• Title/Summary/Keyword: Reaction monitoring

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Detection and Quantification of Fusarium oxysporum f. sp. niveum Race 1 in Plants and Soil by Real-time PCR

  • Zhong, Xin;Yang, Yang;Zhao, Jing;Gong, Binbin;Li, Jingrui;Wu, Xiaolei;Gao, Hongbo;Lu, Guiyun
    • The Plant Pathology Journal
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    • v.38 no.3
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    • pp.229-238
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    • 2022
  • Fusarium wilt caused by Fusarium oxysporum f. sp. niveum (Fon) is the most serious soil-borne disease in the world and has become the main limiting factor of watermelon production. Reliable and quick detection and quantification of Fon are essential in the early stages of infection for control of watermelon Fusarium wilt. Traditional detection and identification tests are laborious and cannot efficiently quantify Fon isolates. In this work, a real-time polymerase chain reaction (PCR) assay has been described to accurately identify and quantify Fon in watermelon plants and soil. The FONRT-18 specific primer set which was designed based on identified specific sequence amplified a specific 172 bp band from Fon and no amplification from the other formae speciales of Fusarium oxysporum tested. The detection limits with primers were 1.26 pg/µl genomic DNA of Fon, 0.2 pg/ng total plant DNA in inoculated plant, and 50 conidia/g soil. The PCR assay could also evaluate the relationships between the disease index and Fon DNA quantity in watermelon plants and soil. The assay was further used to estimate the Fon content in soil after disinfection with CaCN2. The real-time PCR method is rapid, accurate and reliable for monitoring and quantification analysis of Fon in watermelon plants and soil. It can be applied to the study of disease diagnosis, plant-pathogen interactions, and effective management.

State Machine design to support behavioral response in DTT protocol (불연속 개별시도 훈련에서 행동 반응을 지원하는 상태머신 설계)

  • Yun, Hyuk;Yun, Sang-Seok
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2022.05a
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    • pp.147-149
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    • 2022
  • This paper proposes a state machine design methodology in which an interactive robot that mimics discrete trial training (DTT protocol) can support social interaction training for children with autism. The robot applied to social interaction training uses the response to the provided training stimulus as a quantitative indicator by processing the data received from the sensors measuring the behavioral response of the child. In this process, the state machine is used as information that classifies the state of the acquired data and provides the subsequent stimulus for DTT protocol. Through the joint attentional training, it can be used as evidence-based treatment information by quantitatively classifying the data on the number of sustainable and DTT protocol and the child's response, as well as the current reaction status of the child to the observer performing remote monitoring. At the same time, it was confirmed that it is possible to properly respond to misrecognition situations.

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Multiple effects of nano-silica on the pseudo-strain-hardening behavior of fiber-reinforced cementitious composites

  • Hossein Karimpour;Moosa Mazloom
    • Advances in nano research
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    • v.15 no.5
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    • pp.467-484
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    • 2023
  • Despite the significant features of fiber-reinforced cementitious composites (FRCCs), including better mechanical, fractural, and durability performance, their high content of cement has restricted their use in the construction industry. Although ground granulated blast furnace slag (GGBFS) is considered the main supplementary cementitious material, its slow pozzolanic reaction stands against its application. The addition of nano-sized mineral modifiers, including nano-silica (NS), is an alternative to address the drawbacks of using GGBFS. The main object of this empirical and numerical research is to examine the effect of NS on the strain-hardening behavior of cementitious composites; ten mixes were designed, and five levels of NS were considered. This study proposes a new method, using a four-point bending test to assess the use of nano-silica (NS) on the flexural behavior, first cracking strength, fracture energy, and micromechanical parameters including interfacial friction bond strength and maximum bridging stress. Digital image correlation (DIC) was used for monitoring the initiation and propagation of the cracks. In addition, to attain a deep comprehension of fiber/matrix interaction, scanning electron microscope (SEM) analysis was used. It was discovered that using nano-silica (NS) in cementitious materials results in an enhancement in the matrix toughness, which prevents multiple cracking and, therefore, strain-hardening. In addition, adding NS enhanced the interfacial transition zone between matrix and fiber, leading to a higher interfacial friction bond strength, which helps multiple cracking in the composite due to the hydrophobic nature of polypropylene (PP) fibers. The findings of this research provide insight into finding the optimum percent of NS in which both ductility and high tensile strength of the composites would be satisfied. As a concluding remark, a new criterion is proposed, showing that the optimum value of nano-silica is 2%. The findings and proposed method of this study can facilitate the design and utilization of green cementitious composites in structures.

