• Title/Summary/Keyword: Clinical Laboratory Science

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Detection of Dopamine and Serotonin by Competitive Enzyme-Linked Immunosorbent Assay (경쟁적 ELISA를 이용한 도파민과 세로토닌의 검출)

  • Namkung, Su Min;Choi, Jeong Su;Park, Ji Hyang;Yang, Man Gil;Lee, Min Woo;Kim, Suhng Wook
    • Korean Journal of Clinical Laboratory Science
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    • v.49 no.3
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    • pp.220-226
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    • 2017
  • Dopamine (DA) and serotonin (5-Hydroxytryptamine, 5-HT) are neurotransmitters and hormones that exist in small amounts but have important role in the body. Serum and 24-hour urine are used as specimens, and are usually examined by HPLC-MS. In this study, we tried to detect DA and 5-HT by competitive ELISA using antigen-antibody (Ab) reaction. After immobilizing $5{\mu}g/mL$ BSA conjugate on a 96-well surface, hormone and primary Ab, which are respectively diluted to different concentrations, were treated. Then, HRP-conjugated secondary Ab and TMB were added to measure absorbance. The regression equation and $R^2$ value were calculated based on absorbance, and sensitivity of Ab to hormone as well as the correlation between hormone concentration and absorbance were determined. In DA ELISA, $R^2$, the correlation between the concentration of hormone and absorbance, was the highest by 0.91 when anti-dopamine Ab was diluted 6,000 times and 7,000 times. In 5-HT ELISA, $R^2$ was bigger than 0.90 in every concentration except 3,000 times and 6,000 times. Both DA and 5-HT were not effectively detected at low concentrations (less than $1.0{\times}10^{-7}M$); and because reference value of serum DA is lower than this, HPLC-MS was required to detect serum DA. However, competitive ELISA may be effective in detecting 24-hour urine DA, serum, and 24-hour 5-HT. Further studies are needed to detect hormones more accurately at lower concentrations.

Prevalence of CTX-M-type Extended-Spectrum $\beta$-Lactamases Producing Escherichia coli and Klebsieilla pneumoniae Isolates in General Hospitals in 2005 (임상에서 분리된 CTX-M형 Extended-Spectrum $\beta$-Lactamases를 생산하는 Escherichia coli와 Klebsiella pneumoniae의 유행)

  • Kim, Yun-Tae;Kim, Tae-Un
    • Microbiology and Biotechnology Letters
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    • v.34 no.4
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    • pp.342-351
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    • 2006
  • The aim of this study was to survey susceptibilities of Escherichia coli and Klebsiella pneumoniae isolates against cefotaxime and to determine the prevalences of CTX-M type extended-spectrum $\beta$-lactamases (ESBLs) producing E. coli and K. pneumoniae in Korea. During the period of February to July, 2005, 153 E. coli and 52 K. neumoniae isolates were collected from 2 hospitals in Busan. Antimicrobial susceptibilities to cefotaxime were tested by the disk diffusion method. ESBL production of E. coli and K. pneumoniae was determined by the double disk synergy test. MICs of $\beta$-lactam antibiotics were determined by the agar dilution method. Blac$_{CTX-M}$ genes of the organism were detected by PCR. Among 153 isolates of E. coli and 52 isolates of K. neumoniae, 27 (17.6%) and 25 (48.0%) were intermediate or resistant to cefotaxime, respectively. Twenty-three (15.0%) isolates out of 153 E. coli and 13 (25.0%) out of 52 K. neumoniae isolates showed positive results for ESBL by the double disk synergy test. Twenty isolates out of 23 ESBL producing E. coli and 12 out of 13 ESBL producing K. neumoniae isolates harbored biacTx-M gene,11 of ESBL producing E. coli and 12 of ESBL producing K. neuinoniae isolates harbored bla$_{CTX-M}$ gene, 11 of the ESBL producing E. coli and 2 of ESBL producing K. neumoniae isolates harbored bla$_{TEM}$ gene, and 1 of the ESBL producing E. coli and 12 of ESBL producing K. neumoniae isolates harbored bla$_{SHV}$ gene. E. coli and K. neumoniae isolates producing CTX-M-type ESBLs were not uncommon in Korea. It is thought that continuous survey are necessary for inspecting the spread and novel variants of CTX-M-type ESBL genes. Further me]'e investigation and research on ESBL producing strains are needed in order to prevent the spread of resistant bacteria.

