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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.

A basic research for evaluation of a Home Care Nursing Delivery System (가정간호 서비스 질 평가를 위한 도구개발연구)

  • Kim, Mo-Im;Cho, Won-Jung;Kim, Eui-Sook;Kim, Sung-Kyu;Chang, Soon-Bok;Ryu, Ho-Sihn
    • Journal of Home Health Care Nursing
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    • v.6
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    • pp.33-45
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    • 1999
  • The purpose of this study was to develop a basic framework and criteria for evaluation of quality care provided to patients with the attributes of disease in the home care nursing field, and to provide measurement tools for home health care in the future. The study design was a developmental study for evaluation of hospital-based HCN(home care nursing) in Korea. The study process was as follows: a home care nursing study team of College of Nursing. Yonsei University reviewed the nursing records of 47 patients who were enrolled at Yonsei University Medical Center Home Care Center in March, 1995. Twenty-five patients were insured at that time, were selected from 47 patients receiving home care service for study feasibility with six disease groups; Caesarean Section (C/S), simple nephrectomy, Liver cirrhosis(LC), chronic obstructive pulmonary disease(COPD), Lung cancer or cerebrovascular accident(CVA). In this study, the following items were selected : First step : Preliminary study 1. Criteria and items were selected on the basis of related literature on each disease area. 2. Items were identified by home care nurses. 3. A physician in charge reviewed the criteria and content of selected items. 4. Items were revised through preliminary study offered to both HCN patients and discharged patients from the home care center. Second step : Pretest 1. To verify the content of the items, a pretest was conducted with 18 patients of which there were three patients in each of the six selected disease groups. Third step : Test of reliability and validity of tools 1. Using the collected data from 25 patients with either cis, Simple nephrectomy, LC, COPD, Lung cancer, or CVA. the final items were revised through a panel discussion among experts in medical care who were researchers, doctors, or nurses. 2. Reliability and validity of the completed tool were verified with both inpatients and HCN patients in each of field for researches. The study results are as follows: 1. Standard for discharge with HCN referral The referral standard for home care, which included criteria for discharge with HCN referral and criteria leaving the hospital were established. These were developed through content analysis from the results of an open-ended questionnaire to related doctors concerning characteristic for discharge with HCN referral for each of the disease groups. The final criteria was decided by discussion among the researchers. 2. Instrument for measurement of health statusPatient health status was measured pre and post home care by direct observation and interview with an open-ended questionnaire which consisted of 61 items based on Gorden's nursing diagnosis classification. These included seven items on health knowledge and health management, eight items on nutrition and metabolism, three items on elimination, five items on activity and exercise, seven items on perception and cognition, three items on sleep and rest, three items on self-perception, three items on role and interpersonal relations, five items on sexuality and reproduction, five items on coping and stress, four items on value and religion, three items on family. and three items on facilities and environment. 3. Instrument for measurement of self-care The instrument for self-care measurement was classified with scales according to the attributes of the disease. Each scale measured understanding level and practice level by a Yes or No scale. Understanding level was measured by interview but practice level was measured by both observation and interview. Items for self-care measurement included 14 for patients with a CVA, five for women who had a cis, ten for patients with lung cancer, 12 for patients with COPD, five for patients with a simple nephrectomy, and 11 for patients with LC. 4. Record for follow-up management This included (1) OPD visit sheet, (2) ER visit form, (3) complications problem form, (4) readmission sheet. and (5) visit note for others medical centers which included visit date, reason for visit, patient name, caregivers, sex, age, time and cost required for visit, and traffic expenses, that is, there were open-end items that investigated OPD visits, emergency room visits, the problem and solution of complications, readmissions and visits to other medical institution to measure health problems and expenditures during the follow up period. 5. Instrument to measure patients satisfaction The satisfaction measurement instrument by Reisseer(1975) was referred to for the development of a tool to measure patient home care satisfaction. The instrument was an open-ended questionnaire which consisted of 11 domains; treatment, nursing care, information, time consumption, accessibility, rapidity, treatment skill, service relevance, attitude, satisfaction factors, dissatisfaction factors, overall satisfaction about nursing care, and others. In conclusion, Five evaluation instruments were developed for home care nursing. These were (1)standard for discharge with HCN referral. (2)instrument for measurement of health status, (3)instrument for measurement of self-care. (4)record for follow-up management, and (5)instrument to measure patient satisfaction. Also, the five instruments can be used to evaluate the effectiveness of the service to assure quality. Further research is needed to increase the reliability and validity of instrument through a community-based HCN evaluation.

