• Title/Summary/Keyword: Marker enzyme

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Induction of the Anticarcinogenic Marker Enzyme, Quinone Reductase, by Dalbergiae Lignum

  • Yin, Hu-Quan;Lee, Bang-Wool;Kim, Youn-Chul;Sohn, Dong-Hwan;Lee, Byung-Hoon
    • Archives of Pharmacal Research
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    • v.27 no.9
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    • pp.919-922
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    • 2004
  • The effect of an extract of Dalbergiae Lignum and four components that were isolated from the extract on the anticarcinogenic phase II marker enzyme, quinone reductase (QR), was investi-gated. Of the solvent extracts of Dalbergiae Lignum, the CH$_2$CI$_2$ fraction was the most potent in inducing QR activity, with a CD value (the concentration required to double the QR activity) of 29.5 $\mu$/mL. The CH$_2$CI$_2$ extract was further separated into six compounds, four of which were identified as 4-methoxydalbergione, latifolin, 4',6-dihydroxy-7-methoxyflavanone, and obtusafu-ran. Obtusafuran [CD = 1.1 $\mu$M; chemopreventive index (CI) = 101.9] and latifolin (CD = 1.7 $\mu$M; CI = 154.6) displayed potent QR inducing activity and high chemopreventive indices. Lati-folin and 4-methoxydalbergione were identified as strong DPPH-scavengers with half-maximal free radical scavenging concentrations of 15.9 and 17.2 $\mu$M, respectively.

Association of SNP Marker in the Thyroglobulin Gene with Carcass and Meat Quality Traits in Korean Cattle

  • Shin, S.C.;Chung, E.R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.2
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    • pp.172-177
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    • 2007
  • Thyroid hormones play an important role in regulating metabolism and can affect homeostasis of fat depots. The gene encoding thyroglobulin (TG), producing the precursor for thyroid hormones, has been proposed as a positional and functional candidate gene for a QTL with an effect on fat deposition. The SNP occurs in the 5' promoter region of the TG gene and is widely used in marker assisted selection (MAS) programs to improve the predictability of marbling level and eating quality in beef cattle. In this study, we identified three SNPs at the 5' promoter region of the TG gene in Korean cattle. Of the three SNPs identified in TG gene, the C257T and A335G were previously unreported new SNPs. The sequence data were submitted to GenBank (GenBank accession number: AY615525). The previously reported C422T SNP showed three genotypes, CC, CT and TT, by digestion with the restriction enzyme MflI using the PCR-RFLP method. A new allelic variant corresponding to the C${\rightarrow}$T and A${\rightarrow}$G mutations at positions 257 and 335, respectively, could be detected by the SSCP analysis. The gene-specific SNP marker association analysis indicated that the C422T SNP marker was significantly associated (p<0.05) with marbling score. Animals with the CC and CT genotypes had higher marbling score than those with the TT genotype. Results from this study suggest that TG gene-specific SNP may be a useful marker for meat quality traits in future MAS programs in Korean cattle.

A Biochemical Study for the Development of Genetic Marker on Salmonids in Korea (한국산 연어류에서 Genetic Marker 개발을 위한 생화학적 연구)

  • HONG Kyung-Pyo;MYOUNG Jung-Goo;SON Jin-Ki;PARK Chul-Won
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.27 no.1
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    • pp.83-88
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    • 1994
  • For the purpose of genetic stock indentification of three species of salmonid fishs and their hybrid, lactate dehydrogenase(LDH), malate dehydrogenase(MDH), isocitrate dehydrogenase(IDH), a-gylycerophosphate dehydrogenase(a-GPDH), malic enzyme(ME), 6-phospho-gluconate dehydrogenase(6-PGD), phosphoglucose isomerase(PGI) and phospho-glucomutase(PGM) from skeletal muscle, liver, heart and gill tissues in all three species were analyzed. Chum and masu salmon showed no polymorphic patterns in all isozyme loci, however rainbow trout were found to have polymorphic patterns at MDH-B, LDH and IDH loci. Especially, significant differences were found at MDH-B loci between the three species and the IDH patterns of rainbow trout were also different from the other two species. These loci therefore can be utilized as efficient genetic markers for the identification of hybrids and improve the efficiency of fish breeding. There was no difference except PGI between diploid and triploid isozyme patterns but PGI showed some potential as a marker for triploid in masu salmon.