Nutrition Education Performance of Elementary School Dietitians in North Gyeonggi Province (경기 북부 지역 초등학교 영양사의 영양 교육 실시 현황)

  • Min Kyung-Chan;Park Young-Sim;Park Hae-Won;Lee Myung-Ho;Shin Yong-Chill;Cho Kyu-Bong;Rhie Kyoung-Ik;Jeaung Koang-Ock;Shin Yim-Sook;Yoon Hee-Sun
    • The Korean Journal of Food And Nutrition
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    • v.19 no.2
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    • pp.183-192
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    • 2006
  • The purpose of this study was to investigate the performance of elementary school dietitians in terms of nutrition education in the northern portion of Gyeonggi province. Self-administered questionnaires were given to 50 dietitians who have worked in elementary schools with self-operation food service, and 35(70%) dietitians returned the questionnaires. The results are summarized as follows: no students took part in nutrition education as a regular course, but all dietitians performed nutrition education in passive ways, such as 'using home correspondence'(39.0%), 'bulletin board/poster'(22.0%), 'using the internet'(13.4%) and 'indirectly through a classroom teacher'(12.2%). Most respondents performed nutrition education 'one time/month'(66.0%) or 'one time/week'(20.0%). The respondents thought that suitable teaching times for nutrition education were 'during a related subject'(35.5%), 'during lunch time'(22.6%) rather than 'during an independent subject'(16.1%). Most of the dietitians(94.3%) did not perform nutrition counseling because of 'a lack of opportunity'(72.7%) and 'workload'(27.3%). Additionally 88.6% of respondents did not have the time of for nutrition counseling for parents because 'am not a teacher'(56.7%) and 'workload'(30,0%). Information sources for nutrition education were mainly 'internet'(71.4%) and 're-educationa1 materials'(17.1%). They possessed instructional materials in the forms of 'printed materials'(35.1 %), 'exhibition/bulletin board'(31.2%), and 'electrical materials'(33.8%), 'but did not have 'solid materials' such as food models and dolls. Generally they had mostly 'leaflets'(82.9%), 'bulletins'(68.6%), 'internet'(57.1%), and 'CDs'(57.1%). Preferences for instructional materials used were 'printed materials'(46.2%), 'exhibition/bulletin board'(36.5%), and 'electrical materials'(17.3%) 'Leaflets'(80.0%) were mainly used; 'CD'(17.1 %) use was low compared to the proportion possessing CDs. The topics frequently chosen by the subjects for nutrition education were 'table manners'(82.9%), 'basic concepts of food and nutrition'(80.0%), and 'proper food habits'(80.0%), but the topics helpful for practical use, such as 'how much do I eat'(20.0%) and 'nutrition labeling'(37.1%), were not included frequently. The respondents thought that 'eating only what they like'(60.0 %), 'intake of processed foods'(17.8%), and 'obesity'(17.8%) were the most common nutritional problems among elementary school children. They also thought that establishing a regular course for nutrition education was an effective way to cut down on these nutritional problems. In conclusion, nutrition education programs that are combined with effective instructional materials and practical topics should be developed. Additionally, it is recommended that dietitians act as teachers who participate in regular courses as soon as possible.

Cerebral-perfusion Reserve after Carotid-artery Stenting: Relationship with Power Spectrum of Electroencephalography (경동맥스텐트삽입술 후의 뇌관류예비능: 뇌파파워스펙트럼과의 연관성)

