• Title/Summary/Keyword: Marker enzyme

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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.

Molecular Cloning of a $\beta$-D-Galactosidase Gene from Lactococcus lactis subsp. lactis 7962

  • CHANG, HAE-CHOON;YANG-DO CHOI;HYONG-JOO LEE
    • Journal of Microbiology and Biotechnology
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    • v.6 no.6
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    • pp.386-390
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    • 1996
  • The ${\beta}$-galactosidase gene from Lactococcus lactis subsp. lactis ATCC 7962 was cloned and its enzymatic properties were characterized, with a view to assessing its potential use as a selection marker in the food-grade cloning vector. Chromosomal DNA from L. lactis subsp. lactis 7962 was cleaved with PstI and ligated into pBR322 for transformation into Escherichia coli TGl. Transformants showing ${\beta}$-galactosidase activity possessed the pBR322 plasmid containing a 10 kilobase (kb) PstI fragment and this plasmid was named pCKL11. The cloned ${\beta}$-galactosidase gene came from the chromosomal DNA of L. lactis subsp. lactis 7962 was confirmed by Southern hybridization. A restriction map of pCKL11 was constructed from the cleavage of both pCKL11 and the purified 10kb insert fraqment. The. optimum pH of the ${\beta}$-galactosidase determined with the E. coli harboring the pCKL11 was 7.0. The optimum temperature was $50^{\circ}C$, while the pI of the enzyme was 7.4. These values were the same as those of the enzyme from the parent strain.

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Generation of a Transformant Showing Higher Manganese Peroxidase (Mnp) Activity by Overexpression of Mnp Gene in Trametes versicolor

  • Yeo, Su-Min;Park, Nam-Mee;Song, Hong-Gyu;Choi, Hyoung-T.
    • Journal of Microbiology
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    • v.45 no.3
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    • pp.213-218
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    • 2007
  • Trametes versicolor has a lignin degrading enzyme system, which is also involved in the degradation of diverse recalcitrant compounds. Manganese-dependent peroxidase (MnP) is one of the lignin degrading enzymes in T. versicolor. In this study, a cDNA clone of a putative MnP-coding gene was cloned and transferred into an expression vector (pBARGPE1) carrying a phosphinothricin resistance gene (bar) as a selectable marker to yield the expression vector, pBARTvMnP2. Transformants were generated through genetic transformation using pBARTvMnP2. The genomic integration of the MnP clone was confirmed by PCR with bar-specific primers. One transformant showed higher enzyme activity than the recipient strain did, and was genetically stable even after 10 consecutive transfers on non-selective medium.

Effect of Dietary Minerals and Ca-Regulating Hormones on Bone Enzyme Alkaline Phosphatase Activity

  • Chung, Cha-Kwon;Ha, Kyung-Sun;Sohn, Jeong-In
    • Preventive Nutrition and Food Science
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    • v.1 no.1
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    • pp.80-86
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    • 1996
  • Parathyroid hormone(PTH) is known to stimulate bone resorption and to inhibit bone collagen synthesis. In contrast, as the evidence of stimulation of bone formation by PTH has recently been observed, the study on the role of PTH involved in osteoporosis draws remarkable attention. This study has dealt with the role of alkaline phoshatase(AP), a marker enzyme for bone formation and osteoblast action, Animals(BALS/cmice) were divided into three dietary groups(high and medium Ca and Ca-free) and hormones including PTH, calcitonin(CT), cholecalciferol(citamin D) were i.p. injected. AP in the serum and liver was measured using Sigma 221 alkaline buffer solutions containing 9mM of p-nitrophenyl phoshate. Enzume was reacted at 37$^{\circ}C$ for 10 minutes and the reaction was stopped by 1.8ml of 0.1N NaOH and measured at 410nm. We found that serum and liver AP activity was increased by low dietary Ac. Compared to the control, and serum Ap activity was enhanced by PTH and vitamin D regardless of the dietary Ca. On the other hand, liver AP activity was inhibited by OTH and vitamin D at all levels of dietary Ca. CT inhibited the action of PTH and vitamin D in the serum. But, the inhibition of PTH and vitamin D action by CT was not observed in the liver, unlike in the case of serum.

