• BMB Reports
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• 제30권6호
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• pp.403-409
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• 1997

Studies were undertaken to develop a competitive radioimmunoassay (RIA) and indirect antigen capture enzyme-linked immunosorbent assay (ELISA) for the determination of 5'-deoxy-5'-methylthioadenosine (MTA), which is formed from decarboxylated S-adenosylmethionine by spermidine and spermine synthase. Specific antiserum against MTA was raised in rabbits by immunization with MTA-BSA which was prepared by coupling BSA to oxidized MTA with periodate. Since MTA is oxidized easily to the sulfoxide, the sulfhydryl reagent, DTT. was added to the immunogen. For RIA, immunocomplexes were separated from free MTA by using ammonium sulfate precipitation. The antiserum showed almost no cross-reactivity with a variety of other nucleotides and riboses. But, the level of cross-reactivity of 5'-isobutylthioadenosine (SIBA) was high. These results showed the importance of hydrophobicity adjacent to the 5'-OH for determining antigenicity. The lower limit of detection by this assay was 100 fmol of MTA per tube. Using this assay. MTA levels were more easily and precisely determined in biological samples when compared with HPLC analysis. The RIA procedure is less time consuming. More than 24 analyses can be carried out in 2 h and required only a very small amount of sample ($20{\mu}l$ serum). In ELISA, biotin conjugated MTA-BSA was used as the labelled MTA. The sensitivity limit of this assay was lower than 100 pmol.

• Asian Pacific Journal of Cancer Prevention
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• 제16권7호
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• pp.2601-2611
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• 2015

Prostate cancer, with a lifetime prevalence of one in six men, is the second cause of malignancy-related death and the most prevalent cancer in men in many countries. Nowadays, prostate cancer diagnosis is often based on the use of biomarkers, especially prostate-specific antigen (PSA) which can result in enhanced detection at earlier stage and decreasing in the number of metastatic patients. However, because of the low specificity of PSA, unnecessary biopsies and mistaken diagnoses frequently occur. Prostate cancer has various features so prognosis following diagnosis is greatly variable. There is a requirement for new prognostic biomarkers, particularly to differentiate between inactive and aggressive forms of disease, to improve clinical management of prostate cancer. Research continues into finding additional markers that may allow this goal to be attained. We here selected a group of candidate biomarkers including PSA, PSA velocity, percentage free PSA, $TGF{\beta}1$, AMACR, chromogranin A, IL-6, IGFBPs, PSCA, biomarkers related to cell cycle regulation, apoptosis, PTEN, androgen receptor, cellular adhesion and angiogenesis, and also prognostic biomarkers with Genomic tests for discussion. This provides an outline of biomarkers that are presently of prognostic interest in prostate cancer investigation.

• Journal of Yeungnam Medical Science
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• 제31권1호
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• pp.52-55
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• 2014

Protein S deficiency is one of the several risk factors for thrombophilia and can cause blood clotting disorders such as deep vein thrombosis and pulmonary embolism. A 54-year-old man was admitted with the complaint of dyspnea and was diagnosed with pulmonary embolism. The patient had very low level of free protein S, total protein S antigen, and protein S activity (type I protein S deficiency). In history taking, we found that his mother, 78 year old, had a history of same disease 10 years ago, and confirmed the pronounced low level of protein S. The patient's son also had very low level of protein S, however there had not been any history of pulmonary embolism yet. This case study suggests that asymptomatic persons with a family history of protein S deficiency and pulmonary embolism should be checked regularly for early detection of the disease, as protein S deficiency can be suspected.

• Asian Pacific Journal of Cancer Prevention
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• 제15권14호
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• pp.5909-5916
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• 2014

Human papillomavirus infection (HPV) and HPV related immune perturbation play important roles in the development of cervical cancer. Since mature dendritic cells (DCs) are potent antigen-presenting cells (APC), they could be primed by HPV antigens against cervical cancers. In this study we were able to generate, maintain and characterize, both phenotypically and functionally, patient specific dendritic cells in vitro. A randomized Phase I trial with three arms - saline control (arm I), unprimed mature DC (arm II) and autologous tumor lysate primed mature DC (arm III) and fourteen patients was conducted. According to WHO criteria, grade 0 or grade one toxicity was observed in three patients. One patient who received tumor lysate primed dendritic cells and later cis-platin chemotherapy showed a complete clinical response of her large metastatic disease and remained disease free for more than 72 months. Our findings indicate that DC vaccines hold promise as adjuvant sfor cervical cancer treatment and further studies to improve their efficacy need to be conducted.

