• Title/Summary/Keyword: in vitro detection

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THE EFFECT OF GENISTEIN IN ORAL SQUAMOUS CELL CARCINOMA WITH RESPECT TO THE ANGIOGENESIS AND BASEMENT MEMBRANE INVASION (구강편평세포암종에서 신생혈관화와 기저막침습에 미치는 제니스타인의 효과)

  • Kim, Yong-Hun;Yun, Pil-Young;Myoung, Hoon;Kim, Myung-Jin
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.28 no.6
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    • pp.434-439
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    • 2002
  • Oral squamous cell carcinoma (OSCC) is one of the most common head and neck cancers. OSCC generally has a poor prognosis due to its tendency towards a local invasion and subsequent metastasis, which is mediated by multiple proteolytic enzymes and angiogenesis. Soy products contain high levels of isoflavonoids, including the tyrosine kinase inhibitor, genistein, which has been identified as a potent inhibitor of cell proliferation and in vitro angiogenesis. The purpose of this in vitro study is to evaluate the anti-cancer effect of genistein with respect to the angiogenesis and basement membrane invasion in OSCC. The highly invasive OSCC cell line, HSC-3 cells were cultured in the presence of $10{\mu}M$ genistein for 24h. To evaluate the effects of genistein on the invasiveness and the gelatinolytic activity, in vitro invasion assay and zymography were performed. In order to evaluate the effect on the VEGF and bFGF mRNA expression, RT-PCR and northern hybridization reaction, and chemiluminescence detection were applied. The in vitro invasion assay showed that the genistein treatment reduced the cellular invasion through the artificial basement membrane and significant difference between the control group and the genistein treated group was shown in MMP-2 activity. Especially, the 62 kDa activated form of MMP-2 in the control group was 1.8 times higher than that in the genistein treated group. The results of the northern blot analyses indicated that VEGF mRNA expression in the genistein treated group was significantly down regulated. This study showed that genistein inhibits angiogenesis and reduces basement membrane invasion in OSCC. It seems to support the possibility of genistein as an anti-cancer agent.

Changes in microbial population and chemical composition of corn stover during field exposure and effects on silage fermentation and in vitro digestibility

  • Sun, Lin;Wang, Zhijun;Gentu, Ge;Jia, Yushan;Hou, Meiling;Cai, Yimin
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.6
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    • pp.815-825
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    • 2019
  • Objective: To effectively use corn stover resources as animal feed, the changes in microbial population and chemical composition of corn stover during field exposure, and their silage fermentation and in vitro digestibility were studied. Methods: Corn cultivars (Jintian, Jinnuo, and Xianyu) stovers from 4 random sections of the field were harvested at the preliminary dough stage of maturity on September 2, 2015. The corn stover exposed in the field for 0, 7, 15, 30, 60, 90, and 180 d, and their silages at 60 d of ensiling were used for the analysis of microbial population, chemical composition, fermentation quality, and in vitro digestibility. Data were analyzed with a completely randomized $3{\times}6$ [corn stover cultivar $(C){\times}exposure$ d (D)] factorial treatment design. Analysis of variance was performed using SAS ver. 9.0 software (SAS Institute Inc., Cary, NC, USA). Results: Aerobic bacteria were dominant population in fresh corn stover. After ensiling, the lactic acid bacteria (LAB) became the dominant bacteria, while other microbes decreased or dropped below the detection level. The crude protein (CP) and water-soluble carbohydrate (WSC) for fresh stover were 6.74% to 9.51% and 11.75% to 13.21% on a dry matter basis, respectively. After exposure, the CP and WSC contents decreased greatly. Fresh stover had a relatively low dry matter while high WSC content and LAB counts, producing silage of good quality, but the dry stover did not. Silage fermentation inhibited nutrient loss and improved the fermentation quality and in vitro digestibility. Conclusion: The results confirm that fresh corn stover has good ensiling characteristics and that it can produce silage of good quality.

