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Susceptibility tests of oral antibiotics including cefixime against Escherichia coli, isolated from pediatric patients with community acquired urinary tract infections (소아 원외 요로감염 환아에서 분리된 E. coli에 대한 cefixime을 포함한 경구 항생제의 감수성 연구)

  • Lee, Soo Young;Lee, Jung Hyun;Kim, Jong Hyun;Hur, Jae Kyun;Kim, Sun Mi;Ma, Sang Hyuk;Kang, Jin Han
    • Clinical and Experimental Pediatrics
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    • v.49 no.7
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    • pp.777-783
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    • 2006
  • Purpose : Urinary tract infection(UTI) is one of the most frequent infections in children. E. coli is the most frequent etiological micropathogen in pediatric community UTI, and E. coli has developed resistance to many antibiotics, highlighting the need for regular surveys of this organism resistant patterns in the community. The aim of this study was to determine the oral antibiotic susceptibility patterns of E. coli, isolated from pediatric patients with uncomplicated community acquired UTI. Methods : E. coli isolates, obtained from pediatric patients with uncomplicated community acquired UTI between October in 2004 to September in 2005. And minimal inhibitory concentrations(MICs) of oral aminopenicillins and beta-lactamase inhibnitors(ampicillin, amoxacillin, ampicillin-sulbactam), oral cephalosporins(cefaclor, cefixime) and sulfa drug(trimethoprime-sulfamethoxazole) were performed according to the National Committee for Clinical Laboratory Standards(NCCLS) guide line. Results : Total 211 organisms were isolated from pediatric out-patients with community UTI. E. coli was the most common organism(89 percent), followed by E. fecalis, Proteus species, S. aureus, M. morganii, and P. aeruginosa. The resistant rates of aminopenicillins and beta-lactamase inhibitors, cefaclor and sulfa drug to E. coli were very high. But, the resistant rate of cefixime was markedly low, and ESBL strains were isolated with small rates. Conclusion : Our study results suggest that aminopenicillins, cefaclor and sulfa drug may not be useful as first line empirical antibiotics to treat pediatric patients with community UTI in Korea. But, 3rd generation cephalosporin such as cefixime can be used as effective second line antibiotics after primary treatment failure, also may be useful as an empirical first line antibiotic. Finally, we conclude that a continuous surveillance study to monitor susceptibility patterns of E. coli in community UTI will be needed for the standard guide lines of empirical oral antibiotic treatment.

Convenient Nucleic Acid Detection for Tomato spotted wilt virus: Virion Captured/RT-PCR (VC/RT-PCR) (Tomato spotted wilt virus를 위한 간편한 식물바이러스 핵산진단법: Virion Captured/RT-PCR (VC/RT-PCR))

  • Cho Jeom-Deog;Kim Jeong-Soo;Kim Hyun-Ran;Chung Bong-Nam;Ryu Ki-Hyun
    • Research in Plant Disease
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    • v.12 no.2
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    • pp.139-143
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    • 2006
  • Virion captured reverse transcription polymerase chain reaction (VC/RT-PCR) could detect plant virus quickly and accurately. In the VC/RT-PCR, no antibody is needed unlike immuno-captured RT-PCR (IC/RT-PCR) which had been improved method of RT-PCR for plant viruses, and virus nucleic acids can be obtained easily within 30minutes by property of polypropylene PCR tube which is hold and immobilized viral particles on its surface. For the virion capture of Tomato spotted wilt virus (TSWV), the extraction buffer was tested. The optimum macerating buffer for TSWV was 0.01M potassium phosphate buffer, pH 7.0, containing 0.5% sodium sulfite. The viral crude sap was incubated for 30 min at $4^{\circ}C$. The virions in the PCR tubes were washed two times with 0.01M PBS containing 0.05% Tween-20. The washed virions were treated at $95^{\circ}C$ immediately for 1 min containing RNase free water and chilled quickly in the ice. Disclosed virions' RNAs by heat treatment were used for RT-PCR. Dilution end point of $10^{-5}$ from plant's crude sap infected with TSWV showed relatively higher detection sensitivity for VC/RT-PCR. During multiple detection using two or more primers, interference was arisen by interactions between primer-primer and plant species. The result of multiplex RT-PCR was influenced by combinations of primers and the kind of plant, and the optimum extraction buffer for the multiplex detection by VC/RT-PCR should be developed.

