• Title/Summary/Keyword: laboratory method

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Determination of homogentisic acid in human plasma by GC-MS for diagnosis of alkaptonuria (GC-MS를 이용한 혈장 중 호모겐티식산의 분석;알캅톤뇨증의 진단)

  • Thapa, Maheshwor;Yu, Jundong;Lee, Wonjae;Islam, Fokhrul;Yoon, Hye-Ran
    • Analytical Science and Technology
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    • v.28 no.5
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    • pp.323-330
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    • 2015
  • Alkaptonuria, a rare inherited metabolic disease, is characterized by a lack of homogentisate dioxygenase and accumulation of homogentisic acid (HGA), leading to homogentisic aciduria, arthritis, and ochronosis. In this study, a rapid analytical method, without an expensive and tedious solid phase extraction step, was developed to quantify HGA in plasma using GC-MS. HGA-spiked pooled plasma samples were subjected to liquid-liquid extraction (LLE) with ethyl acetate, followed by trimethylsilyl derivatization (TMS) and GC-MS quantification using selected ion monitoring. The formation of TMS derivative of the 1 carboxylic and 2 hydroxyl functional groups was performed by reacting BSTFA (with 10% TMCS) for 5 min at 80 ℃. For selected ion monitoring, quantification and confirmation ions were determined based on specific ions (m/z 384, m/z 341 and m/z 252) of the TMS derivative of HGA. Calibration curves of pooled normal plasma specimens showed a linear relationship in the range of 1-100 ng/µL. The precision and accuracy were within a relative standard deviation (RSD) of 1 to 15% and a bias of -5 to 25%. Recoveries were obtained in the range of 99-125% and 95-115% for intra-day and inter-day assay, respectively, at 2, 20 and 80 ng/µL. The limit of detection (LOD) and limit of quantification (LOQ) were 0.4 ng/µL and 4 ng/µL, respectively. No homogentisic acid was excreted from normal Korean plasma samples. Collectively, the results from the present study suggest that this method could be useful for routine diagnosis and therapeutic monitoring of alkaptonuria patients with excellent sensitivity and rapidity.

Textbook Analysis of Middle School-Home Economics and Survey on Consumption Status and Nutritional Knowledge of Milk and Dairy Products of Middle School Students in Gongju City, Chungnam Province (중학교 가정교과서의 우유 교육 내용 분석과 중학생의 우유·유제품 섭취 실태 및 영양지식 조사 - 충남 공주시 중학생을 중심으로 -)

  • Kim, Sun Hyo
    • Journal of Korean Home Economics Education Association
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    • v.29 no.4
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    • pp.117-131
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    • 2017
  • This study was performed to analyze education contents related to milk in current home economics textbooks of middle school, and to investigate intake status, consumption behaviors, perception and nutritional knowledge of milk and dairy products among 364 middle school students in Gongju for improvement of milk education in home economics subjects and milk intake of adolescents. As a result, education contents of milk and dairy products in home economics textbooks currently applied in middle school were major nutrients, consumption method for balanced diet, and selection and storage of milk and dairy products, thus it tended not to match current food trend. Only 30.5% of subjects met 2 cups of milk a day, the recommended level. The main reason for drinking milk was to 'be taller' and 'to quench thirst' and there was a difference by gender(p<0.01). The rate of not participating in school milk program was 23.1% of total and its satisfaction was moderate. The most popular dairy products by subjects were ice cream, followed by yogurt and cheese, and the choice of milk was focused on 'taste' or 'expiration date'. The rate of knowing certification mark of K-MILK was low at 28.8%, and most subjects knew as 'domestic milk use'. In home economics class, experience-based learning such as cow ranch experience was the most preferred instruction method for milk followed by laboratory practice and lecture, and there was a difference by gender(p<0.001). Perception degree of milk and dairy products was moderate and male subjects were more positively perceived than female subjects(p<0.01). Nutritional knowledge level of milk and dairy products was moderate and female subjects were higher than male subjects(p<0.01). Therefore, education contents of milk and dairy products of home economics textbooks of middle school should be centered on real life in accordance with food trend, and applied student participation-based instruction methods such as experience-based learning. In addition, it is necessary to enhance taste and merchandise of milk and to provide them with preferred milk and dairy products in school milk program for improvement of milk intake of adolescents.

