• Title/Summary/Keyword: in vitro detection system

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Recent Development in Biocompatible Biosensors

  • Yongju Lee;Swarup Biswas;Minsuk Koo;Hyeok Kim
    • Journal of Sensor Science and Technology
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    • v.32 no.6
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    • pp.403-411
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    • 2023
  • The shift in the medical paradigm from treatment to prevention and diagnosis has underscored the growing significance of biosensors. Notably, the recent COVID-19 pandemic has spurred the widespread adoption of biosensors for the detection of viral genes and antigens. Consequently, there has been a substantial increase in both the demand for biosensors and the industries associated with their production. Furthermore, biosensors find applications not only in healthcare but also in diverse fields such as environmental monitoring, food quality control, military defense, and industrial processes. In this brief review, we delve into the essential attributes of biosensors, namely sensitivity, selectivity, and stability. We provide an overview of the latest research trends aimed at improving these attributes. Additionally, we introduce recent research cases in which these attributes are being applied both in vivo and in vitro.

In Vitro Regeneration of Lycium chinense Miller and Detection of Silent Somaclones with RAPD Polymorphisms

  • Ahn, In-Suk;Park, Young-Goo;Shin, Dong-Ill;Sul, Ill-Whan
    • Journal of Plant Biotechnology
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    • v.6 no.3
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    • pp.157-163
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    • 2004
  • An efficient system for the regeneration of adventitious shoots from in vitro cultured leaf sections of Lycium chinense Miller was developed and silent somaclones from the regenerants detected with RAPD method. Among the eight media tested (B5, SH, N&N, 1/2MS, MS, 3/2MS, GD and WPM), and four cytokinins (BA, kinetin, 2ip and zeatin) with different concentrations (1, 5, 10, 20, 30 and 40 $\mu{M}$), 1/2 MS medium supplemented with 20 and 30 $\mu{M}$ zeatin showed the best regeneration frequency (100% and 93.7%) and higher average number of shoots (9.0 and 9.4). All regenerants easily elongated after subculturing on 1/4MS without growth stimulants and produced spontaneous adventitious roots from their basal parts. With phenotypically normal 40 regenerants, RAPD analysis with 15 different random primers was performed to examine the cryptic somaclonal variants. No substantial differences in banding patterns were found in the amplified polymorphic DNAs implying no DNA changes during dedifferentiation into adventitious shoots. However, one (OPF-4) of the 15 primers detected silent somaclonal variation in one regenerant in which two different polymorphic bands did not appear when compared with the rest regenerants. The results indicate that regenerantion via intervening callus phase can be used to establish true-to-type planting stocks for homogeneous population.

Quantitative Assessment of Xenoestrogenic Environmental Pollutants using E-SCREEN Assay (E-SCREEN Assay를 이용한 내분비계 장애물질의 정량적 평가)

  • 오승민;이상기;정규혁
    • YAKHAK HOEJI
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    • v.44 no.5
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    • pp.416-423
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    • 2000
  • There is a growing concern that a wide variety of chemicals released into the environment can disrupt the endocrine system of fish, wildlife and humans. Endocrine disrupting chemicals (EDCs) include pesticides such as DDT lindane and atrazine, the food packaging chemicals, phthalates and bisphenol A, alkylphenol ethoxylate detergents and the chemical industry by-products, dioxins. Xenoestrogens in the environment have been argued about health risk, because of estrogen mimetic chemicals are exposed only small amounts to human. A number of in vivo and in vitro assays are now in use to assess the activity of xenoestrogens in the environment. A human breast cancer cell line (MCF-7) was used to develop in vitro screening assay for the detection of xenoestrogenic environmental pollutants. The E-SCREEN (MCF7-BUS) assay is proposed as a reliable, easy and rapid-to-perform method. To optimize and validate this method before it can be used routinely, several phenol compounds and pesticides suspected to be estrogenic were tested using I-SCREEN assay. The results showed that this method is a valuable tool for screening potential estrogen-mimicking environmental pollutants and quantitative determination of estrogeniciy.

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Verification of Estrogen Like Activities of Herbal Medicines Using an In Vitro Detection System (In vitro 검출 시스템을 이용한 한약재 추출물들의 에스트로겐 유사활성 검증)