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Tissue Culture Studies of Anthranilate Synthase the Tryptophan Biosynthetic Control Enzyme

  • Widholm, Jack.M.
    • Journal of Plant Biotechnology
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    • v.2 no.2
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    • pp.55-60
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    • 2000
  • Experiments initiated 30 years ago to obtain selectable markers have led to a series of studies of Trp biosynthesis and anthranilate synthase (AS) the control enzyme using largely plant tissue cultures since they have experimental properties that can be readily exploited. Enzymological and compound feeding studies provided evidence that AS is the control point in the Trp biosynthesis branch and that altering the AS feedback control by the selection of mutants resistant to the Trp analog 5-methyl-tryptophan (5MT) can lead to the overproduction of this important amino acid. Plants regenerated from these Trp overproducing lines of most species also had high free Trp levels but Nicotiana tabaum (tobacco) plants expressed the feedback altered AS only in cultured cells and not in the regenerated plants. further tests by transient and stable expression of the cloned promoter for the naturally occurring tobacco feedback-insensitive AS, denoted ASA2, confirmed the tissue culture specific nature of the expression control. The 5MT caused by the expression of a feedback-insensitive AS from tobacco has been used to select protoplast fusion hybrids with several species since the resistance is expressed dominantly. Recently the ASA2 gene has been used successfully as a selectable marker to select transformed Astragalus sinicus and Glycine max hairy roots induced by Agrobactetium rhizogenes. These results show that the ASA2y-subunit can interact with the y-subunit of another species to form active feedback-insensitive enzyme that may be useful for selecting transformed cells. Plastid DNA transformation of tobacco has also effectively expressed ASA2 in the compartment in which Trp biosynthesis is localized in the cell.

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Study of plasma transforming growth factor-β1 level as a useful tumor marker in various cancers (종양 표지 인자로서 혈장 Transforming Growth Factor-β1에 대한 연구)

  • Shin, Hoon;Lim, Chang Ki;Choi, In Young;Lee, Doo Yun;Noh, Dong Yong;Ryu, Min Hee;Lee, Hyo Suk;Bang, Yung Jue;Park, Jong Sup;Jin, Seung Won
    • IMMUNE NETWORK
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    • v.1 no.2
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    • pp.143-150
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    • 2001
  • Background : Many investigators have found transforming growth factor-${\beta}1$ (TGF-${\beta}1$) to be elevated in tumors. Changes in responsiveness to TGF-${\beta}1$ have been linked to malignant transformation, tumor progression and tumor regression. Many malignant cell lines of epithelial or hematopoietic origin are refractory to the antiproliferative effects of TGF-${\beta}1$. However, a little is known about the association of TGF-${\beta}1$ with progression of malignant tumor. Methods : In this study, we measured the plasma level of TGF-${\beta}1$ in various cancer patients and evaluated the utility of plasma TGF-${\beta}1$ as a possible tumor marker. Plasma TGF-${\beta}1$ levels were measured using enzyme-linked immunosorbent assay in cancer patients and normal controls. Carcinoembryonic antigen (CEA) and alpha-fetoprotein (AFP) as tumor marker were compared with TGF-${\beta}1$ in the aspects of sensitivity and specificity. Results : The mean of plasma TGF-${\beta}1$ levels was $1.219{\pm}0.834ng/ml$ in normal controls, $5.491{\pm}3.598ng/ml$ in breast cancer, $12.670{\pm}10.386ng/ml$ in lung cancer, $5.747{\pm}3.228ng/ml$ in hepatocellular carcinoma and $10.854{\pm}7.996ng/ml$ in cervical cancer. In comparison with CEA and AFP, TGF-${\beta}1$ is more sensitive. Conclusion : We conclude that the high levels of TGF-${\beta}1$ are common in the plasma of cancer patients. These results suggest that the plasma TGF-${\beta}1$ level can be a potent tumor marker in various cancer patients.

