• 제목/요약/키워드: Quantitative screening

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Use of DNA-Specific Anthraquinone Dyes to Directly Reveal Cytoplasmic and Nuclear Boundaries in Live and Fixed Cells

  • Edward, Roy
    • Molecules and Cells
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    • 제27권4호
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    • pp.391-396
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    • 2009
  • Image-based, high-content screening assays demand solutions for image segmentation and cellular compartment encoding to track critical events - for example those reported by GFP fusions within mitosis, signalling pathways and protein translocations. To meet this need, a series of nuclear/cytoplasmic discriminating probes have been developed: DRAQ5$^{TM}$ and CyTRAK Orange$^{TM}$. These are spectrally compatible with GFP reporters offering new solutions in imaging and cytometry. At their most fundamental they provide a convenient fluorescent emission signature which is spectrally separated from the commonly used reporter proteins (e.g. eGFP, YFP, mRFP) and fluorescent tags such as Alexafluor 488, fluorescein and Cy2. Additionally, they do not excite in the UV and thus avoid the complications of compound UV-autofluorescence in drug discovery whilst limiting the impact of background sample autofluorescence. They provide a convenient means of stoichiometrically labelling cell nuclei in live cells without the aid of DMSO and can equally be used for fixed cells. Further developments have permitted the simultaneous and differential labelling of both nuclear and cytoplasmic compartments in live and fixed cells to clearly render the precise location of cell boundaries which may be beneficial for quantitative expression measurements, cell-cell interactions and most recently compound in vitro toxicology testing.

A High-speed Miniature Screening Gaschromatograph with Flame Ionization Detector

  • Banik Rahul;Lee Dong-Yeon;Gweon Dae-Gab
    • Journal of Mechanical Science and Technology
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    • 제19권12호
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    • pp.2197-2204
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    • 2005
  • The combination of Gas chromatography (GC) for separation and Flame Ionization Detection (FID) for detection and identification of the components of a mixture of compounds is a fast and strongly proved method of analytic chemistry. The objective of this research was to design a combined High-speed miniature screening Gas chromatograph along with a Flame Ionization Detector for quick, quantitative and qualitative analysis of gas components. This combined GC-FID system is suitable to detect the volatile and semi-volatile hydrocarbons present in a gas mixture. The construction made it less expensive, easy to use and movable. The complete gas path was developed. On/off valves, temperature and flow sensors and their interface electronics were used for controlling purpose. A Microcontroller was programmed to measure the temperature and gas flow using the sensors and to control and regulate them using the electronics and valves. A pocket PC with its touch screen served as a user interface for the system. Software was developed for the pocket PC, which makes the communication possible with the Microcontroller. The system parameters can be indicated in the Pocket PC as simple text and also the analysis result can be displayed.

위험 선별 및 분석 통합 자동화 시스템 개발에 대한 연구 (Study on Development of Automated System for Hazard Screening at Analysis)

  • 한의진;김용하;최승준;김구회;윤인섭
    • 한국화재소방학회논문지
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    • 제17권4호
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    • pp.20-27
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    • 2003
  • 화학공장은 위험한 물리 화학적 특성을 가진 많은 물질들을 다루기 때문에 다른 어떤 공장보다 더 사고의 위험성이 크다. 띠라서 화학공장에서의 각 장치에 대한 위험성 평가는 반드시 필요하다. 현재 사용되고 있는 위험성 평가 방법은 정성적 또는 정량적인 방법으로 각각 개별적으로 적용되는데 정성적, 정량적의 통합된 방법을 적용하면 빠르고 쉽게 적용할 수 있다는 정성적인 방법의 장점과 정밀하게 평가될 수 있다는 정량적인 방법의 장점을 모두 갖출 수 있다. 이러한 통합된 위험성 평가방법을 적용하고 자동화한 ASCA(Automated System for Hazard Screening & Analysis)라는 S/W를 개발하여 실제 공정에 적용하였다. 본 연구에서는 방향족 긍정 중 H.T.U(Hydrotreating Unit)에 대하여 ASCA를 적용하여 각 장치의 상대 순위를 알아보고 D-101이라는 저장탱크에서 단열 실패로 온도 상승이 일어난 경우의 장치고장으로 인한 변수 이상과 발생 가능한 사고의 영향을 알아보았다. 이러한 통합 위험성 평가 시스템을 화학공장에 적용하여 봄으로써 사고 시나리오를 세워서 사고에 대한 비상사태를 대처할 수 있는 능력을 키울 수 있으며 사고를 예방할 수 있을 것이다.