  • Park, Seong Hwan;Jeon, Myong Je;Jang, Min Kyung;Lee, Sol Jee;Kim, Bo Kyung;Jeon, Myeong Jeong;Kim, Seo Yeon;Kim, Mihyang;Lee, Dong Geun;Lee, Tae Ho;Nam, Jae Sub;Hur, Ji Won;Lee, Sang Hyeon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.27 no.6
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    • pp.752-758
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    • 2013
  • Estrogen like activities were evaluated using ethanol and hot water extracts of herbal medicines by using an in vitro detection system. Bokryung (Poria cocos), Sanyak (root of Dioscorea batatas) and Mokdanpi (root skin of Paeonia suffruticosa) represented statistically significant estrogen-like activities (p<0.001), while Omija (fruit of Schizandra chinensis), Taeksa (root of Alisma canaliculatum A. BR.), Jihwang (root of Rhemannia glutinosa), and Sansuyu (fruit of Cornus officinalis) did not. Estrogen-like activities of Bokryung hot water extract (500 ${\mu}g/ml$) and ethanol extract (50 ${\mu}g/ml$) were almost same as that of a C M $17{\beta}$-estradiol. Furthermore, estrogen-like activities of ethanol extracts (500 ${\mu}g/ml$) of Bokryung and Mokdanpi were stronger than that of $10^{-7}$ M $17{\beta}$-estradiol. These results suggest that Bokryung, Sanyak and Mokdanpi show estrogen-like activities. Especially, Sanyak and Mokdanpi represented promotive effect on the proliferation of MC3T3-E1 osteoblastic cells. Bokryung, Sanyak and Mokdanpi also exhibited superior inhibitory effect on the viability of RAW 264.7 cells. In conclusion, these three herbal medicines might be interpreted as candidates for the further study or development of functional foods or medicine to prevent or avoid postmenopausal symptoms of women.

COMPARATIVE STUDY ON THE EARLY DETECTION OF ENAMEL LESIONS USING DIFOTI AND LASER FLUORESCENCE (Digital Imaging Fiber-Optic Trans-Illumination과 Laser Fluorescence를 이용한 법랑질 우식증의 조기 진단에 관한 비교 연구)

  • Maeng, Myoung-Ho;Kim, Seung-Oh;Kim, Jong-Soo
    • Journal of the korean academy of Pediatric Dentistry
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    • v.33 no.2
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    • pp.207-220
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    • 2006
  • The newly developed equipments for the early detection of carious lesion are LFD (laser fluorescence device), Ultrasonic diagnostic system, CLSM(confocal laser scanning microscopy), QLF(quantitative light-induced fluorescence) and DIFOTI (digital imaging fiber-optic trans-illumination) system. In this study, DIFOTI system and LFD were used for the detection of early enamel caries. Twenty five primary teeth extracted from twenty one children at around the dentitional exchanging period were selected as samples. The results obtained from DIFOTI imaging and LFD measurement were compared with those of CLSM and comprehensive evaluations were made for the diagnostic capacity of each device. In vitro test, 40 sample teeth with their buccal & lingual surface formed by a window of $2{\times}3mm$ in diameter were immersed in artificial demineralizing solution for the period of 4, 8, 12 and 16 days. The results obtained from the experimental groups (DIFOTI, LFD) were compared to control group (CLSM) and we have reached to the following conclusions. 1. The sensitivity and specificity of DIFOTI system operated in oral environment was 88.2% and 76.9% respectively. 2. The sensitivity and specificity of LFD measured in oral environment was 76.5% and 69.2% respectively. 3, Regression analysis on the light transparent rate of DIFOTI showed its decrease according to the length of primary enamel decalcification performed in vitro(r=-0.96, p<0.05). 4. No statistically significant difference between LFT measurement and the length of in vitro decalcification was found in regression analysis (p>0.05). 5. The correlation coefficient of DIFOTI image transparent rate and the lesion depth of CLMS was -0.6988 (p<0.05), whereas no statistically significant difference was found for LFD measurement.

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Visible and Fast Assay System for Tobacco Transformant Introduced with Adenosine Deaminase Marker Gene (Adenosine Deaminase 표지유전자로 형질전환된 연초의 신속한 Assay 방법)

  • 양덕춘;김용환;임학태;방극수;배창휴
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.3
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    • pp.165-171
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    • 2001
  • New visible and fast assay system have been developed for tobacco transformant introduced with adenosine deaminase (ADA) marker gene, which converts cytotoxic adenosine analogues to non-toxic inosine analogues and ammonia. Ammonia was changed to blue color in the solution of phenol-nitoprusside and alkaline-hypochlorite. It was possible to detect activity of ADA visibly on the holes of 96 well plate using tiny explant of transgenic tobacco leaves within 1 hour incubation time. As substrates of ADA enzyme from transgenic plant on the plate, a number of adenosine analogues such as 9-D-arabinofuranosyl adenine, cordycepin, 2'-deoxyadenosine, adenosine and xylofuranosyl adenine were possible for detection of ADA activity. Optimal condition of substrate for ADA enzyme was each 10 mM and pH 7.5 in adenosine solution. Especially, transgenic plant did not convert adenosine to inosine and ammonia in the presence of ADA inhibitor deoxycoformycin, which means that ammonia produced from transgenic plant is due to expression of ADA gene. Now, we show that this detection system can be easily, sensitively, fast and cheaply as well as visibly assayed in vitro as GUS gene system with very small size of transformant explant.