The Fate of Intractable Tuberculosis Cases Under National Tuberculosis Programme (국가결핵관리 체계내의 난치성 결핵환자(만성 배균자)의 운명)

  • Lew, Woo-Jin;Lee, Eun-Gyu;Kwon, Dong-Won;Kim, Sang-Jae;Hong, Yong-Pyo;Kim, Jeong-Bae
    • Tuberculosis and Respiratory Diseases
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    • v.42 no.1
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    • pp.11-18
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    • 1995
  • Background: The natural history of bacillary tuberculosis was studied in India and results showed that at the end of the 5-year period, 49% of the patients were dead, 33% were cured and 18% remained sputum-positive. The aim of this survey is to observe the natural course of the patients with intractable tuberculosis disease who were incurable with all drug regimens of the national tuberculosis programme(NTP). Method: Of the patients who have been found as intractable cases in Kang-Weon Province by the supervisory medical officer during the period from January 1,1987 to December 31,1992, 179 were eligible for this study. Sputum examination was done for those who were survived until October in 1993 at the Kang-Weon provincial laboratory of KNTA. 49 out of 179 patients were transferred to the private sectors and retreated with the combination of prothionamide, cycloserine, ofloxacin, enviomycin, etc. They seemed to have been bacteriologically cured, and so they were excluded from the study. Finally 130 patients were analyzed by modified life table method to calculate the fatality rate and the survival rate during the period of 7 years. Results: 1) 80.8% of intractable cases were male and 19.2%, female. 2) More than 94% of intractable cases showed moderately or far advanced Tb findings on their X-rays at the time of registration at health centres. 3) The cumulative case-fatality rate was 19.74% at the end of 1-year period and has risen to 34.55% by the end of 4-year period(increasing by 4.9% a year on an average). The case-fatality rate has shown no appreciable rise since then until the end of 7-year period. 4) The case-survival rate was 80.26% at the end of 1-year period and has decreased to 65.45% by the end of 4-year period. And then there was no appreciable change in the survival rate until the end of 7-year observation. Conclusion: The case-survival rate of intractable cases was higher than that of untreated pulmonary tuberculosis patients and they may have risk of spreading multidrug resistant organisms. It is time we made an effort to improve case-management qualitatively.

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Bionomics and a Lana Sampling Method of Coppery Wireworm, Selatosomus puncticollis (Motschulsky), (Coleoptera: Elateridae) in Potato Field (청동방아벌레(Selatosomus puncticollis Motschulsky)의 생태적 특성 및 감자포장내 유충밀도 조사법)

  • Kwon, Min;Park, Cheon-Soo;Lee, Seung-Hwan
    • Korean journal of applied entomology
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    • v.43 no.3 s.136
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    • pp.195-200
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    • 2004
  • The occurrence pattern and bionomics of Selatosomus puncticollis (Motschulsky) were surveyed in three net house (6$\times$70m) in the field and in a laboratory ($20^{\circ}C$, RH$75\%$, L16/DB). Emergence of hibernated adults of S. puncricollis started from early May, reached peak at early June and diminished in late June. Occurrence rate to total number was $42.1\%$ in early June and $31.7\%$ in late May, when most of hibernated adults emerged at these periods. Adults started to oviposit from mid June, and eggs hatched from early July Larvae turned into pupae from mid July and emerged to adults from mid August. Egg-period was 23 days and pupal period was 21 days. It is estimated that preovipositional period and larval period were approximately 10 months and 30 months, respectively. Wireworms were distributed at the soil depth of 10-5 cm, $56.8\%$ in 1997 and $45.8\%$ in 1998. To establish bait techniques to attract wireworms in the soil, six baits: pieces of potato, carrot, and sweet potato, wheat grains, corns, and flour dough wrapped with gauze, were buried at 15 cm of soil depth, and collected after 5 and 10 days. The numbers of wireworms attracted by potato pieces, wheat grains and sweet potato pieces after 10 days were 1.8, 1.6, and 1.4/bait, respectively. Therefore, burying potato pieces at 15 cm of soil depth and collected after 10 days could be recommended as a wireworm baiting technique.

