• Title/Summary/Keyword: Marker enzyme

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Detection of Serum Anti-Extracellular Protein Kinase a Autoantibodies as a Potential Tumor Marker

  • Lee, Seung-Ho;Kim, Ki-Nam;Seo, Sang-Hui;Sohn, Sung-Hwa;Kim, Yu-Ri;Kim, Hye-Won;Choi, Chul-Won;Kim, Jun-Suk;Kim, Meyoung-Kon
    • Molecular & Cellular Toxicology
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    • v.2 no.1
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    • pp.67-73
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    • 2006
  • In previous studies, it has been discovered that cancer cells not only overexpress regulatory subunit I (Rl)/protein kinase type I (PKA-I) but also secrete outside the cell an extracellular form of PKA (ECPKA) and that the ECPKA secretion detected in patients' serum is obviously greater than that found in non-cancer patients or healthy subjects. We now found that ECPKA elicits the formation of serum autoantibodies that can serve as a cancer diagnostic and prognostic marker. To measure the presence of anti-ECPKA autoantibody in the human sera, basic methodology for ECPKA assay was established an enzyme-linked immunosorbent assay (ELISA). We obtained serum samples from 199 patients with different types of cancer, and also obtained 31 serum samples to compare with ECPKA concentrations from non-cancer patients and 119 normal volunteers. Compared with normal or non-cancer patient sera, we found that the frequency of anti-ECPKA autoantibody was significantly higher in cancer patients (88%) than in those without cancer (17%). Furthermore the presence of anti-ECPKA autoantibodies in the serum of cancer patients was highly correlated with the site of metastasis. The immunoassay developed for anti-ECPKA antibodies is highly sensitive and specific. Therefore, this discovery of an autoantibody-based cancer diagnostic may have serious clinical application and may become an important advance over current technology.

Identification of Korean Native Pork Using Breed-Specific DNA Marker of KIT Gene

  • Chung, Eui-Ryong;Chung, Ku-Young
    • Food Science of Animal Resources
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    • v.30 no.3
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    • pp.403-409
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    • 2010
  • Accurate methods for the identification of closely related species or breeds in raw and processed meats must be developed in order to protect both consumers and producers from mislabeling and fraud. This paper describes the development of DNA markers for the discrimination and improvement of Korean native pig (KNP) meat. The KIT gene is related to pig coat color and is often used as a candidate marker. A 538 bp fragment comprising intron 19 of the pig KIT gene was amplified by PCR using specific primers, after which the PCR amplicons of a number of meat samples from KNP and three major improved breeds (Landrace, Duroc and Yorkshire) were sequenced in order to find a nucleotide region suitable for PCR-RFLP analysis. Sequence data showed the presence of two nucleotide substitutions, g.276G>A and g.295A>C, between KNP and the improved pig breeds. Digestion of KIT amplicons with AccII enzyme generated characteristic PCR-RFLP profiles that allowed discrimination between meats from KNP and improved pig. KNP showed three visible DNA bands of 264/249, 199, and 75 bp, whereas DNA bands of 249, 199, and 90 bp were detected in the three improved pig breeds. Therefore, the 75 bp DNA fragment was specific only to KNP, whereas the 90 bp DNA fragment was specific to the improved breeds. The breed-specific DNA markers reported here that target the KIT gene could be useful for the identification of KNP meat from improved pig meats, thus contributing to the prevention of falsified breed labeling.

