• Title/Summary/Keyword: Reaction monitoring

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Utility of a multiplex reverse transcriptase-polymerase chain reaction assay (HemaVision) in the evaluation of genetic abnormalities in Korean children with acute leukemia: a single institution study

  • Kim, Hye-Jin;Oh, Hyun Jin;Lee, Jae Wook;Jang, Pil-Sang;Chung, Nack-Gyun;Kim, Myungshin;Lim, Jihyang;Cho, Bin;Kim, Hack-Ki
    • Clinical and Experimental Pediatrics
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    • v.56 no.6
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    • pp.247-253
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    • 2013
  • Purpose: In children with acute leukemia, bone marrow genetic abnormalities (GA) have prognostic significance, and may be the basis for minimal residual disease monitoring. Since April 2007, we have used a multiplex reverse transcriptase-polymerase chain reaction tool (HemaVision) to detect of GA. Methods: In this study, we reviewed the results of HemaVision screening in 270 children with acute leukemia, newly diagnosed at The Catholic University of Korea from April 2007 to December 2011, and compared the results with those of fluorescence in situ hybridization (FISH), and G-band karyotyping. Results: Among the 270 children (153 males, 117 females), 187 acute lymphoblastic leukemia and 74 acute myeloid leukemia patients were identified. Overall, GA was detected in 230 patients (85.2%). HemaVision, FISH, and G-band karyotyping identified GA in 125 (46.3%), 126 (46.7%), and 215 patients (79.6%), respectively. TEL-AML1 (20.9%, 39/187) and AML1-ETO (27%, 20/74) were the most common GA in ALL and AML, respectively. Overall sensitivity of HemaVision was 98.4%, with false-negative results in 2 instances: 1 each for TEL-AML1 and MLL-AF4. An aggregate of diseases-specific FISH showed 100% sensitivity in detection of GA covered by HemaVision for actual probes utilized. G-band karyotype revealed GA other than those covered by HemaVison screening in 133 patients (49.3%). Except for hyperdiplody and hypodiploidy, recurrent GA as defined by the World Health Organizationthat were not screened by HemaVision, were absent in the karyotype. Conclusion: HemaVision, supported by an aggregate of FISH tests for important translocations, may allow for accurate diagnosis of GA in Korean children with acute leukemia.

Oral tissue response to soft tissue expanders prior to bone augmentation: in vitro analysis and histological study in dogs

  • Yoo, Jung Min;Amara, Heithem Ben;Kim, Min Kyoung;Song, Ju Dong;Koo, Ki-Tae
    • Journal of Periodontal and Implant Science
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    • v.48 no.3
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    • pp.152-163
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    • 2018
  • Purpose: To determine whether the swelling and mechanical properties of osmotic self-inflating expanders allow or not the induction of intraoral soft tissue expansion in dogs. Methods: Three different volumes (0.15, 0.25, and 0.42 mL; referred to respectively as the S, M, and L groups) of soft tissue expanders (STEs) consisting of a hydrogel core coated with a silicone-perforated membrane were investigated in vitro to assess their swelling behavior (volume swelling ratio) and mechanical properties (tensile strength, tensile strain). For in vivo investigations, the STEs were subperiosteally inserted for 4 weeks in dogs (n=5). Soft tissue expansion was clinically monitored. Histological analyses included the examination of alveolar bone underneath the expanders and thickness measurements of the surrounding fibrous capsule. Results: The volume swelling ratio of all STEs did not exceed 5.2. In tensile mode, the highest mean strain was registered for the L group ($98.03{\pm}0.3g/cm$), whereas the lowest mean value was obtained in the S group ($81.3{\pm}0.1g/cm$), which was a statistically significant difference (P<0.05). In addition, the S and L groups were significantly different in terms of tensile strength ($1.5{\pm}0.1g/cm$ for the S group and $2.2{\pm}0.1g/cm$ for the L group, P<0.05). Clinical monitoring showed successful dilatation of the soft tissues without signs of inflammation up to 28 days. The STEs remained volumetrically stable, with a mean diameter in vivo of 6.98 mm, close to the in vitro post-expansion findings (6.69 mm). Significant histological effects included highly vascularized collagen-rich fibrous encapsulation of the STEs, with a mean thickness of $0.67{\pm}0.12mm$. The bone reaction consisted of resorption underneath the STEs, while apposition was observed at their edges. Conclusions: The swelling and mechanical properties of the STEs enabled clinically successful soft tissue expansion. A tissue reaction consisting of fibrous capsule formation and bone loss were the main histological events.