Determination of methamphetamine, 4-hydroxymethamphetamine, amphetamine and 4-hydroxyamphetamine in urine using dilute-and-shoot liquid chromatography-tandem mass spectrometry (시료 희석 주입 LC-MS/MS를 이용한 소변 중 메스암페타민, 4-하이드록시메스암페타민, 암페타민 및 4-하이드록시암페타민 동시 분석)

  • Heo, Bo-Reum;Kwon, NamHee;Kim, Jin Young
    • Analytical Science and Technology
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    • v.31 no.4
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    • pp.161-170
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    • 2018
  • The epidemic of disorders associated with synthetic stimulants, such as methamphetamine (MA) and amphetamine (AP), is a health, social, legal, and financial problem. Owing to the high potential of their abuse and addiction, reliable analytical methods are required to detect and identify MA, AP, and their metabolites in biological samples. Thus, a dilute-and-shoot liquid chromatography-tandem mass spectrophotometry (LC-MS/MS) was developed for simultaneous determination of MA, 4-hydroxymethamphetamine (4HMA), AP, and 4-hydroxyamphetamine (4HA) in urine. Urine sample ($100{\mu}L$) was mixed with $50{\mu}L$ of mobile phase consisting of 0.4 % formic acid and methanol and $50{\mu}L$ of working internal-standard solution. Aliquots of $8{\mu}L$ diluted urine was injected into the LC-MS/MS system. For all analytes, chromatographic separation was performed using a C18 reversed-phase column with gradient elution and a total run time of 5 min. The identification and quantification were performed by multiple reaction monitoring (MRM). Linear least-squares regression was conducted to generate a calibration curve, with $1/x^2$ as the weighting factor. The linear ranges were 2.0-200, 1.0-800, and 10-2500 ng/mL for 4HA and 4HMA, AP, and MA, respectively. The inter- and intraday precisions were within 6.6 %, whereas the inter- and intraday accuracies ranged from -14.9 to 11.3 %. The low limits of quantification were 2.0 ng/mL (4HA and 4HMA), 1.0 ng/mL (AP), and 10 ng/mL (MA). The proposed method exhibited satisfactory selectivity, dilution integrity, matrix effect, and stability, which are required for validation. Moreover, the purification efficiency of high-speed centrifugation was clearly higher than 6-15 % for QC samples (n=5), which was higher than that of the membrane-filtration method. The applicability of the proposed method was tested by forensic analysis of urine samples from drug abusers.

Determination of finasteride in human serum by LC-MS/MS (LC-MS/MS를 이용한 혈청 중 finasteride 분석)

  • Nam, Hye-Seon;Nam, Kyong-Hee;Jung, Su-Hee;Lee, Jang-Woo;Kang, Jin-Yeong;Hong, Soon-Keun;Kim, Tae-Sung;Kang, Tae-Seok;Yoon, Hae-Jung;Lee, Kwang-Ho;Rhee, Gyu-Seek
    • Analytical Science and Technology
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    • v.24 no.5
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    • pp.345-351
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    • 2011
  • A liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI/MS/MS) method was developed and validated for the determination of finasteride in human serum. Beclomethasone was used as internal standard (IS) and liquid-liquid extraction (LLE) using methyl tert-butyl ether (MTBE) was carried out to isolate analyte. The mass transitions monitored in multiple reaction monitoring (MRM) in positive ion mode were m/z 373.2${\rightarrow}$305.2 for finasteride and m/z 409.3${\rightarrow}$391.2 for IS. Retention times of finasteride and IS were 5.81 and 5.46 min, respectively. The limit of quantitation (LOQ) was 0.1 ng/mL and the calibration curve showed good linearity in the range of 0.1~20.0 ng/mL ($R^2$=0.9997). The intra-day assay precision and accuracy were in the range 6.3~10.6% and 97.3~103.6%, respectively, and the inter-day assay precision and accuracy were in the range 0.8~5.2% and 99.8~102.5%, respectively. The sample extract recovery of the method was 80~83%.