  • Jeong, Da-hye;Jung, Seokwon;Kwak, Byeonggeun;Kim, Young-Soo;Kim, Soo-kyoung;Kwon, Oh-Young
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.2
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    • pp.144-152
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    • 2016
  • Carotid-artery stenosis may reduce cerebral perfusion, and affect cerebral neuronal activities. We examined the question of whether the recovery of cerebral-perfusion reserve after carotid-artery stenting (CAS) can affect the EEG power-spectrum. Nineteen candidates for CAS were initially recruited. Subtraction imaging of single photon emissary computerized tomography (SPECT) and an electroencephalogram (EEG) were taken twice, before and 1 month after CAS. At each time point, the EEGs were recorded before and after injection of acetazolamide (pre-ACZ EEG and post-ACZ EEG). Finally, 7 patients were enrolled after exclusion of incomplete studies. We obtained the spectral ratio (SR) of each hemisphere. SR was defined as the divided value of the power-spectrum sum of fast activities by that of slow activities. The power-spectrum values between hemispheres were compared using the inter-hemispheric index of spectral ratio (IHISR), and we examined the correlation between the power-spectrum and the cerebral-perfusion reserve. Cerebral-perfusion reserve improved after CAS on the stent side in 6 of 7 patients. In 3 patients with unilateral carotid-artery stenosis, CAS increased SR on the pre-ACZ EEGs, and IHISR on the post-ACZ EEGs. The increases of SR and IHISR were concordant with the increment of cerebral-perfusion reserve. In contrast, the results in the other patients with bilateral stenosis showed complex patterns. The SR of pre-ACZ EEGs and IHISR of post-ACZ EEGs may be useful electrophysiological markers for the blood-flow reserve after CAS in patients with unilateral carotid-artery stenosis, but not in those with bilateral stenosis.

Population-Based Newborn Hearing Impairment Screening Test Using GJB2 Mutation Analysis

  • Lee, Kyung-Ok;Jeong, Su-Jin;Byun, Ji-Young;Kim, Jeong-Sook;Lee, Hye-Jung;Seong, Hye-Soon;Kim, Kyung-Tae
    • Korean Journal of Clinical Laboratory Science
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    • v.39 no.2
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    • pp.113-121
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    • 2007
  • Hearing loss is a common congenital disorder that is frequently associated with mutations in the Cx26 gene (GJB2). Recently, the mutation analysis of GJB2 has been used in a newborn screening test for the detection of hearing impairment. Population-based studies should be performed before the application of genetic testing for the identification of deaf newborns. In this study, 8 positions of GJB2 mutations-including 35delG, 167delT, 235delC, V27I, V37I, M34T, E114G, and I203T-were analyzed using PCR-direct sequencing in a total of 437 healthy Korean neonates. DNAs from dried blood spots were extracted using a commercial DNA extraction kit. The PCR-amplified products (783 bps) of the GJB2 gene were detected using 2% agarose gel electrophoresis and subjected to direct sequencing. The sequences were compared with those in the GenBank database by using the BLAST program. In this study, 5 GJB2 mutations -including V27I (79G>A), V37I (109G>A), E114G (341A>G), I203T (608T>C), and 235delC- were found. Of the 437 neonate samples, 301 subjects showed GJB2 mutations (68.9%, 301/437). The V27I mutation was found in 271 subjects and was the most frequent (62.0%, 271/437). The E114G, I203T and V37I mutations were shown in 146, 17 and 14 subjects, respectively. The 235delC mutation was found in 1 subject. The E114G mutation was frequently accompanied by the V27I mutation. V27I/E114G (97.2%, 143/147) was the most common double mutation and 3 subjects had the double mutation V27I/I203T. A triple mutation, V27I/E114G/I203T, was found in 1 subject. In conclusion, PCR-direct sequencing is a convenient tool for the rapid detection of GJB2 mutations and this data might provide information for the genetic counseling of the GJB2 gene.

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Evaluation of p16INK4a/Ki-67 Dual Immunostaining in Liquid-based Cytology for Diagnosis of Uterine Cervical Dysplasia and Cancer (자궁경부 이형성증과 암의 진단을 위한 액상세포 검체에서 p16INK4a/Ki-67 이중면역염색의 평가)

  • Sung, Mi Hee;Lee, Hoon Taek;Shin, Min Shik;Oh, Seo Young;Kim, Wook Youn
    • Korean Journal of Clinical Laboratory Science
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    • v.47 no.3
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    • pp.132-139
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    • 2015
  • Recently, $p16^{INK4a}$/Ki-67 dual immunostaining has been introduced as a new biomarker protocol for early detection of uterine cervical dysplasia and cancer in liquid-based cytology (LBC). We performed the $p16^{INK4a}$/Ki-67 dual immunostaining using a CINtec$^{(R)}$ PLUS kit in a total of 109 LBC cases of cervicovaginal smear and compared its results with those from LBC, HPV hybrid capture II (HC II) test and histological diagnosis. Expression of $p16^{INK4a}$ and Ki-67 was significantly associated with cases of LSIL or higher in cytological diagnosis and cases of cervical intraepithelial neoplasia (CIN) 1 or higher in histological diagnosis (p<0.001 and p<0.001, respectively). Among forty-six cases of atypical squamous cells of undetermined significance (ASCUS) in LBC, $p16^{INK4a}$ and Ki-67 was expressed in 31 (67.4%), which were positively associated with cases of CIN I lesion or higher in histology. The sensitivity of $p16^{INK4a}$/Ki-67 dual immunostaining for finding lesions of CIN 1 or higher was 89.0%, which was higher than LBC. The specificity was 73.5%, which was higher than that of the HC II test. Based on these results, the $p16^{INK4a}$/Ki-67 dual immunostaining method can be a useful diagnostic marker for improving the sensitivity of LBC and the specificity of HC II test.