Molecular Detection and Epidemiology of Etiologic Agents among Children with Acute Gastroenteritis at a Secondary Hospital from 2015 to 2018 (2015년부터 2018년까지 일개 이차병원에서 동정된 소아 급성 위장염 원인 병원체의 분자진단과 역학의 임상적 연구)

  • Kim, Young Sang;Chung, Ju Young
    • Pediatric Infection and Vaccine
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    • v.27 no.2
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    • pp.90-101
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    • 2020
  • Purpose: The multiplex reverse transcriptase-polymerase chain reaction (RT-PCR) test developed recently can help detect enteric pathogens of acute gastroenteritis (AGE). This study aimed to investigate the epidemiology of pathogens in children with AGE using the multiplex RT-PCR. Methods: From May 2015 to June 2018, multiplex RT-PCR tests were performed to identify pathogens in the feces of pediatric patients diagnosed with AGE at a secondary hospital in Seoul, Korea. Results: Of the 1,366 stool samples examined for viral pathogens, 483 (35.3%) tested positive for ≥1 pathogen. Group A rotavirus (RV) was detected in 106 cases (7.8%). The positivity rate increased annually from 3.0% (8/263) to 16.7% (48/288) and surged in 2018 (P<0.001). Norovirus (NoV) GII was the most common viral pathogen (263/1,366, 19.3%), and the positivity rate did not increase during the 3 years. Of the 304 stool samples tested for bacterial pathogens, Campylobacter spp. was the most common bacterial pathogen (32/304, 10.5%), followed by Clostridium difficile (22/304, 7.2%) and Salmonella spp. (17/304, 5.6%). The positivity rate of these bacterial pathogens did not change significantly during the study period. Conclusions: NoV GII is the main pathogen in childhood AGE since the introduction of RV vaccine, yet the number of rotavirus-infected patients increased during our study, especially in 2018. Therefore, further research is needed including the possibility of emergence of novel RV strains. Campylobacter spp. is the predominant cause of bacterial AGE in children. For proper treatment, the clinical characteristics of the bacteria should be taken into consideration, and continuous monitoring is necessary.

The Prevalence and Distribution of the P and G Genotypes of a Group A Rotavirus Detected in Acute Gastroenteritis Patients from Incheon (인천지역 급성 설사환자의 group A rotavirus 감염 실태 및 P와 G 유전자형 분포)

  • Choi, Hye-Jin;Oh, Bo-Young;Lee, Mi-Yeon;Koh, Yeon-Ja;Gong, Young-Woo;Hur, Myung-Je;Lee, Jea-Mann;Kim, Young-Hee;Jeong, Hye-Sook;Cheon, Doo-Sung
    • Journal of Life Science
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    • v.22 no.5
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    • pp.600-604
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    • 2012
  • Rotavirus is the main cause of severe diarrhea in infants and young children of the world. However, the frequency of genetic alterations makes it hard to control the prophylaxis. Therefore, continuous monitoring of the rotavirus's genetic change is inevitable to prevent disease prevalence and is useful in inventing an efficient vaccine. From January 2005 to December 2010, we investigated 11,607 stool samples of acute gastroenteritis patients in the Incheon metropolitan area. About 13.18% (1,530 stool samples) of all samples had a positive reaction against rotavirus using an antigen capture enzyme-linked immunosorbent assay (ELISA). Then, the 160 stool samples were searched for subtypes of group A rotavirus by using a reverse transcription polymerase chain reaction (RT-PCR) and a nested multiplex RCR. In P sub-typing, P8 (56.3%) was an extremely prevalent genotype, followed by P6 (21.3%), and P1A (10.0%). G1 (39.4%) was most widespread in the G subtype, followed by G4 (25.0%) and G3 (18.8%). G1P8 (35.5%) was the most common G and P subtype combination, followed by G4P6 (19.3%) and G3P8 (13.1%). These results might be useful data for understanding the epidemiological status of group A-rotavirus dispersion in the Incheon metropolitan area.