Lymphocyte Subpopulations and Proliferation of T cells, Phagocytic Activity of Leukocytes on Alcoholics (알코올중독자의 백혈구탐식능, 림프구아형 및 증식능)

  • 김용호;서병배;이정녀;김영훈
    • Biomedical Science Letters
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    • v.2 no.2
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    • pp.167-174
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    • 1996
  • Alcoholics increased susceptibility to microbial infection that is associated with decreased immunity. but there has been little experimental evidence to support alcoholics-induced increase of microbial infection directly in non-specific immunity. Therefore, we were used the method of phagocytic-plaque including all the stimulating factors for the phagocytosis, subtypes of lymphocytes and T-lymphocyte proliferation. The experimental groups were divided into 3 groups: (1) alcoholics who were hospitalized less than 1 week (newly hospitalized alcoholics), (2) alcoholics who were hospitalized more than 2 weeks (old hospitalized alcoholics), (3) healthy blood donors. We have studied 98 alcoholics and 35 healthy blood donors and control groups. A physician has checked the biological markers and diagnosed the body-condition alcoholics. The immunity and non-specific immunity on the alcoholics were analyzed by using the simultest kit and flow cytometry. Proliferation of the lymphocytes was analyzed by the phytohemmagglutinine mitogen. Phagocytosis and migration properties of leukocytes were identified on the layer formed by Staphylococcus aureus Cowan I strain. Biological markers of alcoholics and control groups, by such as blood glucose, ${\gamma}$-glutamyl transpeptidase and mean corpuscular volumes of red blood cells, were determined by biochemical and hematological methods. Compared with control groups, cluster of differentiation (CD)3+, CD8+ and CD19+ in alcoholic were more decreased except CD4+/CD8+ ratio. Proliferation of the T-lymphocytes, phagocytosis and migration properties of the leukocytes in alcoholics were decreased compared with those of control groups. According to the results observed in our experiment, they can be summerized as follows: 1, Cellular, humoral and non-specific immunities, are markedly decreased in alcoholics than those in control groups. 2. It is inferred that Phagocytic plaque formation is a very useful method to evaluate phagocytosis and migration properties of the alcoholic leukocytes 3. It is thought that the subtypes of lymphocytes, especially CD4+/CD8+ ratio, are essential methods to analyzed the alcoholic immunity. 4. Specific and non-specific immunity on the old hospitalized alcoholics was slightly increased, which depends upon the alcoholic medication.

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Comparison of C-reactive Protein between Capillary and Venous Blood in Children (소아에 있어서 C-반응성 단백의 모세혈 및 정맥혈 검사의 비교평가)

  • Jin, Ji Hoon;Jung, Soo Ho;Hong, Young Jin;Son, Byong Kwan;Kim, Soon Ki
    • Pediatric Infection and Vaccine
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    • v.17 no.2
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    • pp.101-107
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    • 2010
  • Purpose : In evaluation of patients, laboratory results are crucial in determination of a treatment plan. Obtaining venous blood from infants and children is a difficult procedure. Substitution of a capillary blood sample for a venous blood sample has been suggested. However, there are few studies showing mutual correlation between C-reactive protein (CRP) results in capillary and venous blood. This study was designed to determine whether the result of the capillary sample is the same as the result of the venous blood sample. Methods : After informed consent, a pair of venous and fingertip capillary blood samples were simultaneously collected from 100 children. The LC-178CRPTM was used for analysis of capillary blood and the Hitachi 7180 automatic hematology analyzer was used for analysis of venous blood. We compared CRP of both venous and capillary blood samples. Results were analyzed by crosstabulation analysis, simple regression analysis and the Bland Altman Plot method. Results : A close correlation (90.63%) was observed between capillary and venous blood analyzed by crosstabulation analysis. CRP results were similar between the two groups and showed a high coefficient correlation ($\beta$=1.3434, $R^2$=0.9888, P<0.0001) when analyzed by a simple regression model. The average value in venous blood was also higher compared to capillary blood. According to Bland Altman Plot analysis, lab results were measured at a 95% confidence interval. Conclusion : CRP results from capillary blood showed close correlation with venous blood sampling. At present, venous blood sampling is the preferred method. However, due to difficulty in venous blood sampling, capillary sampling could be considered as an alternative technique for use with children.

