative food frequency questionnaires (SFFQ). Plasma vitamin C level was measured by 2,4-dinitrophenylhydrazine method and plasma levels of vitamin E, A and ${\beta}$-carotene were measured by HPLC. Plasma levels of IL-2 and IL-6 were determined with a solid phase sandwich enzyme linked-immuno-sorbent assay (ELISA) using commercial kits. The average intakes of antioxidant vitamins were 96.3mg (137.5% of RDA) for vitamin C and 523.3 ${\mu}$gRE (74.8% of RDA) for vitamin A in elderly women. All of the average plasma levels of antioxidant vitamins were within normal range. However the percentage of the elderly women with deficiency plus marginal values were 7.9% in vitamin C, 9.2% in vitamin A and 7.9% in vitamin E. Plasma levels of IL-2 and IL-6 were 27.1${\pm}$7.1pg/ml and 5.9${\pm}$5.3pg/ml in elderly women. Correlation data showed that plasma IL-2 level was negatively correlated with plasma vitamin C level. In addition, IL-6 level was also negatively correlated with plasma vitamin C, A and E levels, respectively. There was a significant positive correlation between erythrocyte thiobarbituric acid-reactive substance(TB-ARS) level and plasma IL-2 or IL-6 levels. In addition, erythrocyte TBARS level showed a significant positive correlation with plasma total antioxidant status (TAS) level and a significant negative correlation with plasma vitamin C level. Overall results might imply that the decreased levels of antioxidant vitamins result in an increase in oxidative stress and thereby increase cytokine production such as IL-2 and IL-6. However further research is required to elucidate these relationships.

Seroprevalence of Coxiella burnetii in bulk-tank milk and dairy cattle in Gyeongbuk province, Korea (경북지역 집합유와 젖소에서 큐열 항체 보유율 조사)

  • Ouh, In-Ohk;Seo, Min-Goo;Do, Jae-Cheul;Kim, In-Kyoung;Cho, Min-Hee;Kwak, Dong-Mi
    • Korean Journal of Veterinary Service
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    • v.36 no.4
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    • pp.243-248
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    • 2013
  • Q fever is a rickettsial infection caused by Coxiella burnetii that is transmitted from animals to humans. Modes of transmission to humans include inhalation, tick bites and ingestion of unpasteurized milk or dairy products. This survey was aimed at monitoring the seroprevalence of C. burnetii in bulk-tank milk (BTM) in Gyeongbuk province. In addition, the seroprevalence of C. burnetii was investigated at the herd level of dairy cattle in eastern Gyeongbuk province in which many dairy cattle are reared. Among 324 BTM samples collected from 20 country areas, 175 (54%) BTM samples from 15 (75%) country areas were positive for C. burnetii by ELISA. By regions, the seroprevalence of BTM samples in eastern, central, western and northern areas of Gyeongbuk province were 62.7%, 48.4%, 45.1%, and 41.4%, respectively. When analyzed in the dairy cattle reared in the eastern area where high seroprevalence occurred in BTM samples, 119 (24.2%) out of 492 dairy cattle were positive for C. burnetii. Seroprevalence of C. burnetii in dairy cattle was increased with daily milk yield of farm (P<0.05) and age (P<0.001). Since seroprevalence of C. burnetii is relatively high in both BTM samples and dairy cattle reared in Gyeongbuk province, further studies on the high risk farms and herds are needed to evaluate infection status and appropriate control programs in this region.

Kyungheechunggan-tang suppresses inflammatory cytokines and fibrotic genes in LPS-induced RAW 264.7 cells and LX-2 cells

  • Bae, Junghan;Jang, Eungyeong;Lee, Jang-Hoon
    • The Journal of Korean Medicine
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    • v.39 no.4
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    • pp.40-50
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    • 2018
  • Objectives: The aim of this study is to investigate anti-inflammatory effects of Kyungheechunggan-tang (KHCGT) on LPS- induced RAW 264.7 cells and LX-2 cells and anti-fibrotic effects of KHCGT on LX-2 cells. Materials and Methods: Three types of KHCGTs (KHCGT-A, -B, and -C) by narrowing down the number of constituent herbs from 9 (KHCGT-A) to 5 (KHCGT-B) and to 3 (KHCGT-C) were developed. To understand pharmacological effects of KHCGT, three types of KHCGTs were treated on RAW 264.7 cells and LX-2 cells. Anti-inflammatory activities of KHCGT were evaluated by ELISA assay for pro-inflammatory cytokines, IL-6, $TNF-{\alpha}$ and IL-10, in LPS-stimulated RAW 264.7 cells and for IL-6 production in LPS-induced LX-2 cells. In addition, anti-fibrotic effects of KHCGT were determined by quantitative real-time PCR assay for fibrosis-related genes, ${\alpha}-SMA$, collagen1A1, TIMP1, MMP-2, in LX-2 cells. Results: KHCGT-A and KHCGT-C showed inhibitory effects on secretion of IL-6 in LPS-stimulated RAW 264.7 cells and LX-2 cells. KHCGT-B and KHCGT-C exhibited inhibitory effects on the expression of pro-inflammatory cytokines such as IL-6, $TNF-{\alpha}$, and IL-10 in LPS-stimulated RAW 264.7 cells. The mRNA expression levels of collagen1A1 and MMP-2 were significantly reduced by KHCGT-C whereas TIMP-1 was suppressed by KHCGT-A and KHCGT-B in LX-2 cells. Among three different formulas, KHCGT-C demonstrated the most remarkable effects on inflammation and fibrosis. Conclusions: In this study, KHCGT showed both anti-inflammatory and anti-fibrotic effects which make it to be a prospective agent for chronic liver diseases with inflammation and fibrosis.