The influence of misinformation on memory: detection of original memory using concealed information test (CIT) (기억에 대한 오정보의 영향: 숨긴정보검사를 이용한 원기억의 탐지)

  • Han, Yuhwa;Park, Kwangbai
    • Science of Emotion and Sensibility
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    • v.18 no.2
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    • pp.85-100
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    • 2015
  • This study aimed at examining if the original memory remains after a misinformation is presented, using Event-Related Potential based Concealed Information Test (ERP-based CIT). In the first stage of the study, the participant was presented with either the original information or a misleading information after experiencing an event (Post-information). The second stage was to measure brain wave and reaction time on the original, misleading, and irrelevant information (CIT-Stimulus). P300 amplitude, P300 area, P300 latency, and reaction time were used as dependant variables. In the result, a significant Post-information ${\times}$ CIT-Stimulus interaction effect was found on the P300 area measured at Cz, Pz, and Oz area. This interaction effect implied the possibility that the original information could be partially impaired in memory by misleading information presented afterward. P300 amplitude at Pz area did not differ between the accurate and the misleading stimuli in the condition in which a misleading information was presented. This result can be explained by source monitoring error. In discussion, the limitations of this study and directions of future studies were discussed.

Pretreatment method of urinary thiodiglycolic acid as metabolite of vinyl chloride (염화비닐의 요중 대사물질인 thiodiglycolic acid의 분석을 위한 전처리 조건)

  • Hong, Joo Youn;Kim, Chi Nyon;Jung, Jae Hoon;Chang, Jung Hwan;Roh, Jaehoon
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.9 no.1
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    • pp.23-40
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    • 1999
  • The analysis of thiodiglycolic acid in urine has been used as an index of biological exposure to vinyl chloride. Unfortunately thiodiglycolic acid has a strong hydrophilic character, because it has two carboxylic groups, so that it can only be extracted with organic solvent with a great difficulty. Underivatized thiodiglycolic acid tends to tail because of non-specific interaction with the inert support. Therefore, esterification is the obvious first choice for derivatization of thiodiglycolic acid, particularly for gas chromatography. In this study, the focus of interest is to compare two method of esterifications (methylation and silylation). Methylation is to make the methyl ester of thiodiglycolic acid by reaction with diazomethane. Silylation is to make the trimethylsilyl ester of thiodiglycolic acid by reaction with N-trimethylsily-ldiethylamine. The results and conclusions are as the following: 1. The detection limit (sensitivity) of methylated thiodiglycolic acid was $5.00{\mu}g/m{\ell}$ and silylated thiodiglycolic acid was $3.07{\mu}g/m{\ell}$ by gas chromatography with flame ionization detector. 2. The optimal liquid-liquid extraction of thiodiglycolic acid was as following: To each of the tubes, $15m{\ell}$ of urine, concentrated sulfuric acid (pH 1 - 2) and 5 gsodium sulfate were added. The samples was extracted three times with $5m{\ell}$ ethylacetate each time. 3. The methylated thiodiglycolic acid was more stable than silylated thiodiglycolic acid in extractional solvent which contained humidity. 4. The precision (pooled coefficient of variation for 4 days) of the analysis was 0.07324 in methylated thiodiglycolic acid with external standard calibration, and 0.07033 in methylated thiodiglycolic acid with internal standard calibration. 5. The precision (pooled coefficient of variation for 4 days) of the analysis was 0.10914 in silylated thiodiglycolic acid with external standard calibration, and 0.13602 in silylated thiodiglycolic acid with internal standard calibration. From the above results, the analysis of methylated thiodiglycolic acid was more sensitive (limit of detection) than silylated thiodiglycolic acid by gas chromatography. However, the methylated thiodiglycolic acid was stable in the humidity and was separated sharply on chromatogram. Also, analysis of methylated thiodiglycolic acid was more precise (pooled coefficient of variation for 4 days) than silylated thiodiglycolic acid. In conclusion, it is established that the analysis of methylated thiodiglycolic acid is appropriate for biological monitoring of exposure to vinyl chloride.