Analysis of dutasteride in human serum by LC-MS/MS (LC-MS/MS를 이용한 혈청 중 dutasteride 분석)

  • Nam, Hye-Seon;Nam, Kyong-Hee;Jung, Su-Hee;Lee, Jang-Woo;Kang, Jin-Yeong;Hong, Soon-Keun;Kim, Tae-Sung;Jung, Ki-Kyung;Kang, Tae-Seok;Yoon, Hae-Jung;Lee, Kwang-Ho;Rhee, Gyu-Seek
    • Analytical Science and Technology
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    • v.25 no.1
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    • pp.76-82
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    • 2012
  • The determination and confirmation of dutasteride in human serum was performed by a liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI/MS/MS). Beclomethasone as an internal standard (I.S.) was added to the serum and the mixed sample was pretreated by liquid-liquid extraction (LLE) with methyl tert-butyl ether (MTBE). The mass transitions of dutasteride and I.S. monitored in multiple reaction monitoring (MRM) were m/z 529.6${\rightarrow}$461.5 and m/z 409.3${\rightarrow}$391.2, respectively, and the retention times were 6.45 and 5.46 min, respectively. The calibration curve was linear in the concentration range of 0.5~30.0 ng/mL ($R^2$= 0.9999) and the limit of quantitation (LOQ) was found to be 0.5 ng/mL. The recovery of dutasteride was shown to be 66~72%. The intra-day assay precision and accuracy were in the range 3.5~5.5% and 85.7~89.9%, respectively, and the interday assay precision and accuracy were in the range 4.2~5.8% and 90.8~95.8%, respectively.

Development and Validation of the Determination of Sorafenib in Human Plasma using Tandem Mass Spectrometry Coupled with Liquid Chromatography (고속액체크로마토그래피 텐덤질량분석기법을 이용한 사람 혈장 내 소라페닙 농도분석법의 개발 및 검정)

  • Park, Daejin;Lee, Sunggon;Kim, Woomi
    • Journal of Life Science
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    • v.22 no.11
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    • pp.1456-1462
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    • 2012
  • Sorafenib is a multikinase inhibitor and an oral anticancer drug approved for the treatment of patients with advanced renal cell carcinoma and those with unresectable hepatocellular carcinoma. The purpose of this study was to develop an efficient method of the determination of sorafenib in human plasma using tandem mass spectrometry coupled with liquid chromatography (LC/MS/MS) and validate the method by the guidelines of the Korean Food and Drug Administration (KFDA). Plasma samples ($100{\mu}l$) were added with chlorantraniliprole as an internal standard and then mixed with the 0.1% formic acid-containing extraction solution composed of isopropyl alcohol and ethyl acetate (1:4, v/v). After centrifugation, the supernatant was concentrated at $45^{\circ}C$ under negative pressure and centrifugal force. The residue was reconstituted with a mobile phase and injected into the HPLC instrument using a reverse phase Waters XTerra$^{TM}$ C18 column (particle size $3.5{\mu}m$). Liquid chromatography was carried out within the run time of 5 min using a mobile phase composed of buffer (0.1% formic acid and 10 mM ammonium formate), methanol, and acetonitrile (1:6:3, v/v/v). The analytes were monitored by tandem mass spectrometry in the multiple reaction monitoring method programmed to detect sorafenib at 'm/z 465.2 ${\rightarrow}$ 252.5' and chlorantraniliprole at 'm/z 484.4 ${\rightarrow}$ 286.2' with positive electrospray ionization mode ($ES^+$). The result showed the proper linearity ($r^2$ > 0.99) over the range of 2,000-5,000 ng/ml with good accuracy (90.7-103.9%) and precision (less than 10%). The newly developed method using LC/MS/MS was validated by the guideline of KFDA and identified as more sensitive compared to the previous methods.

Determination of plasma C16-C24 globotriaosylceramide (Gb3) isoforms by tandem mass spectrometry for diagnosis of Fabry disease (패브리병(Fabry) 진단을 위한 혈장 중 Globotriaosylceramide (Gb3)의 탠덤매스 분석법 개발과 임상 응용)