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Evaluation of the Herbal Extract Mixture for the Effects of Hair-Regrowth Compared to 3% Minoxidil; Elongation of Anagen Period on C3H Mice (모발생장기 유도 C3H 생쥐에 있어서 미녹시딜과 생약추출 혼합 조성물의 모발 재성장 유도 효능)

  • 이계호;한선일;박길흥;권영이
    • YAKHAK HOEJI
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    • v.47 no.1
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    • pp.14-19
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    • 2003
  • The hair cycle consists of three phases, growth (anagen), involution (catagen) and quiescence (telogen) phases. In order to evaluate hair re-growth effect of herbal extracts mixture containing the 70% ethanol extracts of Polygoni Multiflori Radix, Mori Cortex Radicis, Gingko Biloba Folium and Pine bud, we have examined the induction of the anagen phase and/or elongation of the anagen period using C3H mice. Morphological examination was done by Hattori' and Ogawa's method. Enzyme activities of ${\gamma}$-glutamyl transpeptidase (${\gamma}$-GT) and alkaline phosphatase (ALP) was detected by Bessey-Lovry-Brock's method. Enzyme activity as a biochemical marker of hair cycle was investigated in the third hair cycle period of C3H mice after depilation. 3% Minoxidil treated group and herbal extract mixture treated group were shown 3 days earlier initiation of anagen than control group. In cycling mouse skin, ${\gamma}$-GT activity is pronounced during anagen and greatly diminished during telogen. Herbal extract mixture has shown promising hair re-growth effect on hair follicular cycles of C3H mice.

Evaluation of the inhibitory effect of Gynostemma pentaphyllum extracts on CYP450 enzyme activities using LC-MS/MS

  • Jun Sang Yu;Young Seok Ji;So Young Jo;Xiang-Lan Piao;Hye Hyun Yoo
    • Mass Spectrometry Letters
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    • v.14 no.3
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    • pp.116-119
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    • 2023
  • Gynostemma pentaphyllum (Thunb.) Makino extract, a natural product with a history of traditional use, has gained attention for its potential health benefits. This study aimed to investigate its effects on key cytochrome P450 (CYP) enzymes using LC-MS/MS. Human liver microsomes and cDNA-expressed CYP2C8, CYP2C9, CYP2C19, and CYP3A4 supersomes were employed. Enzyme activity was assessed based on the formation of CYP-specific marker metabolites. The resulting data showed that the extract exhibited inhibitory effects on CYP2C8, CYP2C9, CYP2C19, and CYP3A4. Thus, G. pentaphyllum extract may influence the pharmacokinetics of drugs metabolized by CYP2C8, CYP2C9, CYP2C19, and CYP3A4. These findings emphasize the importance of considering potential herb-drug interactions when incorporating this extract into therapeutic regimens or dietary supplements.

Effects of Relative Lysyl Oxidase and Hydrogen Peroxide on Odontoblastic Differentiation (인간치수세포 분화과정에서 과산화수소에 대한 Lysyl Oxidase의 역할)

  • Lee, Hwa-Jeong
    • Journal of dental hygiene science
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    • v.13 no.3
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    • pp.321-329
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    • 2013
  • Although it has been reported that lysyl oxidase (LOX) is involved in odontoblastic differentiation, the role of LOX on odontoblastic differentiation by hydrogen peroxide ($H_2O_2$) have not been clarified. In the present study, we investigated whether $H_2O_2$, reactive oxygen species (ROS), is modulated the messenger RNA (mRNA) expression and activity of LOX during odontoblastic differentiation of human dental pulp (HDP) cells. The mRNA expression was quantified by reverse transcriptase polymerase chain reaction (RT-PCR) analysis, and LOX enzyme activity was measured by high sensitive fluorescent assay. Expression of the odontoblastic differentiation marker genes were assessed in the presence and absence of specific small interfering RNAs (siRNAs) of the LOX and LOXL. The $H_2O_2$-induced mRNA expression of LOX family was significant reduction of LOX, LOXL, and LOXL3 mRNA levels in HDP cells. LOX enzyme activity was increased at $H_2O_2$ 0.3 mM for 24 hours. The mRNA expression of alkaline phosphatase (ALP), osteopontin (OPN), and osteocalcin (OCN) was inhibited by LOX- and LOXL-specific siRNAs whereas the mRNA expression of dentin matrix protein1 (DMP1), and dentin sialophosphoprotein (DSPP) was inhibited by LOX-specific siRNA. In LOX enzyme activity, siRNA-induced knockdown of both LOX and LOXL inhibited the total amine oxidase activity in HDP cells, as in the case of mRNA expression. In conclusion, the essential role of $H_2O_2$ on odontoblastic differentiation suggests that its regulation by LOX may have pharmacologic importance in HDP cells.

