• Title/Summary/Keyword: Quantitative Screening Method

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Validation of QF-PCR for Rapid Prenatal Diagnosis of Common Chromosomal Aneuploidies in Korea

  • Han, Sung-Hee;Ryu, Jae-Song;An, Jeong-Wook;Park, Ok-Kyoung;Yoon, Hye-Ryoung;Yang, Young-Ho;Lee, Kyoung-Ryul
    • Journal of Genetic Medicine
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    • v.7 no.1
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    • pp.59-66
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    • 2010
  • Purpose: Quantitative fluorescent polymerase chain reaction (QF-PCR) allows for the rapid prenatal diagnosis of common aneuploidies. The main advantages of this assay are its low cost, speed, and automation, allowing for large-scale application. However, despite these advantages, it is not a routine method for prenatal aneuploidy screening in Korea. Our objective in the present study was to validate the performance of QF-PCR using short tandem repeat (STR) markers in a Korean population as a means for rapid prenatal diagnosis. Material and Methods: A QF-PCR assay using an Elucigene kit (Gen-Probe, Abingdon, UK), containing 20 STR markers located on chromosomes 13, 18, 21, X and Y, was performed on 847 amniotic fluid (AF) samples for prenatal aneuploidy screening referred for prenatal aneuploidy screening from 2007 to 2009. The results were then compared to those obtained using conventional cytogenetic analysis. To evaluate the informativity of STR markers, the heterozygosity index of each marker was determined in all the samples. Results: Three autosomes (13, 18, and 21) and X and Y chromosome aneuploidies were detected in 19 cases (2.2%, 19/847) after QF-PCR analysis of the 847 AF samples. Their results are identical to those of conventional cytogenetic analysis, with 100% positive predictive value. However, after cytogenetic analysis, 7 cases (0.8%, 7/847) were found to have 5 balanced and 2 unbalanced chromosomal abnormalities that were not detected by QF-PCR. The STR markers had a slightly low heterozygosity index (average: 0.76) compared to those reported in Caucasians (average: 0.80). Submicroscopic duplication of D13S634 marker, which might be a unique finding in Koreans, was detected in 1.4% (12/847) of the samples in the present study. Conclusion: A QF-PCR assay for prenatal aneuploidy screening was validated in our institution and proved to be efficient and reliable. However, we suggest that each laboratory must perform an independent validation test for each STR marker in order to develop interpretation guidelines of the results and must integrate QF-PCR into the routine cytogenetic laboratory workflow.

Arrangement of Disposal Holes According to the Features of Groundwater Flow (지하수 유동 특성을 이용한 심층처분의 처분공 배치 방안)

  • Ko, Nak-Youl;Baik, Min-Hoon
    • Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
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    • v.14 no.4
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    • pp.321-329
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    • 2016
  • Based on the results of groundwater flow system modeling for a hypothetical deep geological repository site, quantitative and spatial distributions of groundwater flow rates at the positions of deposition holes, groundwater travel length and time from the positions to the surface environment were analyzed and used to suggest a method for determining locations of deposition holes. The hydraulic head values at the depth of the deposition holes and a particle tracking method were used to calculate the ground-water flow rates and groundwater travel length and time, respectively. From the results, an approach to designing a layout of deposition holes was suggested by selecting relatively favorable positions for maintaining performance of the disposal facility and screening some positions of deposition holes that did not comply with specific constraints for the groundwater flow rates, travel length and time. In addition, a method for determining a geometrical direction for extension of the disposal facility was discussed. Designing the layout of deposition holes with the information of groundwater flow at the disposal depth can contribute to secure performance and safety of the disposal facility.

Evaluation and Selection Method of Best Available Techniques for Integrated Environmental Management System (통합환경관리제도 운영을 위한 최적가용기법 평가·선정기법 연구)