자동화 매독검사 키트의 임상적 유용성 및 생물학적 위양성률의 평가 (Evaluation of Clinical Utility and Biologic False Positive (BFP) Rates in Automated Syphilis Test Kits for Syphilis Screening)

  • 김성만;이제훈
    • 대한임상검사과학회지
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    • 제41권1호
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    • pp.42-46
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    • 2009
  • Unlike most bacteria, Treponema pallidum subspecies cannot be readily isolated or sustained in cell culture for numerous generations. In korea, two non treponemal tests are currently considered as standard; the VDRL slide test and RPR card test. These tests are based on an antigen composed of an alcoholic solution containing measured amount of cardiolipin, cholesterol, and sufficient purified lecithin to produce reactivity. The nontreponemal reagin tests measure immunoglobulin M (IgM) and IgG to lipoidal material released from damaged host cells as well as to lipoprotein-like material and possibly by cardiolipin released from the treponemes. The object of the evaluation was to evaluate the performance of the Mediace RPR kit on the automated biochemistry analyzer system as a method for screen method of syphilis as well as to identify BFP possibility. For evaluation of routine screening test, a total 2,380 specimens tested by Mediace RPR from 28th Oct, 2007 to 22th Feb, 2008. For evaluation of BFP possiblility, we measured samples which have potential BFP reaction in Syphilis test such as ANA (anti-nuclear antibody) positive (135 samples), CRP (C-reactive protein) positive (100 samples), RF (Rheumatoid factor) positive (26 samples), and other potential BFP cases (17 samples) including total 278 samples. These samples were tested quantitative test Mediace RPR with Hitachi 7600 P module. For comparison with current manual test, VDRL slide test were performed. Of these 2380 specimens, 2350 were negative, 30 were positive, and one were positive with TPHA. Both methods agreed for 2356 (98.9%) samples. Of the 30 samples showed positive results over 1.0 R.U, 6 samples showed positive results with VDRL test. Of these 6 samples, 1 samples showed positive with TPHA test. The combination of the Automated Biochemistry analyzer and VDRL test for retest can be increase efficiency of syphilis screening test.

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피부의 탄성변화에 따른 피부 가압과 복귀시간 측정을 통한 정량적 림프부종 진단 시스템 개발 (Development of Quantitative Lymphedema Screening System to Monitor Change in Skin Elasticity through the Measurement of Indentation Force and Return Time)

  • 서종현;조창노;김성천;정승현;고은실;김광기
    • 대한의용생체공학회:의공학회지
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    • 제34권4호
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    • pp.170-176
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    • 2013
  • Lymphedema is a phenomenon in which fluid is accumulated inside tissues due to the damaged lymphatic systems. Lymphedema can cause complications such as lymphangitis, infection, changes in skin texture, fibrosis, and lymphangiosarcoma. In this study, a lymphedema screening system based on the elasticity of the skin is proposed to easily quantify lymphedema. The developed probe consists of touch sensors, a load cell and hall-effect sensors to measure the indentation force on the skin and the return time of the skin. The developed system can be used to estimate the change in the elasticity of the skin to quantify lymphedema. The system was tested with a thyroid phantom and gelatin phantoms of different concentrations and the resulting force and the time were recorded. It was found that the increase in the elasticity leads to a higher indentation force and shorter return time. This shows that the developed system can monitor the change in the skin elasticity by measuring the return time and the indentation force. The feasibility of the system in clinical applications will be evaluated in the future study.

토픽 모델링을 활용한 광범위 선천성 대사이상 신생아 선별검사 관련 온라인 육아 커뮤니티 게시 글 분석: 계량적 내용분석 연구 (Analysis of online parenting community posts on expanded newborn screening for metabolic disorders using topic modeling: a quantitative content analysis)

