• Title/Summary/Keyword: cell enzyme-linked immunosorbent assay

검색결과 297건 처리시간 0.023초

비뇨생식기로부터 Chlamydia trachomatis의 세포배양 및 효소면역학적동정 (Identification of Chlamydia trachomatis from the Urethral Specimens by McCoy Cell Culture and Enzyme-Linked Immunosorbent Assay)

  • 이재상;이연태
    • 대한미생물학회지
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    • 제21권2호
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    • pp.261-270
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    • 1986
  • A total of 339 urethral, vaginal swab and eye discharge materials from the out-patients in the hospitals of Seoul area was microbiologically collected for the detection of Chlamydia trachomatis infection during May through August, 1985. McCoy cell culture system (MCC) and enzyme-likned immunosorbent assay (EIA) methods were employed in this study as the tools for the detection of C. trachomatis, and the detectabilities of two methods were compared. The results obtained in this study are summarized as follows: 1. The positive rate of C. trachomatis in 339 swab specimens was 18.6%, and the rate in females (20.1%) was much higher than that in males (7.1%). 2. The positive rate of C. trachomatis infection the prostitutes was the highest (24.2%), and the rate in the eye discharge specimens obtained from the new barns was 12.8%. 3. The positive rates of C. trachmoatis infection detected in the specimens from the patients with vaginitis and leucorrhea, with infertility, with cystitis and with nongonococcal urethritis were 17.2%, 21.9%. 18.0% and 7.1%, respectively. 4. The positive rate of C. trachomatis infection in 20-25 age group was 30.5%. This rate was the highest among the other age groups. 5. The positive rate of C. trachomatis infection in the randomly screened 89 swab specimens by EIA (30.3%) was much higher than the rate detected by MCC (18.6%). 6. The positive rate of C. trachomatis infection in females detected by EIA was also much higher than in males, and the 20-25 age group showed the highest positive rate as compared to the other age groups. 7. Sensitivity and specificity of EIA for the detection on C. trachomatis were 100% and 88.6%, respectively, in case that MCC was regarded as perfect method. In summarizing the above results, it is known that considerable cases with genital diseases and with eye discharges were associated with C. trachomatis, and that EIA method is recommendable for the detection of C. trachomatis especially in the specimens swabed from the genital tracts.

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High Expression of MICA in Human Kidney Cancer Tissue and Renal Cell Carcinoma Lines

  • Jia, Hong-Ying;Liu, Jun-Li;Zhou, Cheng-Jun;Kong, Feng;Yuan, Ming-Zhen;Sun, Wen-Dong;Wang, Jue;Liu, Ling;Zhao, Jing-Jie;Luan, Yun
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권4호
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    • pp.1715-1717
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    • 2014
  • The overall incidence and mortality of renal cell carcinoma (RCC), the most common kidney cancer, are steadily increasing for reasons that are not fully explained. Our aim was to explore the expression of membrane MHC class I chain-related gene A (mMICA) in human RCC cell lines and tissue specimens, and to determine expression of soluble MICA (sMICA) in serum of patients with renal cell carcinoma, we used flow cytometry (FCM) and immunohistochemistry as well as an enzyme linked immunosorbent assay (ELISA). The results showed that percentage of mMICA expression was significantly increased in human kidney cancer tissues and RCC cell lines (786-O and Ketr-3) than that in healthy adults and human embryonic kidney 293 (HEK293) cell line individuality (P<0.05). sMICA content in healthy adults was negative, but in renal cancer patients was significantly elevated (P<0.05). Our research showed that high expression of MICA in human kidney cancer, this results show that MICA might serve as potential tumor-associated antigen (TAA) in RCC.

Immune Reconstitution of CD4+T Cells after Allogeneic Hematopoietic Stem Cell Transplantation and its Correlation with Invasive Fungal Infection in Patients with Hematological Malignancies