Effect of Taeeumjoweetang on the Body Composition, Serum Lipid Level and Antioxidant Enzyme Activity of Obese Female College Students (태음조위탕의 적용이 태음인 비만여대생의 신체조성, 혈청지질농도 및 혈중 항산화 효소에 미치는 효과)

  • Kim, Hye-Ju;Ahn, Hong-Seok;Oh, Eun-Ha;Kim, Young-Locke
    • Journal of Sasang Constitutional Medicine
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    • v.23 no.3
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    • pp.391-401
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    • 2011
  • 1. Objectives: Though the eating habits have improved and the living method has become convenient according to the economic growth thanks to the modern industrialization, because of the lack of exercise, obesity, wrong eating habits and stress etc, various symptoms of disease of adults are on the rise. This is the phenomenon that happens as the eating life has become life in the West along with the inundation of the western culture in our society. In this perspective, there has been many various clinical research that's been proceeded so far about the physical constitution and obesity, but there has been little research on the objective analysis of the clinical research about the alimentotherapy using taeeumjoweetang. 2. Methods: In this research we have checked the weights, fat rates, fat weight, abdominal fat rate, blood pressure, and BMI over the objects of the women that were diagnosed as lunisolar system as their physical constitution, and assessed the paramecium lipid, in-blood antioxidation enzyme and the damage of oxidization in the urine by physical constitution-specific of the body shapes that were determined by BMI. The statistical analysis of the current research was processed by using of SPSS 17.0 program. We have figure out statistical amounts such as the arithmetic average, average deviation rate and percentage number. Fro the verification of he significancy of each elements, we have used the paired t-test, ANOVA, Chi-square test at the level of p<0.05. 3. Results and Conclusions: Their characteristics are age $21.20{\pm}1.35$, height $160.30{\pm}6.11cm$, weight $64.66{\pm}8.72kg$, fat rates are $35.97{\pm}4.87%$, fat amount $23.40{\pm}5.48$, abdominal fat rate $0.823{\pm}0.03$, BMI $25.12{\pm}2.79kg/m^2$, and systolic blood pressure $111.60{\pm}10.28mmHg$ and diastolic blood pressure $68.60{\pm}7.43mmHg$ and we have let them take the medicine twice a day for 8 weeks. The clinical result for the Cholesterol, Triglyceride, HDL-cholesterol, LDL-cholesterol, SGOT, SGPT, of the object people was that for the function of the liver, the result of the SGOT and SGPT test was $17.16{\pm}3.05$, $15.00{\pm}2.99IU/L$, which was a decrease, and had statistical meaning, but for the SGOT, though the figure reduced to $11.92{\pm}4.61$, $10.80{\pm}3.07$, it had no statistical meaning. For the whole cholesterol level, the figure reduced to $169.00{\pm}19.95$, $160.08{\pm}22.52$ mg/dL and had statistical significance(p<0.05). Neuter fat number, Triglyceride has slightly increased to $67.52{\pm}36.32$, $68.08{\pm}47.33$ mg/dL but did not have any statistical meaning. The antioxdant enzyme marker standard marker, SOD has increased to $2.52{\pm}0.73$, $2.86{\pm}0.60U/ml$, and had statistical significance(p<0.01). Catalase also increased by $0.63{\pm}0.18$, $1.07{\pm}0.25mmol/ml$ and had statistical meaning(p<0.01). GPx also increased to $204.76{\pm}32.64$ nmol/ml and had statistical meaning(p<0.01). But, for the Total antioxidant, though it has raised to $1.51{\pm}0.26$, $1.57{\pm}0.17nmol/{\mu}l$, it did not have any statistical meaning. MDA of oxidative stress marker has decreased to $1.70{\pm}0.68{\mu}g/ml$, $1.21{\pm}0.50{\mu}g/m$ and had statistical significance(p<0.01). 8-OHdG also decreased $3.35{\pm}0.95ng/ml$, $2.21{\pm}0.50ng/ml$ and had statistical meaning(p<0.01). In this research, we have analyzed the various markers relating to BFM and changes in oxidative enzyme in blood by takingtaeeumjoweetang. Taeeumjoweetang has the positive effect on inbody antioxidant system and reducing the content of cholesterol, which is proven to help losing weight and improving hyperlipidemia statistically. With this research, we hope to improve the lifestyle of those who are either obese or need to manage their dietary habits, and also to become the touchstone of integrating Oriental Medicine with the science of food & nutrition.