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Estimation of Glomerular Filtration Rate(GFR) Using $^{99m}Tc$-DTPA Renal Scan and the Parameters for Renal Function ($^{99m}Tc$-DTPA를 이용한 신장스캔에서 사구체 여과율의 측정방법과 영상분석에서 구한 지표들에 의한 신장기능의 평가)

  • Cho, Ihn-Ho;Yoon, Hyun-Dae;Won, Kyu-Chang;Lee, Chan-Woo;Lee, Hyoung-Woo;Lee, Hyun-Woo
    • Journal of Yeungnam Medical Science
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    • v.11 no.1
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    • pp.101-108
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    • 1994
  • Many previously described nuclear medicine procedures to assess glomerular filtration rate have some problems because numerous blood sample is to be taken and they don't measure each separate renal function. Gates described isotopic method for the measurement of global and unilateral GFR based on the fractional renal uptake of $^{99m}Tc$-DTPA 2 to 3 minutes after its intravenous injection. We evaluated GFR using $^{99m}Tc$-DTPA in 57 people according to Gates method and compared with creatinine clearance. A good correlation was observed between creatinine clearance and GFR calculated by Gates' formula with an r value of 0.9(P<0.05). And also the relationship between parameters of $^{99m}Tc$-DTPA renal scan images and GFR was taken. They were significantly correlated with GFR calculated by Gates' formula : r value 0.66 between relative intensity of peak renal to peak aortic activity(pK/pA) and GFR, -0.42 between time between aortic and kidney peak(A-K) and GFR and -0.48 between parenchymal renal activity at 25 min compared to peak kidney activity(25K/pK) and GFR. In conclusion, the determination of GFR according to the Gates' formula shows good and reproducible of GFR with rapidity and simplicity. And the parameters from the renal scan images can use to estimate the renal function.

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Phase Image Analysis in Conduction Disturbance Patients (심실내 전도장애 환자에서의 $^{99m}Tc$-RBC Gated Blood-Pool Scintigraphy을 통한 Phase Image Analysis)

  • Kwak, Byeng-Su;Choi, Si-Wan;Kang, Seung-Sik;Park, Ki-Nam;Lee, Kang-Wook;Jeon, Eun-Seok;Park, Chong-Hun
    • The Korean Journal of Nuclear Medicine
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    • v.28 no.1
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    • pp.44-51
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    • 1994
  • It is known that the normal His-Purkinje system provides for nearly synchronous activation of right (RV) and left (LV) ventricles. When His-Purkinje conduction is abnormal, the resulting sequence of ventricular contraction must be correspondingly abnormal. These abnormal mechanical consequences were difficult to demonstrate because of the complexity and the rapidity of it's events. To determine the relationship of the phase changes and the abnormalities of ventricular conduction, we performed phase image analysis of $^{99m}Tc$-RBC gated blood pool scintigrams in patients with intraventricular conduction disturbances (24 complete left bundle branch block (C-LBBB), 15 complete right bundle branch block (C-RBBB), 13 Wolff-Parkinson-White syndrome (WPW), 10 controls). The results were as follows; 1) The ejection fraction (EF), peak ejection rate (PER), and peak filling rate (PFR) of LV in gated blood pool scintigraphy (GBPS) were significantly lower in patients with C-LBBB than in controls ($44.4{\pm}13.9%$ vs $69.9{\pm}4.2%,\;2.48{\pm}0.98$ vs $3.51{\pm}0.62,\;1.76{\pm}0.71$ vs $3.38{\pm}0.92$, respectively, p<0.05). 2) In the phase angle analysis of LV, Standard deviation (SD), width of half maximum of phase angle (FWHM), and range of phase angle were significantly increased in patients with C-LBBB than in controls ($20.6{\pm}18.1$ vs $8.6{\pm}1.8,\;22.5{\pm}9.2$ vs $16.0{\pm}3.9,\;95.7{\pm}31.7$ vs $51.3{\pm}5.4$, respectively, p<0.05). 3) There was no significant difference in EF, PER, PFR between patients with the Wolff-parkinson-White syndrome and controls. 4) Standard deviation and range of phase angle were significantly higher in patients with WPW syndrome than in controls ($10.6{\pm}2.6$ vs $8.6{\pm}1.8$, p<0.05, $69.8{\pm}11.7$ vs $51.3{\pm}5.4$, p<0.001, respectively), however, there was no difference between the two groups in full width of half maximum. 5) Phase image analysis revealed relatively uniform phase across the both ventricles in patients with normal conduction, but markedly delayed phase in the left ventricle of patients with LBBB. 6) In 13 cases of WPW syndrome, the site of preexcitation could be localized in 10 cases (77%) by phase image analysis. Therefore, it can be concluded that phase image analysis can provide an accurate noninvasive method to detect the mechanical consequences of a wide variety of abnormal electrical activation in ventricles.

