• Title/Summary/Keyword: five dose assay

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Comparison of 5 Assays for Quantification of Antibody to Hepatitis B Virus Surface Antigen with Immunoglobulin G Preparations (면역글로불린제제 효능평가를 위한 5종 B형간염 표면항원항체검출법의 비교)

  • Shin, In-Soo;Lee, Yoo-Kyoung;Kim, Oh-Jung;Ban, Sang-Ja
    • KSBB Journal
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    • v.26 no.2
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    • pp.157-164
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    • 2011
  • Five assays for anti-HBs were compared to improve potency test of Human lgG preparations. The three commercial EIA kits were optimized including dose response curve ranges and compared by conducting a co-laboratory study. After selecting the most reproducible EIA kit, methods comparison was performed with 22 samples in 5 different days. As a result, EIA (7.7 ${\pm}$ 5.3%) and MEIA (AxSYM: 3.7 ${\pm}$ 1.9%, IMx: 1.6 ${\pm}$ 0.8%) showed precision and accuracy (100.1 ${\pm}$ 12.6%). Therefore, the validated EIA assay was established and it is believed to be comparable to current MEIA.

Construction of a Thermotolerant Saccharomyces cerevisiae Strain for Bioethanol Production with Reduced Fermentation Time and Saccharifying Enzyme Dose

  • Lim, Ji Sung;Jang, You Ri;Lim, Young Hoon;Kim, Keun
    • Journal of Microbiology and Biotechnology
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    • v.22 no.10
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    • pp.1401-1405
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    • 2012
  • A thermotolerant Saccharomyces cerevisiae mutant strain, TT6, was constructed after multi-parental hybridization of five mutant strains obtained by UV or NTG treatment of the original strain, S. cerevisiae KV1. When incubated at $40^{\circ}C$ in YPD broth, TT6 began to grow exponentially in 10 h, but KV1 did not show any noticeable growth even after 22 h. The thermotolerant growth of TT6 was confirmed by serial dilution assay at $42^{\circ}C$; TT6 grew at a cell concentration ($10^{-5}$) 10,000 times lower than that of KV1 ($10^{-1}$). Whereas ethanol production from YP containing 23% (w/v) glucose by KV1 decreased with increasing temperature from $30^{\circ}C$ to $36^{\circ}C$, ethanol production by TT6 did not decrease at temperatures up to $37^{\circ}C$. When TT6 was tested for ethanol production at $36^{\circ}C$ by simultaneous saccharification and fermentation (SSF) from 23% corn, 24 h of fermentation time or 50% of the glucoamylase dose was saved when compared with KV1 at $30^{\circ}C$. The ethanol yield from corn by SSF with TT6 at $36^{\circ}C$ was 91.7% of the theoretical yield, whereas that of KV1 at $30^{\circ}C$ was 90.6%.

Short-Term Changes in Gut Microflora and Intestinal Epithelium in X-Ray Exposed Mice

  • Tsujiguchi, Takakiyo;Yamaguchi, Masaru;Yamanouchi, Kanako
    • Journal of Radiation Protection and Research
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    • v.45 no.4
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    • pp.163-170
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    • 2020
  • Background: Gut microflora contributes to the nutritional metabolism of the host and to strengthen its immune system. However, if the intestinal barrier function of the living body is destroyed by radiation exposure, the intestinal bacteria harm the health of the host and cause sepsis. Therefore, this study aims to trace short-term radiation-induced changes in the mouse gut microflora-dominant bacterial genus, and analyze the degree of intestinal epithelial damage. Materials and Methods: Mice were irradiated with 0, 2, 4, 8 Gy X-rays, and the gut microflora and intestinal epithelial changes were analyzed 72 hours later. Five representative genera of Actinobacteria, Firmicutes, and Bacteroidetes were analyzed in fecal samples, and the intestine was pathologically analyzed by Hematoxylin-Eosin and Alcian blue staining. In addition, DNA fragmentation was evaluated by the TdT-mediated dUTP nick-end labeling (TUNEL) assay. Results and Discussion: The small intestine showed shortened villi and reduced number of goblet cells upon 8 Gy irradiation. The large intestine epithelium showed no significant morphological changes, but the number of goblet cells were reduced in a radiation dose-dependent manner. Moreover, the small intestinal epithelium of 8 Gy-irradiated mice showed significant DNA damaged, whereas the large intestine epithelium was damaged in a dose-dependent manner. Overall, the large intestine epithelium showed less recovery potential upon radiation exposure than the small intestinal epithelium. Analysis of the intestinal flora revealed fluctuations in lactic acid bacteria excretion after irradiation regardless of the morphological changes of intestinal epithelium. Altogether, it became clear that radiation exposure could cause an immediate change of their excretion. Conclusion: This study revealed changes in the intestinal epithelium and intestinal microbiota that may pave the way for the identification of novel biomarkers of radiation-induced gastrointestinal disorders and develop new therapeutic strategies to treat patients with acute radiation syndrome.

