• Title/Summary/Keyword: Colorimetric assay

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Simple, Rapid and Sensitive Portable Molecular Diagnosis of SFTS Virus Using Reverse Transcriptional Loop-Mediated Isothermal Amplification (RT-LAMP)

  • Baek, Yun Hee;Cheon, Hyo-Soon;Park, Su-Jin;Lloren, Khristine Kaith S.;Ahn, Su Jeong;Jeong, Ju Hwan;Choi, Won-Suk;Yu, Min-Ah;Kwon, Hyeok-il;Kwon, Jin-Jung;Kim, Eun-Ha;Kim, Young-il;Antigua, Khristine Joy C.;Kim, Seok-Yong;Jeong, Hye Won;Choi, Young Ki;Song, Min-Suk
    • Journal of Microbiology and Biotechnology
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    • v.28 no.11
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    • pp.1928-1936
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    • 2018
  • Recently, human infections caused by severe fever with thrombocytopenia syndrome virus (SFTSV), which can lead to fatality, have dramatically increased in East Asia. With the unavailability of vaccines or antiviral drugs to prevent and/or treat SFTSV infection, early rapid diagnosis is critical for prevention and control of the disease. Here, we report the development of a simple, rapid and sensitive portable detection method for SFTSV infection applying reverse transcription-loop mediated isothermal amplification (RT-LAMP) combined with one-pot colorimetric visualization and electro-free reaction platform. This method utilizes a pocket warmer to facilitate diagnosis in a resource-limited setting. Specific primers were designed to target the highly-conserved region of L gene of SFTSV. The detection limit of the RT-LAMP assay was approximately $10^0$ viral genome copies from three different SFTSV strains. This assay exhibited comparable sensitivity to qRT-PCR and 10-fold more sensitivity than conventional RT-PCR, with a rapid detection time of 30 to 60 minutes. The RT-LAMP assay using SFTSV clinical specimens has demonstrated a similar detection rate to qRT-PCR and a higher detection rate compared to conventional RT-PCR. Moreover, there was no observed cross-reactive amplification of other human infectious viruses including Japanese Encephalitis Virus (JEV), Dengue, Enterovirus, Zika, Influenza and Middle East Respiratory Syndrome Coronavirus (MERS-CoV). This highly sensitive, electro- and equipment-free rapid colorimetric visualization method is feasible for resource-limited SFTSV field diagnosis.

Cytotoxic activity of processing the traditional drug on monkey kidney cell (Vero) and human liver cell (WRL68) (수치(修治) 한약재가 사람의 간세포 WRL68와 원숭이의 신장세포 Vero에 미치는 영향)

  • Ju Young-Sung;Kim Ho-Kyoung;Ko Byoung-Seob
    • Journal of Society of Preventive Korean Medicine
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    • v.4 no.2
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    • pp.258-272
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    • 2000
  • The cytotoxic activities of Pinellia ternate, Aconitum carmichaeli, Arisaema amurense, Aconitum kusnezoffii and Scutellaria baicalensis on monkey kidney cell (Vero) and human liver cell (WRL68) were evaluated by Sulforhodamine B Protein (SRB) and Tetrazolium-based (MTT) colorimetric assay methods The results were as fellows : 1 The Pinellia ternate and Arisaema amurense did not show the cytotoxic activities at any concentration without processing or natural drugs. 2 The extracts of Aconitum carmichaeli, Aconitum kusnezoffii and Scutellaria baicalensis showed cytotoxic activities. However, the cytotoxic activities of processing drugs were less effective than the natural drugs. 3. The cytotoxic activities on monkey kidney cell (Vero) and human liver cell (WRL68) of Aconitum carmichaeli was determined by MTT assay. The Kyungpo(京?) of Aconitum carmichaeli showed less concentrate than that of the Dangpo(唐?). The $IC_{50}$ value on monkey kidney cell (Vero) and human liver cell (WRL68) of Kyungpo was $937{\pm}29\;and\;731{\pm}31{\mu}g/ml$, respectively 4. The cytotoxicity on monkey kidney cell (Vero) and human liver cell (WRL68) of Scutellaria baicalensis showed strong activities without processing or natural drugs.

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Effect of Cadmium on $C_6$ Glioma Cells in Culture

  • Son Young-Woo;Lee Kang-Chang
    • Biomedical Science Letters
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    • v.12 no.3
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    • pp.275-279
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    • 2006
  • It is demonstrated that cadmium has cytotoxic effect on glial cells, oxygen radicals are involved in cadmium-induced cytotoxicity. However, the toxic mechanism of cadmium is left unknown so far. The purpose of this study was to examine the cytotoxicity of $CdCl_2$ on $C_6$ glioma cells. The cytotoxicy was measured by cell viability via XTT assay in $C_6$ glioma cells. Colorimetric assay such as XTT assay is regarded as a very sensitive screening method for the determination of the cell viability on a lots of chemicals. In this study, $CdCl_2$ decreased cell viability according to the dose- and time dependent manners after $C_6$ glioma cells were treated with various concentrations of $CdCl_2$ for 48 hours. $IC_{90}\;and\;IC_{50}$ values for XTT assay was determined at $5{\mu}M\;and\;55{\mu}M$ of $CdCl_2$, respectively. These results suggest that $CdCl_2$ has highly cytotoxic effect on $C_6$glioma cells by the decrease of cell viability.

