• Title/Summary/Keyword: NO assay

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Effects of Anemarrhenae Rhizoma on LPS-stimulated Expression of COX-2 and iNOS in mouse BV2 microglial cells

  • Seo, Bong-Won;Kim, Ee-Hwa;Park, Se-Keun;Jang, Mi-Hyeon;Choi, Sun-Mi;Lee, Eun-Yong
    • Journal of Acupuncture Research
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    • v.22 no.2
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    • pp.155-162
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    • 2005
  • Anemarrhenae Rhizoma (AR) has been widely used for the treatment of various diseases in Oriental medicine. To investigate whether AR possesses anti-inflammatory effects against lipopolysaccharide (LPS)-induced inflammation in the BV2 microglial cells, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide) assay. reverse transcription -polymerase chain reaction (RT-PCR), and prostaglandin E2 (PGE2) assay, and nitric oxide (NO) detection assay were performed. From the present results, AR was shown to suppress PGE2 synthesis and NO production by inhibiting the LPS-stimulated enhancement of cyclooxygenase-2 and inducible nitric oxide synthase expression in BV2 microglial cells. These results suggest that AR may offer a valuable means of therapy in the treatment of inflammatory diseases by attenuating LPS-induced PGE2 synthesis and NO production.

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Neuroprotective Effect of Insamyangyung-tang (인삼양영탕(人蔘養營湯)의 산화적 stress에 대한 뇌세포 보호효과)

  • Kim, Seung-Hyun;Lee, Chang-Hoon;Lee, Jin-Moo;Cho, Jung-Hoon;Jang, Jun-Bock;Lee, Kyung-Sub
    • The Journal of Korean Obstetrics and Gynecology
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    • v.22 no.1
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    • pp.1-14
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    • 2009
  • Purpose: Oxidative stress was thought to play a critical role in neurodegenerative disease. Many in vivo and in vitro reports explained the possible pathway of human aging. But in therapeutic aspects, there was no clear answers to prevent aging associated with neural diseases. In this study, we investigated the antioxidant and neuroprotective effects of the Insamyangyung-tang (IYT). Methods: To estimate the antioxidant effects, we carried out 1.1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging assay, 2,2'-azinobis-(3- ethylbenzothiazoline-6- sulfonic acid (ABTS) radical cation decolorization assay, and measurement of total polyphenolic content. To evaluate neuroprotective effect of IYT in vitro. We performed thiazolyl blue tetrazolium bromide (MTT) assay, reactive oxygen species (ROS) creation in SH-SY5Y. Tyrosine hydroxylase (TH) immunocytochemistry, nitric oxide (NO) assay, and TNF-${\alpha}$ assay in primary rat mesencephalic dopaminergic neurons. Results: The $IC_{50}$ values were $571.6{\mu}g/m{\ell}$ and $202.3{\mu}g/m{\ell}$ in DPPH and ABTS assay respectively. Total polyphenolic content was 1.05%. In SH-SY5Y culture, IYT significantly increased the decreased cell viability by 6-OHDA at the concentrations of $10{\mu}g/m{\ell}$ in pre-treatment group, $10-100{\mu}g/m{\ell}$ in post-treatment group, and $100{\mu}g/m{\ell}$ in co-treatment group. The production of ROS induced by 6-OHDA was significantly inhibited in IYT treated group. In mesencephalic dopaminergic cell culture, the IYT group reduced the dopaminergic cell loss against 6-OHDA toxicity and the production of No and TNF-${\alpha}$ at the concentration of $0.2{\mu}g/m{\ell}$. Conclusion: These results showed that IYT has antioxidant and neuroprotectctive effects in the dopaminergic cells through decreasing the production of ROS, NO and TNF-${\alpha}$ which can cause many neurodegenerative changes in brain cell.

