• Title/Summary/Keyword: bio assay

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Biochemical Study of Recombinant PcrA from Staphylococcus aureus for the Development of Screening Assays

  • Dubaele, Sandy;Martin, Christophe;Bohn, Jacqueline;Chene, Patrick
    • BMB Reports
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    • v.40 no.1
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    • pp.7-14
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    • 2007
  • Helicases are ubiquitous enzymes, which utilize the energy liberated during nucleotide triphosphate hydrolysis to separate double-stranded nucleic acids into single strands. These enzymes are very attractive targets for the development of new antibacterial compounds. The PcrA DNA helicase from Staphylococcus aureus is a good candidate for drug discovery. This enzyme is unique in the genome of S. aureus and essential for this bacterium. Furthermore, it has recently been published that it is possible to identify inhibitors of DNA helicases such as PcrA. In this report, we study the properties of recombinant PcrA from S. aureus purified from Escherichia coli to develop ATPase and helicase assays to screen for inhibitors.

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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Pinus densiflora Gnarl Inhibits Migration through Suppression of Protein Kinase C in C6 Glioma Cells (C6 Glioma 세포에서 Protein Kinase C Alpha 발현 저해를 통한 송절 약침액의 이주 억제 효과)

  • Min, Ilguk;Lee, Kangpa;Chang, Haeryong;Moon, Jinyoung
    • Korean Journal of Acupuncture
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    • v.32 no.2
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    • pp.51-58
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    • 2015
  • Objectives : Pinus densiflora gnarl, called Song-Jeol in Korean medicine, has been used to cure inflammatory diseases such as arthritis. In the present study, we evaluated inhibitory property of Song-Jeol pharmacopuncture(SJ) on C6 glioma cell migration. Methods : To evaluate cell viability on C6 glioma cells of SJ, the viability was assessed by using Ez-cytox assay kit. The cell migration was assessed by wound-healing assay and Boyden chamber assay, respectively. LPS-induced NO productions were determined by using the Griess reagent. The expression of iNOS and protein kinase $C(PKC)-{\alpha}$ were estimated by western blotting assay. Results : In the wound-healing assay and Boyden chamber assay, SJ showed a significant inhibition on serum-induced C6 glioma cell migration. In addition, NO production was decreased by SJ through suppression of iNOS expression in LPS-stimulated C6 glioma cell. Futhermore, LPS-induced protein kinase $C(PKC)-{\alpha}$ expression was effectively inhibited by SJ. Conclusions : These results demonstrated that SJ was useful for the suppression of the C6 glioma cell migration.

TWO COLORIMETRIC ASSAYS VERIFY THAT CALCIUM SULFATE PROMOTES PROLIFERATING ACTIVITY OF HUMAN GINGIVAL FIBROBLASTS

  • Chae, Min;Kim, Su-Yeon;Kim, Soo-Yeon;Lee, Suk-Won
    • The Journal of Korean Academy of Prosthodontics
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    • v.45 no.3
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    • pp.382-388
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    • 2007
  • Statement of problem. The role of calcium sulfate in stimulating the growth of gingival soft tissue has been reported in few studies. Such a unique property of calcium sulfate could serve as a trouble-solving broker in compensating for the lack of soft tissues in various oral surgeries. Purpose. The purpose of this study was to compare the proliferating activities of human gingival fibroblasts seeded on various bone graft barrier materials of calcium sulfate, collagen, and polytetrafluorethylene (PTFE). Material and methods. Two calcium sulfates ($CAPSET^{(R)}$. and $CalForma^{(R)}$, Lifecore Biomedical Inc., St. Paul, Minnesota, USA), a resorbable natural collagen ($Bio-Gide^{(R)}$, Geistlich Pharma Ag., Wolhusen, Switzerland), and a non-resorbable PTFE ($TefGen-FD^{(R)}$, Lifecore Biomedical Inc., St. Paul, Minnesota, USA) served as the human gingival fibroblasts' substrates and comprised the four experimental groups, whereas the untreated floors of culture plastics were used in the control group, in this study. Cells were trypsinized, seeded, and incubated for 48 h. The proliferating activities of fibroblasts were determined by XTT and SRB assay and absorbance (optical density, OD) was measured. One-way ANOVA was used to analyze the differences in the mean OD values between the groups of CAPSET, CalForma, Bio-Gide, TefGen, and the control (p<0.05). Results. From the XTT assay, the mean OD value of the control group, the highest, was significantly greater than that of any of the four experimental groups followed by CalForma, CAPSET, TefGen, and Bio-Gide. Further, the mean OD value of CalForma, was significantly greater compared to that of Bio-Gide. From the SRB assay, Calforma showed the highest mean OD value, which was significantly greater than that of any other groups, followed by the control, CAPSET, Bio-Gide, and TefGen. The mean OD values of both the control and CAPSET were significantly greater compared to that of TefGen (p<0.05). Conclusion. Assessment of the viability and proliferation of cultured fibroblasts seeded and incubated for 48 h on various barrier-material substrates using XTT and SRB assay showed that calcium sulfate $CalForma^{(R)}$ promotes the proliferating activity of human gingival fibroblasts.

