• Title/Summary/Keyword: bio assay

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The Effect of Pinus Densiflora Gnarl Extract for Pharmacopuncture on Human LDL Oxidation Induced by Free Radical and Metal Ion (송절(松節) 약침액이 자유기와 금속 이온으로 유도된 인체 저밀도 지단백질의 산화 반응에 미치는 효과)

  • Leem, Sun-Hee;Lee, Kang-Pa;Moon, Jin-Young
    • Korean Journal of Acupuncture
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    • v.28 no.2
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    • pp.23-36
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    • 2011
  • 목적 : 이 연구는 관절 및 심혈관계 질환 치료에 사용되는 송절(松節)(Pinus densiflora Gnarl)을 약침용 시료로 조제하여 본 약물의 항산화 효능을 규명하고자 하였으며 이를 다양한 시스템에서 검토하였다. 방법 : $FeCl_2$-ascorbic acid system에서 흰쥐 간조직의 지질과산화 반응을 관찰하였고, Fenton reaction system에서 자유기에 의한 plasmid DNA 분절을 유도하였다. 또한 deoxyribose assay를 통해 hydroxyl radical 소거능을 관찰하였고, NBT reduction assay로 superoxide radical 소거능을 검토하였다. 또한 human low-density lipoprotein(LDL)의 산화를 유도하기 위해 $CuSO_4$와 AAPH를 사용하였으며 relative electrophoretic mobility (REM) assay로 LDL 산화 억제 효능을 대조 항산화물질과 비교 검토하였다. 결과 : 송절 약침액은 자유기에 의한 간조직의 지질과산화(p < 0.01)및 DNA 분절을 현저하게 억제하였으며, hydroxyl radical, superoxide radical (p < 0.01), nitric oxide 및 peroxynitrite를 강하게 소거하였다. 또한 $CuSO_4$ ($IC_{50}=9.2{\pm}0.2\;{\mu}g/ml$)와 AAPH ($IC_{50}=34.8{\pm}5.1\;{\mu}g/ml$)에 의해 유도된 human LDL의 산화를 억제하였고, REM assay에서도 산화 억제 효능을 재확인할 수 있었다. 결론 : 송절 약침액은 활성산소종 및 활성질소종를 소거하였고, 지질과산화를 억제하였으며, 특히 human LDL의 산화적 손상을 방어하였다. 이에 본 약물은 자유기에 의한 심혈관의 산화적 손상을 효과적으로 보호할 것으로 판단된다.

Quality Assessment and Comparison of Several Radioimmunoassay Kits and Chemiluminescence Immunoassay Methods for Evaluating Serum Estradiol (혈중 Estradiol 농도 측정을 위한 여러 방사면역측정 검사키트 및 화학면역발광 검사법의 성능평가 및 상호비교)

