• Title/Summary/Keyword: marker substances

Search Result 57, Processing Time 0.031 seconds

An in vitro vitellogenin bioassay for estrogenic substances in the Rhynchocypris oxycephalus (버들치(Rhynchocypris oxycephalus)에서 에스트로겐류 물질에 의한 vitellogenin의 유도 및 단일클론항체 제작)

  • Shin, Jung-A;Song, Jae-Young;Lim, Hee-Young;Chang, Min-Ho;Lee, Hye-Min;Chung, Kyu-Hoi;Lim, Yoon-Kyu;Yoon, Byoung-Su
    • Korean Journal of Veterinary Research
    • /
    • v.46 no.1
    • /
    • pp.35-41
    • /
    • 2006
  • Vitellogenin (Vtg), a phospholipoglycoprotein precursor of egg yolk is synthesized and secreted from the liver in response to estrogens in female fish. Vtg is normally undetectable in the blood of male fish, but can be induced by exposure to chemicals possessing estrogenic activity. Thus, the presence of Vtg in blood of male fish can serve as a useful biomarker for assessing previous exposure to estrogenic compounds. In the present study, Vtg was abnormally expressed in Rhynchocypris oxycephalus using estradiol benzoate ($E_2$). As the result, it was found that the level of Vtg in blood from R. oxycephalus was increased by treated quantity of $E_2$ with dose-effect manner. Monoclonal antibodies were generated against Vtg of R. oxycephalus. The hybridoma were screened with an enzyme immunoassay for the production of specific anti-Vtg antibodies. Five positive cell lines with a high specificity were selected. Monoclonal antibodies against vtg of R. oxycephalus that was developed in this study, may be a useful bio-indicator for the detection of estrogenic contamination in the aquatic ecosystem.

New Method for Simultaneous Quantification of 12 Ginsenosides in Red Ginseng Powder and Extract: In-house Method Validation

  • In, Gyo;Ahn, Nam-Geun;Bae, Bong-Seok;Han, Sung-Tai;Noh, Kil-Bong;Kim, Cheon-Suk
    • Journal of Ginseng Research
    • /
    • v.36 no.2
    • /
    • pp.205-210
    • /
    • 2012
  • For quality control of components in Korean red ginseng powder and extract, a new method for simultaneous quantification of 12 ginsenosides ($Rg_1$, Re, Rf, $Rh_1$, $Rg_2$[S], $Rg_2$[R], $Rb_1$, Rc, $Rb_2$, Rd, $Rg_3$[S], and $Rg_3$[R]) was studied. Compared to the official method for quantification of marker substances (ginsenosides $Rg_1$ and $Rb_1$), the proposed methods were guaranteed by in-house method validation. Several criteria such as linearity, specificity, precision and accuracy were evaluated. For red ginseng powder, recovery (averaging 95% to 105%) was calculated, and analysis of variance was carried out to estimate the relative standard deviation (0.20% to 2.12%). For red ginseng extract, the average recovery rate was 90% to 99% and the relative standard deviation was 0.39% to 2.40%. These results indicate that the proposed method could be used in the laboratory for determination of 12 ginsenosides in red ginseng powder and extract. In addition, this method was found to be suitable for quality control of ginseng products and potentially offer time and cost benefits.

Quantitative Analysis and Antioxidant Effects of Gyejibokryeong-hwan (LC-MS/MS를 이용한 계지복령환(桂枝茯苓丸)의 동시분석 및 항산화 효능 연구)

