• Title/Summary/Keyword: Colorimetric Detection

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Sensitivity Enhancement of Polydiacetylene Vesicles through Control of Particle Size and Polymerization Temperature (입자크기와 중합온도 제어를 통한 폴리다이아세틸렌의 센싱감도 향상)

  • Lee, Gil Sun;Oh, Jae Ho;Ahn, Dong June
    • Korean Chemical Engineering Research
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    • v.49 no.4
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    • pp.400-404
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    • 2011
  • Many studies on polydiacetylene(PDA) have been investigated to apply to chemical and biological sensors due to their unique optical properties of color change from blue to red and fluorescence change from non-fluorescence to red fluorescence. Especially, high sensitivity against specific molecules is very important to apply polydiacetylenes to various sensors. In this study, we examined the effect of sensitivity enhancement of 10,12-pentacosadynoic acid(PCDA) vesicles in detection ${\alpha}$-cyclodextrin(CD) according to control of vesicle size by filters with different pore sizes and polymerization temperature. Colorimetric response(CR) was calculated using visible spectrometer. In order to investigate the effect of vesicle size on sensitivity of PDA vesicles, two PCDA vesicles were filtered without filtration and with 0.22 ${\mu}m$ filter. The two PCDA vesicles were polymerized at $25^{\circ}C$ and were incubated with ${\alpha}$-CD(5 mM) for 30 min. The CRs of the former and latter vesicles were 31.4% and 74.0%, respectively. Then, two PCDA vesicles filtered with 0.22 ${\mu}m$ filter were polymerized at $25^{\circ}C$ and $5^{\circ}C$ and were reacted with ${\alpha}$-CD(5 mM) for 30 min to examine the effect of polymerization temperature. The CRs of the former and latter vesicles were 74.0 and 99.2%, respectively. This suggests that vesicle sizes and polymerization temperature are key factors in enhancing the sensitivity of PDA vesicles. In addition, these results are expected to be useful to apply the PDA vesicles as biosensors to detect DNA, protein, and cells.

Synthesis and physicochemical characterization of NixZnx-Fe2O4/MWCNT nanostructures as enzyme mimetics with peroxidase-like catalytic activity

  • Salarizadeh, Navvabeh;Sadri, Minoo;Hosseini, Hassan;Sajedi, Reza. H.
    • Carbon letters
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    • v.24
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    • pp.103-110
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    • 2017
  • Carbon-based magnetic nanostructures in several instances have resulted in improved physicochemical and catalytic properties when compared to multi-wall carbon nanotubes (MWCNTs) and magnetic nanoparticles. In this study, magnetic MWCNTs with a structure of $Ni_xZn_xFe_2O_4/MWCNT$ as peroxidase mimics were fabricated by the one-pot hydrothermal method. The structure, composition and morphology of the nanocomposites were characterized with X-ray diffraction (XRD), Fourier transform infrared spectroscopy and transmission electron microscopy. The magnetic properties were investigated with a vibrating sample magnetometer. The peroxidase-like catalytic activity of the nanocomposites was investigated by colorimetric and electrochemical tests with 3,3',5,5'-tetramethylbenzidine (TMB) and $H_2O_2$ as the substrates. The results show that the synthesis of the nanocomposites was successfully performed. XRD analysis confirmed the crystalline structures of the $Ni_xZn_xFe_2O_4/MWCNT$ nanohybrids and MWCNTs. The main peaks of the $Ni_xZn_xFe_2O_4/MWCNT$s crystals were presented. The $Ni_{0.25}Zn_{0.25}Fe_2O_4/MWCNT$ and $Ni_{0.5}Zn_{0.5}Fe_2O_4/MWCNT$ nanocatalysts showed nearly similar physicochemical properties, but the $Ni_{0.5}Zn_{0.5}Fe_2O_4/MWCNT$ nanocatalyst was more appropriate than the $Ni_{0.25}Zn_{0.25}Fe_2O_4/MWCNT$ nanocatalyst in terms of the magnetic properties and catalytic activity. The optimum peroxidase-like activity of the nanocatalysts was obtained at pH 3.0. The $Ni_{0.5}Zn_{0.5}Fe_2O_4/MWCNT$ nanocatalyst exhibited a good peroxidase-like activity. These magnetic nanocatalysts can be suitable candidates for future enzyme-based applications such as the detection of glucose and $H_2O_2$.