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Detection of Lily symptomless virus, Lily mottle virus, and Cucumber mosaic virus from Lilium Grown in Korea by RT-PCR (RT-PCR법을 이용한 백합 바이러스 LSV, LMoV, CMV의 검출)

  • Lim, Ji-Hyun;Bae, Eun-Hye;Lee, Yong-Jin;Park, Sung-Han;Lee, Kyu-Jun;Kim, Sae-Ro-Mi;Jung, Yong-Tae
    • Korean Journal of Microbiology
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    • v.45 no.3
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    • pp.251-256
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    • 2009
  • Leaf samples and bulbs showing characteristic symptoms of virus infection were collected from Gang-won, Chung-nam, and Jeju Province of Korea in 2008-2009. Three viruses, Lily symptomless virus (LSV), Lily mottle virus (LMoV), and Cucumber mosaic virus (CMV) were detected by RT-PCR. Virus-infected plant samples were identified; 12 plants with LSV, 20 plants with LMoV, and 1 plant with CMV. Of the twelve LSV infected samples, seven samples were found to be mix-infected with LMoV and LSV. Symptoms of LMoV and LSV mixed infection were fairly severe, like as vein clearing, leaf curling, leaf mottling, leaf mosaic, and yellow streaking. Mixed infection with LMoV and LSV was also found in lily bulbs which have been stored under unfavorable environmental conditions. LMoV predominated in our tests, whereas spread of Lilyvirus X (LVX) was not found. The nucleotide sequences of coat protein (CP) region of seven isolates (4 LMoV, 2 LSV, and 1 CMV) were compared with the corresponding regions of LMoV (AJ564636), LSV (AJ516059) and CMV(AJ296154). The nucleotide sequence homologies between reference viruses and seven isolates were 95-99%. Complete sequencing of seven isolates is necessary to obtain more information on the molecular characteristics of these viruses as well as to increase sensitivity and rapidity of viral detection.

Perspective on Rapid and Selective Method for Detecting Microbiology in Dairy Industry: A Review (낙농산업에 필요한 미생물 검사방법과 전망: 총설)