  • Seo, Chang-Seob;Kim, Ohn Soon;Shin, Hyeun-Kyoo
    • Korean Journal of Pharmacognosy
    • /
    • v.45 no.3
    • /
    • pp.240-248
    • /
    • 2014
  • Gyejibokryeong-hwan (GJBRH) has been used for treatment of patients with climacteric syndrome. In this study, an ultra-performance liquid chromatography-electrospray ionization-mass spectrometer (UPLC-ESI-MS) method was established for the simultaneous quantification of seven marker compounds in GJBRH extract. In addition, we assessed the antioxidant effects of GJBRH. All analytes were separated by gradient elution using two mobile phases on a UPLC BEH $C_{18}$ column and maintained at $45^{\circ}C$. The antioxidant activities of GJBRH were evaluated by measuring free radical scavenging activities on 2,2'-azinobis-3-ethyl-benzothiazoline-6-sulfonic acid (ABTS) and 1-1-diphenyl-2-picrylhydrazyl (DPPH). The inhibitory effects on low-density lipoprotein (LDL) oxidation were evaluated by the formation of thiobarbituric acid relative substances (TBARS) and relative electrophoretic mobility (REM). Regression equations of the seven compounds were acquired with $r^2$ values ${\geq}0.9988$. The amounts of the seven compounds, amygdalin, albiflorin, paeoniflorin, coumarin, cinnamic acid, cinnamaldehyde, and paeonol in GJBRH water extract were 21.71, 2.16, 17.17, 1.97, 0.40, 0.78, and 3.42 mg/g, respectively. The GJBRH showed the radical scavenging activity in a dose-dependent manner. The concentration required for 50% reduction ($RC_{50}$) against ABTS and DPPH radicals were $54.18{\mu}g/mL$ and $79.53{\mu}g/mL$. Furthermore, GJBRH reduced the oxidation properties of LDL induced by $CuSO_4$.

Pre- and post-initiation modulating effects of green tea ingestion on rat hepatocarcinogenesis

  • Kim, Hyung-Sook;Kim, Hee-Seon;Choi, Hay-Mie
    • Nutrition Research and Practice
    • /
    • v.2 no.4
    • /
    • pp.234-239
    • /
    • 2008
  • The purpose of this study was to investigate the effects of green tea ingestion on hepatocarcinogenesis before and after its initiation. Male Sprague-Dawley rats were fed an AIN76A diet with or without green tea. Initiation was induced by a single dose (200 mg/kg) of diethylnitrosamine at week 4 and 0.02% (w/w) 2-acetylaminofluorene was supplied in the diets. The control group had free access to water for 13 weeks (CTR13). Tea infusion was provided from the beginning of the experiment for 13 weeks (PRE13) or from the post-initiation stage until week 13 (POST13). Three other groups (CTR24, PRE24 and POST24) were added to examine the longer-term effects (24 weeks) with the same experimental design. The percentage area of liver sections that were positive for hepatic placental glutathione S-transferase (GST-P), which was used as a marker of preneoplastic lesions, was smaller in PRE13 ($20.2{\pm}5.0%$, $mean{\pm}SD$) and POST13 ($26.0{\pm}4.8%$) than in CTR13 ($33.2{\pm}5.8%$, p<0.05). Over the longer period, the GST-P lesions were significantly smaller for both PRE24 and POST24 ($21.6{\pm}8.5%$ and $22.2{\pm}4.0%$, respectively) than for CTR24 ($28.6{\pm}5.1%$, p<0.05), but there was no significant difference between PRE24 and POST24. The liver content of thiobarbituric acid reactive substances was significantly lower in the tea groups than in the controls (p<0.05). However, no significant differences were observed among groups of GST activity. The results show that tea consumption exhibits a stronger short-term initiation-inhibiting ability in liver carcinogenesis, but over a longer period, the preventive effects of green tea ingestion do not differ in post- and pre-initiation.

A new validated analytical method for the quality control of red ginseng products