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Development of a Competitive Direct Enzyme-Linked Immunosorbent Assay for Teicoplanin

  • Lee, Hyang-Burm;Kwak, Bo-Yeon;Lee, Jae-Chan;Kim, Chang-Jin;Shon, Dong-Hwa
    • Journal of Microbiology and Biotechnology
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    • v.14 no.3
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    • pp.612-619
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    • 2004
  • A competitive direct enzyme-linked immunosorbent assay (cdELISA) was developed for selective and rapid detection of a glycopeptide antibiotic, teicoplanin (TP). TP was conjugated to bovine serum albumin (BSA) for use as an immunogen. Repeated subcutaneous injections of 0.5 mg of the conjugate was effective in generating specific polyclonal antibody (PAb) toward TP in rabbits, as determined by cdELISA. TP-horseradish peroxidase conjugate (TP-HRP) was used as an enzyme marker. The cdELISA was developed based on a competition reaction between TP-BSA PAb and TP-HRP conjugate. The TP-BSA PAb was highly sensitive (detection limit, 0.3 ng/ml and specific toward teicoplanin, showing no cross-reactivity to other glycopeptide antibiotics including vancomycin. There were good correlations ($r^2$=0.84 and 0.76, respectively) between cdELISA and microbiological assay, and high-performance liquid chromatography. The cdELISA system developed in this work is expected to be useful not only for selective and rapid monitoring of TP but also for study of TP pharmacokinetics.

Sulforaphane is Superior to Glucoraphanin in Modulating Carcinogen-Metabolising Enzymes in Hep G2 Cells

  • Abdull Razis, Ahmad Faizal;Noor, Noramaliza Mohd
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.7
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    • pp.4235-4238
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    • 2013
  • Glucoraphanin is the main glucosinolate found in broccoli and other cruciferous vegetables (Brassicaceae). The objective of the study was to evaluate whether glucoraphanin and its breakdown product sulforaphane, are potent modulators of various phase I and phase II enzymes involved in carcinogen-metabolising enzyme systems in vitro. The glucosinolate glucoraphanin was isolated from cruciferous vegetables and exposed to human hepatoma cell line HepG2 at various concentrations (0-25 ${\mu}M$) for 24 hours. Glucoraphanin at higher concentration (25 ${\mu}M$) decreased dealkylation of methoxyresorufin, a marker for cytochrome P4501 activity; supplementation of the incubation medium with myrosinase (0.018 U), the enzyme that converts glucosinolate to its corresponding isothiocyanate, showed minimal induction in this enzyme activity at concentration 10 ${\mu}M$. Quinone reductase and glutathione S-transferase activities were unaffected by this glucosinolate; however, supplementation of the incubation medium with myrosinase elevated quinone reductase activity. It may be inferred that the breakdown product of glucoraphanin, in this case sulforaphane, is superior than its precursor in modulating carcinogen-metabolising enzyme systems in vitro and this is likely to impact on the chemopreventive activity linked to cruciferous vegetable consumption.