Establishment of In Vitro Test System for the Evaluation of the Estrogenic Activities of Natural Products

  • Kim, Ok-Soo;Choi, Jung-Hye;Soung, Young-Hwa;Lee, Seon-Hee;Lee, Jae-Hwa;Ha, Jong-Myung;Ha, Bae-Jin;Heo, Moon-Soo;Lee, Sang-Hyeon
    • Archives of Pharmacal Research
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    • v.27 no.9
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    • pp.906-911
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    • 2004
  • In order to evaluate estrogenic compounds in natural products, an in vitro detection system was established. For this system, the human breast cancer cell line MCF7 was stably trans-fected using an estrogen responsive chloramphenicol acetyltransferase (CAT) reporter plas-mid yielding MCF7/pDsCAT-ERE119-Ad2MLP cells. To test the estrogenic responsiveness of this in vitro assay system, MCF7/pDsCAT-ERE119-Ad2MLP cells were treated with various concentrations of 17f3-estradiol. Treatments of 10$^{-8}$ to 10$^{-12}$ M 17$\beta$-estradiol revealed significant concentration dependent estrogenic activities compared with ethanol. We used in vitro assay system to detect estrogenic effects in Puerariae radix and Ginseng radix Rubra extracts. Treat-ment of 500 and 50 $\mu\textrm{g}$/ml of Puerariae radix extracts increased the transcriptional activity approximately 4- and 1.5-fold, respectively, compared with the ethanol treatment. Treatment of 500, 50, and 5 $\mu\textrm{g}$/ml of Ginseng radix Rubra extracts increased the transcriptional activity approximately 3.2-,2.7, and 1.4-fold, respectively, compared with the ethanol treatment. These observations suggest that Puerariae radix and Ginseng radix Rubra extracts have effective estrogenic actions and that they could be developed as estrogenic supplements.

Artifical Neural Network for In-Vitro Thrombosis Detection of Mechanical Valve

  • Lee, Hyuk-Soo;Lee, Sang-Hoon
    • Proceedings of the Korean Institute of Intelligent Systems Conference
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    • 1998.06a
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    • pp.762-766
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    • 1998
  • Mechanical valve is one of the most widely used implantable artificial organs, Since its failure (mechanical failures and thrombosis to name two representative example) means the death of patient, its reliability is very important and early noninvasive detection is essential requirement . This paper will explain the method to detect the thrombosis formation by spectral analysis and neural network. In order quantitatively to distinguish peak of a normal valve from that of a thrombotic valve, a 3 layer backpropagation neural network, which contains 7,000 input nodes, 20 hidden layer and 1output , was employed. The trained neural network can distinguish normal and thrombotic valve with a probability that is higher than 90% . In conclusion, the acoustical spectrum analysis coupled with a neural network algorithm lent itself to the noninvasive monitoring of implanted mechanical valves. This method will be applied to be applied to the performance evaluation of other implantable rtificial organs.

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Allergy, hypersensitivity and cosmetics (화장품에 있어서 엘러지, 민감성에 대하여)

  • Hardy, Joan
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.3 no.1
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    • pp.40-84
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    • 1973
  • Synopsis-The difficulties of immunological nonienclature are discussed, the term ALLERGY defined and the various types of HYPERSENSITIVITY reactions are listed and characterized. Evidence for the association of Type I and Type 11 hypersensitivity reactions with COSMETICS is discussed. A table of cosmetic ingredients which have been implicated as SENSITIZERS are given. PREDICTIVE PATCH TESTS for contact sensitizers on GUINEA-PIGS and man are evaluated. The difficulties of testing for ALLERGENS likely to produce Type I hypersensitivity are discussed. IN VITRO tests for sensitizers are mentioned. The failure of all standard tests in the detection of weak sensitizers is emphasized.asized.

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Validation and Application of a Real-time PCR Protocol for the Specific Detection and Quantification of Clavibacter michiganensis subsp. sepedonicus in Potato

  • Cho, Min Seok;Park, Duck Hwan;Namgung, Min;Ahn, Tae-Young;Park, Dong Suk
    • The Plant Pathology Journal
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    • v.31 no.2
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    • pp.123-131
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    • 2015
  • Clavibacter michiganensis subsp. sepedonicus (Cms) multiplies very rapidly, passing through the vascular strands and into the stems and petioles of a diseased potato. Therefore, the rapid and specific detection of this pathogen is highly important for the effective control of the pathogen. Although several PCR assays have been developed for detection, they cannot afford specific detection of Cms. Therefore, in this study, a computational genome analysis was performed to compare the sequenced genomes of the C. michiganensis subspecies and to identify an appropriate gene for the development of a subspecies-specific PCR primer set (Cms89F/R). The specificity of the primer set based on the putative phage-related protein was evaluated using genomic DNA from seven isolates of Cms and 27 other reference strains. The Cms89F/R primer set was more specific and sensitive than the existing assays in detecting Cms in in vitro using Cms cells and its genomic DNA. This assay was also able to detect at least $1.47{\times}10^2copies/{\mu}l$ of cloned-amplified target DNA, 5 fg of DNA using genomic DNA or $10^{-6}$ dilution point of 0.12 at $OD_{600}$ units of cells per reaction using a calibrated cell suspension.