  • Kim, Il-Woung;Cha, Kyu-Min;Wee, Jae Joon;Ye, Michael B.;Kim, Si-Kwan
    • Journal of Ginseng Research
    • /
    • v.37 no.4
    • /
    • pp.475-482
    • /
    • 2013
  • The main active components of Panax ginseng are ginsenosides. Ginsenoside Rb1 and Rg1 are accepted as marker substances for quality control worldwide. The analytical methods currently used to detect these two compounds unfairly penalize steamed and dried (red) P. ginseng preparations, because it has a lower content of those ginsenosides than white ginseng. To manufacture red ginseng products from fresh ginseng, the ginseng roots are exposed to high temperatures for many hours. This heating process converts the naturally occurring ginsenoside Rb1 and Rg1 into artifact ginsenosides such as ginsenoside Rg3, Rg5, Rh1, and Rh2, among others. This study highlights the absurdity of the current analytical practice by investigating the time-dependent changes in the crude saponin and the major natural and artifact ginsenosides contents during simmering. The results lead us to recommend (20S)- and (20R)-ginsenoside Rg3 as new reference materials to complement the current P. ginseng preparation reference materials ginsenoside Rb1 and Rg1. An attempt has also been made to establish validated qualitative and quantitative analytical procedures for these four compounds that meet International Conference of Harmonization (ICH) guidelines for specificity, linearity, range, accuracy, precision, detection limit, quantitation limit, robustness and system suitability. Based on these results, we suggest a validated analytical procedure which conforms to ICH guidelines and equally values the contents of ginsenosides in white and red ginseng preparations.

Effects of Ecklonia cava Extract on Neuronal Damage and Apoptosis in PC-12 Cells against Oxidative Stress

  • Shin, Yong Sub;Kim, Kwan Joong;Park, Hyein;Lee, Mi-Gi;Cho, Sueungmok;Choi, Soo-Im;Heo, Ho Jin;Kim, Dae-Ok;Kim, Gun-Hee
    • Journal of Microbiology and Biotechnology
    • /
    • v.31 no.4
    • /
    • pp.584-591
    • /
    • 2021
  • Marine algae (seaweed) encompass numerous groups of multicellular organisms with various shapes, sizes, and colors, and serve as important sources of natural bioactive substances. The brown alga Ecklonia cava Kjellman, an edible seaweed, contains many bioactives such as phlorotannins and fucoidans. Here, we evaluated the antioxidative, neuroprotective, and anti-apoptotic effects of E. cava extract (ECE), E. cava phlorotannin-rich extract (ECPE), and the phlorotannin dieckol on neuronal PC-12 cells. The antioxidant capacities of ECPE and ECE were 1,711.5 and 1,050.4 mg vitamin C equivalents/g in the ABTS assay and 704.0 and 474.6 mg vitamin C equivalents/g in the DPPH assay, respectively. The dieckol content of ECPE (58.99 mg/g) was approximately 60% higher than that of ECE (36.97 mg/g). Treatment of PC-12 cells with ECPE and ECE increased cell viability in a dose-dependent manner. Intracellular oxidative stress in PC-12 cells due to ECPE and ECE decreased dose-independently by up to 63% and 47%, respectively, compared with the stress control (323%). ECPE reduced the production of the pro-apoptotic proteins Bax and caspase-3 more effectively than ECE. Early and late apoptosis in PC-12 cells were more effectively decreased by ECPE than ECE treatments. From the results obtained in this study, we concluded that ECPE, which is rich in phlorotannins, including the marker compound dieckol, may be applied to the development of functional materials for improving cognition and memory.

Effects of Green Tea Extract on Acute Ethanol-induced Hepatotoxicity in Rats (녹차추출물이 에탄올 투여에 의한 초기 간 손상에 미치는 영향)

  • Jin, Dong-Chun;Jeong, Seung-Wook;Park, Pyoung-Sim
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.39 no.3
    • /
    • pp.343-349
    • /
    • 2010
  • The liver is the major target of ethanol toxicity and oxidative stress plays a role in development of alcoholic liver disease. This study was performed to investigate the effects of green tea extracts (GTE) on acute ethanol-induced hepatotoxicity in rats. Experimental animals were divided into 4 groups, control, GTE, ethanol, and GTE+ethanol treatment, with 5 rats in each group. Ethanol (6 g/kg body weight (BW)) and GTE (200 mg/kg BW) were treated by gavage. At 1 hour, 3 hours and 20 days (6 g/kg BW every 2 days for total 10 doses) after ethanol and/or GTE treatments, animals were killed; hepatic tumor necrosis factor-alpha (TNF-$\alpha$) and glutathione level, serum aspartate aminotransferase (AST), serum alanine aminotransferase (ALT), hepatic antioxidant enzymes (SOD, CAT, GPx) activities and hepatic thiobarbituric acid reactive substances (TBARS) were measured. At 1 hour and 3 hours, hepatic TNF-$\alpha$ levels were increased significantly in ethanol group and ethanol+GTE group but that levels was significantly lower in ethanol+GTE group compared with ethanol group. Hepatic glutathione level was decreased by ethanol treatment but GTE prevented the ethanol-induced glutathione decrement. The levels of liver marker enzymes (AST, ALT), liver antioxidant enzymes (SOD, CAT, GPx) and lipid peroxidation marker (TBARS) were not changed in rats of 1 and 3 hours after ethanol treatment. After 20 days, GTE decreased the changes of liver marker enzymes (AST, ALT) activities and TBARS level by ethanol. This study shows that GTE beneficially modulates TNF-$\alpha$ and glutathione levels in liver of ethanol administered rats. The GTE supplementation could be beneficial to liver by decreasing early changes of biomarkers of liver damage caused by ethanol.