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Development of Anticancer Agents from Korean Medicinal Plants (Part 4). Antitumor Activity of the Butanol Soluble Fraction of Perilla frutescens (한국산 생약으로부터 항암물질의 개발(제4보) 소엽 부탄올 가용분획의 항암활성)

  • 최규은;곽정숙;김영옥;백승화;한두석
    • Toxicological Research
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    • v.13 no.4
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    • pp.311-316
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    • 1997
  • This study was carried out to develop antitumor effect of the n-butanol soluble fraction of Perilla frutescens on (KB cells) human oral epitheloid carcinoma cells. The cytotoxictty of methanollc extract of Perilla frutescens on KB cells was evaluated by 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide(MTT) assay. The antitumor activity of various fractions obtained from n-butanol soluble fraction of Perilla frutescens was evaluated in human oral epithelold carcinoma cells. The antitumor acavity of the n-butanol soluble fraction on human oral epitheloid carcinoma cells was evaluated by MTT assay of colorimetric method. The light microscopic study was carried out to observe morphological changes of cultured human oral epitheloid carcinoma cells. These results were obtained as follows; 1. The fractions 1,2 and 3 of the n-butanol soluble fraction of Perilla frutescens were shown significant antitumor activities. 2. The number of human oral epitheloid carcinoma cells were decreased and tend to form cell cluster by treatment with fractions 1,2,3 and 4 of the n-butanol soluble fraction of Perilla frutescens. 3. The fraction 1 of the n-butanol soluble fraction of Perllla frutescens showed the highest antitumor activity on Perilla frutescens. It has been selected as a lead fraction for further examinations.

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Paper-Based Neuraminidase Assay Sensor for Detection of Influenza Viruses (인플루엔자 바이러스 검출을 위한 종이 기반 neuraminidase 효소 활성 평가 센서 개발)

  • Hwang, Cheol-hwan;Jeong, Seong-Geun;Park, Han-Kyu;Lee, Chang-Soo;Kim, Yun-Gon
    • Korean Chemical Engineering Research
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    • v.54 no.3
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    • pp.380-386
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    • 2016
  • In this study, we described a paper-based neuraminidase assay sensor (PNAS) which can be applied to detect the infection by influenza viruses. The PNAS was designed and manufactured to quantitatively identify the levels of neuraminidase in the sample, which is based on colorimetric analysis using the X-Neu5Ac substrate. The limit of detection of the PNAS was determined as 0.004 U/mL of neuraminidase. According to the amount of neuraminidase in human serum, the PNAS could monitor the enzyme activity with a good linearity ($R^2$ > 0.99). In addition, the initial performance of the PNAS has been maintained up to 70 days in the $4^{\circ}C$. Finally, we demonstrated whether the Michaelis-Menten kinetics is applied to the PNAS, which can show the reliability of the enzyme reactions. The kinetic studies indicated that the PNAS provides the good condition for enzyme reactions ($K_m=8.327{\times}10^{-3}M$), but they were performed on paper chip nonetheless. The paper-based neuraminidase assay sensor may be useful in a wide range of rapid and safe detection of influenza virus.

A novel cis/trans-diaminocyclohexane platinum coordination complexes possessing in vitro and in vivo antitumor activity

  • Jung, Jee-Chang;Chang, Sung-Goo;Lee, Kyung-Tae;Park, Young-Soo;Lee, Joo-Han;Lee, Kyou-Heung;Kim, Sang-Lin
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1997.04a
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    • pp.101-101
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    • 1997
  • As part of a drug discovery program to develope more effective platinum-based anticancer drugs, a series of platinum complexes trans -diaminocyclohexane platinum bi sdiphenylphosphino -ethane (KHPC-002) cis-diaminocyclohexane platinum bisdiphenylphosphino-ethane (KHPC-006) has been evaluated in vitro against 4 human carcinoma cell lines with those of cisplatin using a tetrazolium-based colorimetric assay (MTT assay). The cell lines were two human bladder carcinoma cell lines, HT-1197 and HT-1376, human colon carcinoma cell line, HCT-116, and prostate cancer cell line DU-145.