Effects of Five Fractions of Artemisia capillaris THUNB on $TGF{\beta}1-induced$ Apoptosis in HepG2 Cells (인진분획물이 인체간세포의 $TGF{\beta}1-induced$ Apoptosis에 미치는 영향)

  • 이지현;이장훈;우홍정
    • The Journal of Korean Medicine
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    • v.21 no.1
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    • pp.53-61
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    • 2000
  • Objectives: This study was camed out to examine the effect of five fractions of aqueous extract from Artemisia capillaris THUNB(ACT), on TGF, 1-induced apoptosis, cell viability, cell cycle progression and mRNA expression of apoptosis-related genes in human hepatocyte cell line HepG2. Methods: This study employed Tryphan blue exclusion assay, DNA fragmentation assay, Cpp32 protease activity assay and Quantitative RT-PCR analysis. Results: In the Tryphan blue exclusion assay, the butanol fraction of ACT with $TGF{\beta}$, l showed magnificent (Nice word, ut is it appropriate in a medical abstract\ulcorner) viability and the H2O fraction of ACT with $TGF{\beta}$, l also showed higher viability than only $TGF{\beta}$, l-treated group. DNA fragmentation assay showed that the butanol fraction and the H2O fraction carried inhibitory effects on apoptosis induction, with the butanol fraction displaying greater effects. The Cpp32 protease activity assay showed that the butanol fraction decreased Cpp32 protease activity. The H2O fraction of ACT had no significant effect on the Cpp32 protease activity. Quantitative RT-PCR showed that the butanol fraction suppressed Bax, p 15/INK4B, p21/Waf1, PAI-1 and increased Bcl-2 gene. Conclusions: The data shows that butanol fraction of ACT increases the hepatocyte viability and has the hepatocellular protective effect by the suppression of $TGF{\beta}$, l induced-apoptosis through gene regulation.

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The Influence of Assay Error Weight on Gentamicin Pharmacokinetics Using the Bayesian and Nonlinear Least Square Regression Analysis in Appendicitis Patients

  • Jin, Pil-Burm
    • Archives of Pharmacal Research
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    • v.28 no.5
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    • pp.598-603
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    • 2005
  • The purpose of this study was to determine the influence of weight with gentamicin assay error on the Bayesian and nonlinear least squares regression analysis in 12 Korean appen dicitis patients. Gentamicin was administered intravenously over 0.5 h every 8 h. Three specimens were collected at 48 h after the first dose from all patients at the following times, just before regularly scheduled infusion, at 0.5 h and 2 h after the end of 0.5 h infusion. Serum gentamicin levels were analyzed by fluorescence polarization immunoassay technique with TDxFLx. The standard deviation (SD) of the assay over its working range had been determined at the serum gentamicin concentrations of 0, 2, 4, 8, 12, and 16 ${\mu}g$/mL in quadruplicate. The polynominal equation of gentamicin assay error was found to be SD (${\mu}g$/mL) = 0.0246-(0.0495C)+ (0.00203C$^2$). There were differences in the influence of weight with gentamicin assay error on pharmacokinetic parameters of gentamicin using the nonlinear least squares regression analysis but there were no differences on the Bayesian analysis. This polynominal equation can be used to improve the precision of fitting of pharmacokinetic models to optimize the process of model simulation both for population and for individualized pharmacokinetic models. The result would be improved dosage regimens and better, safer care of patients receiving gentamicin.

Thermal-and Bio-degradation of Starch-Polyethylene Films Containing High Molecular Weight Oxidized-Polyethylene

  • Kim, Mee-Ra;Pometto, Anthony-L.
    • Preventive Nutrition and Food Science
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    • v.3 no.1
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    • pp.27-35
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    • 1998
  • Starch-polyethylene films containing high molecular weight(NW) oxidized-polyethylene and prooxidant were prepared , and thermal -and bio-degradability of the films were determined. Increased levels of starch resulted in a corresponding reduction in mechanical strength of the films. However, the addition of high MW oxidized-polyethylene did not significantly reduce the percent elongation of the films. Thefilms containing high MW oxidized-polyethylene andproosicant were degreaded faster than those containing no aadditive during the heat treatment. The films lost their measureable mechanical properties when their weight-average MW(Mw) fell below 50,000. Biodegradability of the films was determined by a pure culture assay with either Streptomyces badius 252.S. setonii 75Vi2 or S. viridosporous T7A, and by an extracellulr enzyme assay using S. setonii 75vi2. The results from pure culture assay indicated that biomass accumulation on the film surface inhibited chemical and biological degradation of the films. The extracellular enzyme assay demonstrated decrease of percent elongation and increase of carbonyl index of the films. Therefore, extracellular enzyme assay could be used as a good method to evaluate biodegradability of the films.