Antioxidant action of soy isoflavones on oxidative stress and antioxidant enzyme activities in exercised rats

  • Yoon, Gun-Ae;Park, Sunmin
    • Nutrition Research and Practice
    • /
    • v.8 no.6
    • /
    • pp.618-624
    • /
    • 2014
  • BACKGROUND/OBJECTIVES: Isoflavones are widely believed to be beneficial to human health, in relation to their antioxidant potentials. Exercise can cause an imbalance between reactive oxygen species (ROS) and antioxidants. This study was conducted in order to investigate the ability of isoflavones in amelioration of oxidative stress induced by exercise. MATERIALS/METHODS: Male Sprague-Dawley rats were assigned to one of four groups: isoflavone-free with no exercise (CON-sd), isoflavone-free with exercise (CON-ex), isoflavone-supplemented with no exercise (ISF-sd), and isoflavone-supplemented with exercise (ISF-ex). Animals exercised on the treadmill for 30 minutes per day, five days per week. TBARS as a marker of oxidative stress and antioxidant enzyme activity, including SOD, GSH-px, and catalase were determined in liver tissue. Serum lipid profile was also examined. RESULTS: A significant effect of isoflavone alone was observed on abdominal fat pad mass. ISF-ex had significantly less abdominal fat pad than CON-ex. Both exercise and isoflavone treatment had significant effects on lowering plasma triglyceride (TG), thus, the ISF-ex group had a significantly lower TG level than the CON-sd group, by 30.9%. However, no differences were observed in plasma cholesterol, HDL-C, and cholesterol/HDL-C ratio. Exercise, isoflavone, and exercise-isoflavone interaction effects were significant on thiobarbituric acid reactive substances (TBARS) (P = 0.001, 0.002, and 0.005, respectively). The CON-ex group showed a higher TBARS level than the other three groups. By contrast, in the ISF-ex group, TBARS was restored to the level of the ISF-sd or CON-sd group. Isoflavone had a significant effect on superoxide dismutase (SOD) (P = 0.022) and catalase activities (P = 0.049). Significantly higher SOD and catalase activities were observed in ISF-ex than CON-ex. SOD and catalase activities showed an inverse pattern of TBARS. Taken together, isoflavones increased the activities of SOD and catalase with concomitant decreases in TBARS, indicative of decreased oxidative stress. CONCLUSIONS: Isoflavone supplementation enhances antioxidant action with attenuation of exercise-induced oxidative stress, as measured by decreases in TBARS, and inhibits body fat accumulation and plasma TG increase. Antioxidative effects ascribed to isoflavones may be partially exerted via enhancement of antioxidant enzyme activities.