  • Park, Jae Hong
    • Journal of Korean Society on Water Environment
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    • v.33 no.3
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    • pp.348-358
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    • 2017
  • The process of evaluating and selecting the best available techniques presents various characteristics for each country. In the case of EU, BAT is selected through TWG meeting after first screening, mass and energy balance, impact assessment and decision support process. Korea has proposed four principles to select BAT that can be carbon neutral for each environmental infrastructure in order to reduce greenhouse gas emissions. In order to evaluate and select the best available technique, it is necessary to differentiate the method according to whether it is a technique generally applied at the current workplace, whether it is a single technique or a combination technique, and whether it is a technology technique or management technique. In the case of a single technique, it should be evaluated whether it is a technique applied in the workplace, excessive cost, superior environmental technique over BAT, and secondary environmental pollution. In the case of multiple techniques, it is necessary to examine whether the emission standards are met and whether the pollutants can be treated at the same level as BAT. In the case of BAT candidates for management techniques, whether or not they contribute directly or indirectly to lowering the emission level of pollutants can be an important evaluation item. In the case of environmental techniques that are not generally applied in the workplace, it is recommended that the following 8 steps be carried out, including those prescribed by law. In the first stage, the list of performance evaluation factors is listed. In the second stage, the level of disposal of pollutants and the level of satisfaction with standards are listed. In the third stage, the environmental evaluation elements are listed. In the fourth stage, Is to list the economic evaluation elements, step 6 is to list the pollution and accident prevention evaluation factors, step 7 is the quantitative evaluation of the technical working group, and step 8 is BAT confirmation through deliberation of the central environmental policy committee.

Chemical Composition and Antimicrobial Activity of Essential Oil Extracted from Eucalyptus citriodora Leaf

  • Insuan, Wimonrut;Chahomchuen, Thippayarat
    • Microbiology and Biotechnology Letters
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    • v.48 no.2
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    • pp.148-157
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    • 2020
  • Eucalyptus oil is a rich source of bioactive compounds with a variety of biological activities and is widely used in traditional medicine. Eucalyptus citriodora is cultivated for the production of essential oils. However, the mode of antibacterial action of essential oils from E. citriodora is not well-known. This study aimed to determine the chemical components, microbial inhibitory effect, and mechanism of action of the essential oil from E. citriodora. The oil was extracted from E. citriodora leaves by hydro-distillation and the chemical components were analyzed using gas chromatography-mass spectrometry. The antibacterial activities of eucalyptus oil against gram-positive bacteria (Bacillus subtilis, Staphylococcus aureus, and Staphylococcus intermedius) and gram-negative bacteria (Escherichia coli and Pseudomonas aeruginosa) were screened by disc diffusion method and quantitative analysis was conducted by the microdilution method. The mechanism of action of the extracted essential oil was observed using SEM and analyzed by SDS-PAGE. The major components of E. citriodora oil were citronellal (60.55 ± 0.07%), followed by dl-isopulegol (10.57 ± 0.02%) and citronellol (9.04 ± 0.03%). The antibacterial screening indicated that E. citriodora oil exhibited prominent activity against all tested strains. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against B. subtilis were 0.5% and 1.0%, respectively. The MIC and MBC concentrations against S. aureus, S. intermedius, E. coli, and P. aeruginosa were 1% and 2%, respectively. As observed by SEM, the antibacterial mechanism of E. citriodora oil involved cell wall damage; SDS-PAGE revealed decrease in protein bands compared to untreated bacteria. Thus, E. citriodora oil showed significant antimicrobial properties and caused cellular damage.

Separation Characteristic and Recycling of Excavated Materials Containing Waste (폐기물혼입굴착물의 선별특성과 재활용성 평가)

  • Lee, Suyoung;Kim, Kyuyeon;Jeon, Taewan;Shin, Sunkyoung
    • Journal of the Korea Organic Resources Recycling Association
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    • v.27 no.2
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    • pp.5-12
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    • 2019
  • The study is carried out to survey the proper management and to propose an eco - friendly separation system through efficient screening and resource recovery of excavated materials containing waste from various excavating fields such as reconstruction of landfill sites for reuse, reclamation of unsanitary landfill and residential land development of waste dumping sites. The current status and screening process and analytical characteristics of the excavated materials containing waste were reviewed. Through the analysis of the samples such as separated combustibles, recyclable soils and residues collected from the on-site visits we were able to understand the characteristics of separated materials and excavated materials containing waste such as calorific value, elementary composition, TOC, foreign material content and LOI. It has been found that elimination of the moisture of excavations, removal of attached soil from the surfaces of the excavated combustibles and the quantitative supply method of the input devices are the main operating factors as essential factors for the optimal separation of excavated materials containing waste. For efficient management and recycling of excavated materials containing, it is necessary to set criteria of ash content in separated combustibles and criteria organic matter content in separated soils.