Matrix Metalloproteinase-13 - A Potential Biomarker for Detection and Prognostic Assessment of Patients with Esophageal Squamous Cell Carcinoma

  • Sedighi, Maryam;Aledavood, Seyed Amir;Abbaszadegan, MR;Memar, Bahram;Montazer, Mehdi;Rajabian, Majid;Gholamin, Mehran
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.6
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    • pp.2781-2785
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    • 2016
  • Background: Matric metalloproteinase (MMP) 13 gene expression is increased in esophageal squamous cell carcinomas (ESCCs) and associated with increasing tumor invasion, lymph node involvement and decreased survival rates. Levels of the circulating enzyme may be elevated and used as a marker of tumor progression. In this study, clinical application of MMP-13 serum levels was evaluated for early detection, prediction of prognosis and survival time of ESCC patients. Materials and Methods: Serum levels of MMP13 were determined by ELISA in 66 ESCC patients prior of any treatment and 54 healthy controls for comparison with clinicopathological data through statistical analysis with Man Whitney U and Log-Rank tests. In addition, clinical value of MMP13 levels for diagnosis was evaluated by receiver operating characteristic (ROC) test. Results: The serum level of MMP-13 in patients (>250 pg/ml) was significantly higher than in the control group (<100 pg/ml) (p value=0.004). Also the results showed a significant correlation between MMP-13 serum levels with tumor stage (p value = 0.003), depth of tumor invasion (p value=0.008), involvement of lymph nodes (p value = 0.011), tumor size (p value = 0.018) and survival time. While there were no significant correlation with grade and location of tumors. ROC analysis showed that MMP-13 level is an accurate diagnostic marker especially to differentiate pre-invasive/ invasive lesions from normal controls (sensitivity and specificity: 100%). Conclusions: These findings indicate a potential clinical significance of serum MMP13 measurement for early detection and prognostic assessment in ESCC patients.

Evaluation of Clinical Usefulness of Gamma Glutamyl Transferase as a Surrogate Marker for Metabolic Syndrome in Non Obese Adult Men (비만하지 않은 성인 남성에서 대사증후군의 대리 표지자로서 감마 글루타밀 전이효소의 임상적 유용성 평가)

  • Shin, Kyung-A;Kim, Eun Jae
    • Journal of Convergence for Information Technology
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    • v.10 no.12
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    • pp.146-155
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    • 2020
  • This study was to evaluate the usefulness of gamma glutamyl transferase (GGT) as a surrogate marker predicting metabolic syndrome. 7,155 non obese men over the age of 20 were studied as subjects. The criteria for diagnosing MetS were the National Cholesterol Education Program - Third Adult Treatment Panel (NCEP-ATP III). The risk of developing MetS according to GGT was conducted logistic regression analysis, and the ROC (receiver operating characteristic) curve was obtained to confirm GGT ability to predict the risk of MetS. Regardless of age and body mass index, MetS had a 7.09 times higher risk of onset in the fourth quartile than in the first quartile of GGT (p<0.001). The AUC (area under the curve) of GGT for the diagnosis of MetS was 0.715, and the cutoff value of GGT was 40.0 U/L, the sensitivity was 65.0%, and the specificity was 70.2%. Therefore, GGT is considered to be a useful diagnostic index for diagnosing MetS.

Study of plasma TGF-β1 level as a useful tumor marker in gastric cancer and prostate cancer (위암 및 전립선암의 종양 표지 인자로서 혈장 TGF-β1에 대한 연구)

