• Title/Summary/Keyword: antigen free

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Analysis of 5-aza-2'-deoxycytidine-induced Gene Expression in Lung Cancer Cell Lines (폐암 세포주에서 5-aza-2'-deoxycytidine 처치에 의해 발현되는 암항원 유전자 분석)

  • 김창수;이해영;김종인;장희경;박종욱;조성래
    • Journal of Chest Surgery
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    • v.37 no.12
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    • pp.967-977
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    • 2004
  • Background: DNA methylation is one of the important gene expression mechanisms of the cell. When cytosine of CpG dinucleotide in promotor is hypomethylated, expression of some genes that is controlled by this promoter is altered. In this study, the author investigated the effect of DNA demethylating agent, 5-aza-2'-deoxycytidine (ADC), on the expressions of cancer antigen genes, MHC and B7 in 4 lung cancer cell lines, NCIH1703, NCIH522, MRC-5, and A549. Material and Method: After treatment of cell lines, NCIH1703, NCIH522, MRC-5 and A549 with ADC (1 uM) for 48 hours, RT-PCR was performed by using the primers of MAGE, GAGE, NY-ESO-1, PSMA, CEA, and SCC antigen gene. In order to find the optimal ADC treatment condition for induction of cancer antigen, we studied the effect of ADC treatment time and dose on the cancer antigen gene expression. To know the effect of ADC on the expression of MHC or B7 and cell growth, cells were treated with 1 uM of ADC for 72 hours for FACS analysis or cells were treated with 0.2, 1 or 5 uM of ADC for 96 hours for cell counting. Result: After treatment of ADC (1 uM) for 48 hours, the expressions of MAGE, GAGE, NY-ESO-1, and PSMA genes increased in some cell lines. Among 6 MAGE isotypes tested, and gene expression of MAGE-1, -2, -3, -4 and -6 could be induced by ADC treatment. However, CEA gene expression did not change and SCC gene expression was decreased by ADC treatment. Gene expression was generally induced 24 - 28 hours after ADC treatment and expression of MAGE, GAGE, and NY-ESO-1 was maintained at least 14 days after ADC ADC teatment, and expression of MAGE, GAGE, and NY-ESO-1 was maintained at least 14 days after ADC teatment in ADC-Free medium. Most gene expression could be induced at 0.2 uM of ADC, but gene expression increased dependently on ADC treatment dose. The expression of MHC and B7 was not increased by ADC treatment in all four cell lines, and the growth rate of 4 cell lines decreased significantly with the increase of ADC concentrations. Conclusion: Treatment of lung cancer cell lines with ADC increases the gene expression MAGE, GAGE and NY-ESO-1 that are capable of induction of cytotoxic T lymphocyte response. We suggest that treatment with 1 uM of ADC for 48 hours and then culturing in ADC-free medium is optimal condition for induction of cancer antigen. However, ADC has no effect on MHC and B7 induction, additional modification for increase of expression of MHC, B7 and cytokine will be needed for production of efficient cancer cell vaccine.

Radioimmunoassay Reagent Survey and Evaluation (검사별 radioimmunoassay시약 조사 및 비교실험)