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Performance Evaluation of PCR Kits for Detecting Genetically Modified Crop Ingredients (유전자 변형 작물 성분 검출용 PCR Kit의 성능 평가 연구)

  • 윤시온;정순천;윤원기;박상규;문제선;이정현;김환묵
    • Toxicological Research
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    • v.20 no.2
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    • pp.101-108
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    • 2004
  • The different social reflections about the benefits and the potential risks of genetically modified (GM) crops have evolved with .different reactions in different countries. Many countries including Korea are working toward setting down new guidelines. Korea requires companies to label all food that contains more than 3% GM ingredients. One of the rapid and convenient detection methods of GM ingredients is amplification of the introduced DNAs by polymerase chain reaction (PCR). Many PCR kits for this purpose are commercially available. The objective of this study was to evaluate performance of commercialized GM crop detection kits. The results showed that 6 out of 15 kits tested did not meet the requirements even purposed by the manufacturers themselves in terms of stability, reproducibility, and detection limits, suggesting a potential quality control problem in their design stage or production line. The evaluation also suggests that, although the duplex and triplex detection kits allowed unambiguous detection in a single PCR reaction, the monoplex detection kits were the most sensitive to the detection of GM ingredients. The detection limits also differ between soybean and corn. Results from this study will be useful in the development of sound qualitative tracking systems of GM ingredients for monitoring throughout the cultivation of GM crops, their trans-boundary movement, and food production using GM grains as well as for complying with government guidelines associated with GM crops.

Pan-serotype reverse transcription loop-mediated isothermal amplification (RT-LAMP) for the rapid detection of foot-and-mouth disease virus (구제역바이러스 신속진단을 위한 pan-serotype reverse transcription loop-mediated isothermal amplification (RT-LAMP) 진단법)

  • Lim, Da-Rae;Park, Yu-Ri;Park, Sun-Young;Kim, Hye-Ryung;Park, Min-Ji;Ku, Bok-Kyung;Nah, Jin-Ju;Ryoo, So-Yoon;Wee, Sung-Hwan;Jeon, Hyo-Sung;Kim, Ji-Jeong;Jeon, Bo-Young;Lee, Hyeong-Woo;Park, Choi-Kyu
    • Korean Journal of Veterinary Service
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    • v.41 no.1
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    • pp.29-39
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    • 2018
  • In this study, we developed a sensitive and specific reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for rapid visual detection of foot-and-mouth disease virus (FMDV) circulated in Korea. The RT-LAMP was completed in 40 min at $62^{\circ}C$ and the results of the assay were directly detected by naked eye without any detection process. The assay specifically amplified all 7 serotypes of FMDV RNAs but not amplified other viral and cellular nucleic acids. The sensitivity of the RT-LAMP was $10^2$, $10^3$ and $10^3TCID_{50}/mL$ for serotype O, A and Asia 1 FMDV, respectively, which was comparable to conventional reverse transcription polymerase chain reaction (RT-PCR) and relatively lower than that of real time quantitative RT-PCR (qRT-PCR). Clinical evaluation of the RT-LAMP using different serotypes of Korean and foreign FMDV strains showed a 100% (35/35) agreement with the results of the RT-PCR and qRT-PCR. These results indicated that RT-LAMP assay developed in this study could be a valuable diagnostic method for FMDV monitoring and surveillance.