  • Peng, Xin-Guo;Dong, Yan;Zhang, Ting-Ting;Wang, Kai;Ma, Yin-Jian
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권8호
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    • pp.3137-3140
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    • 2015
  • Objective: To explore the immune reconstitution of $CD4^+T$ cells after allogeneic hematopoietic stem cell transplantation (Allo-HSCT) and its relationship with invasive fungal infection (IFI) in patients with hematological malignancies. Materials and Methods: Forty-seven patients with hematological malignancies undergoing Allo-HSCT in Binzhou Medical University Hospital from February, 2010 to October, 2014 were selected. At 1, 2 and 3 months after transplantation, the immune subpopulations and concentration of cytokines were assessed respectively using flow cytometry (FCM) and enzyme linked immunosorbent assay (ELISA). The incidence of IFI after transplantation and its correlation with immune reconstitution of $CD4^+T$ cells were investigated. Results: The number of $CD4^+T$ cells and immune subpopulations increased progressively after transplantation as time went on, but the subpopulation cell count 3 months after transplantation was still significantly lower than in the control group (p<0.01). In comparison to the control group, the levels of interleukin-6 (IL-6) and IL-10 after transplantation rose evidently (p<0.01), while that of transforming growth factor-${beta}$ (TGF-${beta}$) was decreased (p<0.01). There was no statistically significant difference level of interferon-${\gamma}$ (IFN-${\gamma}$) (p>0.05). The incidence of IFI was 19.2% (9/47), and multivariate logistic regression revealed that IFI might be related to Th17 cell count (p<0.05), instead of Th1, Th2 and Treg cell counts as well as IL-6, IL-10, TGF-${beta}$ and IFN-${\gamma}$ levels (p>0.05). Conclusions: After Allo-HSCT, the immune reconstitution of $CD4^+T$ cells is delayed and Th17 cell count decreases obviously, which may be related to occurrence of IFI.

Induction of Mac-2BP by nerve growth factor is regulated by the PI3K/Akt/NF-κB-dependent pathway in the HEK293 cell line

  • Park, Yuk-Pheel;Choi, Seung-Chul;Kim, Bo-Yeon;Kim, Jong-Tae;Song, Eun-Young;Kang, Seong-Ho;Yoon, Do-Young;Paik, Sang-Gi;Kim, Kwang-Dong;Kim, Jong-Wan;Lee, Hee-Gu
    • BMB Reports
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    • 제41권11호
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    • pp.784-789
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    • 2008
  • Mac-2BP is a ligand of the galectin family that has been suggested to affect tumor proliferation and metastasis formation. We assessed Mac-2BP expression at the transcriptional and translational levels to evaluate nerve growth factor (NGF)-induced Mac-2BP expression. A time kinetic analysis using reverse transcription-polymerase chain reaction showed that NGF-induced Mac-2BP transcript levels were 4-5 times higher than in controls. Mac-2BP enzyme-linked immunosorbent assay and immuno-fluorescence staining showed a 2-3-fold increase in intracellular and secreted Mac-2BP as a result of NGF stimulation. This increase was regulated by Akt activation and NF-${\kappa}B$ binding. p65 and p50-NF-${\kappa}B$ are major transcriptional factors in the Mac-2BP promoter region, and were shown to be regulated in accordance with the Akt activation states. Collectively, these results suggest that NGF induces Mac-2BP expression via the PI3K/Akt/NF-${\kappa}B$ pathway.

Sex hormones alter the response of Toll-like receptor 3 to its specific ligand in fallopian tube epithelial cells

  • Zandieh, Zahra;Amjadi, Fatemehsadat;Vakilian, Haghighat;Aflatoonian, Khashayar;Amirchaghmaghi, Elham;Fazeli, Alireza;Aflatoonian, Reza
    • Clinical and Experimental Reproductive Medicine
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    • 제45권4호
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    • pp.154-162
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    • 2018
  • Objective: The fallopian tubes play a critical role in the early events of fertilization. The rapid innate immune defense is an important part of the fallopian tubes. Toll-like receptor 3 (TLR3), as a part of the innate immune system, plays an important role in detecting viral infections. In this basic and experimental study, the effect of sex hormones on the function of TLR3 in the OE-E6/E7 cell line was investigated. Methods: The functionality of TLR3 in this cell line was evaluated by cytokine measurements (interleukin [IL]-6 and IL-1b) and the effects of sex hormones on TLR3 were tested by an enzyme-linked immunosorbent assay kit. Additionally, TLR3 small interfering RNA (siRNA) and a TLR3 function-blocking antibody were used to confirm our findings. Results: The production of IL-6 significantly increased in the presence of polyinosinic-polycytidylic acid (poly(I:C)) as the TLR3 ligand. Using a TLR3-siRNA-ransfected OE-E6/E7 cell line and function-blocking antibody confirmed that cytokine production was due to TLR3. In addition, 17-${\beta}$ estradiol and progesterone suppressed the production of IL-6 in the presence and absence of poly(I:C). Conclusion: These results imply that sex hormones exerted a suppressive effect on the function of TLR3 in the fallopian tube cell line when different concentrations of sex hormones were present. The current results also suggest that estrogen receptor beta and nuclear progesterone receptor B are likely to mediate the hormonal regulation of TLR3, as these two receptors are the main estrogen and progesterone receptors in OEE6/E7 cell line.

