• Title/Summary/Keyword: Biological assay

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Real-Time Voltammetric Assay of Lead Ion in Biological Cell Systems

  • Ly, Suw-Young
    • Toxicological Research
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    • v.25 no.4
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    • pp.231-235
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    • 2009
  • Trace lead detection for cyclic voltammetry (CV) and square-wave (SW) stripping voltammetry was performed using mercury immobilized onto a carbon nanotube electrode (HNPE). Using the characteristics of mercury and the catalytic carbon nanotube structure, a modified technique, the $0.45{\mu}g/l$ detection limit of lead ion was attained. The developed method can be applied to pond water, fish tissue, plant tissue, and in vivo direct assay.

Novel Cell-based Protease Assay System for Molecular Cell Biology and Drug Discovery

  • Hwang, Hyun-Jin;Kim, Jeong-Hee;Park, Joon-Woo;Kim, Sung-Hee;Lee, Min-Jeon;Jeong, Han-Seung;Hwang, In-Hwan
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.169.1-169.1
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    • 2003
  • Recently development of cell-based assay systems which are useful in molecular cell biology and drug discovery attracts significant attention. Here, we introduce a new technologies for monitoring enzyme activity and its inhibition inside living cells. Among various enzymes, proteases are important targets for studying various biological and disease-related processes such as viral infections, apoptosis and Alzheimer's disease. In this study, a sensitive cell-based protease detection system that enables direct fluorescence detection of a target protease and its inhibition inside living cells is introduced. (omitted)

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Evaluation of genotoxic potentials in diesel exhaust particulate matter with the Ames test, the comet assay and the micronucleus assay

  • Kim, Soung-Ho;Lee, Do-Han;Han, Kyu-Tae;Oh, Seung-Min;Chung, Kyu-Hyuck
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.165.1-165.1
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    • 2003
  • This research was designed to examine the presence of mutagenic/carcinogenic compounds in airborne pollutants in diesel particulate matter using an integrated biological approach. Respirable air borne particulate matter (PM2.5: <2.5mm) was collected from diesel engine exhaust using a high-volume sampler equipped with a cascade impactor. (omitted)

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Stability of the current biological drugs(typhoid vaccine)

  • Lee, Joo-Hyeun;Ham, Myung-Hwa;Lim, So-Hee;Lee, Gil-Woong
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.328.2-328.2
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    • 2002
  • This paper presents stability of tyhoid vaccine -attenuuated vaccine(oral) and killed vaccine{vi polysaccharide)- 5 classes with various temperature(3 points: iced temperature, refrigeration temp.. indoor temp.). Analytical techniques -vi polysaccharide content. pH. sterility. assay - have been used for the quantity of pharmacologically active chemical entries. From this study. we have found the attenuated vaccine is show iced temp. 258% and indoor temp. 0.02% compare with refrigeration temp. in assay examination and the killed vaccine is not found different. (omitted)

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In Vitro Bioassay for Transforming Growth Factor-$\beta$ Using XTT Method

  • Kim, Mi-Sung;Ahn, Seong-Min;Moon, Aree
    • Archives of Pharmacal Research
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    • v.25 no.6
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    • pp.903-909
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    • 2002
  • Research in the cytokine field has grown exponentially in recent years, and the validity of such studies relies heavily on the appropriate measurement of levels of cytokines in various biological samples. Transforming growth factor (TGF)-$\beta$, a hormonally active polypeptide found in normal and transformed tissue, is a potent regulator of cell growth and differentiation. The most widely used bioassay for TGF-$\beta$ is the inhibition of the proliferation of mink lung epithelial cells. Though detection of [$^3$H]thymidine incorporation is more sensitive than the MTT assay, it presents some disadvantages due to the safety and disposal problems associated with radioisotopes. In this study, we attempted to ascertain the experimental conditions which could be used for measuring the in vitro biological activity of TGF-$\beta$ in a safer and more sensitive way compared with the currently available methods. We compared the commonly used method, the MTT assay, to the XTT assay using different parameters including cell number, incubation time and the wave length used for detecting the product. We examined the anti-proliferative activities of TGF-$\beta$ in three different cell lines: Mv-1-Lu mink lung epithelial cells, MCF10A human breast epithelial cells and H-ras-transformed MCF10A cells. Herein, we present an experimental protocol which provides the most sensitive method of quantifying the biological activity of TGF-$\beta$, with a detection limit of as low as 10 pg/ml: Mv-1-Lu or H-ras MCF10A cells ($1{\times}10^5/well$) were incubated with TGF-$\beta$ at $37^{\circ}C$ in a humidified $CO_2$ incubator for 24 hr followed by XTT treatment and determination of absorbance at 450 or 490 nm. Our results may contribute to the establishment of an in vitro bioassay system, which could be used for the satisfactory quantitation of TGF-$\beta$.