Amine functionalized plasma polymerized PEG film: Elimination of non-specific binding for biosensing

  • Park, Jisoo;Kim, Youngmi;Jung, Donggeun;Kim, Young-Pil;Lee, Tae Geol
    • Proceedings of the Korean Vacuum Society Conference
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    • 2016.02a
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    • pp.378.2-378.2
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    • 2016
  • Biosensors currently suffer from severe non-specific adsorption of proteins, which causes false positive errors in detection through overestimation of the affinity value. Overcoming this technical issue motivates our research. Polyethylene glycol (PEG) is well known for its ability to reduce the adsorption of biomolecules; hence, it is widely used in various areas of medicine and other biological fields. Likewise, amine functionalized surfaces are widely used for biochemical analysis, drug delivery, medical diagnostics and high throughput screening such as biochips. As a result, many coating techniques have been introduced, one of which is plasma polymerization - a powerful coating method due to its uniformity, homogeneity, mechanical and chemical stability, and excellent adhesion to any substrate. In our previous works, we successfully fabricated plasmapolymerized PEG (PP-PEG) films [1] and amine functionalized films [2] using the plasma enhanced chemical vapor deposition (PECVD) technique. In this research, an amine functionalized PP-PEG film was fabricated by using the plasma co-polymerization technique with PEG 200 and ethylenediamine (EDA) as co-precursors. A biocompatible amine functionalized film was surface characterized by X-ray photoelectron spectroscopy (XPS) and Fourier-transform infrared spectroscopy (FT-IR). The density of the surface amine functional groups was carried out by quantitative analysis using UV-visible spectroscopy. We found through surface plasmon resonance (SPR) analysis that non-specific protein adsorption was drastically reduced on amine functionalized PP-PEG films. Our functionalized PP-PEG films show considerable potential for biotechnological applications such as biosensors.

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Quantification of Icariin Contents in Epimedium koreanum N. by Using a Near Infrared Reflectance Spectroscopy (NIRS를 이용한 삼지구엽초의 이카린 함량 분석)

  • Kim, Yong-Ho;Choi, Byoung-Ryourl;Baek, Hum-Young;Lee, Young-Sang
    • Korean Journal of Medicinal Crop Science
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    • v.10 no.5
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    • pp.340-343
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    • 2002
  • Near infrared reflectance spectroscopy (NIRS) has become widely accepted for rapid quantitative analysis of components in some crops. Our object was to determine icariin contents in whole plant of Epimedium koreanum by using an NIRS system. Total 150 plant samples previously analyzed by HPLC were scanned by NIRS and 68 samples were selected for calibration and validation equation. A calibration equation calculated by MPLS(modified partial least squares) regression technique was developed and a coefficient of determination in calibration and validation sets were 0.95 and 0.82, respectively. A comparison between NIRS estimation and HPLC value was performed with the remaining samples not included in the calibration and validation sets. Most of samples also showed a positive correlation like a validation set. Our results demonstrate that this developed NIRS equation can be practically used as a mass screening method for rapid quantification of icarin contents in Epimedium koreanum N.

A Study on Penicillin Residue In Market Milk in Seoul (시판 우유중의 잔류 Penicillin에 관한 실험연구)

  • 강윤형;이용욱
    • Journal of Environmental Health Sciences
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    • v.2 no.1
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    • pp.11-15
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    • 1975
  • The study Was carried out to determine the residue concentrations of peniccillin in Merket milk samples collected from some of the retail shops located in Seoul City. During the period from June to September 1974, 160 milk samples were collected and examined, using the Filter Paper Disk Method, for Screening test and Quantitative test of penicillin residue. The results obtained in the study were as follows: 1. Among the 160 Market Milk samples. 28 samples (17.5%) proved to contain some antibiotics residue. 2. In the case of the antibiotics residue positive milk samples, the rate of penicillin residue alone was 46.4% and the rate of the other antibiotics was 53.6%. 3. In the distribution of milk plants examined in relation-to the antibiotics residue positive milk samples, it was shown that S.K.H. and M.Market Milk plants had 15.0, 17.5, 20.0 and 17.5 percent respectively. 4. According to the monthly distribution, the highest antibiotics residue positive rate (25.0% of the samples) was shown in the samples collected in August, and the next (20.0% of the samples) in July. While the samples collected in June and September 1974 Showed lower rates of 7.5% and 17.5% respectively. 5. The range of the residue concentraction of penicillin in the positive milk samples were as follows: a. 53.8% of the total samples were found in the lowest range of 0.02-0.05 IU/ml. b. 30.8% of the total samples were found in the lower range of 0.05-0.1 IU/ml. c. 7.7% of the total samlpes were found in the higher range of 0.1-0.2 IU/ml. d. 7.7% of the total samples were found in the highest range of over 0.2 IU/ml.