  • Kim, Ji-Na;An, Jae-seok;Jeon, Young-woo;Yoon, Sang-hyuk;Kim, Yoon-cheol
    • The Korean Journal of Nuclear Medicine Technology
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    • v.25 no.1
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    • pp.34-40
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    • 2021
  • Purpose If a new test is introduced or reagents are changed in the laboratory of a medical institution, the characteristics of the test should be analyzed according to the procedure and the assessment of reagents should be made. However, several necessary conditions must be met to perform all required comparative evaluations, first enough samples should be prepared for each test, and secondly, various reagents applicable to the comparative evaluations must be supplied. Even if enough comparative evaluations have been done, there is a limit to the fact that the data variation for the new reagent represents the overall patient data variation, The fact puts a burden on the laboratory to the change the reagent. Due to these various difficulties, reagent changes in the laboratory are limited. In order to introduce a competitive bid, the institute conducted a full investigation of Radioimmunoassay(RIA) reagents for each test and established the range of reagents available in the laboratory through comparative evaluations. We wanted to share this process. Materials and Methods There are 20 items of tests conducted in our laboratory except for consignment tests. For each test, RIA reagents that can be used were fully investigated with the reference to external quality control report. and the manuals for each reagent were obtained. Each reagent was checked for the manual to check the test method, Incubation time, sample volume needed for the test. After that, the primary selection was made according to whether it was available in this laboratory. The primary selected reagents were supplied with 2kits based on 100tests, and the data correlation test, sensitivity measurement, recovery rate measurement, and dilution test were conducted. The secondary selection was performed according to the results of the comparative evaluation. The reagents that passed the primary and secondary selections were submitted to the competitive bidding list. In the case of reagent is designated as a singular, we submitted a explanatory statement with the data obtained during the primary and secondary selection processes. Results Excluded from the primary selection was the case where TAT was expected to be delayed at the moment, and it was impossible to apply to our equipment due to the large volume of reagents used during the test. In the primary selection, there were five items which only one reagent was available.(squamous cell carcinoma Ag(SCC Ag), β-human chorionic gonadotropin(β-HCG), vitamin B12, folate, free testosterone), two reagents were available(CA19-9, CA125, CA72-4, ferritin, thyroglobulin antibody(TG Ab), microsomal antibody(Mic Ab), thyroid stimulating hormone-receptor-antibody(TSH-R-Ab), calcitonin), three reagents were available (triiodothyronine(T3), Tree T3, Free T4, TSH, intact parathyroid hormone(intact PTH)) and four reagents were available are carcinoembryonic antigen(CEA), TG. In the secondary selection, there were eight items which only one reagent was available.(ferritin, TG, CA19-9, SCC, β-HCG, vitaminB12, folate, free testosterone), two reagents were available(TG Ab, Mic Ab, TSH-R-Ab, CA125, CA72-4, intact PTH, calcitonin), three reagents were available(T3, Tree T3, Free T4, TSH, CEA). Reasons excluded from the secondary selection were the lack of reagent supply for comparative evaluations, the problems with data reproducibility, and the inability to accept data variations. The most problematic part of comparative evaluations was sample collection. It didn't matter if the number of samples requested was large and the capacity needed for the test was small. It was difficult to collect various concentration samples in the case of a small number of tests(100 cases per month or less), and it was difficult to conduct a recovery rate test in the case of a relatively large volume of samples required for a single test(more than 100 uL). In addition, the lack of dilution solution or standard zero material for sensitivity measurement or dilution tests was one of the problems. Conclusion Comparative evaluation for changing test reagents require appropriate preparation time to collect diverse and sufficient samples. In addition, setting the total sample volume and reagent volume range required for comparative evaluations, depending on the sample volume and reagent volume required for one test, will reduce the burden of sample collection and planning for each comparative evaluation.

Evaluation of Biochemical Recurrence-free Survival after Radical Prostatectomy by Cancer of the Prostate Risk Assessment Post-Surgical (CAPRA-S) Score

  • Aktas, Binhan Kagan;Ozden, Cuneyt;Bulut, Suleyman;Tagci, Suleyman;Erbay, Guven;Gokkaya, Cevdet Serkan;Baykam, Mehmet Murat;Memis, Ali
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.6
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    • pp.2527-2530
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    • 2015
  • Background: The cancer of the prostate risk assessment (CAPRA) score has been defined to predict prostate cancer recurrence based on the pre-clinical data, then pathological data have also been incorporated. Thus, CAPRA post-surgical (CAPRA-S) score has been developed based on six criteria (prostate specific antigen (PSA) at diagnosis, pathological Gleason score, and information on surgical margin, seminal vesicle invasion, extracapsular extension and lymph node involvement) for the prediction of post-surgical recurrences. In the present study, biochemical recurrence (BCR)-free probabilities after open retropubic radical prostatectomy (RP) were evaluated by the CAPRA-S scoring system and its three-risk level model. Materials and Methods: CAPRA-S scores (0-12) of our 240 radical prostatectomies performed between January 2000-May 2011 were calculated. Patients were distributed into CAPRA-S score groups and also into three-risk groups as low, intermediate and high. BCR-free probabilities were assessed and compared using Kaplan-Meier analysis and Cox proportional hazards regression. Ability of CAPRA-S in BCR detection was evaluated by concordance index (c-index). Results: BCR was present in 41 of total 240 patients (17.1%) and the mean follow-up time was $51.7{\pm}33.0$ months. Mean BCR-free survival time was 98.3 months (95% CI: 92.3-104.2). Of the patients in low, intermediate and high risk groups, 5.4%, 22.0% and 58.8% had BCR, respectively and the difference among the three groups was significant (P = 0.0001). C-indices of CAPRA-S score and three-risk groups for detecting BCR-free probabilities in 5-yr were 0.87 and 0.81, respectively. Conclusions: Both CAPRA-S score and its three-risk level model well predicted BCR after RP with high c-index levels in our center. Therefore, it is a clinically reliable post-operative risk stratifier and disease recurrence predictor for prostate cancer.