Studies on Constituents of Higher Fungi of Korea (LXXIII) Antitumor Components of the Cultured Mycelia of Paxillus atrotomentosus (한국산 고등 균류의 성분 연구(제73보) -좀우단버섯 배양 균사체의 항암 성분-)

  • Kwag, Sang-Deok;Bok, Jin-Woo;Hyun, Jin-Won;Choi, Eung-Chil;Kim, Byong-Kak
    • The Korean Journal of Mycology
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    • v.20 no.3
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    • pp.240-251
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    • 1992
  • To find antitumor components from higher fungi, the cultured mycelia of Paxillus atrotomentosus were extracted with hot water. The water soluble fraction was purified and separated by DEAE-cellulose ion exchange chromatography and Sepharose CL-4B gel filtration method. The separated fractions(Fr.) were designated CR A, B, C and D. Fr. A showed the highest inhibition ratio of 68.51% among the five tractions at a dose of 20 mg/kg/day. When Fr. A was examined for immunopotentiation activity, it increased the amount of the superoxide anion from activated macrophages to 1.1 fold and the number of plaques in hemolytic plaque assay to 2.3 fold, respectively. Otherwise, it did not show direct cytotoxity in sarcoma 180. Delayed type hypersensitiyity reaction showed that the decreased footpad swelling of tumor-hearing was restored to the normal. These results indicate that antitumor activity was exerted through immunopotentiation. Its chemical analysis showed 86.36% polysaccharide, 1.52% protein and 1.64% hexosamine. The polysaccharide consisted of fucose, galactose, glucose, mannose and xylose. This component was named paxillan.

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Effects of Lactic Acid Bacteria on intestinal Microbial Enzyme Activity and Composition in Rats Treated with Azoxymethane

  • Sang-Myeong;Lee, Wan-Kyu
    • Journal of Microbiology
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    • v.39 no.3
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    • pp.154-161
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    • 2001
  • In recent years, colon cancer has been reported to be one of the most important causes of cancer morbidity and mortality in Korea. Epidemiological and experimental studies suggest that lactic acid bacteria (LAB) used to ferment dairy products inhibits colon carcinogenesis. The present study was designed to determine whether the colon cancer inhibitory effect of LAB (Bifidobacterium longum Hy8001; Bif and Lactobacillus acidophilus HY2l04; Lac) of Korean origin, is associated with intestinal microflora composition and certain enzyme activity in rats treated with azoxymethane (AOM). At five weeks of age, SD rats were divided at random into four (AOM alone, Bif, Lac, and Bif+Lac) groups. Oral administration of lactic acid bacteria cultures were performed daily until the termination of the study. Two weeks later all animals were given a subcutaneous injection of AOM dissolved in normal saline at a dose of 15 mg/kg of body weight once weekly for 2 weeks. Every two weeks for 10 weeks, five of the rats in each group were randomly chosen for fecal specimen collection. The fecal specimens were used for assay of $\beta$-glucuronidase and nitroreductase, and analysis of intestinal microflora composition. The activity of $\beta$-glucuronidase which plays an important role in the production of the carcinogenic metabolite of azoxymethane was remarkably increased in the AOM alone group after AOM injection and maintained the high level during the experiment. However, LAB inhibited the AOM-induced increase in $\beta$-glucuronidase activity. Nitroreductase activity decreased by 30-40% in LAB treated groups in comparison with that of the AOM alone group. The results of the present study suggest that LAB inhibits colon carcinogenesis by modulating the metabolic activity of intestinal micro-flora and improving the composition of intestinal microflora.

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Gossypii Semen oil alleviates memory dysfunction in scopolamine-treated mice (면화자 정유의 기억력 손상 완화 효과)