  • Choi, Sung Hee;Noh, Gyeong Woon;Kim, Jin Eui;Song, Yoo Sung;Paeng, Jin Chul;Kang, Keon Wook;Lee, Dong Soo
    • The Korean Journal of Nuclear Medicine Technology
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    • v.19 no.1
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    • pp.72-80
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    • 2015
  • Purpose Serum estradiol ($E_2$) measurement is requested for diagnosing menstrual cycles, ovulation induction, infertility, and menopause. $E_2$ is measured using several methods and kits including radioimmunoassay (RIA) and chemiluminescece immunoassay (CLIA). The purpose of this study was to evaluate quality of these methods and to compare them with each other. Materials and Methods Seven radioimmunoassay kits and two CLIA methods were included in the analysis. Using standard samples and patient samples, intra-assay precision, inter-assay precision, correlation between other methods, sensitivity, and recovery rate were evaluated. Results For all tested kits and methods, coefficients of variance (CVs) of intra-assay precision test were 10.9~13.6% in low-level samples and less than 10% in medium and high-level samples. CVs of inter-assay precision test were 10.8~12.3% in low-level samples and less than 10% in medium and high-level samples with all tested kits and methods. Recovery rates were $92.7{\pm}12.4%$ for SIEMENS, $101.4{\pm}18.4%$ for DIAsource, $95.1{\pm}11.5%$ for AMP, $108.4{\pm}18.5%$ for BECKMAN COULTER, $104.2{\pm}13.5%$ for BECKMAN COULTER Ultra Sensitive, $101.3{\pm}11.6%$ for CIS Bio, and $93.1{\pm}13.2%$ for MP kits. Sensitivity was 7.5, 6.2, 5.7, 6.2, 5.3, 4.5, and 5.5 pg/mL for SIEMENS, DIAsource, AMP, BECKMAN COULTER, BECKMAN COULTER Ultra Sensitive, CIS Bio, and MP kits, respectively. The measurement by MP kit was slightly higher than those by other kits in low-level samples, and the measurement by E170 was slightly higher than those of other kits in medium and high-level samples. In the measurement of standard sample for external quality control, SIEMENS kit produced relatively lower values whereas E170, Architect, and MP kits produced relatively higher values compared with other kits. Conclusion All tested kits for $E_2$ measurement have satisfactory performance for clinical use. However, correlation between kits should be considered when test kits are to be changed, because some pairs of kits do not have correlations with each other.

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Protective Effect of the Ethyl Acetate-fraction of Methanol Extract of Ophiophogon japonicus on Amyloid beta Peptide-induced Cytotoxicity in PC12 Cells (소엽맥문동-에틸아세테이트 분획물의 아밀로이드 베타단백질-유발 세포독성에 대한 억제 효능)

  • Moon, Ja-Young;Kim, Eun-Sook;Choi, Soo-Jin;Kim, Jin-Ik;Choi, Nack-Shik;Lee, Kyoung;Park, Woo-Jin;Choi, Young-Whan
    • Journal of Life Science
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    • v.29 no.2
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    • pp.173-180
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    • 2019
  • Amyloid ${\beta}$-protein ($A{\beta}$) is the principal component of senile plaques characteristic of Alzheimer's disease (AD) and elicits a toxic effect on neurons in vitro and in vivo. Many environmental factors, including antioxidants and proteoglycans, modify $A{\beta}$ toxicity. It is worthwhile to isolate novel natural compounds that could prove therapeutic for patients with AD without causing detrimental side effects. In this study, we investigated the in vitro neuroprotective effects of the ethyl acetate fraction of methanol extract of Ophiophogon japonicas (OJEA fraction). We used an MTT reduction assay to detect protective effects of the OJEA fraction on $A{\beta}_{25-35}$-induced cytotoxicity to PC12 cells. We also used a cell-based ${\beta}$-secretase assay system to investigate the inhibitory effect of the OJEA fraction on ${\beta}$-secretase activity. In addition, we performed an in vitro lipid peroxidation assay to evaluate the protective effect of the OJEA fraction against oxidative stress induced by $A{\beta}_{25-35}$ in PC12 cells. The OJEA fraction had strong protective effects against $A{\beta}_{25-35}$-induced cytotoxicity to PC12 cells and was strongly inhibitory to ${\beta}$-secretase activity, which resulted in the attenuation of $A{\beta}$ generation. In addition, the OJEA fraction significantly decreased malondialdehyde (MDA) content, which is induced by the exposure of PC12 cells to $A{\beta}_{25-35}$. Our results suggested that the OJEA fraction contained active compounds exhibiting a neuroprotective effect on $A{\beta}$ toxicity.