Establishment of Reference Intervals of Osteocalcin according to Age in Women for Health Promotion Center (건강검진이 의뢰된 여성의 연령에 따른 Osteocalcin의 참고범위 설정)

  • Kang, Ji-Soon;Yoo, Byoung-Joo;Oh, Jung-Eun;Kim, Geon-Jae
    • The Korean Journal of Nuclear Medicine Technology
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    • v.13 no.1
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    • pp.104-111
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    • 2009
  • Purpose: Osteocalcin is also known as the bone gamma-carboxyglutamic acid (Gla) protein (BGP), is noncollagenous bone protein synthesized by osteoblasts. Serum concentrations of Osteocalcin have been used as a biochemical marker of bone turnover. The reference intervals of Osteocalcin is categorized by kit corporation according to the age. However, each laboratory should establish its own reference intervals. In this study, the variation in the serum Osteocalcin level were used to find actual standard age-specific Osteocalcin reference intervals. Materials and Methods: We have selected 864 healthy females aged 20~80 years who visited a health promotion center between Aug. 2007 and Sep. 2008. The Osteocalcin IRMA Kit (OSTEO-RIACT, CIS Bio international, Gif-sur-Yvette, France) was used for the quantification. Each results were analyzed with the SPSS 12.0 statistical software. Results: The analyzed reference intervals of Osteocalcin by using Hoffmann method are from 8.8~39.4 ng/mL to 6.3~28.8 ng/mL for the case of the age from 20 to 30, from 7.7~31.9 ng/mL to 5.9~17.4 ng/mL for the case of the age from 31 to 40, and from 8.0~36.0 ng/mL to 5.5~20.1 ng/mL for the case of the age from 41 to 50, and from 8.0~50.5 ng/mL to 6.7~27.0 ng/mL for the case of the age from 51 to 60, and from 12.9~55.9 ng/mL to 7.5~27.5 ng/mL for the case of the age from 61 to 80. Reference intervals of Osteocalcin were not in agreement with those recommended by the manufacturers. Conclusions: Osteocalcin is used as an indication of metabolic bone diseases. So in our study we wanted to provide reference intervals of Osteocalcin that can be useful to a clinical decisions. Also, previous reference intervals should not be re-used and new intervals should be set by continuous analyzing.

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Single Laboratory Validation and Uncertainty Estimation of a HPLC Analysis Method for Deoxynivalenol in Noodles (면류에서 HPLC를 이용한 데옥시니발레놀 분석법의 검증과 불확도 산정)

  • Ee, Ok-Hyun;Chang, Hyun-Joo;Kang, Young-Woon;Kim, Mee-Hye;Chun, Hyang-Sook
    • Journal of Food Hygiene and Safety
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    • v.26 no.2
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    • pp.142-149
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    • 2011
  • An isocratic high performance liquid chromatography (HPLC) method for routine analysis of deoxynivalenol in noodles was validated and estimated the measurement uncertainty. Noodles (dried noodle and ramyeon) were analyzed by HPLC-ultraviolet detection using immunoaffinity column for clean-up. The limits of detection (LOD) and quantification (LOQ) were 7.5 ${\mu}g$/kg and 18.8 ${\mu}g$/kg, respectively. The calibration curve showed a good linearity, with correlation coefficients $r^2$ of 0.9999 in the concentration range from 20 to 500 ${\mu}g$/kg. Recoveries and Repeatabilities expressed as coefficients of variation (CV) spiked with 200 and 500 ${\mu}g$/kg were $82{\pm}2.7%$ and $87{\pm}1.3%$% in dried noodle, and $97{\pm}1.6%$ and $91{\pm}12.0%$ in ramyeon, respectively. The uncertainty sources in measurement process were identified as sample weight, final volume, and sample concentration in extraction volume as well as components such as standard stock solution, working standard solution, 5 standard solutions, calibration curve, matrix, and instrument. Deoxynivalenol concentration and expanded uncertainty in two matrixes spiked with 200 ${\mu}g$/kg and 500 ${\mu}g$/kg were estimated to be $163.8{\pm}52.1$ and $435.2{\pm}91.6\;{\mu}g$/kg for dried noodle, and $194.3{\pm}33.0$ and $453.2{\pm}91.1\;{\mu}g$/kg for ramyeon using a coverage factor of two which gives a level of statistical confidence with approximately 95%. The most influential component among uncertainty sources was the recovery of matrix, followed by calibration curve.