  • Lee, Jihye;Jung, Eun Mi;Lee, Eunhong;Jang, Gwi Yeong;Seo, Kyung Hye;Kim, Mi Ryeo;Jung, Ji Wook
    • The Korea Journal of Herbology
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    • v.36 no.2
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    • pp.1-9
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    • 2021
  • Objectives : Gossypium arboreum (cotton) is traditionally used to treat various health disorders. However, anti-amnesic effect of G. arboreum has not been reported. The objective of this study was to investigate in-vivo the anti-amnesic effects along with in vitro antioxidant and acetylcholinesterase (AChE) inhibition potential in G. arboreum seed essential oil. Methods : The essential oil of G. arboreum obtained by solid phase microextraction (SPME) techniques were identified by gas chromatography-mass spectroscopy (GC-MS). 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assay were performed to determine the antioxidant activity at various concentrations (312.5, 625, 1250, 2500, 5000, 10000 ㎍/㎖. Y-maze, passive avoidance and Morris water maze tests were carried out to evaluate improved effect on scopolamine (1 mg/kg)-induced memory dysfunction at the dose level of 50, 100 and 200 mg/kg. Donepezil (5 mg/kg) was used as a positive drug control. We performed acetylcholinesterase (AChE) activity assay in ex vivo. Results : Five volatile compounds were identified in G. arboreum. The assays of DPPH and ABTS revealed that G. arboreum increased antioxidant activity in a dose-dependent manner. G. arboreum ameliorated the percent of spontaneous alternation in the Y-maze test, shortened step-through latency in the passive avoidance test, and increased swimming time in the target zone in the Morris water maze test. In addition, G. arboreum inhibited the AChE activity. Conclusions : Based on these findings, G. arboreum may aid in the prevention and treatment of learning and memory-deficit disorders through antioxidant and AChE inhibitory activities.

Anti-oxidative Activity of Five Plant Extracts including Apios fortune, Colubrina arborescens, Croton caudatus, Osmanthus matsumuranus and Schima noronhae (Apios fortunei, Colubrina arborescens, Croton caudatus, Osmanthus matsumuranus, 그리고 Schima noronhae를 포함하는 5종 식물 추출물의 항산화 활성)

  • Lee, Su Hyeon;Jin, Kyong-Suk;Kwon, Hyun Ju;Kim, Byung Woo
    • Journal of Life Science
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    • v.28 no.9
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    • pp.1092-1099
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    • 2018
  • This study was orchestrated with the purpose of uncovering new nutraceutical resources possessing biological activities in the plant kingdom. To fulfill our objective, we analyzed several plant extracts and selected five species possessing powerful anti-oxidative activity. The anti-oxidative effect of these five plants, Apios fortunei Maxim., Colubrina arborescens Sarg., Croton caudatus Geiseler, Osmanthus matsumuranus Hayata and Schima noronhae Reinw. ethanol extracts were then evaluated by using in vitro assay, cell model system, and Western blot analysis of target proteins. As the results, all of them possessed the potent scavenging activity against 1,1-diphenyl-2-picryl hydrazyl (DPPH), similar with that of ascorbic acid, used as a common positive control. Moreover, they strongly inhibited hydrogen peroxide ($H_2O_2$)-induced reactive oxygen species (ROS), in a dose-dependent manner, in RAW 264.7 murine macrophage cells. Furthermore, they induced the protein expression of an anti-oxidative enzyme, heme oxygenase 1 (HO-1), and its upstream transcription factor, nuclear factor-E2-related factor 2 (Nrf2). Taken together, these results indicate that these five plants possess potent anti-oxidative activity and thus appear to be useful sources as potential anti-oxidant agents. Therefore, they might be utilized as promising materials in the field of nutraceuticals.

Activation of Antioxidant-Response Element (ARE), Mitogen- Activated Protein Kinases (MAPKs) and Caspases by Major Green Tea Polyphenol Components during Cell Survival and Death

  • Chen, Chi;Yu, Rong;Owuor, Edward D.;Kong, A.NTony
    • Archives of Pharmacal Research
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    • v.23 no.6
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    • pp.605-612
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    • 2000
  • Green tea polyphenols (GTP) have been demonstrated to suppress tumorigenesis in several chemical-induced animal carcinogenesis models, and predicted as promising chemopreventive agents in human. Recent studies of GTP extracts showed the involvement of mitogen-activated protein kinases (MAPKs) in the regulation of Phase II enzymes gene expression and induction of apoptosis. In the current work we compared the biological actions of five green tea catechins: (1) induction of ARE reporter gene, (2) activation of MAP kinases, (3) cytotoxicity in human hepatoma HepG2-C8 cells, and (4) caspase activation in human cervical squamous carcinoma HeLa cells. For the induction of phase IIgene assay, (-)-epigallocatechin-3-gallate (EGCG) and (-)-epicatechin-3-gallate (ECG) potently induced antioxidant response element (ARE)-mediated luciferase activity, with induction observed at 25 $\mu\textrm{m}$with EGCG. The induction of ARE reporter gene appears to be structurally related to the 3-gallate group. Comparing the activation of MAPK by the five polyphenols, only EGCG showed potent activation of all three MAPKs (ERK, JNK and p38) in a dose- and time-dependent manner, whereas EGC activated ERK and p38. In the concentration range of 25 $\mu\textrm{m}$ to 1 mM, EGCG and ECG strongly suppressed HepG2-ARE-C8 cell-growth. To elucidate the mechanisms of green tea polyphenol-induced apoptosis, we measured the activation of an important cell death protein, caspase-3 induced by EGCG, and found that caspase-3 was activated in a dose- and time-dependent manner. Interestingly, the activation of caspase-3 was a relatively late event (peaked at 16 h), whereas activation of MAPKs was much earlier (peaked at 2 h). It is possible, that at low concentrations of EGCG, activation of MAPK leads to ARE-mediated gene expression including phase II detoxifying enzymes. Whereas at higher concentrations of EGCG, sustained activation of MAPKs such as JNK leads to apoptosis. These mechanisms are currently under investigation in our laboratory. As the most abundant catechin in GTP extract, we found that EGCG potently induced ARE-mediated gene expression, activated MAP kinase pathway, stimulated caspase-3 activity, and induced apoptosis. These mechanisms together with others, may contribute to the overall chemopreventive function of EGCG itself as well as the GTP.