I. Primary cultured hepatocytes as a key in vitro model to improve preclinical drug development (간세포 배양-약물대사를 위한 모델 연구)

  • 이경태
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1994.11a
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    • pp.135-140
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    • 1994
  • Over past decades, numerous in vitro model has been developed to investigate drug metabolism. In the order of complexity we found the isolated perfused liver, hepatocytes in co-culture with epithelial cells, hepatocytes in suspension and in primary culture and subcellular hepatic microsomal fractions. Because they can be easily prepared from both animals (pharmacological and toxicological species) and humans (whole livers as well as biopsies obtained during surgery) hepatocytes in primary culture provide the most powerful model to better elucidate drug behavior at an early stage of preclinical development such as : 1. the characterization of main biotransformation reactions. 2. the identification of phase I and phase II isozymes involved in such reactions 3. the evaluation of interspecies differences allowing the selection of a second toxicological animal species more closely related to man on the basis of metabolic profiles 4. the detection of the inducing and/or inhibitory effects of a drug on metabolic enzymes, the prediction of drug interactions 5. the estimation of inter-individual variability in biotransformation reactions. The use of hepatocytes, and in particular those obstained from humans, at an early stage of drug development allows the obtention of more predictive preclinical data and a better knowledge of drug behavior in humans before the first administration of the drug in healthy volunteers.

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A New Multiplex-PCR for Urinary Tract Pathogen Detection Using Primer Design Based on an Evolutionary Computation Method

  • Garcia, Liliana Torcoroma;Cristancho, Laura Maritza;Vera, Erika Patricia;Begambre, Oscar
    • Journal of Microbiology and Biotechnology
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    • v.25 no.10
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    • pp.1714-1727
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    • 2015
  • This work describes a new strategy for optimal design of Multiplex-PCR primer sequences. The process is based on the Particle Swarm Optimization-Simplex algorithm (Mult-PSOS). Diverging from previous solutions centered on heuristic tools, the Mult-PSOS is selfconfigured because it does not require the definition of the algorithm's initial search parameters. The successful performance of this method was validated in vitro using Multiplex-PCR assays. For this validation, seven gene sequences of the most prevalent bacteria implicated in urinary tract infections were taken as DNA targets. The in vitro tests confirmed the good performance of the Mult-PSOS, with respect to infectious disease diagnosis, in the rapid and efficient selection of the optimal oligonucleotide sequences for Multiplex-PCRs. The predicted sequences allowed the adequate amplification of all amplicons in a single step (with the correct amount of DNA template and primers), reducing significantly the need for trial and error experiments. In addition, owing to its independence from the initial selection of the heuristic constants, the Mult-PSOS can be employed by non-expert users in computational techniques or in primer design problems.

A Study on the Optical Influence by Photosensitizer in Vitro (In Vitro에서 광증감제에 의한 광학적 영향에 관한 연구)

  • Kim, Ki-Jun;Sung, Ki-Chun
    • Journal of the Korean Applied Science and Technology
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    • v.22 no.2
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    • pp.182-190
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    • 2005
  • The propagation of light radiation within tissues is an important problem that confronts the dosimetry of therapeutic laser delivery and the development of diagnostic spectroscopy. In the clinical application of photodynamic therapy(PDT) and in photobiology, the photon deposition within a tissue determines the spatial distribution of photochemical reactions. Scattered light is measured as a function of the distance (r) between the axis of the incident beam and the detection spot. Consequently, knowledge of the photosensitizer(Chlorophyll-a) function that characterizes a phantom is important. To obtain the results of scattering coefficients(${\mu}s$) of a turbid material from diffusion described by experimental approach. It was measured the energy fluency of photon radiation at the position of penetration depth. From fluorescence experimental method obtained the analytical expression for the scattered light as the values of $(I\;/I_o)_{wavelength}$ vs the distance between the center of the incident beam and optical fiber in terms of the condition of "in situ spectroscopy(optically thick)" and real time by fluorometric measurements.