Effect of Prostaglandin F2 Alpha on E-cadherin, N-cadherin and Cell Adhesion in Ovarian Luteal Theca Cells (난소의 황체협막세포에서 E-cadherin, N-cadherin과 세포부착에 미치는 Prostaglandin F2 Alpha의 영향)

  • Lee, Sang-Hee;Jung, Bae Dong;Lee, Seunghyung
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.3
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    • pp.360-369
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    • 2019
  • Cadherins are essential transmembrane proteins that promote cell-cell adhesion and maintain the corpus luteum structure in the ovary. This study examined the influence of prostaglandin F2 alpha ($PGF2{\alpha}$) on E-cadherin, N-cadherin, and adhesion in luteal theca cells (LTCs). The luteal cells were isolated from the mid-phase corpus luteum, and the LTCs were cultured separately from the luteal heterogeneous cells according to the morphology of the mesenchymal cells and to determine if steroidogenic and endothelial cells of LTCs, 3beta-hydroxysteroid dehydrogenase ($3{\beta}$-HSD), and vascular endothelial growth factor receptor 2 (VEGFR2) mRNA were used. The LTCs were then incubated in the culture medium supplemented with 0.01, 0.1, and 1.0 mM $PGF2{\alpha}$ for 24 h, and the E-cadherin and N-cadherin proteins in the LTCs were detected by confocal laser scanning microscopy. The results revealed $3{\beta}$-HSD mRNA expression in the LTC but no VEGF2R mRNA expression. The E-cadherin and N-cadherin proteins of the LTCs were damaged in the 0.01, 0.1, and 1.0 mM $PGF2{\alpha}$ treatment groups, and the expression of the N-cadherin protein was reduced significantly in 0.01 mM $PGF2{\alpha}$ compared to the 0 mM $PGF2{\alpha}$ treatment groups (P<0.05). In addition, the number of attached LTCs were significantly lower in the 0.01 mM $PGF2{\alpha}$ treatment group than in the 0 mM $PGF2{\alpha}$ treatment group (P<0.05). In conclusion, $PGF2{\alpha}$ affected the disruption of cadherin proteins and cell adhesion in LTCs. These results may help better understand the cadherin and adhesion mechanism during corpus luteum regression in the ovary.

Expression of the Brother of the Regulator of Imprinted Sites Gene in the Sputum of Patients with Lung Cancer

  • Lee, Hae Young;Kim, Jong In;Cho, Sung Ho;Ko, Taek Yong;Kim, Hyun Su;Park, Sung Dal;Cho, Sung Rae;Chang, Hee Kyung;Hwang, Guk Jin;Jung, Sang Bong
    • Journal of Chest Surgery
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    • v.47 no.4
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    • pp.378-383
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    • 2014
  • Background: Brother of the regulator of imprinted sites (BORIS) is a putative new oncogene that is classified as a cancer germline gene; however, its role in the development of cancer is unclear. This study investigated the expression of BORIS in lung cancer and its clinical implications. Methods: The expression of BORIS messenger ribonucleic acid (mRNA) in the sputum of 100 patients with lung cancer (50 with squamous cell carcinoma, 36 with adenocarcinoma, and 14 with small-cell carcinoma) was evaluated by reverse transcription polymerase chain reaction. Results: The overall expression rate of BORIS in patients with lung cancer was 36.0%: 19 of 50 squamous cell carcinomas (38.0%), 13 of 36 adenocarcinomas (36.1%), and 4 of 14 (28.6%) small-cell carcinomas. There was no significant difference in the BORIS expression according to age, gender, or histologic type. However, the mRNA expression of BORIS was significantly related to the pathologic cancer stage (p=0.004) and lymph node metastasis (p=0.001). The expression of the melanoma antigen gene family A1-6 was not associated with the expression of BORIS. Conclusion: Our results suggest that the expression of BORIS might be a negative prognostic factor in lung cancers and implicate BORIS as a molecular target for immunotherapy.