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A novel cis/trans-diaminocyclohexane platinum coordination complexes possessing in vitro and in vivo antitumor activity

  • Jung, Jee-Chang;Chang, Sung-Goo;Lee, Kyung-Tae;Park, Young-Soo;Lee, Joo-Han;Lee, Kyou-Heung;Kim, Sang-Lin
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1997.04a
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    • pp.111-111
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    • 1997
  • As part of a drug discovery program to develope more effective platinum-based anticancer drugs, a series of platinum complexes trans-diaminocyclohexane platinum bi sdiphenylphosphino - ethane ( KHPC- 002) cis-diaminocyclohexane platinum bi sdiphenylphosphino - ethane ( KHPC- 006) has been evaluated in vitro against 4 human carcinoma cell lines with those of cisplatin using a tetrazolium-based colorimetric assay (MTT assay). The cell lines were two human bladder carcinoma cell lines, HT-1197 and HT-1376, human colon carcinoma cell line, HCT-116, and prostate cancer cell line DU-145. in vitro cytotoxic potential of each platinum complex was expressed as the cytotoxicity index (Cl, %).

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Effect of Allopurinol on Methylmercuric Chloride-Induced Cytotoxicity in $C_6$ Cultured Glioma Cells

  • Oh, Yong-Leol;Son, Byoung-Kwan
    • Biomedical Science Letters
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    • v.12 no.4
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    • pp.451-455
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    • 2006
  • It is demonstrated that inorganic mercury has cytotoxic effect on glial cells. Recently, oxygen radicals is involved in methylmercuric chloride (MMC)-induced cytotoxicity. But, the toxic mechanism of MMC is left unknown. The purpose of this study was to examine the cytotoxicity of MMC on $C_6$ glioma cells. The cytotoxicy was measured by cell viability using XTT assay in $C_6$ glioma cells. Colorimetric assay is regarded as a very sensitive screening method for the determination of the cell viability on various agents. In this study, MMC decreased cell viability according to the dose- and time dependent manners after $C_6$ glioma cells were grown with various concentrations of MMC for 48 hours. In the protective effect of allopurinol on MMC-induced cytotoxicity, allopurinol was effective in the prevention of MMC-induced cytotoxicity in these cultures. These results suggest that MMC has highly cytotoxic effect on $C_6$ glioma cells by the decrease of cell viavility, and free radical scavenger such as allopurinol was effective on organic mercury-induced cytotoxicity in these cultures.

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EFFECT OF CULTURE SUPERNATANT OF BACTERIA ISOLATED FROM INFECTED ROOT CANALS ON CELL LINES (감염 근관에서 분리한 세균 배양액이 배양된 세포에 미치는 영향)

  • Im, Mi-Kyung
    • Restorative Dentistry and Endodontics
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    • v.22 no.2
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    • pp.761-768
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    • 1997
  • Microorganisms and their by-products are considered to be the major causes of pulpal and periapical pathosis. The role of microorganisms in endodotic infection has been studied regarding the prevalence of particular organisms found in root canal and periapical lesions. The aim of this study was to investigate the effect of culture supernatants of several oral microorganims isolated from infected root canals on the viability of cultured cell lines using colorimetirc assay. S. simulans, S. sciuri, E. faecium, S. intermedius, S. mitis, S. sanguis and S. uberis were incubated in Todd-Hewitt broth for 16 hours. 20 and 100ul of filtered bacterial cell culture supernatants were added to MK and Hep-2 cells. Cell viability was measured using MIT colorimetric assay. 20ul and 100u1 of S. sanguis supernatant showed significant cytotoxicity compared to control on MK cells. 100ul of S. sanguis supernatant significantly depressed viability of HEp-2 cells. E. faecium and S. intermedius did not affect the viability of MK and HEp-2 cells.

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The Effects of Taklysodoksan Extract on the Anti-cancer and Immunity (抗癌 및 免疫에 대한 托裡消毒散의 效果;萬病回春方을 중심으로)

  • Kim, Hong-Jin;Choi, Jung-hwa
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.13 no.1
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    • pp.100-115
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    • 2000
  • Taklysodoksan(TSS) was a drug used in the treatment of carbuncle and cellulitis in oriental medicine. The purpose of this Study is to investigate the anti-cancer effect of TSS, the proliferation of immunocytes and nitric oxide(NO) production from peritoneal macrophages of mice. This Study estimated the proliferation of Ll210 cell lines, mouse thymocytes and splenocytes and NO production from peritoneal macrophages in vitro and vivo. The proliferation of cells was tested using a colorimetric tetrazoliun assay(MTT assay). From the in vitro and vivo Study of TSS treatment, it did not effect the proliferation of L1210 cells. It also did not have any positive prodution of NO in peritoneal macrophages. This results suggest that TSS treatment in WanBingHuiChun(萬病回春) did not have significant anti-cancer effect and immuno-action comparing with TSS treatment of WaiKeZhengZhong(外科正宗).

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