  • 이명선;정현숙;김진선
    • 여성건강간호학회지
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    • 제29권1호
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    • pp.20-31
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    • 2023
  • Purpose: As more newborns have received expanded newborn screening (NBS) for metabolic disorders, the overall number of false-positive results has increased. The purpose of this study was to explore the psychological impacts experienced by mothers related to the NBS process. Methods: An online parenting community in Korea was selected, and questions regarding NBS were collected using web crawling for the period from October 2018 to August 2021. In total, 634 posts were analyzed. The collected unstructured text data were preprocessed, and keyword analysis, topic modeling, and visualization were performed. Results: Of 1,057 words extracted from posts, the top keyword based on 'term frequency-inverse document frequency' values was "hypothyroidism," followed by "discharge," "close examination," "thyroid-stimulating hormone levels," and "jaundice." The top keyword based on the simple frequency of appearance was "XXX hospital," followed by "close examination," "discharge," "breastfeeding," "hypothyroidism," and "professor." As a result of LDA topic modeling, posts related to inborn errors of metabolism (IEMs) were classified into four main themes: "confirmatory tests of IEMs," "mother and newborn with thyroid function problems," "retests of IEMs," and "feeding related to IEMs." Mothers experienced substantial frustration, stress, and anxiety when they received positive NBS results. Conclusion: The online parenting community played an important role in acquiring and sharing information, as well as psychological support related to NBS in newborn mothers. Nurses can use this study's findings to develop timely and evidence-based information for parents whose children receive positive NBS results to reduce the negative psychological impact.

확률적 정량모델을 이용한 토양에서의 바이러스 저감 평가 (Assessment of Viral Attenuation in Soil Using Probabilistic Quantitative Model)

  • 박정안;김재현;이인;김성배
    • 대한환경공학회지
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    • 제33권7호
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    • pp.544-551
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    • 2011
  • 본 연구의 목적은 미국 환경보호청에서 개발한 확률적 정량모델인 VIRULO 모델의 구성과 특성을 분석하는 것이다. 이 모델은 몬테카를로 방법을 이용하여 수리지질학적 차단벽으로써 토양의 바이러스 저감능을 평가할 수 있는 모델이다. 모델에 사용된 지배방정식은 크게 불포화 지역에서의 물의 흐름식과 바이러스의 이동식으로 구성되어 있다. 사용되는 파라미터들 중, 물의 흐름과 관련된 파라미터는 11종류의 토양에 대하여 UNSODA 데이터베이스로부터 얻어진 것 들이며, 바이러스와 관련된 파라미터 값들은 다섯 종류의 바이러스에 대하여 문헌조사를 통해 정리된 것이다. 모델은 목표로 하는 바이러스 저감 역치값과 특정 조건에서 몬테카를로 모사를 통해 얻어진 토양의 바이러스 저감인자를 비교하여, 목표로 하는 바이러스 저감 역치값에 도달하지 못하는 확률을 결정한다. 그리고, 몬테카를로 모사횟수와 목표 역치값에 도달하지 못한 횟수를 결과물로 제시한다. 11개의 USDA 토양을 대상으로 바이러스 저감을 평가한 결과, 양질사토와 모래의 바이러스 저감능이 점토나 미사 계열의 토양에 비하여 상당히 떨어지는 것으로 평가되었다. 5종의 바이러스를 대상으로 저감을 비교한 결과, 바이러스 간에 저감 정도에 차이가 있는 것으로 나타났으며, 그 중 폴리오바이러스의 저감 정도가 가장 큰 것으로 분석되었다. 그리고, 토양 함수량이 증가함에 따라 토양의 바이러스 저감능이 급격하게 감소하였으며, 토양의 깊이가 증가함에 따라 바이러스 저감능이 비선형적으로 증가하였다. 본 연구에 의하면, VIRULO 모델은 지중환경에서의 바이러스 위해성 평가에 사용될 수 있는 유용한 스크리닝 도구로 판단된다.

Alternative Carcinogenicity Screening Assay Using Colon Cancer Stem Cells: A Quantitative PCR (qPCR)-Based Prediction System for Colon Carcinogenesis