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Assay Error for Improved Pharmacokinetic Modeling and Simulation of Vancomycin (반코마이신의 약물동태학적 모델링과 시뮬레이션의 향상을 위한 분석오차)

  • Burm, Jin Pil
    • YAKHAK HOEJI
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    • v.57 no.1
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    • pp.32-36
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    • 2013
  • The purpose of this study was to determine the influence of assay error for improved pharmacokinetic modeling and simulation of vancomycin on the Bayesian and nonlinear least squares regression analysis in 24 Korean gastric cancer patients. Vancomycin 1.0 g was administered intravenously over 1 hr every 12 hr. Three specimens were collected at 72 hr after the first dose from all patients at the following times, at 0.5 hr before regularly scheduled infusion, at 0.5 hr and 2 hr after the end of 1 hr infusion. Serum vancomycin levels were analyzed by fluorescence polarization immunoassay technique with TDX-FLX. The standard deviation (SD) of the assay over its working range had been determined at the serum vancomycin concentrations of 0, 20, 40, 60, 80 and $120{\mu}g/ml$ in quadruplicate. The polynomial equation of vancomycin assay error was found to be SD $({\mu}g/ml)=0.0224+0.0540C+0.00173C^2$ ($R^2=0.935$). There were differences in the influence of weight with vancomycin assay error on pharmacokinetic parameters of vancomycin using the nonlinear least squares regression analysis but there were no differences on the Bayesian analysis. This polynomial equation can be used to improve the precision of fitting of pharmacokinetic models to optimize the process of model simulation both for population and for individualized pharmacokinetic models. The result suggests the improvement of dosage regimens for the better and safer care of patients receiving vancomycin.

Rapid and sensitive detection of Salmonella species targeting the hilA gene using a loop-mediated isothermal amplification assay

  • Chu, Jiyon;Shin, Juyoun;Kang, Shinseok;Shin, Sun;Chung, Yeun-Jun
    • Genomics & Informatics
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    • v.19 no.3
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    • pp.30.1-30.8
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    • 2021
  • Salmonella species are among the major pathogens that cause foodborne illness outbreaks. In this study, we aimed to develop a loop-mediated isothermal amplification (LAMP) assay for the rapid and sensitive detection of Salmonella species. We designed LAMP primers targeting the hilA gene as a universal marker of Salmonella species. A total of seven Salmonella species strains and 11 non-Salmonella pathogen strains from eight different genera were used in this study. All Salmonella strains showed positive amplification signals with the Salmonella LAMP assay; however, there was no non-specific amplification signal for the non-Salmonella strains. The detection limit was 100 femtograms (20 copies per reaction), which was ~1,000 times more sensitive than the detection limits of the conventional polymerase chain reaction (PCR) assay (100 pg). The reaction time for a positive amplification signal was less than 20 minutes, which was less than one-third the time taken while using conventional PCR. In conclusion, our Salmonella LAMP assay accurately detected Salmonella species with a higher degree of sensitivity and greater rapidity than the conventional PCR assay, and it may be suitable for point-of-care testing in the field.

Imported Intraocular Gnathostomiasis with Subretinal Tracks Confirmed by Western Blot Assay

  • Yang, Ji-Ho;Kim, Moo-Sang;Kim, Eung-Suk;Na, Byoung-Kuk;Yu, Seung-Young;Kwak, Hyung-Woo
    • Parasites, Hosts and Diseases
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    • v.50 no.1
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    • pp.73-78
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    • 2012
  • We report a case of intraocular gnathostomiasis diagnosed by western blot assay in a patient with subretinal tracks. A 15-year-old male patient complained of blurred vision in the right eye, lasting for 2 weeks. Eight months earlier, he had traveled to Vietnam for 1 week and ate raw wild boar meat and lobster. His best-corrected visual acuity was 20/20 in both eyes and anterior chamber examination revealed no abnormalities. Fundus examination showed subretinal tracks in the right eye. Fluorescein angiography and indocyanine green angiography showed linear hyperfluorescence of the subretinal lesion observed on fundus in the right eye. Ultrasound examination revealed no abnormalities. Blood tests indicated mild eosinophilia (7.5%), and there was no abnormality found by systemic examinations. Two years later, the patient visited our department again for ophthalmologic evaluation. Visual acuity remained 20/20 in both eyes and the subretinal tracks in the right eye had not changed since the previous examination. Serologic examination was performed to provide a more accurate diagnosis, and the patient's serum reacted strongly to the $Gnathostoma$ $nipponicum$ antigen by western blot assay, which led to a diagnosis of intraocular gnathostomiasis. This is the first reported case of intraocular gnathostomiasis with subretinal tracks confirmed serologically using western blot in Korea.