The Production of HBsAg in the Recombinant Yeast Cells (재조합 효모 세포내에서의 간염백신 생산)

  • Park, Cha-Yong;Lee, Hei-Chan
    • Microbiology and Biotechnology Letters
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    • v.14 no.6
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    • pp.455-460
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    • 1986
  • Dane particle was prepared from the plasma of chronic HBsAg carrier with high levels of HBsAg activity. DNA extracted front Dane particle core after a DNA polymerase reaction with $\alpha$-($^{32}$P) dNTP, was identified as HBV DNA by liquid scintillation counter and agarose gel electrophoresis-G.M. counting. To produce Hepatitis B surface antigen for use as a vaccine against Hepatitis B virus infection, yeast strains harboring recombinant plasmid with Apase promoter was used. Recombinant plasmid was construced from pHBV 130 and pAN 82, transformed into E coli, and then transferred into yeast strains. HBsAg was produced by derepression in Burkholder minimal medium with controlled inorganic phosphate concentration. The kinetics of HBsAg production was also investigated. Total HBsAg activity increased rapidly between 3 and 6 hours after transfer to phosphate-free medium and reached a maximum at around 9th hour. The transfer into phosphate-free medium after 6 hours in high phosphate cell growth medium gave maximum activity.

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Studies on Sexing of Bisected Mouse Embryos by Rat H-Y Antibody (Rat H-Y 항체에 의한 생쥐 분할란의 성 조절에 관한 연구)

  • 정장용;박희성;박충생
    • Korean Journal of Animal Reproduction
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    • v.15 no.3
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    • pp.179-187
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    • 1991
  • This experiment was carried out to develop a new technique of identifying XX of XY-bearing bisected embryos prior to implantation by immunological method. H-Y antiserum prepared in inbred Wastar female rats by repeated immunization with spleen cells from males of the same strain. The reactivity of H-Y antibody was confirmed by culturing mouse embryos in the medium containing H-Y antiserum and complement obtained from the guinea pig. The optimal condition for the activity of H-Y antibody was also investigated by culturing embryos under the concentraton or affected H-Y antibody was also investigated by culturing embryos under the concentration or affected H-Y antibody and culture rate. However, production of live young or sex rates of male and female from embryos transferred with psudopregnant. The biological test with the morula stage embryos showed that H-Y antibody was formed in all female rats immunized with spleen cell, but it was formed only in 80% female rats immunized with the antigen. When the bisected mouse embryos were cultured in vitro for 5~6 hours in morula stage, of 457 bisected embryos 81.4% of then were developed to the blastocyst stage. When the concentration rate of complement to H-Y antiserum varied from 1.0~5.0${mu}ell$, the lysis-rate of embryo was 19.5 to 67.3%. The concentration rate of complement did not influence the lysis-rate of embryos(P<0.05). The morphology embryos of bisected, zona-free and intact embryos showed the embryos lysis rate of 58.6, 42.7 and 48.5% respectively(P<0.05). Pregnancy rate were 50.0, 45.5 and 57.1% in psudopregnant recipient transferred with bisected, zona-free and intact blastocyst embryos. However, production of live youngs, sexual rate of male or female was 24(50.0:50.0), 22(45.5:55.5) and 36(58.3:41.7)mice, but affected and non affected half embryos with H-Y antiserum treatment was 23.1 and 26.7%. Also production of live youngs and sexual rate was 14(92.9:7.1) and 17(17.6:82.4)mice in affected and non affected half embryos in H-Y antiserum treatment(P<0.05).