Effect of Sasamsaengmaek-san Extract on Immunity Activity in Balb/c Mice (사삼생맥산 추출물이 Balb/c mice의 면역활성에 미치는 효과)

  • Chae Hyun, Park;Jae Hui, Kang;Hyun, Lee
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.36 no.6
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    • pp.235-241
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    • 2022
  • The purpose of this study was to investigate the immune activation effect of Sasamsaengmaek-san (SSSMS) consisted of a mixture of Adenophora triphylla var. japonica, Liriope platyphylla, Panax ginseng C. A. Meyer and Schisandra chinensis. in Balb/c mice. Measuring alanine aminotransferase (ALT) and aspartic acid transaminase (AST) levels in Balb/c mice was performed to analyze the cytotoxicity. Cytokines (IFN-γ, IL-2, IL-12) which regulate the immune activation in Balb/c mice were measured by enzyme-linked immunosorbent assay (ELISA). Activated T lymphocytes in peripheral blood mononuclear cell (PBMC), spleen, lymph nodes were analyzed by flow cytometry using percentages. All tests were compared with red ginseng 100 ㎍/mL (RG 100), which is the most used for immune activity. As a result, cytokine activity was significantly increased at SSSMS 300 group. Activated T lymphocytes in PBMC, spleen, lymph nodes were significantly increased at SSSMS 300 group. These results suggest that there is a possibility of SSSMS activating an immune system by activating the cytokines, and it is confirmed that SSSMS also effective for generation and differentiation of T, B lymphocytes which activate the immune response.

Identification of Potato mop-top virus from Solanum tuberosum cv. Gawon in Korea

  • Lee, Young-Gyu;Park, Jong-A;Yoon, Young-Nam;Cheon, Jeong-Uk;Lee, Key-Woon
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.138.1-138
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    • 2003
  • Potato mop-top virus(PMTV) was identified from Solanum tuberosum cv. Gawon showing bright chlorotic mottle symptom in Namwon, Korea. Samples were collected green-house in February, 2003. Electron microscopic examination of negatively stained preparation revealed that PMTV were rigid-rod shaped particles about 100-150, 250-300 nm x 18-20 nm in length. In ultrathin sections of leaf tissue from diseased potato plants, cluster of viruses particles were observed in the cytoplasm. TAS-ELISA determined that the virus was serologically related to PMTV. PMTV produced double ring necrotic local lesion in inoculated leaf of Chenopodium amaranticolor in incubated at 15$^{\circ}C$. The PMTV could be detected with RT-PCR using PMTV detectable primer set designed to amplify about 540 bp of the partial CP gene of PMTV

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발암물질의 조기검색법 개발 및 Chemoprevention에 관한 연구

  • 이병무;윤여표
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1994.04a
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    • pp.193-193
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    • 1994
  • 발암물질의 조기검색법 개발 및 chemoprevention연구의 일환으로 발암물질과 DNA 및 단백질의 공유결합체인 발암물질-DNA 및 -단백질 adduct를 연구하였다. 발암물질(예, 밴조피렌)-단백질 adduct에 관한 연구에서는 시료(단백질)에 soluble protease를 이용하는 간편하고 손쉬운 ELISA(Enzyme Linked Immunosorbent Assay)분석법을 확립했다. 발암물질(예,벤조피렌,아플라톡신 B1) -DNA 및 -단백질 adduct를 이용한 발암성 조기검색법의 개발을 Ames test 및 염색체이상시험과 비교 연구한 결과 본 연구에서 새로이 개발한 DNA 및 Protein-adduct형성 시험법은 저농도에서 고농도에 이르기까지 뚜렷한 용량-반응 관계를 나타냈으며 Ames test 및 Chromosomal test에서 일어날 수 있는 false positive나 false negative의 결과를 나타낼 우려가 없었다. 벤조피렌-DNA adduct를 이용한 chemoprevention 연구에서는 항산화제로 알려진 비타민 E,C 및 $\beta$-carotene을 시험한 결과 용량의존적으로 벤조피렌-DNA adduct 형성을 억제하였다.

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