Mutagenic Assessment of Olmesartan Cilexetil by Bacterial Mutation Assay

  • Kim, Ji Won;Ahn, Ilyoung;Ryu, Sung Ha;Jeon, Hong Ryeol;Lee, Bong Sang;Kim, Kyu-Bong
    • Toxicological Research
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    • v.29 no.3
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    • pp.217-219
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    • 2013
  • Hypertension is a serious health problem due to high frequency and concomitant other diseases including cardiovascular and renal dysfunction. Olmesartan cilexetil is a new antihypertensive drug associated with angiotensin II receptor antagonist. This study was conducted to evaluate the mutagenicity of olmesartan cilexetil by bacterial reverse mutation test using Salmonella typhimurium (TA100, TA1535, TA98, and TA1537) and Escherichia coli (WP2 uvrA). At the concentrations of 0, 62, 185, 556, 1667, and 5000 ${\mu}g$/plate, olmesartan cilexetil was negative in both Salmonella typhimurium and Escherichia coli regardless of presence or absence of metabolic activation system (S9 mix). These results demonstrate that olmesartan cilexetil does not induce bacterial reverse mutation.

Antimicrobial effect of Australia propolis on cariogenic and periodontopathic bacteria

  • Lim, Yun Kyong;Yoo, So Young;Lee, Dae Sung;Kook, Joong-Ki
    • International Journal of Oral Biology
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    • v.44 no.4
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    • pp.191-194
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    • 2019
  • The purpose of this study was to investigate the antimicrobial effects of Australia propolis against cariogenic and periodontopathic bacteria. Antimicrobial activity was determined by evaluating the minimal bactericidal concentration (MBC). Cell cytotoxicity of propolis extract on normal human gingival fibroblast (HGF-1) cells was observed using the methylthiazolyldiphenyl-tetrazolium bromide assay. The data indicated that, with the exception of Aggregatibacter actinomycetemcomitans (KCOM 1306), the MBC values of the propolis strains were 0.25-1% without HGF-1 cell cytotoxicity. These results suggest that propolis can be used to develop oral hygiene products for the prevention of oral infectious disease.

Bio-Rex 젤을 이용한 난백으로 부터의 Lysozyme의 분리

  • Kim, Hyeong-Won;Park, In-Gyu;Song, Jae-Yang;Kim, In-Ho
    • 한국생물공학회:학술대회논문집
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    • 2002.04a
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    • pp.493-496
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    • 2002
  • Lysozyme is an enzyme which has the ability to lyse bacteria such as Micrococcus lysodeikticus or gram positive and gram negative bacteria by hydrolyzing in the peptidoglycan layer of the bacterial cell wall. Lysozyme is abundantly contained in an egg white. In order to obtain lysozyme from egg white, we used Bio-rex ion exchange chromatography and can identify the exist of lysozyme by SDS-PAGE and protein assay.

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Gustatory Receptors Required for Avoiding the Toxic Compound Coumarin in Drosophila melanogaster

  • Poudel, Seeta;Lee, Youngseok
    • Molecules and Cells
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    • v.39 no.4
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    • pp.310-315
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    • 2016
  • Coumarin is a phenolic compound that mainly affects the liver due to its metabolization into a toxic compound. The deterrent and ovicidal activities of coumarin in insect models such as Drosophila melanogaster have been reported. Here we explore the molecular mechanisms by which these insects protect themselves and their eggs from this toxic plant metabolite. Coumarin was fatal to the flies in a dosage-dependent manner. However, coumarin feeding could be inhibited through activation of the aversive gustatory receptor neurons (GRNs), but not the olfactory receptor neurons. Furthermore, three gustatory receptors, GR33a, GR66a, and GR93a, functioned together in coumarin detection by the proboscis. However, GR33a, but not GR66a and GR93a, was required to avoid coumarin during oviposition, with a choice of the same substrates provided as in binary food choice assay. Taken together, these findings suggest that anti-feeding activity and oviposition to avoid coumarin occur via separate mechanisms.