Detection of Circulating Melanoma Cells by a Two-marker Polymerase Chain Reaction Assay in Relation to Therapy

  • Bitisik, Ozlem;Camlica, Hakan;Duranyildiz, Derya;Tas, Faruk;Kurul, Sidika;Dalay, Nejat
    • BMB Reports
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    • v.36 no.2
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    • pp.173-178
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    • 2003
  • Malignant melanoma is one of the most rapidly increasing cancer types, and patients with metastatic disease have a very poor prognosis. Detection of metastatic melanoma cells in circulation may aid the clinician in assessing tumor progression, metastatic potential, and response to therapy. Tyrosinase is a key enzyme in melanine biosynthesis. The gene is actively expressed in melanocytes and melanoma cells. Melan A is a differentiation antigen that is expressed in melanocytes. The presence of these molecules in blood is considered a marker for circulating melanoma cells. In this study, we analyzed the usefulness of this marker combination I evaluating the response to therapy in the blood of 30 patients with malignant melanoma. Circulating cells were detected by a reverse-transcriptase-polymerase-chain reaction. The tyrosinase expression was observed in 9 (30%) patients and Melan A in 19 (63.3%) patients before therapy. Following treatment, the tyrosinase mRNA was detected in only one patient, while Melan A transcripts were still present in 14 patients. We suggest that this molecular assay can identify circulating melanoma cells that express melanoma-associated antigens and may provide an early indication of therapy effectiveness.

Feasibility of Serum Pentosidine Level as a Potential Risk Factor for Osteoporotic Vertebral Compression Fracture

  • Choi, Dong-Hyuk;Lee, Sang-Min;Lim, Sung-An;Choi, Yong-Soo
    • Asian Spine Journal
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    • v.12 no.6
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    • pp.992-997
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    • 2018
  • Study Design: Feasibility study. Purpose: To evaluate the feasibility of using serum pentosidine level as a potential marker for osteoporotic vertebral compression fracture (OVCF). Overview of Literature: A review of previous studies suggests a negative correlation between serum pentosidine concentration and bone strength. However, it is unclear whether serum pentosidine level might be a potential marker of OVCF in Koreans. Methods: Forty patients who underwent bone mineral density examination were included in this study, and their serum pentosidine levels were prospectively analyzed. Serum pentosidine level was evaluated using enzyme-linked immunosorbent assay. Among all the patients, 11 with OVCF were assigned to the vertebral fracture group and 29 who did not have vertebral fracture were included in the non-fracture group. In addition, we used the Fracture Risk Assessment (FRAX) tool Korean version for assessing the 10-year probability of fracture. Results: There was a statistically significant difference in the mean serum pentosidine level (p=0.04) of the vertebral fracture group (110.8 ng/mL) and the non-fracture group (64.3 ng/mL). Logistic regression analyses showed that serum pentosidine was significantly associated with OVCF. The vertebral fracture group had significantly higher 10-year probability of major osteoporotic fracture as per FRAX than the non-fracture group. There was a positive correlation between pentosidine level and FRAX results (r=0.35, p=0.02). Conclusions: These results suggest that increased serum pentosidine level could be a potential marker for OVCF.

Induction of Quinone Reductase , an Anticarcinogenic Marker Enzyme, by Vitamin E in Both Hepalclc7 Cells and Mice

  • Kwon, Chong-Suk;Kim, Jong-Sang
    • Preventive Nutrition and Food Science
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    • v.4 no.2
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    • pp.122-124
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    • 1999
  • Induction of NAD(P)H : (quinone-acceptor) oxidoreductase (QR) which obligatory two electron reduction of quinones and prevents their participation in oxidative cycling and thereby the depletion of intracellular glutathione, has been used as a marker for chemopreventive agents. We postulated that vitamin E, an antioxidant, which induces QR as the gene of QR was reported to contain antioxidant reponsive element in the 5'-flanking region. Vitamin E resulted in significant induction of QR in both hepalclc7 cells and mouse tissues. QR induction was observed; to be maximal at 25uM vitamin E for hepalclc7 cells while it was maximal in the level of 2.5∼5 μmoles vitamin E/㎏ BW for mouse tissues. Thus the cancer-preventive effect of vitamin E may be exerted by it induction of intracellular detoxifying enzymes.

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