• Title/Summary/Keyword: Cell-based assay

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The Effect of the Long-Term Ginseng Intakes on Serum Lipids Profile and Hemostatic Factors in Human (장기적인 인삼섭취가 인체의 혈전 관련 요인에 미치는 영향)

  • 이정희
    • Journal of Nutrition and Health
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    • v.28 no.9
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    • pp.862-871
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    • 1995
  • This study was designed to investigate the effectiveness of ginseng intakes in modifying serum lipid profiles and plasma clotting factors. The participants in this study were 47 normal healthy volunteers(men 24, women 23) with an age range of 35-49 years and a mean age of 41 years residing in Taejon. Based on the diet intakes, subjects were classed into one of three groups : control, vegetarian, and ginseng consumed over 3-4 years. There was no significant difference in their physical characteristics. Dietary calorie intakes were not significantly different in subjects. The ratio of energy intake in the control and ginseng consumed group was 63-64% : 20-21% : 15-16%(Cho : Fat : Pro), but 70-73% : 13-14% :14-15%(Cho : Fat : Pro) in the vegetarians. The intakes of animal food in the vegetarian was significantly lower than the control and ginseng consumed group in men. The ratio of P/S(1.27) was the highest in the vegetarians. Venous blood samples were taken for serum lipid profiling, plasma clotting assay and platelet function. The concentration of serum triglyceride in the men ginseng group is significantly lower than those of the men control group. Serum lipid profiles values of the men ginseng group, such as total cholesterol and phospholipid were lower those of the men control group, but higher those of the men vegetarian group. the serum lipid profile in the women were not significant, but total cholesterol, triglyceride and LDL cholesterol levels in the ginseng groups were low. The concentration of HDL cholesterol was not significantly different. Platelet cell count and platelet aggregation were low in the ginseng groups. APTT(Activated Partial thromboplastin time) was significantly elongated in ginseng groups in the normal range. In seems that the major beneficial effects of ginseng intakes in especially men were on the blood concentrations of triglyceride, total cholesterol and elongation of plasma clotting time.

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Anti-oxidative, Nitric Oxide Inhibitory Activities and Irritation Test of the Fermented Opuntia humifusa Cladodes

  • Chang, Zhi-Qiang;Hwang, Mi-Hyun;Oh, Byung-Chul;Lee, Sam-Pin;Rhee, Man-Hee;Kim, Kil-Soo;Jeong, Kyu-Shik;Kim, Jong-Choon;Park, Seung-Chun
    • Toxicological Research
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    • v.22 no.3
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    • pp.229-235
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    • 2006
  • Opuntia humifusa is a member of the Cactaceae family. In the present study, the antioxidant, nitric oxide(NO) inhibitory activities and potential irritation response of the fermented Opuntia humifusa cladodes(FOH) were investigated for cosmetic use. Antioxidant activities were tested using 1,1-diphenyl-2-picrylhydrazyl(DPPH) and xanthine oxidase assay, we found that FOH could scavenge DPPH free radicals and inhibit xanthine oxidase activity in a dose dependent manner, with $IC_{50}$ of 2599.46${\mu}g/ml$ and 721.38${\mu}g/ml$, respectively. To investigate the possible anti-inflammatory effects of FOH, RAW 264.7 macrophages were pretreated with FOH($0{\sim}400{\mu}g/ml$) for 30 min and then treated with LPS for 24 h. We found that cell number did not vary significantly with the treatment of FOH, and FOH did not show any inhibitory effects on LPS-induced NO production. After application of FOH to rabbits for skin and eye irritation test, the experimental sites did not show any response compared to the control. FOH were considered to be a non-irritant to the skin and eye. Based on the above information, we suggest that FOH can be considered to be a non-irritant base cosmetic material for safely use.