  • Chon, Jung-Whan;Kim, Hyun-Sook;Kim, Hong-Seok;Kim, Dong-Hyeon;Song, Kwang-Young;Yim, Jin-Hyuk;Choi, Dasom;Lim, Jong-Soo;Jeong, Dong-Gwan;Kim, Soo-Ki;Seo, Kun-Ho
    • Journal of Dairy Science and Biotechnology
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    • v.33 no.2
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    • pp.119-127
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    • 2015
  • To date, detection of microbial populations in dairy products has been performed using culture media, which is a time-consuming and laborious method. The recently developed chromogenic media could be more rapid and specific than classical culture media. However, the newly developed molecular-based technology can detect microbial populations with greater rapidity and sensitivity than the classical method involving culture media and chromogenic media. This molecular-based technology could provide various options for monitoring the characterization of different states of bacteria and cells. Thus, it could help upgrade the processing system of the dairy industry so as to maintain the safety and quality of dairy foods. Among the various newly developed molecular-based technologies, flow cytometry can potentially be used for monitoring microbiological populations in the dairy industry if official international standards are available for this purpose. When omics technology would have biomarker identification, it could be regarded as the rapid and sensitive analytical methods. Methods based on PCR, which has become a basic technique in microbiological research, can be developed and validated as alternative methods for quantification of dairy microorganisms. This review discusses methods for monitoring microbiological populations in dairy foods and the limitations of these studies, as well as the need for further research on such methods in the dairy industry.

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A Study on the Forest Vegetation of Deogyusan National Park (덕유산 국립공원 삼림식생에 관한 연구)

  • Kim, Chang-Hwan;Oh, Jang-Geun;Lee, Nam-Sook
    • Korean Journal of Ecology and Environment
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    • v.46 no.1
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    • pp.33-40
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    • 2013
  • From March 2012 to January 2013, this study was conducted as a part of the project for making a precise electronic ecological zoning map of vegetation on a highly reduced scale of 1 to 5,000 with a view to improving management efficiency of national parks and enlarging the availability of the data produced from the basic research monitoring the resources of national parks. For the research accuracy and rapidity, a vegetation map was specially created for the on-the-site-vegetation research. To make the map more meticulous, we categorized the vegetation database into five groups: broadleaved forest, coniferous forest, mixed forest, rock vegetation and miscellaneous one. After comparing the results of the data built for the vegetation research and the actual research findings, it was made clear that vegetation of both categories was almost the same in case of broad-leaved forest with 72.20% and 78.45% respectively, and also equivalent in other groups like, for example, coniferous forest (16.70%, 13.41%), mixed forest (9.50%, 7.49%) and rock vegetation (0.60%, 0.15%). According to the precise vegetation map produced from the research, the deciduous broad-leaved forest was the most widely prevalent type in the correlated hierarchical classification of vegetation, occupying 65.78% of the overall vegetation. It was followed by mountain valley forest (15.17%), coniferous forest (10.90%), and plantation forest (7.00%) in order. It is particularly noteworthy that Mt. Deogyusan national park has retained a very stable and versatile forest vegetation in the outstanding state since approximately 20% of the mountain turns out to belong to the I grade vegetation conservation classification which contains climax forests, unique vegetation, subalpine vegetation, matured stands which are older than 50 years and etc.

Comparative efficacy of 5% EMLA cream and 20% Benzocaine gel during topical anesthesia (5% EMLA cream과 20% Benzocaine gel의 도포마취 효능 비교)

  • Lee, Dae-Woo;Baik, Byeong-Ju;Kim, Jae-Gon;Yang, Yeon-Mi;So, Yu-Ryeo
    • Journal of the korean academy of Pediatric Dentistry
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    • v.38 no.1
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    • pp.1-8
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    • 2011
  • The objective of this study was to determine the effectiveness of topical 5% EMLA cream versus 20% Benzocaine gel in reducing pain from intra oral needle insertion alone as well as injection of anesthetic. The 2 topical anesthetics were tested against each other bilaterally using a randomized, controlled, single blinded, split mouth design. Phase I was conducted to find out the rapidity of onset action of the two agents on anterior/posterior vestibules and anterior/posterior palatal mucosa. Phase II was conducted to evaluate the efficacy of the two topical anaesthetic agents in reducing the pain of intraoral injections. The agents were left in anterior/posterior vestibules and anterior/posterior palatal mucosa for the amount of time recorded in phase I. Subjects recorded pain on a 100-mm modified visual analog scale(VAS). A pulse oximeter was used to recorded the preoperative and postoperative pulse rates. In phase I of the study, two topical agents showed the longer onset of action at anterior part and vestibules than posterior part and palatal mucosa. EMLA cream showed the rapidest onset of action compared to benzocaine gel except on anterior palatal mucosa. In phase II of the study, the VAS grading of the pain for anesthetic administration showed EMLA cream was significantly(P<.05) better in elimination or reducing the pain on the anterior/posterior palatal mucosa. In conclusion, EMLA cream showed the rapidest onset of action compared to benzocaine gel except on anterior palatal mucosa. EMLA cream was found to be superior to Benzocaine gel with regards to pain reduction for anesthetic administration especially on anterior and posterior palatal mucosa.