Detection of Toxoplasma antigens and antibodies in mice infected with different strains of Toxoplasma gonnii (톡소포자충의 충주에 따른 항원과 항체의 검출 시기 및 양상)

  • 이영하;김재영
    • Parasites, Hosts and Diseases
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    • v.33 no.3
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    • pp.201-210
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    • 1995
  • This study aims to assess the possible strain-dependent variations in detection of ToxopLosmn antigens and antibodies. The virulent RH strain or avirulent Beverley strain of T gondii were injected into mice, intraperitoneally, and their antigens, antibodies and parasites were identified from the blood or tissues: liver, brain and spleen by ELISA, Western blot and PCR. In mice infected with RH strain, circulating antigens and parasitemia were first detected from 2 days after infection, and ToxopIasma DNA were found in the blood, liver, brain and spleen from 3 days after infection. It was impossible to detect specific IgM and IgG antibodies to T gondij and any specific band was not found by Western blot. In mice infected with Beverley strain, circulating antigens were detected between day 10 and day 35. The Toxoplusma DNA was found in the blood and liver from day 15 until day 60, and in the brain from day 20. But Toxoplosma DNA in the spleen were mainly detected between day 10 and day 30. The IgM antibodies were first appeared on day 10 post-infection, and were noted obviously increased between day 15 and 25. The IgG antibodies were first detected on day 15, and showed progressively increased titers. The antibody binding bands were specific according to infection period. Sera from mice infected with Beverley strain reacted mainly with the antigen of 27.5-kDa and 32.5-kDa. In conclusion, mice infected with RH strain revealed Toxoplosma antigens strongly, but not antibodies. However. mice infected with Beverley strain revealed both the Toxoplasma antigens and antibodies. The present results showed that immune responses are different between avirulent and virulent T gonnii.

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Development of an in vitro culture method for harvesting the free-living infective larvae of Strongyloides venezuelensis (베네수엘라분선충 (Strongyloides venezuelensis Brumpt, 1934) 자유생활형 유충의 시험관 내 배양 기술 개발)

  • ;M.
    • Parasites, Hosts and Diseases
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    • v.36 no.1
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    • pp.15-22
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    • 1998
  • An in uitro culture technique was established for harvesting Strongwloides venezuelensis free-living infective larvae using a nutrient broth medium as a substitute for rat-feces in polyvinyl culture bags ($10{\;}{\times}{\;}12{\;}cm$). The egg hatch rate V) in sterile saline at different incubation temperatures (X) was expressed as the quadratic function, Y = $-0.192X^2$ + 8.673x - 19.550 (r = 0.901). The highest (100%) egghatch rate was observed at $25^{\circ}C$. A significant difference (p<0.05) in development rate W) of free-living infective larvae was observed between different concentrations of nutrient broth (X) which was highest (20.6%1 in 0.12% nutrient broth concentrations, incubated at $20^{\circ}C$ for 5 days [Y = $-864.032X^2$ + 245.995X- 0.560 (r = 0.875)]. Yields (Y) of infective larvae were observed relatively high when the culture medium was incubated at higher temperatures (X) which peaked at $25^{\circ}C$ (20.0%) than at lower temperatures. $15^{\circ}C$M (10.9%) and $20^{\circ}C$ (18.1%) [Y = $-0.189X^2$ + 8.387x- 72.795 (r = 0.981)]. The period W) required for the development of infective larvae decreased with higher incubation temperatures (X) [Y = $0.035X^2$ - 2.025X + 32.375 (r = 0.995)] The highest yield (19.2%) of infective larvae was obtained from culture bag inoculated with 15.000 eggs than with below and over 15,000 eggs in 0.12% nutrient broth and incubated at $25^{\circ}C$ for 4 days. The newly adapted culture method (from egg to third-stage larva) may be useful as a bio-bar/bioassay system for screening new chemical products, anthelmintics and pesticides, as well as for parasito immunological studies with Strongwloides species.