Analyses of single nucleotide polymorphisms and haplotypes of BoLA-DRB3 gene in Holstein and Hanwoo (홀스타인종과 한우에 있어서 BoLA-DRB3 유전자의 단일염기다형과 반수체 분석)

  • Jeong, Hang-Jin;Yu, Seong-Lan;Hoque, M.R.;Lee, Jun-Heon;Do, Chang-Hee;Ryoo, Seung-Heui;Sang, Byung-Chan
    • Korean Journal of Agricultural Science
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    • v.38 no.1
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    • pp.51-63
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    • 2011
  • BoLA (bovine leukocyte antigens) have been known as gene complex related with bovine diseases and immunological traits. This study was conducted to find out the characteristics of BoLA-DRB3 gene related to mastitis and BL(bovine leukocyte) from 280 cattle [193 animals of Holstein cattle and 87 animals of Hanwoo]. As a result, five PCR-RFLP types (b, d, e, f and g) using HaeIII restriction enzyme, three BstYI restriction patterns (b, d and e) and eight RsaI restriction types(b, d, f, I, j, n, o and w) were identified. Moreover, we identified new d' type ($197{\rightarrow}175$/22), having one more cutting site by BstYI enzyme than d type allele and n' type ($180{\rightarrow}169$/11) having one more cutting site by RsaI enzyme than n allele was additionally identified. Next, we identified 53 SNPs in BoLA-DRB3 exon2 from 280 cattle. SNP frequency and heterozygosity of Holstein and Hanwoo were investigated in all the SNP genotype. These results might be based on research for identifying marker associated with bovine diseases.

Measurement of DNA Damage with Fpg/Endo III FLARE Assay and Real Time RT-PCR in SD Rats Exposed to Cumene

  • Kim, Soo-Jin;Rim, Kyung-Taek;Lee, Seong-Bae;Kim, Hyeon-Yeong
    • Molecular & Cellular Toxicology
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    • v.4 no.3
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    • pp.211-217
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    • 2008
  • To clarify the DNA damage from reactive oxygen species, we measured the DNA damage through Fpg/Endo III FLARE (Fragment Length Analysis with Repair Enzyme) assay and real time RT-PCR. The 80 SD rats assigned to 4 dose groups exposed to cumene vapor for 90 days. With Fpg/Endo III FLARE assay in hepatocytes, we found the OTM (Olive Tail Moment) and TL (Tail Length) significantly increased in no-enzyme treated and Fpg-treated control and 8 ppm groups with 28 days exposure. In Endo III-treated 8 ppm group, significantly increased the values with 90 days exposure. With lymphocytes, it was founded the values significantly increased in no-enzyme treated 800 ppm group in 28 and 90 days. It was significantly increased in Endo III-treated 80 ppm for 28 days and 800 ppm for 90 days. From the above findings, FLARE assay was suggested as being available as a biological marker for DNA damage induced by cumene exposure in SD rats. And we used real time RT-PCR for the OGG1 mRNA expression, it had dose-dependent biologic effects in 1 day exposure, but decrease the levels of rOGG1 mRNA. Our findings provide evidence that cumene exposure may cause suppression of rOGG1 in the rat hepatocytes or lymphocytes.