Generation and Expression in Plants of a Single-Chain Variable Fragment Antibody Against the Immunodominant Membrane Protein of Candidatus Phytoplasma Aurantifolia

  • Shahryari, F.;Safarnejad, M.R.;Shams-Bakhsh, M.;Schillberg, S.;Nolke, G.
    • Journal of Microbiology and Biotechnology
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    • v.23 no.8
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    • pp.1047-1054
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    • 2013
  • Witches' broom of lime is a disease caused by Candidatus Phytoplasma aurantifolia, which represents the most significant global threat to the production of lime trees (Citrus aurantifolia). Conventional disease management strategies have shown little success, and new approaches based on genetic engineering need to be considered. The expression of recombinant antibodies and fragments thereof in plant cells is a powerful approach that can be used to suppress plant pathogens. We have developed a single-chain variable fragment antibody (scFvIMP6) against the immunodominant membrane protein (IMP) of witches' broom phytoplasma and expressed it in different plant cell compartments. We isolated scFvIMP6 from a naïve scFv phage display library and expressed it in bacteria to demonstrate its binding activity against both recombinant IMP and intact phytoplasma cells. The expression of scFvIMP6 in plants was evaluated by transferring the scFvIMP6 cDNA to plant expression vectors featuring constitutive or phloem specific promoters in cassettes with or without secretion signals, therefore causing the protein to accumulate either in the cytosol or apoplast. All constructs were transiently expressed in Nicotiana benthamiana by agroinfiltration, and antibodies of the anticipated size were detected by immunoblotting. Plant-derived scFvIMP6 was purified by affinity chromatography, and specific binding to recombinant IMP was demonstrated by enzyme-linked immunosorbent assay. Our results indicate that scFvIMP6 binds with high activity and can be used for the detection of Ca. Phytoplasma aurantifolia and is also a suitable candidate for stable expression in lime trees to suppress witches' broom of lime.

Neuroprotective effect of caffeic acid phenethyl ester in 3-nitropropionic acid-induced striatal neurotoxicity

  • Bak, Jia;Kim, Hee Jung;Kim, Seong Yun;Choi, Yun-Sik
    • The Korean Journal of Physiology and Pharmacology
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    • v.20 no.3
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    • pp.279-286
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    • 2016
  • Caffeic acid phenethyl ester (CAPE), derived from honeybee hives, is a bioactive compound with strong antioxidant activity. This study was designed to test the neuroprotective effect of CAPE in 3-nitropropionic acid (3NP)-induced striatal neurotoxicity, a chemical model of Huntington's disease (HD). Initially, to test CAPE's antioxidant activity, a 2,2'-azino-bis-3-ethylbenzthiazoline-6-sulfonic acid (ABTS) antioxidant assay was employed, and CAPE showed a strong direct radical-scavenging effect. In addition, CAPE provided protection from 3NP-induced neuronal cell death in cultured striatal neurons. Based on these observations, the in vivo therapeutic potential of CAPE in 3NP-induced HD was tested. For this purpose, male C57BL/6 mice were repeatedly given 3NP to induce HD-like pathogenesis, and 30 mg/kg of CAPE or vehicle (5% dimethyl sulfoxide and 95% peanut oil) was administered daily. CAPE did not cause changes in body weight, but it reduced mortality by 29%. In addition, compared to the vehicle-treated group, robustly reduced striatal damage was observed in the CAPE-treated animals, and the 3NP-induced behavioral deficits on the rotarod test were significantly rescued after the CAPE treatment. Furthermore, immunohistochemical data showed that immunoreactivity to glial fibrillary acidic protein (GFAP) and CD45, markers for astrocyte and microglia activation, respectively, were strikingly reduced. Combined, these data unequivocally indicate that CAPE has a strong antioxidant effect and can be used as a potential therapeutic agent against HD.

Synthesis, Characterization and in vitro Anti-Tumoral Evaluation of Erlotinib-PCEC Nanoparticles