  • Lim, Chang Ki;Shin, Hoon;Choi, In Young;Chung, Byung Ha;Ryu, Min Hee;Bang, Yung Jue;Jin, Seung Won
    • IMMUNE NETWORK
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    • v.1 no.3
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    • pp.260-265
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    • 2001
  • Transforming growth $factor-{\beta}1$ ($TGF-{\beta}1$) is a multipotent growth factor affecting development, homeostasis and tissue repair. Many kinds of malignant tissues were reported to overexpress transforming growth $factor-{\beta}1$ ($TGF-{\beta}1$) gene. However, a little work has been done on the circulating $TGF-{\beta}1$ and the association of $TGF-{\beta}1$ with progression in patients with malignant tumors. In this study, we measured the plasma level of $TGF-{\beta}1$ in gastric cancer and prostate cancer patients and evaluated the utility of plasma $TGF-{\beta}1$ as a possible tumor marker. We used Enzyme-linked immunosorbent assay (ELISA) system in order to measure plasma $TGF-{\beta}1$ level in 134 gastric cancer patients, 50 prostate cancer patients and 290 normal controls. And the tumor marker, carcinoembryonic antigen (CEA), prostate-specific antigen (PSA), was compared with $TGF-{\beta}1$ in the aspects of sensitivity and specificity. The mean plasma $TGF-{\beta}1$ levels were $1.219{\pm}0.834$ (0.272-5.772) ng/mL in normal controls, $5.964{\pm}3.218$ (0.845-18.124) ng/mL in gastric cancer and $4.140{\pm}2.345$ (1.108-13.302) ng/mL in prostate cancer. In gastric cancer patients difference in plasma $TGF-{\beta}1$ level was not detected according to cancer stage. In comparison with other tumor marker (CEA, PSA) $TGF-{\beta}1$ is more potent in sensitivity. These results indicate that the plasma $TGF-{\beta}1$ level can be a potent tumor marker in gastric cancer and prostate cancer.

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Inhibitory Effect of Jeju Tea Extracts and Vanadate on Postprandial Hyperglycemia and Hypertension, and In Vitro Study (제주산 녹차, 발효차 추출물과 바나듐의 식후 혈당강하 및 항고혈압 기능에 미치는 In Vitro 효과)

  • Park, Shin Young
    • Korean Journal of Clinical Laboratory Science
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    • v.52 no.4
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    • pp.398-407
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    • 2020
  • The inhibitory effect on α-glucosidase, a marker of postprandial hyperglycemia, and angiotensinconverting enzyme (ACE), a marker of hypertension, was analyzed using non-fermented green tea and three different types of fermented tea, which are popular beverages in modern life. Green tea was mixed with trace amounts of vanadate (50 ㎍/mL), which has insulin-mimetic effects, to investigate the synergistic effect of vanadate on the inhibition of α-glucosidase. The concentration of epigallocatechin gallate (EGCG) and caffeine was also checked. The extracts of green tea and fermented teas showed clear inhibition on α-glucosidase, which caused a decrease in the postprandial glucose levels. The inhibitory effect was most prominent in the 20% fermented tea. Trace amounts of vanadate (50 ㎍/mL)-mixed green tea extract had twice the inhibitory effect on α-glucosidase than the pure tea extract. All teas showed inhibitory effects on ACE. Among those, the effect was most prominent in green tea, which had higher concentrations of EGCG. In contrast, the postprandial glucose-lowering effect and ACE inhibition of the fermented teas, which have a lower level of EGCG, was attributed to some other different functional substances.

The Efficient Transformation of Pleurotus ostreatus using REMI Method

  • Joh, Joong-Ho;Kim, Beom-Gi;Chu, Kyo-Sun;Kong, Won-Sik;Yoo, Young-Bok;Lee, Chang-Soo
    • Mycobiology
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    • v.31 no.1
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    • pp.32-35
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    • 2003
  • Restriction enzyme-mediated integration(REMI) was used to transform uracil auxotrophs of Pleurotus ostreatus to prototrophy. When protoplasts of Pleurotus ostreatus were treated by the reaction mixture containing 10 units of BamHI, the frequency of REMI was about 64 transformants per 1 ${\mu}g$ of DNA. This efficiency was increased by 14.2 times compared with that of the conventional PEG transformation. The optimal condition for REMI of P. ostreatus was achieved when 1 ${\mu}g$ of linearized pTRura3-2 DNA was added into $1{\times}10^7$ protoplasts along with 10 units BamHI. Southern blot analysis revealed that about 50% of transformants examined were caused by REMI event and 30% carried single copy insertion at the genome. This suggested that the REMI method might be a useful tool for efficient transformation and tagging mutagenesis of P. ostreatus.