Characterizing a full spectrum of physico-chemical properties of (20S)-and (20R)-ginsenoside Rg3 to be proposed as standard reference materials

  • Kim, Il-Woung;Sun, Won Suk;Yun, Bong-Sik;Kim, Na-Ri;Min, Dongsun;Kim, Si-Kwan
    • Journal of Ginseng Research
    • /
    • v.37 no.1
    • /
    • pp.124-134
    • /
    • 2013
  • The authentication of the physico-chemical properties of ginsenosides reference materials as well as qualitative and quantitative batch analytical data based on validated analytical procedures is a prerequisite for certifying good manufacturing practice (GMP). Ginsenoside Rb1 and Rg1, representing protopanaxadiol and protopanaxatriol ginsenosides, respectively, are accepted as marker substances in quality control standards worldwide. However, the current analytical methods for these two compounds recommended by Korean, Chinese, European, and Japanese pharmacopoeia do not apply to red ginseng preparations, particularly the extract, because of the relatively low content of the two agents in red ginseng compared to white ginseng. In manufacturing fresh ginseng into red ginseng products, ginseng roots are exposed to a high temperature for many hours, and the naturally occurring ginsenoside Rb1 and Rg1 are converted to artifact ginsenosides such as Rg3, Rg5, Rh1, and Rh2 during the heating process. The analysis of ginsenosides in commercially available ginseng products in Korea led us to propose the inclusion of the (20S)- and (20R)-ginsenoside Rg3, including ginsenoside Rb1 and Rg1, as additional reference materials for ginseng preparations. (20S)- and (20R)-ginsenoside Rg3 were isolated by Diaion HP-20 adsorption chromatography, silica gel flash chromatography, recrystallization, and preparative HPLC. HPLC fractions corresponding to those two ginsenosides were recrystallized in appropriate solvents for the analysis of physico-chemical properties. Documentation of those isolated ginsenosides was achieved according to the method proposed by Gaedcke and Steinhoff. The ginsenosides were subjected to analyses of their general characteristics, identification, purity, content quantification, and mass balance tests. The isolated ginsenosides showed 100% purity when determined by the three HPLC systems. Also, the water content was found to be 0.534% for (20S)-Rg3 and 0.920% for (20R)-Rg3, meaning that the net mass balances for (20S)-Rg3 and (20R)-Rg3 were 99.466% and 99.080%, respectively. From these results, we could assess and propose a full spectrum of physico-chemical properties of (20S)- and (20R)-ginsenoside Rg3 as standard reference materials for GMP-based quality control.

Prognostic Value of Blood Lactate for Mortality of Acutely Poisoned Patients in Emergency Department (응급실내 급성 중독 환자들의 예후 예측에 대한 혈액 젖산 수치의 유용성)

  • Kim, Hye Ran;Kang, Mun Ju;Kim, Yong Hwan;Lee, Jun Ho;Cho, Kwang Won;Hwang, Seong Youn;Lee, Dong Woo
    • Journal of The Korean Society of Clinical Toxicology
    • /
    • v.14 no.1
    • /
    • pp.16-25
    • /
    • 2016
  • Purpose: Patients suffering from acute poisoning by different substances often visit the emergency department (ED) and receive various prognoses according to the toxic material and patients' condition. Hyperlactatemia, which is an increased blood lactate level that generally indicates tissue hypoperfusion, is commonly utilized as a prognostic marker in critically ill patients such as those with sepsis. This study was conducted to investigate the relationships between blood lactate and clinical prognosis in acute poisoned patients. Methods: This retrospective study was conducted from January 2013 to June 2014 at a single and regional-tertiary ED. We enrolled study patients who were examined for blood test with lactate among acute intoxicated patients. The toxic materials, patient demographics, laboratory data, and mortalities were also reviewed. Additionally, we analyzed variables including blood lactate to verify the correlation with patient mortality. Results: A total of 531 patients were enrolled, including 24 (4.5%) non-survivors. Patient age, Glasgow coma scale (GCS), serum creatinine (Cr), aspartate transaminase (AST), and serum lactate differed significantly between survivors and non-survivors in the binary logistic regression analysis. Among these variables, GCS, AST, and lactate differed significantly. The median serum lactate levels were 2.0 mmol/L among survivors and 6.9 mmol/L among non-survivors. The AUC with the ROC curve and odds ratio of the initial serum lactate were 0.881 and 3.06 (0.89-8.64), respectively. Conclusion: Serum lactate was correlated with fatalities of acute poisoning patients in the ED; therefore, it may be used as a clinical predictor to anticipate their prognoses.

  • PDF