Artemisia capillaris Thunb. inhibits cell growth and induces apoptosis in human hepatic stellate cell line LX2

  • Kim, Young-Il;Lee, Jang-Hoon;Park, Seung-Won;Choi, In-Hwa;Friedman, Scott L.;Woo, Hong-Jung;Kim, Young-Chul
    • Advances in Traditional Medicine
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    • v.10 no.4
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    • pp.254-262
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    • 2010
  • Artemisia capillaris (A. capillaries) is known to play roles in many cellular events, such as cell proliferation, differentiation, and apoptosis. We investigated the antifibrogenic efficacy of A. capillaris in the immortalized human hepatic stellate cell line LX2. Cell proliferation was determined by the MTT assay. Cell cycle was analyzed by the flow cytometry. Apoptotic cells were measured using a cell death detection ELISA. Caspase activity was detected by a colorimetric assay. The mRNA level of Bcl-2 and Bax mRNA were measured by real-time PCR. MEK and ERK protein were detected by Western blot analysis. We provide evidence that A. capillaris induces cell cycle arrest, apoptosis, and potently inhibits the mitogen-activated protein kinase pathway. A. capillaris inhibited cell proliferation of LX2 cells in a dose- and time-dependent manner, increased the apoptosis fraction at cell cycle analysis with an accompanying DNA fragmentation, and resulted in a significant decrease in Bcl-2 mRNA levels and an increase in Bax expression. Exposure of LX2 cells to A. capillaris induced caspase-3 activation, but co-treatment of A. capillaris with the pan-caspase inhibitor Z-VAD-FMK, and the caspase-3 inhibitor Z-DEVE-FMK, blocked apoptosis. A. capillaris down-regulated Mcl-1 protein levels and inhibited phosphorylation of MEK/ERK, suggesting that it mediates cell death in LX2 cells through the down-regulation of Mcl-1 protein via a MEK/ERK-independent pathway.

Techniques for Evaluation of LAMP Amplicons and their Applications in Molecular Biology

  • Esmatabadi, Mohammad javad Dehghan;Bozorgmehr, Ali;zadeh, Hesam Motaleb;Bodaghabadi, Narges;Farhangi, Baharak;Babashah, Sadegh;Sadeghizadeh, Majid
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.17
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    • pp.7409-7414
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    • 2015
  • Loop-mediated isothermal amplification (LAMP) developed by Notomi et al. (2000) has made it possible to amplify DNA with high specificity, efficiency and rapidity under isothermal conditions. The ultimate products of LAMP are stem-loop structures with several inverted repeats of the target sequence and cauliflower-like patterns with multiple loops shaped by annealing between every other inverted repeats of the amplified target in the similar strand. Because the amplification process in LAMP is achieved by using four to six distinct primers, it is expected to amplify the target region with high selectivity. However, evaluation of reaction accuracy or quantitative inspection make it necessary to append other procedures to scrutinize the amplified products. Hitherto, various techniques such as turbidity assessment in the reaction vessel, post-reaction agarose gel electrophoresis, use of intercalating fluorescent dyes, real-time turbidimetry, addition of cationic polymers to the reaction mixture, polyacrylamide gel-based microchambers, lateral flow dipsticks, fluorescence resonance energy transfer (FRET), enzyme-linked immunosorbent assays and nanoparticle-based colorimetric tests have been utilized for this purpose. In this paper, we reviewed the best-known techniques for evaluation of LAMP amplicons and their applications in molecular biology beside their advantages and deficiencies. Regarding the properties of each technique, the development of innovative prompt, cost-effective and precise molecular detection methods for application in the broad field of cancer research may be feasible.

Induction of Human Hepatocellular Carcinoma HepG2 Cell Apoptosis by Naringin

  • Banjerdpongchai, Ratana;Wudtiwai, Benjawan;Khaw-on, Patompong
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.7
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    • pp.3289-3294
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    • 2016
  • Naringin, a bioflavonoid found in Citrus seeds, inhibits proliferation of cancer cells. The objectives of this study were to investigate the mode and mechanism(s) of hepatocellular carcinoma HepG2 cell death induced by naringin. The cytotoxicity of naringin towards HepG2 cells proved dose-dependent, measured by MTT assay. Naringin-treated HepG2 cells underwent apoptosis also in a concentration related manner, determined by annexin V-fluorescein isothiocyanate (FITC) and propidium iodide (PI) employing flow cytometry. Mitochondrial transmembrane potential (MTP) measured using 3,3'-dihexyloxacarbocyanine iodide ($DiOC_6$) and flow cytometer was reduced concentration-dependently, which indicated influence on the mitochondrial signaling pathway. Caspase-3, -8 and -9 activities were enhanced as evidenced by colorimetric detection of para-nitroaniline tagged with a substrate for each caspase. Thus, the extrinsic and intrinsic pathways were linked in human naringin-treated HepG2 cell apoptosis. The expression levels of pro-apoptotic Bax and Bak proteins were increased whereas that of the anti-apoptotic Bcl-xL protein was decreased, confirming the involvement of the mitochondrial pathway by immunoblotting. There was an increased expression of truncated Bid (tBid), which indicated caspase-8 proteolysis activity in Bid cleavage as its substrate in the extrinsic pathway. In conclusion, naringin induces human hepatocellular carcinoma HepG2 cell apoptosis via mitochondria-mediated activation of caspase-9 and caspase-8-mediated proteolysis of Bid. Naringin anticancer activity warrants further investigation for application in medical treatment.