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Graves disease following rabbit antithymocyte globulin treatment of severe aplastic anemia in a Korean child

  • Choi, In Su;Kim, Han Kyul;Han, Dong Kyun;Baek, Hee Jo;Jang, Hae In;Kim, Chan Jong;Kook, Hoon
    • Clinical and Experimental Pediatrics
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    • v.58 no.7
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    • pp.267-269
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    • 2015
  • Antithymocyte globulin (ATG) is used as an immunosuppressive treatment (IST) to deplete clonal suppressor T cells in patients with severe aplastic anemia (SAA). The depletion of suppressor T cells by ATG may affect the activation of B cells, which results in an increased risk for autoimmune conditions. A 12-year-old boy was diagnosed with idiopathic SAA. As he did not have an human leukocyte antigen-matched sibling, he was treated with rabbit ATG (3.5 mg/kg/day for 5 days) and cyclosporine. Five months later, he became transfusion independent. However, 23 months after IST, he complained of mild hand tremors, sweating, weight loss, palpitations, and goiter. Results of thyroid function tests revealed hyperthyroidism (free thyroxine, 3.42 ng/dL; thyroid stimulating hormone [TSH], <0.01 nIU/mL; triiodothyronine, 3.99 ng/mL). Results of tests for autoantibodies were positive for the antimicrosome antibody and TSH-binding inhibitory immunoglobulin, but negative for the antithyroglobulin antibody and antinuclear antibody. He was treated with methimazole, and his symptoms improved. The patient has been disease free for 39 months after IST and 9 months after methimazole treatment. This case report suggests that although rare, rabbit ATG may have implications in the pathogenesis of autoimmune hyperthyroidism. Our findings suggest that thyroid function tests should be incorporated in the routine follow-up of SAA patients treated with ATG.

A Survey of the Serotype Distribution of Streptococcus mutans Isolated from Dental Plaques of Caries-active and Caries-free Subjects (치아우식증환자와 치아정상인(齒牙正常人)의 치태(齒苔)에서 분리(分離)한 Streptococcus mutans의 혈청형(血淸型) 분포(分布)에 관(關)한 조사(調査) 연구(硏究))

  • Lee, Jean-Yong;Choi, Eu-Gene;Ha, Youn-Mun
    • The Journal of the Korean Society for Microbiology
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    • v.18 no.1
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    • pp.23-29
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    • 1983
  • An epidemiological survey was carried out to identify the serotypes of Streplococcus mutans isolated from carious lesions of 65 caries-active subjects(CAS) and sound tooth surfaces of 40 caries-free subjects(CFS). The autoclaved antigen extract was performed on each isolate, and then, serotypes of the isolates were determined in agar-gel immunodiffusion test. The results were as follows: 1. S. mutans was found in 78% of the samples of CAS, and of CFS. The difference of isolation frequency between CAS and CFS was not observed. 2. Only one serotype per single subject was detected in 61% of total samples, in remaining 39% of samples two or more serotypes were detected. 3. In 41.2% of CAS samples plural serotypes of S. mutans were found, whereas 35.5% of CFS samples showed plural serotypes distribution. 4. The most frequently identified serotype in each subject was serotype c; 69.5% of subjects harbored serotype c S. mutans. Serotype d was next most frequently isolated from subjects, comprising 23.2%. 5. Serotype c strain was found in 64.7% of CAS, 77.4% of CFS. 6. Of the isolates from CAS and CFS, serotype c was most commonly found, comprising 48.8%, serotype d was found in 16.3%, serotypes f. e, and g comprising 13.2%, 9.3%, and 7.8% respectively. Serotypes a and b were also found but in far lower frequencies(2.3%, 0.8%). 7. Serotype c strains were more found in CFS than in CAS, but serotypes d and e were more found in CAS.