A study on Genotoxicity Test of Hyeong-gae-yeon-gyo-tang extract (형개연교탕(荊芥連翹湯) 추출물의 유전독성(遺傳毒性) 평가)

  • Jee, Seon-Young;Hwang, Sun-Yi;Lee, Jong-Rok;Kim, Sang-Chan
    • The Korea Journal of Herbology
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    • v.22 no.4
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    • pp.287-300
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    • 2007
  • Objectives : The genotoxicity of extract of "Hyeonggaeyeongyotang", a polyherbal formula has been used as a tonic agents in oriental medicine was tested. Methods : Extract of "Hyeonggaeyeongyotang" was tested by In Vitro Chromosome Aberration Test, Bacterial Reverse Mutation Assay and Micronucleus test according to OECD Guidelines and KFDA Guidelines [2005-60]. Results : The obtained results were as follows: 1. Chromosome Aberration Test: No significant changes in the number of aberrant metaphases having structural and number of aberrations were detected in all concentrations of "Hyeonggaeyeongyotang" extracts treated in this study. 2. Bacterial Reverse Mutation Assay: No significant increases in the number of revertant colonies compared to its negative control were detected in all concentrations of "Hyeonggaeyeongyotang" extracts treated in this study against all 5 strains except for $50{\mu}g/ml$ treated group where significantly decreases in colony numbers were detected agains all five strains used in this study as pharmacological effects not genotoxicity. 3. Micronucleus test: No significant changes in the number of micronucleated polychromatic erythrocytes among 2000 polychromatic erythrocytes compared to negative control were detected in all "Hyeonggaeyeongyotang" extracts-dosing groups tested. Conclusions : From above-mentioned results, it is concluded that "Hyeonggaeyeongyotang" extracts have not any genotoxicity against In Vitro Chromosome Aberration Test, Bacterial Reverse Mutation Assay and Micronucleus test.

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Effects of Gamioncheong-decoction Water-extract on Anti-inflammation, Anti-oxidation and Skin Whitening (가미온청음(加味溫淸飮) 추출물이 항염증, 항산화 및 미백에 미치는 효과)

  • Choi, You Jin;Roh, Jeong Du
    • Journal of Acupuncture Research
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    • v.32 no.3
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    • pp.117-126
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    • 2015
  • Objectives : This study was an analysis of the anti-inflammatory, anti-oxidative and skin whitening properties of Gamioncheong-decoctione(GMOCD) extract. Methods : GMOCD(96 g) and 2 L of distilled water were heated at $100^{\circ}C$ for four hours and then concentrated, frozen, freeze-dried, dissolved in distilled water and filtered. The following analysis was completed: cell cytotoxic effect using MTT assay, oxidative products of NO by griess assay, concentration of prostaglandin $E_2(PGE_2)$ by commercially competitive enzyme immunoassay, and cytokines($IL-1{\beta}$, IL-6 and TNF-${\alpha}$) by Bio-Plex$^{(R)}$ Suspension Array System's Bio-Plex Pro$^{TM}$ mouse cytokine, chemokine, and growth factor assay. Anti-oxidative effect was measured using the DPPH method and skin whitening effect using tyrosinase inhibition assay. Results : GMOCD water-extract did not show any toxicity at all doses and cell viability was more than 90 % at all doses. GMOCD water-extract significantly inhibited NO production at doses of 100, 200, $400{\mu}g/ml$, significantly inhibited $PGE_2$ production at doses of 200 and $400{\mu}g/ml$ and reduced the LPS-induced IL-$1{\beta}$, IL-6 and TNF-${\alpha}$ production in a dose-dependent manner. $IL-1{\beta}$ production was significantly reduced at a dose of $400{\mu}g/ml$ and IL-6 production was significantly reduced at doses of 200 and $400{\mu}g/ml$. DPPH free radical scavenging capability had a skin whitening effect rate of more than 50%. Tyrosinase inhibition activity was apparent in a dose-dependent manner. Conclusions : This study suggests that GMOCD water-extract suppressed NO and $PGE_2$ production and inhibited cytokines($IL-1{\beta}$, IL-6 and TNF-${\alpha}$). GMOCD also improved DPPH free radical scavenging capability. GMOCD water-extract increased tyrosinase inhibitory activity in a dose-dependent manner but this was not a statistically significant result.