Molecular Characterization of Bile Salt Hydrolase from Bifidobacterium animalis subsp. lactis Bi30

  • Jarocki, Piotr
    • Journal of Microbiology and Biotechnology
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    • v.21 no.8
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    • pp.838-845
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    • 2011
  • The present work describes the identification, purification, and characterization of bile salt hydrolase (BSH) from Bifidobacterium animalis subsp. lactis. The enzyme was purified to electrophoretic homogeneity by hydrophobic chromatography, ion-exchange chromatography and ultrafiltration. SDS-PAGE analysis of putative BSH and gel filtration revealed that the analyzed protein is presumably a tetramer composed of four monomers each of about 35 kDa. The purified enzyme was analyzed by liquid chromatography coupled to LTQ FT ICR mass spectrometry and unambiguously identified as a bile salt hydrolase from B. animalis. The isoelectric point of the studied protein was estimated to be around pH 4.9. The pH optimum of the purified BSH is between 4.7 to 6.5, and the temperature optimum is around 50oC. The BSH of B. animalis could deconjugate all tested bile salts, with clear preference for glycine-conjugated bile salts over taurine-conjugated forms. Genetic analysis of the bsh showed high similarity to the previously sequenced bsh gene from B. animalis and confirmed the usefulness of bile salt hydrolase as a genetic marker for B. animalis identification.

Development of a Multicopy Integration Vector in Yarrowia lipolytica (Yarrowia lipolytica의 Multicopy Integration Vector 개발)

  • Kim, Jeong-Yoon;Woo, Moon-Hee;Ryu, Dewey D.Y.
    • Microbiology and Biotechnology Letters
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    • v.23 no.5
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    • pp.536-543
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    • 1995
  • Multicopy integration vector is a very useful vector system in that they can be integrated into chromosomal DNA in several copies and stably maintained under non-selective conditions. To develop a multicopy integration vector system in the yeast Yarrowia lipolytica, P-type ribosomal DNA was cloned from Y lipolytica. A HindIII-BglII fragment of the cloned rDNA and a promoterless URA3 gene were inserted into pGEM1, generating multicopy integration vectors, pMIYL-1 and pMIYL-2. The rDNA fragment is for targeted homologous recombination between the vector and the chromosomal DNA of Y. lipolytica, and the promoterless URA3 gene is a defective selection marker for inducing multicopy integration. pMIYL-1 and pMIYL-2 have an unique restriction enzyme site, KpnI, and two unique restriction enzyme sites, KpnI and EcoRI, repectively, which can be used for targeting of the vectors into the rDNA of Y. lipolytica chromosomal DNA. After transformation of the vectors into Y. lipolytica, copy number and stability were analyzed by Southern hybridization. The vectors were found to be present in less than 5 copies per cell and were stably maintained during growth in non-selective media.

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Anti-Proliferative and Anti-Carcinogenic Enzyme-inducing Activities of Delphinidin in Hepatoma Cells

  • Jang, Chan-Ho;Lee, In-Ae;Lim, Hyun-Ae;Kim, Ju-Ryoung;Ha, Young-Ran;Yu, Hoon;Sung, Mi-Kyung;Kim, Jong-Sang
    • Food Science and Biotechnology
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    • v.16 no.4
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    • pp.641-645
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    • 2007
  • Delphinidin, an aglycone form of anthocyanins, was demonstrated to have anti-carcinogenic potential. The compound at $50\;{\mu}g/mL$ caused a significant increase of quinone reductase activity, an anti-carcinogenic marker enzyme, in mouse hepatoma cell lines (Hepa1c1c7 and BPRc1). Delphinidin enhanced the expression of other detoxifying or antioxidant enzymes including glutathione s-transferase, gamma-glutamylcysteine synthetase, heme oxygenase 1, and glutathione reductase. It suppressed the proliferation of murine hepatoma cells in a dose-dependent manner, with approximately $IC_{50}$ of $70\;{\mu}g/mL$. These results suggest that delphinidin might be useful for cancer prevention.