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Recombinant human granulocyte macrophage colony-stimulating factor (rhGM-CSF) could accelerate burn wound healing in hamster skin

  • Heo, Si-Hyun;Han, Kyu-Boem;Lee, Young-Jun;Kim, Ji-Hyun;Yoon, Kwang-Ho;Han, Man-Deuk;Shin, Kil-Sang;Kim, Wan-Jong
    • Animal cells and systems
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    • v.16 no.3
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    • pp.207-214
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    • 2012
  • Burns are one of the most devastating forms of trauma and wound healing is a complex and multicellular process, which is executed and regulated by signaling networks involving numerous growth factors, cytokines, and chemokines. Recombinant human granulocyte macrophage colony-stimulating factor (rhGM-CSF) was specifically produced from rice cell culture through use of a recombinant technique in our laboratory. The effect of rhGM-CSF on promotion of deep second-degree burn wound healing on the back skin of a hamster model was evaluated through a randomized and double-blind trial. As macroscopic results, hamster skins of the experimental groups showed earlier recovery by new epidermis than the control groups. Immunohistochemical reactions of proliferating cell nuclear antigen and transforming growth factor-b1, which are indicators of cell proliferation, were more active in the experimental group, compared with the control group. On electron microscopy, basal cells in the epidermis of the experimental group showed oval nuclei, prominent nucleoli, numerous mitochondria and abundant free ribosomes. In addition, fibroblasts contained well-developed rough endoplasmic reticulum with dilated cisternae. Bundles of collagen fibrils filled the extracellular spaces. Particularly, ultrastructural features indicating active metabolism for regeneration of injured skin at 15 days after burn injury, including abundant euchromatin, plentiful free ribosomes, and numerous mitochondria, were observed. These findings suggest that use of rhGM-CSF could result in accelerated deep second-degree burn wound healing in animal models.

Development of Air Spring Damper System(ASDS) (공기 스프링 댐퍼 시스템(ASDS)의 개발)

  • Kim, Dong Baek;Park, Heung Sik
    • Journal of the Society of Disaster Information
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    • v.17 no.4
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    • pp.829-838
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    • 2021
  • Purpose: The Air Spring Damper System (ASDS) is proposed when existing concrete structures that have not been seismic resistant for economic and technical reasons or low-rise concrete structures that are difficult to earthquake. Method: To conduct a study on the damping force antigen in the kinetic equation of free vibration, we analyze whether this device has damping ability as a damper experimentally and theoretically, and examine the possibility of field application. Result: The air damper system is considered to be more economical than steel hysteresis dampers even if the number of dampers increases due to its easy manufacture and construction and low restrictions on shape, size, material, etc. Conclusion: In an air spring damper system, it is essential to reduce the diameter of the air inlet/outlet hole to improve the damping ratio, and in this case, if the diameter exceeds a certain lower limit, consideration of the compressibility of air is required, so further research is needed.

Surface Plasmon Resonance Immunosensor for Detection of Legionella pneumophila

  • Oh, Byung-Keun;Lee, Woochang;Bae, Young-Min;Lee, Won-Hong;Park, Jeong-Woo
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.8 no.2
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    • pp.112-116
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    • 2003
  • An immunosensor based on surface plasmon resonance (SPR) onto a protein G layer by Self-assembly technique was developed for detection of Legionella pneumophila. The protein G layer by self-assembly technique was fabricated on a gold (Au) surface by adsorbing the 11-mercaptoundecanoic acid (MUA) and an activation process for the chemical binding of the free amino (-NH$_2$) of protein G and 11-(MUA) using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDAC) in series. The formation of the protein G layer by self-assembly technique on the Au Substrate and the binding of the antibody and antigen in series were confirmed by SPR spectroscopy. The Surface topographies of the fabricated thin films on an Au substrate were also analyzed by using an atomic force microscope (AFM). Consequently, an immunosensor for the detection of L. pneumophila using SPR was developed with a detection limit of up to 10$^2$CFU per mL.