The Past, Present, Future of Newborn Screening in Korea (한국 신생아 집단검사의 과거, 현재, 미래)

  • Lee, Dong Hwan
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.14 no.1
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    • pp.1-9
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    • 2014
  • Many inborn errors of metabolism can be completely cured with early detection and early treatment. This is why neonatal screening on inborn errors of metabolism is implemented worldwide. In 1991, the Ministry of Health & Social affairs adopted a nationwide service program for neonatal screening of phenylketonuria, galactosemia, maple syrup urine disease, homocystinuria, histidinemia and congenital hypothyroidism for newborns delivered from low class pregnant women registered in health centers. Government decreased the test items from six to two, PKU and congenital hypothyroidism to increase test numbers with same budget from 1995. 78 laboratories wanted to participate for neonatal screening test in 1999. Government decided to screen six items of PKU, congenital hypothyroidism, maple syrup urine disease, homocystinuria, galactosemia and congenital adrenal hyperplasia from 2006. In 2014, thirteen laboratories are participating. Inter laboratory quality control was started 6 times a year from 1994. In case a patient with an inherited metabolic disease is diagnosed by screening of government program, special milk is provided at government's expense. According to the government project, from 1997 to 2013, 7,080,569 newborns were screened. 144 PKU, 2.451 congenital hypothyroidism were detected. So incidence of PKU is 1/49,170 and congenital hypothyroidism is 1/2,888. The cost benefit of performing screening procedures coupled with treatment has been estimated to be as high as 1.77 times in PKU, 11.11 times in congenital hypothyroidism than cost without screening. By January 2007, many European countries had expanded of their newborn screening programs by inclusion of Tandem mass spectrometry. We are trying to increase the budget to test all newborns for Tandem mass spectrometry from 2016. We are considering four to five central laboratories which cover all newborns and are equipped with tandem mass spectrometer & enzyme immunoassay for TSH, 17OHP & enzyme colorimetric assay for galactose. And I hope to expand test including Wilson disease screening test and lysosomal storage diseases.

Optimization of Analytical Procedure for Hydrogen Cyanide in Mainstream Smoke

  • Lee, John-Tae;Kim, Hyo-Keun;Hwang, Keon-Joong;Jang, Gi-Chul;Lee, Jeong-Min;Kim, Ick-Joong
    • Journal of the Korean Society of Tobacco Science
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    • v.29 no.2
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    • pp.125-131
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    • 2007
  • Hydrogen cyanide(HCN), formed from pyrolysis of various nitrogenous compounds such as protein, amino acids and nitrate in tobacco, is present in both the particulate phase and vapor phase of cigarette smoke. Typically the determination of HCN in cigarette smoke has been done through colorimetric and electrochemical techniques, such as fluorescence spectrometry, UV-spectrophotometry (UV), continuous flow analyzer (CFA), capillary GC-ECD and ion chromatography (IC). Most of these techniques are known to be time-consuming and some of them lack specificity or sensitivity. The available results from both our laboratory and reported literatures for 2R4F Kentucky reference cigarette, smoked under ISO condition, show a relatively wide variation ranging from 100 to 120 ug/cig of HCN. Especially, the precision and accuracy of the analytical results of HCN tend to get worse in low tar cigarettes and under intense smoking condition. In this paper, a more optimized analytical methods than previous ones are suggested. This method shows lower detection limit and has improved precision and accuracy, so it is applicable for wide tar level cigarettes under intense smoking condition as well as under ISO smoking condition. Important features of this method are improved sample collection and quantification systems such as the number of trapping units, volume, temperature and type of trapping solution. To avoid volatilization loss of HCN in analyzing mainstream smoke, it is highly recommended that pH values of trapping solutions should be maintained over 11 and cold traps should be used in collecting mainstream smoke.

