• 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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    • 제40권1호
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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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    • 제3권1호
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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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C6 Glioma 세포에서 Protein Kinase C Alpha 발현 저해를 통한 송절 약침액의 이주 억제 효과 (Pinus densiflora Gnarl Inhibits Migration through Suppression of Protein Kinase C in C6 Glioma Cells)

  • 민일국;이강파;장해룡;문진영
    • Korean Journal of Acupuncture
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    • 제32권2호
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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
    • 대한치과보철학회지
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    • 제45권3호
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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.

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

  • 왕혜영;최재용;이미진;박진호;조매림;한재철;최경성;채준석
    • 대한수의학회지
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    • 제44권4호
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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.

오미약성이론을 활용한 황련, 승마, 백복령 및 복합물의 생리활성 (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)

  • 유화선;오성화;이정노;김희택;하헌용;김용민;박성민
    • 대한화장품학회지
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    • 제45권1호
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    • pp.77-85
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    • 2019
  • 오미약성 이론은 다섯가지 맛 중에서 적어도 하나 이상을 맛을 포함하는 다양한 허브로 구성되어 있으며, 이러한 이론은 인간의 질병을 예방하고 면역 체계를 강화하는데 사용되어왔다. 본 연구의 목적은 오미약성 이론에 근거한 단일 추출물 및 혼합 추출물의 효능 차이와 성분 변화를 확인하는 것이다. 황련, 승마 및 백복령 세가지 약초를 선택하였고 단일 추출물 및 혼합 추출물의 약리학적 효능을 평가하였다. 결과적으로 혼합추출물은 단일추출물과 비교하여 400 ug/mL 농도에서 우수한 세포 이동 효과를 보였다. 또한 혼합추출물은 수지상세포의 활성을 증가시켜 면역을 강화시켰으며 DPPH assay 및 HPLC-ABTS assay를 통해 가장 높은 항산화 활성을 보였다. 이번 연구에서 우리는 한의학적 이론을 접목하여 화장품 및 의약품 분야에 응용 가능한 새로운 소재를 개발하였다.

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

  • 김주석;이예지;양금봉;사여진;김명옥;박종혁;박동식;유창연;김명조
    • 한국자원식물학회지
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    • 제26권1호
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    • pp.111-118
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    • 2013
  • 본 연구에서는 붉은장목수수(Sorglum Bicolor M.) MeOH 추출물과 분획물들의 다양한 생리활성을 비교하기 위하여 항산화(DPPH assay, 환원력, 총 페놀 및 플라보노이드 함량), 항당뇨(${\alpha}$-Glucosidase 저해활성 및 ${\alpha}$-Amlyase 저해활성), 항암(MTT assay) 활성을 관찰 하였다. 항산화활성의 경우 EtOAc fraction이 높은 총 페놀과 플라보노이드 함량을 나타내었으며, DPPH assay와 reducing power 결과 역시 EtOAc fraction과 MeOH fraction이 positive control로 사용한 ${\alpha}$-tocopherol보다 우수한 활성을 나타냈다. ${\alpha}$-Glucosidase, ${\alpha}$-amlyase 저해능 평가 결과 D.W. fraction이 가장 높은 활성을 보였으며 n-BuOH fraction과 MeOH extract도 활성을 나타내었다. 암세포인 AGS, HT29, HCT116세포주에 대한 세포독성 연구인 MTT assay에서는 EtOAc, n-BuOH, D.W. fraction을 처리한 처리구에서 독성을 보였으며 이중 n-BuOH fraction이 모든 세포주에서 농도 의존적으로 세포독성을 가지는 것을 확인할 수 있었다. 연구결과 붉은장목수수의 추출물 EtOAc fraction의 항산화활성, D.W. fraction의 항당뇨활성, n-BuOH fraction의 암세포에 대한 독성과 관련된 화합물의 분리 및 구조규명에 관한 연구가 필요 할 것으로 사료되어지며, normal cell에 대한 MTT assay를 진행하여 항암활성에 관한 연구를 진행하여 붉은장목수수 추출물을 이용한 다양한 건강기능식품과 의약품 개발을 통해 건강 증진과 질병으로부터 보호 받을 수 있을 것으로 기대한다.

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
    • 한국동물위생학회지
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    • 제34권1호
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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.