A Critical Liquefaction Resistible Characteristic of Saturated Sands Based on the Cyclic Triaxial Test Under Sinusoidal Loadings (정현하중재하 진동삼축시험에 기초한 포화사질토의 액상화 한계저항특성)

  • 최재순;김수일
    • Journal of the Korean Geotechnical Society
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    • v.20 no.8
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    • pp.147-158
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    • 2004
  • Laboratory dynamic tests are carried out to assess the liquefaction potential of saturated sands in most countries. However, simple results such as the maximum cyclic shear stress and the number of cycles at initial liquefaction are used in the experimental assessment of liquefaction potential, even though various results can be obtained from the dynamic test. In addition, it seemed to be inefficient because more than three dynamic tests with different stress ratio have to be carried out to draw a liquefaction resistance experimental curve. To improve the present assessment method fur liquefaction potential, a new critical resistible characteristic far soil liquefaction is proposed and verified through conventional cyclic triaxial tests with Jumunjin sand. In the proposed method, various experimental data such as effective stress path, stress-strain relationship, and the change of excess pore water pressure can be used in the determination of cumulative plastic shear strains at every 1/4 cycle. Especially, the critical cumulative plastic shear strain to initiate liquefaction can be defined in a specific point called a phase change point in the effective stress path and it can be calculated from a hysteric curve of stress-strain relationship up to this point. Through this research, it is found that the proposed cumulative plastic shear strain can express the dissipated energy to resist dynamic loads and consider the realistic soil dynamic behavior of saturated sands reasonably. It is also found that the critical plastic shear strain can be used as a registible index of soils to represent the critical soil dynamic state, because it seems to include no effect of large deformation.

Enhancement of the Deformable Image Registration Accuracy Using Image Modification of MV CBCT (Megavoltage Cone-beam CT 영상의 변환을 이용한 변환 영상 정합의 정확도 향상)

  • Kim, Min-Joo;Chang, Ji-Na;Park, So-Hyun;Kim, Tae-Ho;Kang, Young-Nam;Suh, Tae-Suk
    • Progress in Medical Physics
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    • v.22 no.1
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    • pp.28-34
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    • 2011
  • To perform the Adaptive Radiation Therapy (ART), a high degree of deformable registration accuracy is essential. The purpose of this study is to identify whether the change of MV CBCT intensity can improve registration accuracy using predefined modification level and filtering process. To obtain modification level, the cheese phantom images was acquired from both kilovoltage CT (kV CT), megavoltage cone-beam CT (MV CBCT). From the cheese phantom images, the modification level of MV CBCT was defined from the relationship between Hounsfield Units (HUs) of kV CT and MV CBCT images. 'Gaussian smoothing filter' was added to reduce the noise of the MV CBCT images. The intensity of MV CBCT image was changed to the intensity of the kV CT image to make the two images have the same intensity range as if they were obtained from the same modality. The demon deformable registration which was efficient and easy to perform the deformable registration was applied. The deformable lung phantom which was intentionally created in the laboratory to imitate the changes of the breathing period was acquired from kV CT and MV CBCT. And then the deformable lung phantom images were applied to the proposed method. As a result of deformable image registration, the similarity of the correlation coefficient was used for a quantitative evaluation of the result was increased by 6.07% in the cheese phantom, and 18% in the deformable lung phantom. For the additional evaluation of the registration of the deformable lung phantom, the centric coordinates of the mark which was inserted into the inner part of the phantom were measured to calculate the vector difference. The vector differences from the result were 2.23, 1.39 mm with/without modification of intensity of MV CBCT images, respectively. In summary, our method has quantitatively improved the accuracy of deformable registration and could be a useful solution to improve the image registration accuracy. A further study was also suggested in this paper.