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Effects of Oxidative DNA Damage and Genetic Polymorphism of the Glutathione Peroxidase 1 (GPX1) and 8-Oxoguanine Glycosylase 1 (hOGG1) on Lung Cancer (GPX1 및 hOGG1 유전자다형성에 따른 유전자의 산화적 손상 및 폐암 발생 위험도 평가)

  • Lee, Chul-Ho;Lee, Kye-Young;Choe, Kang-Hyeon;Hong, Yun-Chul;Noh, Sung-Il;Eom, Sang-Yong;Ko, Young-Jun;Zhang, Yan-Wei;Yim, Dong-Hyuk;Kang, Jong-Won;Kim, Heon;Kim, Yong-Dae
    • Journal of Preventive Medicine and Public Health
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    • v.39 no.2
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    • pp.130-134
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    • 2006
  • Objectives : Oxidative DNA damage is a known risk factor of lung cancer. The glutathione peroxidase (GPX) antioxidant enzyme that reduces hydrogen peroxide and lipid peroxides plays a significant role in protecting cells from the oxidative stress induced by reactive oxygen species. The aim of this case-control study was to investigate effects of oxidative stress and genetic polymorphisms of the GPX1 genes and the interaction between them in the carcinogenesis of lung cancer. Methods : Two hundreds patients with lung cancer and 200 age- and sex-matched controls were enrolled in this study. Every subject was asked to complete a questionnaire concerning their smoking habits and their environmental exposure to PAHs. The genotypes of the GPX1 and 8-oxoguanine glycosylase 1 (hOGG1) genes were examined and the concentrations of urinary hydroxypyrene (1-OHP), 2-naphthol and 8-hydroxydeoxyguanosine (8-OH-dG) were measured. Results : Cigarette smoking was a significant risk factor for lung cancer. The levels of urinary 8-OH-dG were higher in the patients (p<0.001), whereas the urinary 1-OHP and 2-naphthol levels were higher in the controls. The GPX1 codon 198 polymorphism was associated with an increased risk of lung cancer. Individuals carrying the Pro/Leu or Leu/Leu genotype of GPX1 were at a higher risk for lung cancer (adjusted OR=2.29). In addition, these individuals were shown to have high urinary 8-OH-dG concentrations compared to the individuals with the GPX1 Pro/Pro genotype. On the other hand, the polymorphism of the hOGG1 gene did not affect the lung cancer risk and the oxidative DNA damage. Conclusions : These results lead to a conclusion that individuals with the GPX1 Pro/Leu or Leu/Leu genotype would be more susceptible to the lung cancer induced by oxidative stress than those individuals with the Pro/Pro genotype.

Effects of Tested Pack Containing Plant Extracts on Elasticity and Size of Women's Breasts (식물추출물 팩의 여성가슴 탄력증진과 크기증대에 관한 연구)

  • Choi, Yea-Hun;Park, Min-Kyung;Kim, Yong-Gyun;Lee, Sang-Mong;Son, Hong-Joo;Park, Hyean-Cheal;Kim, Sun-Tae;Choi, In-Soo;Kim, Keun-Ki
    • Journal of Life Science
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    • v.22 no.3
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    • pp.407-416
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    • 2012
  • In this study, we purified the extracts from the seeds and the roots of various plant species, including Q. acutissima, C. lanceolata, P. mirifica, P. bambusoides, and S. repens, and then investigated the effects of these extracts on cell growth and fat accumulation in adipocytes. We found that the extracts purified from Q. acutissima, C. lanceolata, P. mirifica, P. bambusoides, and S. repens more effectively increased the cell growth, as well as promoting the fat accumulation in adipocytes to a greater extent, than other extracts in vitro. Therefore, we made breast packs containing these effective extracts, and then investigated whether they were effective in enhancing the elasticity and volume of women's breasts. The measurements of breast elasticity and size revealed that the breast packs efficiently increased the elasticity and size of women's breasts. Furthermore, evaluation of the questionnaires related to usage of the breast packs indicated great satisfaction in terms of the lift, firmness, and elasticity of breasts. In conclusion, extracts purified from Q. acutissima, C. lanceolata, P. mirifica, P. bambusoides, and S. repens leading to cell growth and fat accumulation in adipocytes can effectively contribute to improving the elasticity and size of women's breasts.