  • Barghi, Leila;Asgari, Davoud;Barar, Jaleh;Nakhlband, Aylar;Valizadeh, Hadi
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.23
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    • pp.10281-10287
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    • 2015
  • Background: Development of a nanosized polymeric delivery system for erlotinib was the main objective of this research. Materials and Methods: Poly caprolactone-polyethylene glycol-polycaprolactone (PCEC) copolymers with different compositions were synthesized via ring opening polymerization. Formation of triblock copolymers was confirmed by HNMR as well as FT-IR. Erlotinib loaded nanoparticles were prepared by means of synthesized copolymers with solvent displacement method. Results: Physicochemical properties of obtained polymeric nanoparticles were dependent on composition of used copolymers. Size of particles was decreased with decreasing the PCL/PEG molar ratio in used copolymers. Encapsulation efficiency of prepared formulations was declined by decreasing their particle size. Drug release behavior from the prepared nanoparticles exhibited a sustained pattern without a burst release. From the release profiles, it can be found that erlotinib release rate from polymeric nanoparticles is decreased by increase of CL/PEG molar ratio of prepared block copolymers. Based on MTT assay results, cell growth inhibition of erlotinib has a dose and time dependent pattern. After 72 hours of exposure, the 50% inhibitory concentration (IC50) of erlotinib hydrochloride was appeared to be $14.8{\mu}M$. Conclusions: From the obtained results, it can be concluded that the prepared PCEC nanoparticles in this study might have the potential to be considered as delivery system for erlotinib.

Application of the Recombinant Bioluminescence Bacterium on the Toxicity Assessment of the Sole Chemicals and Soil Samples (유전자 재조합 생물 발광 균주를 이용한 순수 오염물과 토양시료의 독성도 평가)

  • Kong, In-Chul;Kim, Jin-Yeong;Lee, Sun-Hee;Ko, Kyung-Seok
    • Journal of Korean Society of Environmental Engineers
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    • v.34 no.2
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    • pp.136-142
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    • 2012
  • Various factors affecting on the bioassay were investigated. Experiments with a low mixture ratio (cell to toxicant solution) of 0.5 : 9.5 (v/v) produced observable bioluminescence intensity for assay. Both sodium lactate and potassium nitrate stimulate bioluminescence activity; 2.6~4.0 times of control. Distilled water and MSM, which gave non significant effects on the bioluminescence activity, were determined as proper diluent or extract solutions. A wide range of toxic responses of metals and organics were observed. In general, organics were much less sensitive than metals. Samples collected from eleven sites showed the bioluminescence activity ranging from 29 to 111% of the control. Significant correlation between toxicity and total metal contents was not observed, but the toxicity of two groups, sorted based on the contaminated arsenic concentration in soils, was 44% and 20%, showing considerable differences.

Molecular Cloning and Characterization of Lysozyme II from Artogeia rapae and its Expression in Baculovirus-infected Insect Cells

  • Bang, In-Seok;Kang, Chang-Soo
    • Animal cells and systems
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    • v.11 no.2
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    • pp.175-182
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    • 2007
  • The lysozyme II gene of cabbage butterfly Artogeia rapae was cloned from fat body of the larvae injected with E. coli and its nucleotide sequence was determined by the RACE-PCR. It has an open reading frame of 414 bp nucleotides corresponding to 138 amino acids including a signal sequence of 18 amino acids. The estimated molecular weight and the isoelectric point of the lysozyme II without the signal peptide were 13,649.38 Da and 9.11, respectively. The A. rapae lysozyme II (ARL II) showed the highest identity (81%) in the amino acid sequence to Manduca sexta lysozyme among other lepidopteran species. The two catalytic residues ($Glu^{32}$ and $Asp^{50}$) and the eight Cys residue motifs, which are highly conserved among other c-type lysozymes in invertebrates and vertebrates, are also completely conserved. A phylogenetic analysis based on amino acid sequences indicated that the ARL II was more closely related to M. sexta, Hyphantria cunea, Heliothis virescens, and Trichoplusia ni lysozymes. The ARL II gene was expressed in Spodoptera frugiperda 21 insect cells and the recombinant ARL II (rARL II) was purified from cell-conditioned media by cation exchange column chromatography and reverse phase FPLC. The purified rARL II was able to form a clear zone in lysoplate assay against Micrococcus luteus. The lytic activity was estimated to be 511.41 U/mg, 1.53 times higher than that of the chicken lysozyme. The optimum temperature for the lytic activity of the rARL II was $50^{\circ}C$, the temperature dependency of the absolute lytic activity of rARL II was higher than that of the chicken lysozyme at low temperatures under $65^{\circ}C$.