Detection of infectious canine hepatitis virus by TaqMan real-time PCR method (TaqMan 실시간 PCR법에 의한 개 전염성 간염 바이러스의 검출)

  • Wang, Hye-young;Choi, Jae-yong;Lee, Mi-jin;Park, Jin-ho;Cho, Mae-Rim;Han, Jae-cheol;Choi, Kyoung-seong;Chae, Joon-seok
    • Korean Journal of Veterinary Research
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    • v.44 no.4
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    • pp.655-662
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    • 2004
  • The aim of this work was the validation of a rapid real-time PCR assay based on TaqMan technology for the unequivocal identification of infectious canine hepatitis (ICH) virus, to be used directly on DNA purified from blood specimens. A real-time PCR system targeting at the E3 ORFA gene sequence of canine adenovirus type 1 was optimized and validated through comparative analysis of samples using conventional PCR system. The real-time PCR assay based on TaqMan technology could disclose 23 (37.7%) out of 61 samples as PCR positive. In contrast, 18 (29.5%) samples were found PCR positive when conventional PCR was applied on these samples. The use of the ABI Prism 7700 sequence detection system allowed the efficient determination of the amplified product accumulation through a fluorogenic probe. The entire real-time TaqMan PCR assay, including DNA extraction, amplification, and detection could be completed within 3 hours. The detection method of real-time TaqMan PCR assay was 1,000 times more sensitive than conventional PCR. Real-time TaqMan probe and primer set developed and optimized in this study is a sensitive, rapid and accurate method for detection of ICH virus and can be effective screening tool for the detection of ICH in a diagnostic laboratory routines.

Physiological Activity of Coptis japonica, White Poria cocos, and Cimicifuga heracleifolia and a Mixture of Their Extracts on Skin Based on The Oriental Medicine OMiYakSung Theory (오미약성이론을 활용한 황련, 승마, 백복령 및 복합물의 생리활성)

  • Ryu, Hwa Sun;Oh, Seong-Hwa;Lee, Jung-No;Kim, Hee-Taek;Ha, Hun-Yong;Kim, Yong-Min;Park, Sung-Min
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.45 no.1
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    • pp.77-85
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    • 2019
  • OMiYakSung theory consists of various herbs that contain at least one or more of the five flavors. This theory has been used to prevent human diseases and enhance the immune system. The main objective of the present study was to investigate efficacy differences and changes in ingredients of blended and single herb extracts based on OMiYakSung theory. We selected three herbs Coptis japonica Makino, Cimicifuga heracleifolia Komarov, and white Poria cocos and assessed their physiological effect. As results, the blended extracts showed excellent cell migration effect at 400 ug/mL concentration, compared to the single extract. In addition, the blended extracts enhanced immune function by increasing the activity of dendritic cells and showed the highest antioxidant activity by DPPH assay and HPLC-ABTS assay. In this study, we developed a new materials that can be applicable to cosmetics and pharmaceuticals field by applying oriental medicine theory.

Biological Activity of Sorghum bicolor M. cv. Bulgeunjangmoksusu Extracts (붉은장목수수 추출물의 생리활성)

  • Kim, Joo-Seok;Lee, Yea-Ji;Yang, Jinfeng;Sa, Yeo-Jin;Kim, Myeong-Ok;Park, Jong-Hyuk;Park, Dong-Sik;Yu, Chang-Yeon;Kim, Myong-Jo
    • Korean Journal of Plant Resources
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    • v.26 no.1
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    • pp.111-118
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    • 2013
  • The objective of this study was to determine the biological activities of Sorglum bicolor extracts. Organic fractions, including n-Hexane, EtOAc, and n-BuOH fractions were obtained from the methanol extract of Sorglum bicolor M.. In DPPH radical scavenging activity, $SC_{50}$ values of methanol extract and EtOAc fraction were exhibited $0.66{\pm}0.26{\mu}g/mL$ and $1.03{\pm}0.02{\mu}g/mL$, respectively. Contents of total polyphenol and flavonoids in EtOAc fraction, which were much higher than those of other fractions, were 58.12 mg/g and 4.79 mg/g respectively. Also, effects of reducing power was strongly showed in EtOAc fraction. ${\alpha}$-Glucosidase and ${\alpha}$-amlyase inhibition activities were showed the higher effect in D.W. fraction ($2.83{\mu}g/mL$, $36.64{\mu}g/mL$). In MTT assay in the AGS, HT29 and HCT116 cell lines were significantly higher in the n-BuOH fraction than in the other fractions at $50{\mu}g/mL$ concentration of extracts.

Development of serodiagnostic surface plasmon resonance imaging assay for the detection of antibodies to porcine circovirus type 2

  • Park, Chul;Kim, Bum-Seok;Kim, Yong-Hwan;Cho, Ho-Seong
    • Korean Journal of Veterinary Service
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    • v.34 no.1
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    • pp.1-4
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    • 2011
  • A surface plasmon resonance imaging (SPRI) assay was developed for measuring porcine circovirus type 2 (PCV2) antibody using a recombinant capsid protein as an antigen. The diagnostic potential of SPRI for detecting antibodies to the PCV2 capsid protein was compared with that of a conventional enzyme-linked immunosorbent assay (ELISA) using 70 pig serum samples taken from 6 pig farms. There was a strong positive correlation between the SPRI and ELISA (n = 70, r = 0.911, P<0.01). Therefore, this recombinant capsid protein can be used as an antigen for serological studies, and the SPRI, a label-free and high-throughput method, is expected to be a valuable tool in the serodiagnosis of PCV2 infection.