  • Bak, Yesol;Jang, Hui-Joo;Shin, Jong-Woon;Kim, Soo-Jin;Chun, Hyun woo;Seo, Ji-Hye;No, Su-Hyun;Chae, Jung-il;Son, Dong Hee;Lee, Seung Yeoun;Hong, Jintae;Yoon, Do-Young
    • Journal of Microbiology and Biotechnology
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    • 제28권4호
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    • pp.645-651
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    • 2018
  • The carcinogenicity of chemicals in the environment is a major concern. Recently, numerous studies have attempted to develop methods for predicting carcinogenicity, including rodent and cell-based approaches. However, rodent carcinogenicity tests for evaluating the carcinogenic potential of a chemical to humans are time-consuming and costly. This study focused on the development of an alternative method for predicting carcinogenicity using quantitative PCR (qPCR) and colon cancer stem cells. A toxicogenomic method, mRNA profiling, is useful for predicting carcinogenicity. Using microarray analysis, we optimized 16 predictive gene sets from five carcinogens (azoxymethane, 3,2'-dimethyl-4-aminobiphenyl, N-ethyl-n-nitrosourea, metronidazole, 4-(n-methyl-n-nitrosamino)-1-(3-pyridyl)-1-butanone) used to treat colon cancer stem cell samples. The 16 genes were evaluated by qPCR using 23 positive and negative carcinogens in colon cancer stem cells. Among them, six genes could differentiate between positive and negative carcinogens with a p-value of ${\leq}0.05$. Our qPCR-based prediction system for colon carcinogenesis using colon cancer stem cells is cost- and time-efficient. Thus, this qPCR-based prediction system is an alternative to in vivo carcinogenicity screening assays.

Application of ELISA for the Detection of Sulfamethazine Residue in Live Cattle

  • Lee, H.J.;Lee, M.H.;Han, In K.
    • Asian-Australasian Journal of Animal Sciences
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    • 제14권3호
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    • pp.378-381
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    • 2001
  • Sulfamethazine has been widely used in swine for prevention or treatment of infections. Recently, the safety of the drug to consumers has been questioned because of carcinogenic effects. To prevent unwanted drug residues entering the human food chain, both government authorities and industries have established extensive control measures. The demands for reliable, simple, sensitive, rapid and low-cost methods for residue analysis of foods are increasing nowadays. In this study, we established a rapid prediction test for the detection of cattle with violative tissue residues of sulfamethazine. The recommended therapeutic dose of sulfamethazine (withdrawal time, 15 days) was administered to each of 10 cattle. Blood was sampled before drug administration and during the withdrawal period. The concentration of sulfamethazine in plasma, determined by a semi-quantitative ELISA, was compared to that of an internal standard (10 ppb). The absorbance ratio of internal standard to sample (B/Bs) was employed as an index to determine whether drug residues in cattle tissues were negative or positive. That is, a B/Bs ratio less than 1 was considered residue positive and if larger than 1 was considered negative. All 10 plasma samples from non-treated cattle showed negative to sulfamethazine. Sulfamethazine was detected in plasmas of treated cattle until Day 7 of withdrawal period. The present study showed that the semi-quantitative ELISA could be easily adapted in predicting residues of sulfamethazine in live cattle.

PARP-1 억제제의 Docking 및 QSAR 연구 (Docking and QSAR studies of PARP-1 Inhibitors)

  • Kim, Hye-Jung;Cho, Seung-Joo
    • 한국생물정보학회:학술대회논문집
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    • 한국생물정보시스템생물학회 2004년도 The 3rd Annual Conference for The Korean Society for Bioinformatics Association of Asian Societies for Bioinformatics 2004 Symposium
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    • pp.210-218
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    • 2004
  • Poly(ADP-ribose)polymerase-1 (PARP-1) is a nuclear enzyme involved in various physical functions related to genomic repair, and PARP inhibitors have therapeutic application in a variety of neurological diseases. Docking and the QSAR (quantitative structure-activity relationships) studies for 52 PARP-1 inhibitors were conducted using FlexX algorithm, comparative molecular field analysis (CoMFA), and hologram quantitative structure-activity relationship analysis (HQSAR). The resultant FlexX model showed a reasonable correlation (r$^{2}$ = 0.701) between predicted activity and observed activity. Partial least squares analysis produced statistically significant models with q$^{2}$ values of 0.795 (SDEP=0.690, r$^{2}$=0.940, s=0.367) and 0.796 (SDEP=0.678, r$^{2}$ = 0.919, s=0.427) for CoMFA and HQSAR, respectively. The models for the entire inhibitor set were validated by prediction test and scrambling in both QSAR methods. In this work, combination of docking, CoMFA with 3D descriptors and HQSAR based on molecular fragments provided an improved understanding in the interaction between the inhibitors and the PARP. This can be utilized for virtual screening to design novel PARP-1 inhibitors.

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