Establishing Quantitative Standards for Residual Alkaline Phosphatase in Pasteurized Milk

  • Kim, Dong-Hyeon;Chon, Jung-Whan;Lim, Jong-Soo;Kim, Hong-Seok;Kang, Il-Byeong;Jeong, Dana;Song, Kwang-Young;Kim, Hyunsook;Kim, Kwang-Yup;Seo, Kun-Ho
    • Food Science of Animal Resources
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    • v.36 no.2
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    • pp.194-197
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    • 2016
  • The alkaline phosphatase (ALP) assay is a rapid and convenient method for verifying milk pasteurization. Since colorimetric ALP assays rely on subjective visual assessments, their results are especially unreliable near the detection limits. In this study, we attempted to establish quantitative criteria for residual ALP in milk by using a more objective method based on spectrophotometric measurements. Raw milk was heat-treated for 0, 10, 20, 30, and 40 min and then subjected to ALP assays. The quantitative criteria for residual ALP in the milk was determined as 2 μg phenol/mL of milk, which is just above the ALP value of milk samples heat-treated for 30 min. These newly proposed methodology and criteria could facilitate the microbiological quality control of milk.

Recent Research Trend in Lateral Flow Immunoassay Strip (LFIA) with Colorimetric Method for Detection of Cancer Biomarkers (암 바이오마커 검출용 비색법 기반 측면 흐름 면역 크로마토그래피 분석법(LFIA) 스트립의 최신 연구 동향)

  • Lee, Sooyoung;Lee, Hye Jin
    • Applied Chemistry for Engineering
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    • v.31 no.6
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    • pp.585-590
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    • 2020
  • Successful early diagnosis of cancer diseases such as lung, prostate, liver and adrenocortical carcinoma is a key step in determining the cost of treatment, survival rate, and cure rate. Most of current cancer diagnosis systems including biopsy, computed tomography (CT), positron emission tomography (PET)-CT, magnetic resonance imaging (MRI), ultrasonography, etc., require expensive and complicated equipment with highly trained human resources. Global medical and scientific communities have thus made numerous efforts on developing effective but rapid disease management system via introducing a wide spectrum of point-of-care medical diagnosis devices. Among them, a lateral flow immunoassay strip technique has gained a great attention due to many advantages such as cost-effectiveness, short inspection time, and user friendly accessibility. In this mini-review, we will highlight recent research trend on the development of colorimetry based LFIA strips for cancer diagnosis and discuss the future research direction and potential applications.

Wheat phytase potentially protects HT-29 cells from inflammatory nucleotides-induced cytotoxicity

  • Jeongmin An;Jaiesoon Cho
    • Animal Bioscience
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    • v.36 no.10
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    • pp.1604-1611
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    • 2023
  • Objective: The aim of this study was to investigate the protective effect of wheat phytase as a structural decomposer of inflammatory nucleotides, extracellular adenosine triphosphate (ATP), and uridine diphosphate (UDP) on HT-29 cells. Methods: Phosphatase activities of wheat phytase against ATP and UDP was investigated in the presence or absence of inhibitors such as L-phenylalanine and L-homoarginine using a Pi Color Lock gold phosphate detection kit. Viability of HT-29 cells exposed to intact- or dephosphorylated-nucleotides was analyzed with an EZ-CYTOX kit. Secretion levels of pro-inflammatory cytokines (IL-6 and IL-8) in HT-29 cells exposed to substrate treated with or without wheat phytase were measured with enzyme-linked immunosorbent assay kits. Activation of caspase-3 in HT-29 cells treated with intact ATP or dephosphorylated-ATP was investigated using a colorimetric assay kit. Results: Wheat phytase dephosphorylated both nucleotides, ATP and UDP, in a dose-dependent manner. Regardless of the presence or absence of enzyme inhibitors (L-phenylalanine and L-homoarginine), wheat phytase dephosphorylated UDP. Only L-phenylalanine inhibited the dephosphorylation of ATP by wheat phytase. However, the level of inhibition was less than 10%. Wheat phytase significantly enhanced the viability of HT-29 cells against ATP- and UDP-induced cytotoxicity. Interleukin (IL)-8 released from HT-29 cells with nucleotides dephosphorylated by wheat phytase was higher than that released from HT-29 cells with intact nucleotides. Moreover, the release of IL-6 was strongly induced from HT-29 cells with UDP dephosphorylated by wheat phytase. HT-29 cells with ATP degraded by wheat phytase showed significantly (13%) lower activity of caspase-3 than HT-29 cells with intact ATP. Conclusion: Wheat phytase can be a candidate for veterinary medicine to prevent cell death in animals. In this context, wheat phytase beyond its nutritional aspects might be a novel and promising tool for promoting growth and function of intestinal epithelial cells under luminal ATP and UDP surge in the gut.