Inhibitory effects of heavy metals on CYP1A expression in eel hepatocyte cultures (뱀장어 배양 간세포에서의 Cytochrome P4501A (CYP1A) 유전자 발현에 대한 중금속들의 억제효과)

  • Kwon, Hyuk-Chu;Maeng, Joon-Ho;Choi, Seong-Hee
    • Journal of fish pathology
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    • v.23 no.2
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    • pp.245-254
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    • 2010
  • Effects of heavy metal ions on the gene expression of cytochrome P4501A (CYP1A) were examined in cultured eel hepatocytes. When the expression of CYP1A mRNA was measured by RT-PCR after incubation of eel hepatocytes with benzo[$\alpha$]pyrene (B[$\alpha$]P) at concentrations of 10-8~10-5 M, the CYP1A expression increased with B[$\alpha$]P treatment in a dose dependent manner, showing significant increase at concentrations more than 10-7 M. When the eel hepatocyte was treated with cadmium (10-6 and 10-5 M), the expression of CYP1A was inhibited and especially at higher concentration (10-5 M). The inhibition of CYP1A expression by cadmium was also observed in cells treated with B[$\alpha$]P. In another study, effects of heavy metal ions on the expression of CYP1A were examined in cultured hepatocytes isolated from eel which was treated previously with B[$\alpha$]P in vivo. Hepatocytes isolated from the liver of eel taken at 48 hours after injection of B[$\alpha$]P (10 mg/kg) were cultured for 2 days with cadmium, copper, lead or zinc (10-6 and 10-5 M). The expression of CYP1A was found to be suppressed by the metal ions compared with the control in which CYP1A was induced with previous treatment of B[$\alpha$]P in vivo. The present results may provide an important basic information for studying the effects of heavy metal ions on CYP1A expression in other species of fish and studying toxicological mechanisms of heavy metal ions in aquatic livings.

Gene analysis of galectin-1, innate immune response gene, in olive flounder Paralichthys olivaceus at different developmental stage (넙치, Paralichthys olivaceus 발생단계별 galectin-1 유전자의 발현 분석)

  • Jang, Min Seok;Lee, Young Mee;Yang, Hyun;Lee, Jeong-Ho;Noh, Jae Koo;Kim, Hyun Chul;Park, Choul-Ji;Park, Jong-Won;Hwang, In Joon;Kim, Sung Yeon
    • Journal of fish pathology
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    • v.26 no.3
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    • pp.255-263
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    • 2013
  • The innate immune response is fundamental defense response of vertebrates and invertebrates. Especially, the innate immune response important for larvae that lack of resistance to infectious diseases in the early stages. Galectin is one of the kinds of lectin and presents in the fish mucous that involves innate immune response. Galectin have been studied from various fishing species, but expression analysis of galectin is still unclear during early developmental stage in olive flounder. In this study, we investigated gene expression of galectin-1 from various developmental stage and tissues. We excised several tissues including the muscle, fin, eye, gill, brain, stomach, intestine, kidney, spleen and liver from adult olive flounder and confirmed gene expression of galectin-1 using RT-PCR and quantitative real-time PCR. Expression of galectin-1 was significantly higher in muscle, stomach and intestinal tissue than other tissue in adult fish (5 and 29 months). Also, galectin-1 gene was detected from 0 DAH and gradually increased to 35 DAH and since then decreased after stomach development period. Induction of galectin-1 during the early developmental stage suggest that muscle, fin and eye tissue is formed and begins the secretion of galectin this period. In addition, increased expression levels at 35 DAH suggest that due to complete formation of stomach and intestine, increase of secretion and activation of enzyme. This study shows that expression of galectin-1 during early developmental stages and adult period in olive flounder and can be expect that galectin-1 play essental role in the innate immune system throughout the whole life time. Galectin-1 is primary barrier such as skin and digestive tissue against pathogen infection, also digestive tract developmental period is important for pathogen invasion can be expected that it will serve. Mass mortality due to the disease in seed production is continuing damage, therefore these result will be meaningful about infectious disease during early developmental stages as a basic data for the study.