The Molecular Biological Study on Anti-Cancer Effects of Sagunjatang plus Cremastrae Appenediculatae Tuber on Human Stomach Cancer Cells (사군자탕가산자고가 위암세포에 미치는 항암효과에 대한 분자생물학적 연구)

  • Ryu, Bong-Ha;Ryu, Ki-Won;Yoon, Sang-Hyub;Kim, Jin-Seong;Kim, Jin-Seok
    • The Journal of Internal Korean Medicine
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    • v.23 no.2
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    • pp.202-211
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    • 2002
  • 1. Background The previous studies on anticancer medicine derived from korean traditional medicine have focused on the life elongation of cancer cell bearing animals. However, it is thought that more molecular biological studies are needed to reveal their mechanism. 2. Objective The aim of this study was to investigate the molecular biological function of Sagunjatang plus Cremastrae Appenediculatae Tuber on cytostaticity, apoptosis and apoptosis related genes revelation against human stomach cancer cells(AGS). 3. Methods After administrating Sagunjatang and Sagunjatang plus Cremastrae Appenediculatae Tuber to human stomach cancer cell. MTT assay was performed to compare and examine the efficacy of each medicine on the cytostaticity of stomach cancer cells in proportion to time and doses, and apoptosis assay was performed to examine their effect on apoptosis by using DAPI dye and counting the number of cells which developed in an apoptotic body. In addition, the quantitative RT-PCR was used to examine their effect on the revelation of Bcl-2, Bax and P53, which are genes related to apoptosis. 4. Result and Conclusion Sagunjatang plus Cremastrae Appenediculatae Tuber demonstrated increased cytostaticity. decreased apoptosis and unremarkable revelation of apoptosis related genes. But in the cytostaticity and apoptosis, Sagunjatang plus Cremastrae Appenediculatae Tuber showed a tendency to control stomach cancer cells. Therefore, we can expect the clinical application to the related diseases. Besides, it needs another experiment on various cancer cells, such as, lung cancer cell and hysterocarcinoma cell.

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Norovirus Targeted Bioreceptor Screening Method based on Lateral Flow Immunoassay (LFIA) (노로바이러스 검출을 위한 측면유동면역분석법 기반의 바이오리셉터 선별기법 개발)

  • Huisoo, Jang;Hyeonji, Cho;Tae-Joon, Jeon;Sun Min, Kim
    • Journal of the Korean Society of Visualization
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    • v.20 no.3
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    • pp.136-145
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    • 2022
  • Later flow immunoassay (LFIA) is a protein analytical method based on immunoreaction. On the LFIA based protein analytical method, bioreceptor molecule plays a key role, and so a system that evaluates and manages the binding affinity of bioreceptor is needed to secure detection reliability. In this study, Lateral Flow Immunoassay based rapid Bioreceptor Screening Method (rBSM) is presented that provide a simple and quick evaluating method for the binding affinity to the target protein of the antibody as model bioreceptor. To verify this evaluation method, Virus-like particles (VLP) and anti-VLP antibodies are selected as a model norovirus, which is target protein, and the candidate bioreceptors respectively. Among the 5 different candidate antibodies, appropriate antibody could be sorted out within 30 minutes through rBSM. In addition, selected antibodies were applied to two representative LFIA based techniques, sandwich assay and competitive assay. Among these methods, sandwich assay showed more effective VLP detection method. Through applying selected antibodies and techniques to the commercialized mass production lines, an VLP detecting LFIA kit was developed with a detection limit of 1012 copies/g of VLPs in real samples. Since this proposed method in this study could be easily transformable into other combinations with bioreceptors, it is expected that this technique would be applied to LFIA kit development system and bioreceptor quality management.

Whitening Effect and Skin Regeneration Effect of Red Sea Cucumber Extract (홍해삼 추출물의 멜라닌 형성 억제를 통한 미백효과 및 피부 재생효과에 관한 연구)

  • Jeon, Mi Ji;Kim, Eun Ji;Kim, Geun Tae;Kim, Ga Yeon;Lee, Seung Jae;Jung, In Cheol;Kim, Sang-Yong;Kim, Young Min
    • Journal of Life Science
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    • v.28 no.6
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    • pp.681-687
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    • 2018
  • Recently, several researchers have been developing cosmetics from natural ingredients for skin whitening and anti-aging products. The red sea cucumber (RSC), Apostichopus japonicas, is a species of sea cucumber in the family stichopodiae, which is widely distributed in China, Japan, and Korea. To use Red Sea Cucumber as a cosmetic ingredient, its inhibitory effects on melanogenesis and the anti-aging effects of RSC extracts were investigated. First, a tyrosinase activity assay was performed, which showed that RSC inhibited tyrosinase activity at a concentration of $200{\mu}g/ml$. An MTT assay was carried out to evaluate cell toxicity, and the results showed that RSC extract has no cytotoxicity in HaCaT cells. Furthermore, the mRNA expression levels of tyrosinase, tyrosinase related protein 1 (TRP-1), tyrosinase related protein 2 (TRP-2), microphthalmia-associated transcription factor (MITF), and matrix metalloproteinase (MMPs) genes treated with RSC extract in B16F10 and HaCaT cells decreased. Moreover, a wound-healing assay was performed to identify the cell regeneration effect of RSC extracts. Also, a skin turnover effect was confirmed by creating a three-dimensional cell culture with HaCaT and human fibroblasts. Altogether, the results suggested that Red Sea Cucumber may possess a high ability to induce whitening and anti-wrinkle effects as a cosmeceutical ingredient.