Antibiotic Resistance of Pectobacterium Korean Strains Susceptible to the Bacteriophage phiPccP-1

  • Vu, Nguyen Trung;Roh, Eunjung;Thi, Thuong Nguyen;Oh, Chang Sik
    • Research in Plant Disease
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    • v.28 no.3
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    • pp.166-171
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    • 2022
  • Commercial products with antibiotics like streptomycin as active ingredients have been used to control soft rot disease caused by Pectobacterium species for a long time. In this study, antibiotic resistance of twenty-seven Korean strains of Pectobacterium species including P. carotovorum, P. odoriferum, P. brasiliense, and P. parmenteri, which were previously shown to be susceptible to the bacteriophage phiPccP-1 was surveyed using a disk diffusion assay. While all strains were highly susceptible to ampicillin, kanamycin, chloramphenicol, tetracycline, and rifampicin, some strains showed weak susceptibility to 300 ㎍/ml of streptomycin. Furthermore, some of them are partially or completely resistant to commercial pesticides-Buramycinand streptomycin at the concentration of 250 ㎍/ml that is recommended by the manufacturer for streptomycin-based pesticides. These results indicate the presence of streptomycin-resistant Pectobacterium strains in South Korea, and the development of antibiotic alternatives to control soft rot is needed.

Isolation and Characterization of Plant Growth Promoting Rhizobacteria From Button Mushroom Compost

  • Oh, Sung-Hoon;Lee, Chang-Jung;Yoon, Min-Ho
    • Korean Journal of Agricultural Science
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    • v.43 no.1
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    • pp.100-108
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    • 2016
  • An auxin-producing bacteria (strain 5-1) was isolated from button mushroom compost in Boryeong-Si, Chungcheongnam-Do. The 5-1 strain was classified as a novel strain of Enterobacter aerogenes based on chemotaxonomic and phylogenetic analyses. The isolated E. aerogenes 5-1 was confirmed to produce indole-3-acetic acid (IAA), one of the auxin hormones, using TLC and HPLC analyses. When the concentration of IAA was assessed by performing HPLC quantitative analysis, a maximum concentration of IAA of $109.9mgL^{-1}$ was detected in the culture broth incubated in R2A medium containing 0.1% L-tryptophan for 24 h at $35^{\circ}C$. Acidification of the culture was deemed caused by an increase of IAA because a negative relationship between IAA production and pH was observed. Supplementation with a known precursor of IAA production, L-tryptophan, appeared to induce maximal production at 0.1% concentration, but it reduced production at concentrations above 0.2%. To investigate the growth-promoting effects to crops, the culture broth of E. aerogenes 5-1 was used to inoculate water cultures and seed pots of mung bean and lettuce. In consequence, adventitious root induction and root growth of mung bean and lettuce were two times higher than those of the control.

Trap identification of the constitutive promoter-like sequences from the bacterial fish pathogen, as exemplified by Edwardsiella tarda

  • Lee, Sang-Yoon;Kim, Ki-Hong;Kim, Dong-Soo;Nam, Yoon-Kwon
    • Journal of fish pathology
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    • v.24 no.3
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    • pp.297-305
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    • 2011
  • A trap identification system for isolating functional sequences to allow the constitutive expression of foreign protein from Edwardsiella tarda was developed. Using the green fluorescent protein (GFP) reporter-based trap system, various functional sequences to drive heterologous expression of the GFP were selectable in Escherichia coli host. However from the bioinformatic sequence analysis, all the segments predicted as regulatory regions were not native promoters actually existing upstream of endogenous E. tarda genes. Instead, a number of non-authentic sequences, possibly resulted from the random shuffling and/or intermolecular ligation were also proven to be able to display a potent GFP expression in the recombinant E. coli. Further analysis with selected clones showed that both authentic and non-authentic sequences could function in as a constitutive promoter, leading quite a consistent and stable GFP expression after repetitive subcultures. Microscopic examination also confirmed the uniform pattern of GFP expression in every host bacterium. Semi-quantitative assay of GFP showed that there was no clear relationship between expression levels and organizational features of the promoters trapped. Functional promoter-like elements achieved in the present study could be a good starting material for multivalent genetic engineering of E. tarda in order to produce recombinant vaccines in a cost-effective fashion.