• Journal of Microbiology and Biotechnology
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• 제12권5호
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• pp.780-786
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• 2002

An immunosensor based on surface plasmon resonance (SPR) with a self-assembled protein G layer was developed for the detection of Escherichia coli O157:H7. A self-assembled protein C layer on a gold (Au) surface was fabricated by adsorbing the mixture of 11-mercaptoundecanoic acid (MUA) and hexanethiol at various molar ratios and by activating chemical binding between free amine (-$NH_2$) of protein G and 11-(MUA) using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDAC) in series. The formation of a self-assembled protein G layer on an Au substrate and the binding of the antibody and antigen in series were confirmed by SPR spectroscopy. The surface morphology analyses of the self-assembled protein G layer on the Au substrate, monoclonal antibody (Mab) against E. coli O157:H7 which was immobilized on protein G, and bound E. coli O157:H7 extracts on Immobilized Mab against E. coii O157:H7 were performed by atomic force microscopy (AFM). The detection limit of the SPR-based immunosensor for E. coli O157:H7 was found to be about $10^4$ cells/ml.

• Toxicological Research
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• 제27권2호
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• pp.111-118
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• 2011

This study was performed to evaluate the toxicity of polysaccharide-based Streptococcus pneumoniae vaccine in Specific Pathogen Free (SPF), Sprague-Dawley (SD) rats. S. pneumoniae vaccine was administrated subcutaneously each dose level of high (560 ${\mu}g$/rat), medium (280 ${\mu}g$/rat) and low (140 ${\mu}g$/rat) on days 0, 14, 28. The rats were observed for 2 weeks or 4 weeks after the final injection. During this test, there were no significant dose-dependent changes in body weight, water and food consumption. In urinalysis and serum chemistry, dose-related changes were not detected. In hematology, the percent of neutrophils and lymphocytes in white blood cells were changed significantly. According to the measurement of organ weight, only spleen weight was significantly increased in all groups of administration compared to the control group. In the histopathological examination, an antigen-deposit, vacuolated macrophages, infiltrated inflammatory cells and a formation of granulation tissue were observed at the site of an administration. These results are considered as an outcome by immune responses through a vaccination. Consequently, the results of this study demonstrated that S. pneumoniae vaccine has no toxicity when it was administrated subcutaneously three times in 2-week interval at a high dose of 560 ${\mu}g$/rat.

• Bulletin of the Korean Chemical Society
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• 제31권11호
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• pp.3259-3264
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• 2010

A label-free amperometric immunosensor has been proposed for the detection of myeloperoxidase (MPO) in human serum. To fabricate such an immunosensor, a composite film consisting of N,N-dimethylformamide (DMF), multiwall carbon nanotubes (MWCNTs) and 1-ethyl-3-methyl imidazolium tetrafluoroborate ($EMIMBF_4$) suspension was initially formed on a glassy carbon electrode (GCE). Then cerium dioxide ($CeO_2$) dispersed by chitosan was coated on the GCE. After that, MPO antibodies (anti-MPO) were attached onto the nano$CeO_2$ surface. With a noncompetitive immunoassay format, the antibody-antigen complex formed between the immobilized anti-MPO and MPO in sample solution. The immunosensor was characterized by cyclic voltammetry, transmission electron microscopy (TEM) and scanning electron microscopy (SEM). The factors influencing the performance of the immunosensor were studied in detail. Under optimal conditions, the current change before and after the immunoreaction was proportional to MPO concentration in the range of 5 to $300\;ng\;mL^{-1}$ with a detection limit of $0.2\;ng\;mL^{-1}$.

• 한국동물위생학회지
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• 제25권1호
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• pp.87-96
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• 2002

In this study, we wanted to determine if the respirotropic JMK strain of infectious bronchitis virus(IBV), which has a spike glycoprotein gene that is 99% similar to the nephropathogenic Gray strain of IBV, could adapt and cause lesions in the kidney following intracloacal passage in chickens. Two day old specific pathogen free(SPF) cchickens were infected with Gray and JMK strains by the intraocular and cloacal route. Several tissue samples were collected at various times. Viruses were recovered from more tissues and earlier in the infection from chickens infected cloacally than chickens infected intraocularly. Virus was isolated from the kidney of chickens infected with Gray by the intraocular route and JMK by the intracloacal route, but not from chicken given JMK the intraocular route. Histopathologically, interstitial nephritis was observed in Gray infected chickens. However, viral RNA or antigen were not detected in the kidney by in situ hybridization and immunohistochemistry. We further passaged the JMK strain ten times in two day old SPF chickens using cloacal inoculation. We examined the virus titer and histopathological change in the kidney at each passage level. The amount of virus recovered from the kidney was stable throughout this serial passage and the passaged virus did not caused renal damage. Further, virus could not be isolated from the kidney when chickens were infected with the passaged virus by the intraocular route. We conclude that the JMK strain has a strict upper respiratory tract tropism since cloacal passage did not produce nephrotropism or nephropathogenicity.