Characterization of Humanized Antibody Produced by Apoptosis-Resistant CHO Cells under Sodium Butyrate-Induced Condition

  • Kim, No-Soo;Chang, Kern-Hee;Chung, Bo-Sup;Kim, Sung-Hyun;Kim, Jung-Hoe;Lee, Gyun-min
    • Journal of Microbiology and Biotechnology
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    • v.13 no.6
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    • pp.926-936
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    • 2003
  • Overexpression of human Bcl-2 protein in recombinant Chinese hamster ovary (rCHO) cells producing humanized antibody (SH2-0.32) considerably suppressed sodium butyrate (NaBu)-induced apoptosis during batch culture by using commercially available serum-free medium, which extended the culture longevity. Due to the extended culture longevity provided by the anti-apoptotic effect of Bcl-2 overexpression, the final antibody concentration of 14C6-bcl-2 culture (Bcl-2 high producer, $23\;\mu\textrm{g}\;ml^{-1}$) was 2 times higher than that of the $SH2-0.32-{\Delta}bcl-2$ culture (cells transfected with bcl-2-deficient plasmid, $10.5\;\mu\textrm{g}\;ml^{-1}$) in the presence of NaBu. To determine the effect of NaBu/Bcl-2 overexpression on the molecular integrity of protein products, antibodies purified from 14C6-bcl-2 and $SH2-0.32-{\Delta}bcl-2$ cultures in the presence of NaBu were characterized by using various molecular assay systems. For comparison, antibody purified from the parental rCHO cell culture (SH2-0.32) in the absence of NaBu was also characterized. No significant changes in molecular weight of antibodies could be observed by SDS-PAGE. From GlycoSep-N column analysis, it was found that the core oligosaccharide structure ($GlcNAc_2Man_3GlcNAc_2$) was not affected by NaBu/Bcl-2 overexpression, while the microheterogeneity of N-linked oligosaccharide structure was slightly affected. Compared with the antibody produced in the absence of NaBu, the proportion of neutral oligosaccharides was increased from 10% (14C6-bcl-2) to 16% ($SH2-0.32-{\Delta}bcl-2$) in the presence of NaBu, which was accompanied by the reduced proportion of acidic oligosaccharides, especially of monosialylated and disialylated forms. The changes in microheterogeneous oligoformal structures of antibody in turn affected the mobility of antibody isoforms in isoelectric focusing (IEF), resulting in the occurrence of some more basic antibody isoforms produced in the presence of NaBu. However, the antigen-antibody binding properties were not changed by alteration of glycosylation pattern. The competitive enzyme-linked immunosorbent assay (ELISA) showed that the antibody produced by NaBu/Bcl-2 overexpression maintained its antigen-antibody binding properties with binding affinity of about $2.5{\times}10^9{\;}M^{-1}$. Taken together, no significant effects of NaBu/Bcl-2 overexpression on the molecular integrity of antibodies, produced by using serum-free medium, could be observed by the molecular assay systems.

BIOLOGICAL HUMAN MONITORING OF CARCINOGEN EXPOSURE: A NEW STRATEGY IN CANCER PREVENTION

  • Lee, Byung-Mu
    • Toxicological Research
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    • v.6 no.1
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    • pp.63-73
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    • 1990
  • Human exposure to environmental carcinogens can be detected by a number of methods including immunoassay, $^{32}P-postlabeling$ assay, and fluorescence technique. These assays have been applied to measure biological markers of carcinogen-adducts formed with macromolecules such as DNA, RNA and protein. In an attempt to investigate causal relationships between carcinogen exposure and tumor formation, specific carcinogen-adducts have been quantitated from human tissues and body fluids of cancer patients, occupational workers heavily exposed to certain carcinogens, smokers and controls. Carcinogens studied for biological human monitoring include benzo(a)pyrene, aflatoxin B1, UV light, ethylene oxide, 8-methoxypsoralen, 4-aminobiphenyl, vinyl choride, N-nitrosamine, cisplatin and other chemotherapeutic agents. Relevance of human monitoring for cancer research, progress in this field, methods to detect carcinogen-adducts are reviewed here. It is hoped that these approaches will be used for the risk assessment of carcinogen exposure, cancer etiology study and cancer prevention in humans.

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