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Differential Effects of Resveratrol and its Oligomers Isolated from Seeds of Paeonia lactiflora (Peony) on Proliferation of MCF-7 and ROS 17/2.8 Cells

  • Kim, Hyo-Jin;Lee, Won-Jung;Park, Yun-Hee;Cho, Sung-Hee;Park, Sang-Won
    • Preventive Nutrition and Food Science
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    • v.8 no.4
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    • pp.356-364
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    • 2003
  • A methanol extract from seeds of Paeonia lactiflora (Paeoniaceae, peony) was found to possess different antiproliferative activities against four different human cancer cell lines: Hela, MCF-7, HepG2 and HT-29. Furthermore, five different methanol (20, 40, 60, 80 and 100 % MeOH) fractions obtained by fractionation of the methanol extract of the seeds on a Diaion HP-20 column exhibited differential antiproliferative effects against the above four cancer cell lines. Among five fractions, the 60 % MeOH fraction showed relatively lower antiproliferative activity on MCF-7 estrogen-sensitive breast cancer cell than the other cancer cell lines. Systematic separation of 60% the MeOH fraction by silica gel and Sephadex LH-20 columns led to the isolation of four known stilbenes, trans-resveratrol (1), trans-(+)- $\varepsilon$ -viniferin (2), gnetin H (3) and suffruticosol B (4). The four stilbenes (1∼4) exerted differential biphasic effects on cell proliferation of MCF-7 cells in a similar manner as genistein, a soybean isoflavone used as a positive reference, in the concentration range from 1.0 to 200 $\mu$M. Three stilbenes (1 ∼ 3) weakly stimulated the proliferation of MCF -7 cells at doses below 10 JIM. However, strong antiproliferative effects on MCF-7 cell were exerted by extract 1 at a dose of 200 JIM, and by 2 and 3 at doses above 25 $\mu$M. In contrast, 4 inhibited the proliferation of MCF-7 cell at a dose below 25 $\mu$M, but stimulated cell proliferation at concentrations of 50 and 100 $\mu$M. All four stilbenes (1∼4) stimulated the proliferation of ROS 17/2.8 osteoblast-like cells in the range of 10$^{-10}$ ∼10$^{-1}$ $\mu$M. Compound 1 exhibited especially potent proliferative activity, although its activity was weaker than that of genistein. Additionally, three resveratrol oligomers (2∼4) also exhibited concentration-dependently moderate proliferative activity, but less than that of 1. These results suggest that resveratrol, and its dimer and trimers from the seeds of Paeonia lactiflora may act as a phytoestrogen, but in a somewhat different manner from that of genistein.

Antioxidant and Whitening Activity of Essential Oils (Essential Oils의 항산화 활성 및 미백작용)

  • Lim, Hye-Won;Kim, Ju-Yeon
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.31 no.3 s.52
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    • pp.265-271
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    • 2005
  • This study is performed to investigate the effects of citrus essential oils on melanin production in B16 melanoma cells and reactive oxygen species (ROS) generation in RBL 2H3 cells. Five kinds of citrus essential oil (bergamot, grapefruit, lemmon, mandarin, petigrain) did not have any influence on DPPH radical scavenger activity, cell growth and cytotoxicity in B16 melanoma cells. In purified tyrosinase assay, both mandarin and petigrain essential oils dose-dependently inhibited its activity, but bergamot did not. In $1{\mu}M\;{\alpha}-MSH-stimulated$ B16 melanoma cells, all of 5 citrus essential oils inhibited melanin production in $\underline{a}$ dose dependent manner. On the other hand, four kinds of citrus essential oil dose-dependently increased ROS generation in RBL 2H3 mast cells, but mandarin did not. From the above results, it is possible that citrus essential oils nay be developed to be anti-melanogenic agent on the basis of their inhibitory effect on MSH-induced melanin production. Hut we can not rule out the possibility of the induction of allergy and inflammation since citrus essential oils caused ROS generation in RBL 2H3 mast cells.