Cell-Surface Loss of Constitutive Activating and Inactivating Mutants of Eel Luteinizing Hormone Receptors

  • Byambaragchaa, Munkhzaya;Choi, Seung-Hee;Kim, Dong-Wan;Min, Kwan-Sik
    • 한국발생생물학회지:발생과생식
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    • 제25권4호
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    • pp.225-234
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    • 2021
  • The present study aimed to investigate the mechanism of cell surface receptor loss by two constitutively activating mutants (designated L469R, and D590Y) and two inactivating mutants (D417N and Y558F) of the luteinizing hormone receptor (LHR) in the Japanese eel Anguilla japonica, known to naturally occur in human LHR transmembrane domains. We investigated cell surface receptor loss using an enzyme-linked immunosorbent assay in HEK 293 cells. The expression level of wild-type eel LHR was considered to be 100%, and the expression levels of L469R and D417N were 97% and 101%, respectively, whereas the expression levels of D590Y and Y558F slightly increased to approximately 110% and 106%, respectively. The constitutively activating mutants L469R and D590Y exhibited a decrease in cell surface loss in a manner similar to that of wild-type eel LHR. The rates of loss of cell surface agonist-receptor complexes were observed to be very rapid (2.6-6.2 min) in both the wild-type eel LHR and activating mutants. However, cell surface receptor loss in the cells expressing inactivating mutants D417N and Y558F was slightly observed in the cells expressing inactivating mutants D417N and Y558F, despite treatment with a high concentration of agonist. These results provide important information on LHR function in fish and the regulation of mutations of highly conserved amino acids in glycoprotein hormone receptors.

Fusarium속이 생성하는 zearalenone 측정을 위한 Indirect Competitive ELISA의 확립 (Establishment of Indirect Competitive ELISA for the Detection of Zearalenone Produced by Fusarium sp.)

  • 강성조;정덕화
    • 한국식품위생안전성학회지
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    • 제13권4호
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    • pp.419-424
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    • 1998
  • Zearalenone 검출을 위하여 Z-M-26 hybridoma cell을 mouse의 복강에 투여한 후 생산, 정제한 항체와 합성한 Zearalenone-oxime-OVA conjugate를 이용하여 ELISA법을 확립하였다. Carbonyl buffer로 희석한 Zearalenone-oxime-OVA conjugate를 4$^{\circ}C$에서 하룻밤 coating 하고 1% BSA용액으로 하룻밤 blocking한 다음, 1,000배 희석한 항체를 Zearalenone 또는 시료와 혼합하여 하룻밤 반응시키는 것이 효과적이었다. 또한 2차 항체와 기질용액의 반응시간은 각각 1시간, 30분이 적당하였고, 발색된 반응액 450nm에서 측정하였다. 이 분석법의 결과 0.1~100 ppb의 Zearalenone이 측정 가능하였으며, 본 실험에서 확립한 indirect competitive ELISA법은 농산물 중 Zearalenone 분석에 효과적으로 활용할 수 있으리라 생각된다.

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RBL-2H3 cells에서 GyoKeisamultang-gagam의 항알레르기 효과 (Anti-allergic Effect of Gyokejsamu1tang-gagam(艽桂四物湯-加減) in the RBL-2H3 Mast Cells)

  • 총배빈;신우진;류지효;김성희;윤화정;김원일
    • 한방안이비인후피부과학회지
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    • 제21권3호
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    • pp.94-103
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    • 2008
  • 비만세포의 한 종류인 rat basophilic leukemia (RBL-2H3) 세포를 이용하여 교계사물탕-가감의 항알레르기 효과를 확인하고자 하였다. Phorbol 12-myristate 13-acetate (PMA)와 calcium ionophore A23187을 이용하여 RBL-2H3 세포를 자극한 후 세포의 탈과립 정도를 $\beta$-hexosaminidase assay로 확인한 결과, 전 처리한 교계사물탕-가감의 농도 의존적으로 탈과립을 억제하였다. Pro-inflammatory cytokines인 tumor necrosis factor (TNF)-alpha와 interleukin(IL)-4의 분비량을 enzyme-linked immunosorbent assay (ELISA)로 확인한 결과 교계사물탕-가감의 농도 의존적으로 감소하였으며, 이들 cytokines와 염증 반응에 주요한 인자인 cyclooxygenase (COX)-2 의 mRNA 발현 정도 역시 교계사물탕-가감에 의해 감소함을 확인할 수 있었다. 이러한 실험 결과로 보아 교계사물탕-가감은 알레르기 관련 질환의 치료에 응용될 수 있을 것으로 사료된다.