Integrated RT-PCR Microdevice with an Immunochromatographic Strip for Colorimetric Influenza H1N1 virus detection

  • Heo, Hyun Young;Kim, Yong Tae;Chen, Yuchao;Choi, Jong Young;Seo, Tae Seok
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.08a
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    • pp.273-273
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    • 2013
  • Recently, Point-of-care (POC) testing microdevices enable to do the patient monitoring, drug screening, pathogen detection in the outside of hospital. Immunochromatographic strip (ICS) is one of the diagnostic technologies which are widely applied to POC detection. Relatively low cost, simplicity to use, easy interpretations of the diagnostic results and high stability under any circumstances are representative advantages of POC diagnosis. It would provide colorimetric results more conveniently, if the genetic analysis microsystem incorporates the ICS as a detector part. In this work, we develop a reverse transcriptase-polymerase chain reaction (RT-PCR) microfluidic device integrated with a ROSGENE strip for colorimetric influenza H1N1 virus detection. The integrated RT-PCR- ROSGENE device is consist of four functional units which are a pneumatic micropump for sample loading, 2 ${\mu}L$ volume RT-PCR chamber for target gene amplification, a resistance temperature detector (RTD) electrode for temperature control, and a ROSGENE strip for target gene detection. The device was fabricated by combining four layers: First wafer is for RTD microfabrication, the second wafer is for PCR chamber at the bottom and micropump channel on the top, the third is the monolithic PDMS, and the fourth is the manifold for micropump operation. The RT-PCR was performed with subtype specific forward and reverse primers which were labeled with Texas-red, serving as a fluorescent hapten. A biotin-dUTP was used to insert biotin moieties in the PCR amplicons, during the RT-PCR. The RT-PCR amplicons were loaded in the sample application area, and they were conjugated with Au NP-labeled hapten-antibody. The test band embedded with streptavidins captures the biotin labeled amplicons and we can see violet colorimetric signals if the target gene was amplified with the control line. The off-chip RT-PCR amplicons of the influenza H1N1 virus were analyzed with a ROSGENE strip in comparison with an agarose gel electrophoresis. The intensities of test line was proportional to the template quantity and the detection sensitivity of the strip was better than that of the agarose gel. The test band of the ROSGENE strip could be observed with only 10 copies of a RNA template by the naked eyes. For the on-chip RT-PCR-ROSGENE experiments, a RT-PCR cocktail was injected into the chamber from the inlet reservoir to the waste outlet by the micro-pump actuation. After filling without bubbles inside the chamber, a RT-PCR thermal cycling was executed for 2 hours with all the microvalves closed to isolate the PCR chamber. After thermal cycling, the RT-PCR product was delivered to the attached ROSGENE strip through the outlet reservoir. After dropping 40 ${\mu}L$ of an eluant buffer at the end of the strip, the violet test line was detected as a H1N1 virus indicator, while the negative experiment only revealed a control line and while the positive experiment a control and a test line was appeared.

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Evaluation of DOM Variations and Reduction Effects in Bioreation Artificial Wetland (생물반응 인공습지 내 DOM 변동 및 저감효과 평가)

  • Joo, Kwangjin;Lee, Jongjun;Kim, Tea-Kyung;Choi, Isong;Chang, Kwang-hyeon;Joo, Jinchul;Oh, Jongmin
    • Journal of Environmental Impact Assessment
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    • v.27 no.6
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    • pp.582-594
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    • 2018
  • In this study, the vertical and horizontal flow wetlands were combined in series to create conditions for flow in the exhalation and anaerobic state with the aim of monitoring the variability and reduction of dissolved organic matterin the bio-reactive artificial wetlands, and the performance assessment was conducted as acrylic reaction groups by designing artificial wetlands that filled the functionalresiduals. In case of artificial wetlands in vertical and horizontal planes, the concentration of dissolved oxygen (DO) in the reaction tank was measured as 2.7 mg/L in the vertical flow wetlands under exhalation, and N.D. in the horizontal flow artificial wetlands under anaerobic conditions. The test was carried out by changing the operation time to 140 min, 80 min, and 60 min. The test was conducted with the same natural operation time of 20 min depending on the operation time. All hours of operation were shown to be due to microbial activity. In 3D-EEM, it was found that the longer the driving time was taken, the more reduction the organic compounds in the areas of insoluble human resources, III and V. Further research on the mechanism analysis of future reduction effects is expected to be carried out, but the findings are expected to contribute to the development of technologies for reducing obfuscated substances using artificial wetlands in the future.