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Effective Application of Multiplex RT-PCR for Characterization of Human Embryonic Stem Cells/ Induced Pluripotent Stem Cells (다중 역전사 중합효소 연쇄 반응(Multiplex RT-PCR)을 이용한 인간배아 줄기세포 및 유도만능 줄기세포의 효과적인 분화 양상 조사)

  • Kim, Jung-Mo;Cho, Youn-Jeong;Son, On-Ju;Hong, Ki-Sung;Chung, Hyung-Min
    • Reproductive and Developmental Biology
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    • v.35 no.1
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    • pp.1-8
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    • 2011
  • Techniques to evaluate gene expression profiling, such as sufficiently sensitive cDNA microarrays or real-time quantitative PCR, are efficient methods for monitoring human pluripotent stem cell (hESC/iPSC) cultures. However, most of these high-throughput tests have a limited use due to high cost, extended turn-around time, and the involvement of highly specialized technical expertise. Hence, there is an urgency of rapid, cost-effective, robust, yet sensitive method development for routine screening of hESCs/hiPSCs. A critical requirement in hESC/hiPSC cultures is to maintain a uniform undifferentiated state and to determine their differentiation capacity by showing the expression of gene markers representing all three germ layers, including ectoderm, mesoderm, and endoderm. To quantify the modulation of gene expression in hESCs/hiPSC during their propagation, expansion, and differentiation via embryoid body (EB) formation, we developed a simple, rapid, inexpensive, and definitive multimarker, semiquantitative multiplex RT-PCR platform technology. Among the 9 gene primers tested, 5 were pluripotent markers comprising set 1, and 3 lineage-specific markers were combined as set 2, respectively. We found that these 2 sets were not only effective in determining the relative differentiation in hESCs/hiPSCs, but were easily reproducible. In this study, we used the hES/hiPS cell lines to standardize the technique. This multiplex RT-PCR assay is flexible and, by selecting appropriate reporter genes, can be designed for characterization of different hESC/hiPSC lines during routine maintenance and directed differentiation.

Clinical Prediction Based on HPV DNA Testing by Hybrid Capture 2 (HC2) in Combination with Liquid-based Cytology (LBC)

  • Junyangdikul, Pairoj;Tanchotsrinon, Watcharaporn;Chansaenroj, Jira;Nilyaimit, Pornjarim;Lursinsap, Chidchanok;Poovorawan, Yong
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.2
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    • pp.903-907
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    • 2013
  • Primary screening by HPV DNA testing is an effective method for reducing cervical cancer and has proven more sensitive than cytology. To advance this approach, many molecular methods have been developed. Hybrid capture 2 provides semi-quantitative results in ratios of relative light units and positive cutoff values (RLU/PC). Twenty-five thousand and five patients were included in this study to analyze the correlation between the ratio of RLU/PC and stage of cervical dysplasia. The results show that the RLU/PC ratios ranged from 0-3500 while almost normal cases, ASC-US and ASC-H, had values below 200. Of those samples negative for cytology markers, 94.6% were normal and their RLU/PC ratios were less than 4. With an RLU/PC ratio greater than 4 and less than or equal to 300, the percentages in all age groups were normal 53.6%, LSIL 20.2%, ASC-US 17.2%, HSIL 6.13%, ASC-H 2.72%, and AGC 0.11%, respectively. In contrast, 64.0% of samples with a RLU/PC ratio greater than 300 and less than or equal to 3500 were LSIL. These results should contribute to cost effective cervical cancer management strategies. Further studies of associations with particular HPV genotypes would be useful to predict the risk of progression to cancer.