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Pregnancy outcomes following the administration of high doses of dexamethasone in early pregnancy

  • Ahmadabad, Hasan Namdar;Jafari, Sabah Kayvan;Firizi, Maryam Nezafat;Abbaspour, Ali Reza;Gharib, Fahime Ghafoori;Ghobadi, Yusef;Gholizadeh, Samira
    • Clinical and Experimental Reproductive Medicine
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    • 제43권1호
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    • pp.15-25
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    • 2016
  • Objective: In the present study, we aimed to evaluate the effects of high doses of dexamethasone (DEX) in early pregnancy on pregnancy outcomes. Methods: Pregnant BALB/c mice were treated with high-dose DEX in the experimental group or saline in the control group on gestational days (GDs) 0.5 to 4.5. Pregnant mice were sacrificed on GDs 7.5, 13.5, or 18.5 and their peripheral blood, placentas, fetuses, and uterine tissue were collected. Decidual and placenta cell supernatants were examined to evaluate the effect of DEX on the proliferation of mononuclear cells, the quantity of uterine macrophages and uterine natural killer (uNK) cells, and levels of progesterone and $17{\beta}-estradiol$, as determined by an 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide assay, immunohistochemistry, and enzyme-linked immunosorbent assay, respectively. We also were measured fetal and placental growth parameters on GD 18.5. Results: We found that high doses of DEX were associated with an increased abortion rate, enhancement of the immunosuppressive effect of the decidua, alterations in placental growth parameters, decreased progesterone and $17{\beta}-estradiol$ levels, and a reduced frequency of macrophages and uNK cells. Conclusion: Our data suggest that the high-dose administration of DEX during early pregnancy negatively affected pregnancy outcomes.

유백피, 토복령 추출물 혼합물의 항산화, 항염, 항균 및 난소세포 보호효과 (Antioxidant, anti-inflammatory, antibacterial and ovoprotective effects of mixture of Ulmi cortex and Smilacis rhizoma extracts)

  • 전상규;안정윤;박수미;박선동;이주희
    • 대한한의학방제학회지
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    • 제28권1호
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    • pp.41-51
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    • 2020
  • Objectives : US extract is a mixture of each extract of Ulmi cortex and Smilacis rhizoma. In this study, we investigated the antioxidant, anti-inflammatory, antibacterial, and ovoprotective effects of US extract in in vitro model to identify potential candidates for improving female reproductive function. Methods : The antioxidant activity of US extract was measured using 1,1-diphenyl- 2-picrylhydrazyl free radical and superoxide anion radical scavenging assays. The anti-inflammatory effect of US extract on lipopolysaccharide (LPS)-stimulated RAW 264.7 cells were determined with a nitric oxide (NO) assay, enzyme linked immunosorbent assays, and western blots analysis. The antibacterial activity of US extract against vaginitis infection microorganisms were determined with disc diffusion and minimum inhibitory concentration assays. The ovoprotective effect of US extract on 4-vinylcyclohexene diepoxide (VCD)-induced ovotoxicity in CHO-K1 cells were evaluated with a cell viability assay. Result : US extract showed good antioxidant capacity and inhibited LPS-induced NO production as well as iNOS and COX-2 expression and secretion of pro-inflammatory cytokine IL-6 without affecting the cell viability. It showed significant clear zones for Staphylococcus aureus and Candida albicans but did not indicate the clear zones for Escherichia coli and Enterococcus faecium. VCD-induced ovotoxicity in CHO-K1 cells was significantly reduced by US extract pre-treatment. Conclusions : These results demonstrate that US extract has antioxidant activity, anti-inflammatory effects on the LPS-stimulated macrophages, antibacterial activity against vaginitis infection microorganisms, and protective effects on the ovarian cells against VCD-induced ovotoxicity. These findings suggest that the US extract can be used as new prescriptions, supplements, functional foods, and cosmetics for improving female reproductive function.