  • Yoon, Hye-Ran;Cho, Kyung-Hee;Yoo, Han-Wook;Choi, Jin-Ho;Lee, Dong-Hwan;Zhang, Kate;Keutzer, Joan
    • Journal of Genetic Medicine
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    • v.4 no.1
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    • pp.45-52
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    • 2007
  • Purpose : A simple, rapid, and highly sensitive analytical method for Gb3 in plasma was developed without labor-ex tensive pre-treatment by electrospray ionization MS/ MS (ESI-MS/MS). Measurement of globotriaosy lceramide (Gb3, ceramide trihex oside) in plasma has clinical importance for monitoring after enzyme replacement therapy in Fabry disease patients. The disease is an X-linked lipid storage disorder that results from a deficiency of the enzyme ${\alpha}$-galactosidase A (${\alpha}$-Gal A). The lack of ${\alpha}$-Gal A causes an intracellular accumulation of glycosphingolipids, mainly Gb3. Methods : Only simple 50-fold dilution of plasma is necessary for the extraction and isolation of Gb3 in plasma. Gb3 in diluted plasma was dissolved in dioxane containing C17:0 Gb3 as an internal standard. After centrifugation it was directly injected and analyzed through guard column by in combination with multiple reaction monitoring mode of ESI-MS/MS. Results : Eight isoforms of Gb3 were completely resolved from plasma matrix. C16:0 Gb3 occupied 50% of total Gb3 as a major component in plasma. Linear relationship for Gb3 isoforms w as found in the range of 0.001-1.0 ${\mu}g$/mL. The limit of detection (S/N=3) was 0.001 ${\mu}g$/mL and limit of quantification was 0.01 ${\mu}g$/mL for C16:0 Gb3 with acceptable precision and accuracy. Correlation coefficient of calibration curves for 8 Gb3 isoforms ranged from 0.9678 to 0.9982. Conclusion : This quantitative method developed could be useful for rapid and sensitive 1st line Fabry disease screening, monitoring and/or diagnostic tool for Fabry disease.

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Estimation of TROPOMI-derived Ground-level SO2 Concentrations Using Machine Learning Over East Asia (기계학습을 활용한 동아시아 지역의 TROPOMI 기반 SO2 지상농도 추정)

  • Choi, Hyunyoung;Kang, Yoojin;Im, Jungho
    • Korean Journal of Remote Sensing
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    • v.37 no.2
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    • pp.275-290
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    • 2021
  • Sulfur dioxide (SO2) in the atmosphere is mainly generated from anthropogenic emission sources. It forms ultra-fine particulate matter through chemical reaction and has harmful effect on both the environment and human health. In particular, ground-level SO2 concentrations are closely related to human activities. Satellite observations such as TROPOMI (TROPOspheric Monitoring Instrument)-derived column density data can provide spatially continuous monitoring of ground-level SO2 concentrations. This study aims to propose a 2-step residual corrected model to estimate ground-level SO2 concentrations through the synergistic use of satellite data and numerical model output. Random forest machine learning was adopted in the 2-step residual corrected model. The proposed model was evaluated through three cross-validations (i.e., random, spatial and temporal). The results showed that the model produced slopes of 1.14-1.25, R values of 0.55-0.65, and relative root-mean-square-error of 58-63%, which were improved by 10% for slopes and 3% for R and rRMSE when compared to the model without residual correction. The model performance by country was slightly reduced in Japan, often resulting in overestimation, where the sample size was small, and the concentration level was relatively low. The spatial and temporal distributions of SO2 produced by the model agreed with those of the in-situ measurements, especially over Yangtze River Delta in China and Seoul Metropolitan Area in South Korea, which are highly dependent on the characteristics of anthropogenic emission sources. The model proposed in this study can be used for long-term monitoring of ground-level SO2 concentrations on both the spatial and temporal domains.

Detection and Classification of Porcine Endogenous Retroviruses by Polymerase Chain Reaction (중합효소 연쇄반응을 이용한 돼지 내인성 레트로 바이러스의 검출과 분류)

  • Lee, D.H.;Lee, J.E.;Kim, H.M.;Kim, G.W.;Park, H.Y.;Kim, Young-Bong
    • Journal of Animal Science and Technology
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    • v.49 no.3
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    • pp.405-414
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    • 2007
  • Pigs have been considered as an ideal source of donor organs because of their plentiful supply and their numerous anatomical and physiological similarities to the human in xenotransplantation. However, for the public health risks associated with the potential for porcine endogenous retrovirus(PERV) infection through xenograft from pig to human, the investigation of methods for elimination and/or control of PERV has been required. In this study we developed the detection and classification methods for PERV based on PCR using specific primers. PERV-A and PERV-B were found in all pigs including Berkshire, Duroc, Landrace, Yorkshire, miniature pig, and Korean native black pig from Jeju by PCR with type-specific primers for PERV. However, PERV-C was detected only from Duroc, miniature pig, and Korean native black pig from Jeju. PERV-A and PERV-B could be distinguished by PCR-RFLP with BamHI. These methods for PERV will be useful in rapid screening of safe organ for xenograft, furthermore, helpful in monitoring of PERV during and after xenotransplantation.