Interspecific Transferability of Watermelon EST-SSRs Assessed by Genetic Relationship Analysis of Cucurbitaceous Crops (박과작물의 유연관계 분석을 통한 수박 EST-SSR 마커의 종간 적용성 검정)

  • Kim, Hyeogjun;Yeo, Sang-Seok;Han, Dong-Yeop;Park, Young-Hoon
    • Horticultural Science & Technology
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    • v.33 no.1
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    • pp.93-105
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    • 2015
  • This study was performed to analyze genetic relationships of the four major cucurbitaceous crops including watermelon, melon, cucumber, and squash/pumpkin. Among 120 EST-SSR primer sets selected from the International Cucurbit Genomics Initiative (ICuGI) database, PCR was successful for 51 (49.17%) primer sets and 49 (40.8%) primer sets showed polymorphisms among eight Cucurbitaceae accessions. A total of 382 allele-specific PCR bands were produced by 49 EST-SSR primers from 24 Cucurbitaceae accessions and used for analysis of pairwise similarity and dendrogram construction. Assessment of the genetic relationships resulted in similarity indexes ranging from 0.01 to 0.85. In the dendrogram, 24 Cucurbitaceae accessions were classified into two major groups (Clade I and II) and 8 subgroups. Clade I comprised two subgroups, Clade I-1 for watermelon accessions [I-1a and I-1b-2: three wild-type watermelons (Citrullus lanatus var. citroides Mats. & Nakai), I-1b-1: six watermelon cultivars (Citrullus lanatus var. vulgaris S chrad.)] a nd C lade I -2 for melon and cucumber accessions [I-2a-1 : 4 melon cultivars(Cucumis melo var. cantalupensis Naudin.), I-2a-2: oriental melon cultivars (Cucumis melo var. conomon Makino.), and I-2b: five cucumber cultivars (Cucumis sativus L.)]. Squash and pumpkin accessions composed Clade II {II-1: two squash/ pumpkin cultivars [Cucurbita moschata (Duch. ex Lam.)/Duch. & Poir. and Cucurbita maxima Duch.] and II-2: two squash/pumpkin cultivars, Cucurbita pepo L./Cucurbita ficifolia Bouche.}. These results were in accordance with previously reported classification of Cucurbitaceae species, indicating that watermelon EST-SSRs show a high level of marker transferability and should be useful for genetic study in other cucurbit crops.

Evaluation of Peptide Nucleic Acid Probe-Based Fluorescence In Situ Hybridization for the Detection of Mycobacterium tuberculosis Complex and Nontuberculous Mycobacteria in Clinical Respiratory Specimens (임상 객담검체에서 Peptide Nucleic Acid Probe를 이용한 결핵과 비결핵 항산균의 구분)

  • Lee, Seung Hee;Kim, Shine Young;Kim, Hyung Hoi;Lee, Eun Yup;Chang, Chulhun L.
    • Annals of Clinical Microbiology
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    • v.18 no.2
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    • pp.37-43
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    • 2015
  • Background: Tuberculosis is globally the most important cause of death from single pathogen. Rapid and accurate identification of mycobacteria is essential for the control of tuberculosis. We evaluated a fluorescence in situ hybridization (FISH) method using peptide nucleic acid (PNA) probes for the differentiation of Mycobacterium tuberculosis complex (MTB) and nontuberculous mycobacteria (NTM) in direct smears of sputum specimens. Methods: The cross-reactivity of MTB- and NTM-specific PNA probes was examined with reference strains of M. tuberculosis ATCC 13950, Mycobacterium kansasii ATCC 12479, Mycobacterium fortuitum ATCC 6841, several clinical isolates of mycobacteria (Mycobacterium abscessus, Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium gordonae and Mycobacterium chelonae), and 11 frequently isolated respiratory bacterial species other than mycobacteria. A series of 128 sputa (89 MTB culture positive, 29 NTM culture positive, and 10 under treatment culture negative) with grades of trace to 4+ were used to evaluate the performance of the method. Results: The MTB- and NTM-specific PNA probes showed specific reactions with the reference strains of MTB and M. kansasii and clinical isolates of mycobacteria except M. fortuitum ATCC 6841, and no cross-reactivity with other tested bacteria. The PNA probe-based FISH assay for detection of MTB had a sensitivity and specificity of 100%, respectively. The sensitivity and specificity of the NTM-specific PNA probe was 100%. The smear grades of the PNA FISH test were same as with those of the fluorescence AFB stain in 2+ or higher grade. Conclusion: Detection and differentiation based on PNA FISH is sensitive and accurate for detecting mycobacteria and for differentiating MTB from NTM in clinical sputum smears.