Clinical Characteristics of Acute Drug Intoxication in Emergency Department

  • Kim In Byung;Chun Sung Pil;Kim Seung Whan
    • Biomedical Science Letters
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    • v.10 no.3
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    • pp.299-303
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    • 2004
  • The number of acute drug intoxication who visiting to emergency department which is located near urban and rural area concomitantly is inclined to increase slightly, The purpose of this study was the investigation of methods and strategies of management of acute drug intoxication in emergency department Clinical trials were done on 92 cases of acute drug intoxication visiting to Emergency Department of Chungnam National University Hospital during 4 months from April to July 2003. 1. The total number of acute drug intoxication during this period was 92 patients, which presented 0.95% of all the emergency department visiting patients during the study. The intentional ingestions were observed in 84 cases (91.3%). 2. The maximal point of age distribution was 4th decade. The number of that was 21 cases (22.8%). 3. The number of patients who had ingested benzodiazepines and doxylamine succinates was 26 cases (28.2%), organophosphorus was 20 (21.7%), paraquat was 10 (10.8%), others were 36 (39.3%). 4. Five kinds of po1ydrug ingestion was observed in 3 cases, four kinds was observed in 3, three kinds was observed in 5 and co-ingestion of alcohol was observed in 28 cases (30.4%). 5. The mortality was occurred in 4 cases, 2 of them were caused by paraquat, 1 of them was caused by organophosphorus (OP), 1 of them was caused by acetic acid. 6. The gastric larvage as a mean of treatment modality was done in 57 cases (61.9%). The use of activated charcoal was done in 8 (8.69%). The maintenance of tracheal intubation was done in 6 (6.52%). 7. The mean observational period in emergency department was 8 hours 42 minutes in benzodiazepine and doxylamine succinate group, 21 hours 46 minutes in OP, 20 hours 39 minutes in other germicidal except OP, 23 hours 9 minutes in paraquat group. Without a drug information and intoxication center in Korea, We should minimize the exhaustion of medical resources by establishment of determinant criteria which can be seen in relatively less-complicated cases of acute drug intoxication. Thus, we should consider the introduction of simple toxicology treatment protocol and toxicologic observation unit in emergency department as possible means to reduce economical and social burdens.

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Detection of Enterohemorrhagic Escherichia coli O157:H7 Strains Using Multiplex Polymerase Chain Reaction (Multiplex PCR을 이용한 장출혈성 대장균 O157:H7의 검출)

  • 엄용빈;김종배
    • Biomedical Science Letters
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    • v.4 no.1
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    • pp.43-56
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    • 1998
  • A multiplex PCR method was designed by employing primers specific for the eaeA gene, conserved sequences of Shiga-like toxins (SLT-I.II), and the 60-MDa plasmid of enterohemorrhagic E. coli (EHEC) O157:H7 strain. A set of six synthetic oligonucleotide primers derived from sequences of the SLT-I.II, eaeA, and 60-MDa plasmid genes of E. coli O157:H7 were used in a multiplex PCR amplification procedure to detect these genes in the same enteric pathogens. In two enterohemorrhagic E. coli O157:H7 (ATCC 35150, ATCC 43894) reference strains, PCR products of 317bps (eaeA), 228bps (SLT-I.II), and 167bps (60-MDa plasmid) were successfully amplified simultaneously in a single reaction. However, the specific PCR products were not amplified in control strains of other enteric bacteria. The sensitivity of the multiplex PCR assay for detection of the SLT-I.II, eaeA, and 60-MDa plasmid genes of E. coli O157:H7 was found to be 2.5$\times$10$^{6}$ of bacteria in diarrheal stool to amplify all three bands. The multiplex PCR technology will allow large-scale screening of many clinical specimens or contaminated foods, and will be a very useful method for the detection of a wide range of microorganisms present in the environment, including EHEC O157:H7 in various types of specimens. The multiplex PCR assay has the potential to be used as a specific and rapid method for clinical diagnosis of disease caused by EHEC O157:H7.

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