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Detection of Ref-1 (Redox factor-1) Interacting Protein Using the Yeast Two-hybrid System (Yeast two-hybrid system을 이용한 Ref-1 (redox factor-1) 결합 단백질의 분리 및 동정)

  • 이수복;김규원;배문경;배명호;정주원;안미영;김영진
    • Journal of Life Science
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    • v.14 no.1
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    • pp.26-31
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    • 2004
  • Redox factor-1 (Ref-1), known as a redox regulator, controls the DNA binding of AP-1 and is activated in HT29 colon cancer cells by hypoxia in vitro. REF-1 also increases tile DNA binding affinity of Hypoxia-inducible Factor-lalpha$ (HIF-lalpha$), HIF-like Factor (HLF) and early growth response-1 (Egr-1) which induce expression of the genes involved in angiogenesis, so that we speculate that REF-1 may play a role in hypoxia-induced angiogenesis. In this research we tried to detect novel proteins interacting with REF-1 using Yeast two-hybrid system using full-length REF-1 cDNA as bait. As result of such screening we detected 3 positive clones. DNA sequencing and GeneBank search revealed that one of the clones contained the same sequences as M.musculus cDNA for tioredoxin.

Effect of maternal gene expression on porcine oocytes in vitro maturation (돼지 미성숙 난자 모계 유전자 발현이 체외성숙에 미치는 영향)

  • Lee, Jae-Dal
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.13 no.8
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    • pp.3532-3536
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    • 2012
  • Understanding of the maternal transcriptome increased to elucidate the underlying molecular mechanism of normal oocyte maturation, which depends on a precise sequence of changes in maternal genes expression. Previous reports that the translational potential of a maternal mRNA is generally determined by the length of the poly(A) tail, and deadenylation is usually the first sign of mRNA degradation. However, in vitro cultured system has the underlying molecular mechanisms remain unclear. We determined whether the role of molecular basis, four important maternal genes, C-mos, cyclin-B1 (regulatory subunit of MPF), BMP15 and GDF9, were selected for detection of their precise mRNA expression patterns by real-time PCR and for determination of their polyadenylation status by poly(A) tail PCR during oocyte maturation. In the present study. the abnormal expression of maternal mRNAs prior to zygotic genome activation, which results in suppression of the corresponding protein level, may be responsible for, at least in part, a profound defect in further embryonic development. Reasonable expression of maternal gene is crucial for proper oocyte maturation and further embryonic development.

Detection of transgene in early developmental stage by GFP monitoring enhances the efficiency of genetic transformation of pepper

  • Jung, Min;Shin, Sun-Hee;Park, Jeong-Mi;Lee, Sung-Nam;Lee, Mi-Yeon;Ryu, Ki-Hyun;Paek, Kee-Yoeup;Harn, Chee-Hark
    • Plant Biotechnology Reports
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    • v.5 no.2
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    • pp.157-167
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    • 2011
  • In order to establish a reliable and highly efficient method for genetic transformation of pepper, a monitoring system featuring GFP (green fluorescent protein) as a report marker was applied to Agrobacteriummediated transformation. A callus-induced transformation (CIT) system was used to transform the GFP gene. GFP expression was observed in all tissues of $T_0$, $T_1$ and $T_2$ peppers, constituting the first instance in which the whole pepper plant has exhibited GFP fluorescence. A total of 38 T0 peppers were obtained from 4,200 explants. The transformation rate ranged from 0.47 to 1.83% depending on the genotype, which was higher than that obtained by CIT without the GFP monitoring system. This technique could enhance selection power by monitoring GFP expression at the early stage of callus in vitro. The detection of GFP expression in the callus led to successful identification of the shoot that contained the transgene. Thus, this technique saved lots of time and money for conducting the genetic transformation process of pepper. In addition, a co-transformation technique was applied to the target transgene, CaCS (encoding capsaicinoid synthetase of Capsicum) along with GFP. Paprika varieties were transformed by the CaCS::GFP construct, and GFP expression in callus tissues of paprika was monitored to select the right transformant.

Development of a Novel ATP Bioluminescence Assay Based on Engineered Probiotic Saccharomyces boulardii Expressing Firefly Luciferase

  • Ji Sun Park;Young-Woo Kim;Hyungdong Kim;Sun-Ki Kim;Kyeongsoon Park
    • Journal of Microbiology and Biotechnology
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    • v.33 no.11
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    • pp.1506-1512
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    • 2023
  • Quantitative analysis of adenosine triphosphate (ATP) has been widely used as a diagnostic tool in the food and medical industries. Particularly, the pathogenesis of a few diseases including inflammatory bowel disease (IBD) is closely related to high ATP concentrations. A bioluminescent D-luciferin/luciferase system, which includes a luciferase (FLuc) from the firefly Photinus pyralis as a key component, is the most commonly used method for the detection and quantification of ATP. Here, instead of isolating FLuc produced in recombinant Escherichia coli, we aimed to develop a whole-cell biocatalyst system that does not require extraction and purification of FLuc. To this end, the gene coding for FLuc was introduced into the genome of probiotic Saccharomyces boulardii using the CRISPR/Cas9-based genome editing system. The linear relationship (r2 = 0.9561) between ATP levels and bioluminescence generated from the engineered S. boulardii expressing FLuc was observed in vitro. To explore the feasibility of using the engineered S. boulardii expressing FLuc as a whole-cell biosensor to detect inflammation biomarker (i.e., ATP) in the gut, a colitis mouse model was established using dextran sodium sulfate as a colitogenic compound. Our findings demonstrated that the whole-cell biosensor can detect elevated ATP levels during gut inflammation in mice. Therefore, the simple and powerful method developed herein could be applied for non-invasive IBD diagnosis.