Purification of Materials Produced by Amylocolatosis sp. and Anticancer Effect in Oral Cancer Model (Amylocolatosis sp.가 생산하는 항암물질의 정제 및 구강암 모델에 미치는 항암 효과)

  • Kim, Jung;Park, Young-Min;Lim, Do-Seon
    • Journal of dental hygiene science
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    • v.3 no.1
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    • pp.11-14
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    • 2003
  • A methylotrophic Actinomycetes strain, which produce the anti-oral cancer activity compound, was isolated from soil and estimated as Amylocolatosis sp. based on taxonomic studies. A methanol didn't have influence on the production of the anticancer compounds. These compound were isolated by ethylacetate extract, silica gel column chromatography, sephadex LH-20 column and reverse phase HPLC. The compounds were very stable under heat ($121^{\circ}C$), acid(pH 2.0) and alkali(pH 11.0) treatment. The cytotoxic effect of isolated anticancer compounds on various cancer cell lines such as A549, SNU-1, KB, L1210, and Sarcoma 180 was investigated by MTT assay method. And these produced compounds also showed the broad antimicrobial spectrum to test strains such as bacteria and yeast.

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Biapigenin, Candidate of an Agonist of Human Peroxisome Proliferator-Activated Receptor γ with Anticancer Activity

  • Kim, Jin-Kyoung;Shin, So-Young;Lee, Jee-Young;Lee, So-Jung;Lee, Eun-Jung;Jin, Qinglong;Lee, June-Young;Woo, Eun-Rhan;Lee, Dong-Gun;Yoon, Do-Young;Kim, Yang-Mee
    • Bulletin of the Korean Chemical Society
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    • v.32 no.8
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    • pp.2717-2721
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    • 2011
  • Peroxisome proliferator-activated receptors (PPARs) are a subfamily of nuclear receptors (NRs). Human peroxisome proliferator-activated receptor gamma (hPPAR${\gamma}$) has been implicated in the pathology of numerous diseases, including obesity, diabetes, and cancer. ELISA-based hPPAR${\gamma}$ activation assay showed that biapigenin increased the binding between hPPAR${\gamma}$ and steroid receptor coactivator-1 (SRC-1) by approximately 3-fold. In order to confirm that biapigenin binds to hPPAR${\gamma}$, fluorescence quenching experiment was performed. The results showed that biapigenin has higher binding affinity to hPPAR${\gamma}$ at nanomolar concentrations compared to indomethacin. Biapigenin showed anticancer activity against HeLa cells. Biapigenin was noncytotoxic against HaCa T cell. All these data implied that biapigenin may be a potent agonist of hPPAR${\gamma}$ with anticancer activity. We will further investigate its anticancer effects against human cervical cancer.

D-Pinitol Promotes Apoptosis in MCF-7 Cells via Induction of p53 and Bax and Inhibition of Bcl-2 and NF-κB

  • Rengarajan, Thamaraiselvan;Nandakumar, Natarajan;Rajendran, Peramaiyan;Haribabu, Lingaiah;Nishigaki, Ikuo;Balasubramanian, Maruthaiveeran Periyasamy
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.4
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    • pp.1757-1762
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    • 2014
  • Development of drugs from natural products has been undergoing a gradual evoluation. Many plant derived compounds have excellent therapeutic potential against various human ailments. They are important sources especially for anticancer agents. A number of promising new agents are in clinical development based on their selective molecular targets in the field of oncology. D-pinitol is a naturally occurring compound derived from soy which has significant pharmacological activitites. Therefore we selected D-pinitol in order to evaluate apoptotic potential in the MCF-7 cell line. Human breast cancer cells were treated with different concentrations of D-pinitol and cytotoxicity was measured by MTT and LDH assays. The mechanism of apoptosis was studied with reference to expression of p53, Bcl-2, Bax and NF-kB proteins. The results revealed that D-pinitol significantly inhibited the proliferation of MCF-7 cells in a concentration-dependent manner, while upregulating the expression of p53, Bax and down regulating Bcl-2 and NF-kB. Thus the results obtained in this study clearly vindicated that D-pinitol induces apotosis in MCF-7 cells through regulation of proteins of pro- and anti-apoptotic cascades.