Development of Prism Dot-sight Combined with Thermal Imaging Camera (열영상 카메라가 결합된 프리즘 도트사이트 개발)

  • Park, Seung-Hwan;Jung, Bo-Seon;Lee, Dong-Hee
    • Journal of Korean Ophthalmic Optics Society
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    • v.19 no.4
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    • pp.479-485
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    • 2014
  • Purpose: This study relates to the development of the prism dot-sight combined with the thermal imaging camera. Methods: We have placed a reflector designed to the doublet type in the front of a BS (beam splitting) prism, have placed an OLED panel and a dot reticle generator to the top and bottom of the reflecting surface of the BS prism, and have placed a detachable magnifier between the BS prism and the observer by which the observer can see the magnified image of the OLED panel. By doing this, we were able to configure the new type prism dot-sight combined with the thermal imaging camera. Results: By placing the removable magnifier designed with a new type between the BS prism and the observer, we could design the new type prism dot-sight which performs the role of the dot sight by removing the magnifier during the day-time, and performs the role of the night scope during the night-time by which we can observe the enlarged image of the thermal imaging camera through the BS prism by attaching the removable magnifier. Conclusions: In this study, we have developed the prism dot-sight combined with the thermal imaging camera which is able to play the role of the day or night scope selectively, by disposing the designed magnifier characterized by the focal length of 44 mm, the viewing angle of ${\pm}7.0^{\circ}$, and the MTF value of 0.5 or more at the criterion of 50 lp/mm and the 0.7 field between the BS prism and the observer. By doing so, we could design and fabricate the new type prism dot-sight combined with the thermal imaging camera which can further increase the rapidity of firing and provide more convenience in the mounting of a firearm than the detachable combination of an existing dot sight and an existing night scope.

Changes in the Physicochemical Characteristics of Green Wheat during Maturation (성숙과정 중 녹색밀의 이화학적 특성변화)

  • Kim, Min-Chul;Lee, Ka-Soon;Lee, Byung-Jin;Kwon, Byung-Gu;Ju, Jung-Il;Gu, Ja-Hyeong;Oh, Man-Jin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.10
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    • pp.1307-1313
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    • 2007
  • In order to evaluate the utilization of immature green wheat kernels as food processing material, these experiments were carried out to analyze general composition, free sugars, free amino acids, mineral content and color of 3 wheat varieties according to their heading dates. Contents such as moisture, crude protein, crude lipid and ash were gradually decreased according to the days after heading of 3 wheat varieties, while crude fiber and starch were gradually increased. Maltose and maltotriose contents in immature green wheat kernels were high but its rapidity decreased by closing to ripeness. Contents of free amino acids showed a tendency to decrease, among which alanine content was the highest around 25 days after heading, and then glutamic acid, GABA, glycine in order. Contents of total free amino acid were decreased by closing to ripeness. Among 3 wheat varieties, its contents of Guru wheat was the highest. As wheat matured, ${\beta}-glucan$ contents were decreased from 1.5% to 0.28% on 43 days after heading. Mineral contents of 3 wheat varieties were generally increased by closing to ripeness after coming into ears, and K, Mg were especially prominent. As wheat matured, lightness was decreased, while redness and yellowness were increased. Considering chemical composition and color, the immature green wheat were produced through blanching the spikes harvested before the yellow ripe stage. The harvested green wheat cereals is able to eat raw wheat or cook it as food processing material.