• 센서학회지
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• 제17권5호
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• pp.380-386
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• 2008

Quartz crystal microbalance immunosensor has a capacity to perform a label-free and real time detection of a trace amount of analyte through the specific interaction between antibody and antigen. However, immobilization of antibody molecules on the sensor surface is a troublesome procedure for researchers who are not experienced in chemistry. Protein G has a specific affinity to antibody and would serve as a capturing agent for antibody when immobilized on the sensor surface. In this work, we prepared a protein G sensor chip by immobilizing protein G on the surface of quartz crystal microbalance and examined its capability to detect human haptoglobin or human transferrin with unpurified corresponding antiserum. Specific and dose dependent response was observed when the protein G chip was used for detection of antigens after saturated with antiserum. We also verified several advantageous aspects of the protein G chip such as improved flexibility and sensitivity.

• Nuclear Medicine and Molecular Imaging
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• 제43권4호
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• pp.323-329
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• 2009

목적: 방사면역측정법 및 면역방사계수법은 매 실험마다 표준용액을 넣고 표준곡선에 대비하여 결과를 산출하는 batch assay가 일반화되어 있다. 비용과 시간에 대한 효율성을 증가 시키기 위해서는 선행 표준 곡선을 이용하여 검사를 진행하는 random assay의 적용이 요구된다. 방사면역측정법 및 면역방사계수법에서 자동분주기를 이용한 random assay적용 가능성을 평가 하기 위해 본 연구를 수행하였다. 대상 및 방법: Triiodothyronine (T3), free thyroxine (fT4), Prostate specific antigen (PSA), Carcinoembryonic antigen (CEA) 4가지 항목을 대상으로 하였으며 각 검사별 20명의 환자 검체를 이용하였다. 각 검사 항목별로 표준곡선을 얻기 위해 3시간 간격으로 4회 측정한 표준용액의 분당계수값(counter per minute, cpm)을 측정하였다. 이를 Friedman test를 이용하여 비교하여 검사시 마다 얻는 표준 곡선간의 차이를 평가하였다. 각 검사별 저, 중, 고농도의 대조 용액을 이용하여 5회 측정하고 이를 Friedman test를 이용하여 비교하여 측정 내 정밀도를 평가하였다. 각 검사별 저, 중, 고농도의 대조용액을 batch assay와 자동분주기를 이용한 random assay로 3시간 간격으로 각각 4회 측정하였다. 측정값을 바탕으로 평균, 표준 편차, 변이계수를 구하고 Wilcoxon test를 이용하여 비교하여 assay 방법에 따른 차이를 비교하였다. 두 assay간에 일치도(agreement)를 평가하기 위해 두 assay의 측정값 사이에 급내상관계수(Intraclass correlation coefficient, ICC)와 상관계수를 계산하였다. 결과: 모든 검사항목에서 표준 용액의 분당계수값은 측정 간에 통계적으로 유의한 차이가 없었다. T3, fT4, PSA, CEA 검사항목마다 측정한 저, 중, 고농도별 측정 내 변이계수는 T3(3.6%, 2.6%, 0.9%), fT4(4.5%, 1.1%. 1.3%), PSA(3.9%, 3.1%, 1.2%), CEA(5.0%, 4.8%, 7.6%)였다. 검사 항목별 측정방법에 따른 대조 용액을 이용한 표준 곡선의 측정 간 정밀도 평가에서 두 검사 방법간에 유의한 차이가 없었다. 각 검사항목별로 20명의 환자 검체에서 측정한 측정 간 변이계수(inter-assay precision)의 평균과 표준편차 값은 batch assay로 시행했을 때 각각 3.2$\pm$1.7%, 3.9$\pm$2.1%. 7.1$\pm$6.2%, 11.2$\pm$7.2%였고 random assay로 측정하였을 때는 2.7$\pm$1.7%, 4.8$\pm$3.1%, 3.6$\pm$4.8%, 7.4$\pm$6.2였다. Batch assay와 random assay는 서로 높은 일치도를 보였다. 측정한 급내상관계수(Intraclass correlation coefficient)는 T3, fT4, PSA, CEA 각각 0.9968, 0.9973. 0.9996, 0.9901였다. Batch assay로 측정한 값과 자등분주기를 이용한 random assay로 측정한 값 사이에 상관 계수(R$^2$)는 T3, fT4, PSA, CEA 각각 0.9924, 0.9974, 0.9994, 0.9989(p<0.005)로 모두 강한 상관관계를 보였다. 결론: T3, fT4, PSA, CEA 4가지 항목의 방사면역측정 법 (Radioimmunoassay)에서 기존의 batch assay와 자동분주기를 이용한 random assay간에 일치도와 상관성이 높았다. 결론적으로 자동분주기를 이용한 random assay를 방사면역측정법 및 면역방사계수법에 적절히 이용할 수 있을 것으로 생각된다.