• Title/Summary/Keyword: Molecular Characteristics

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Genotypic and Phenotypic Characteristics of Staphylococcus aureus Isolates from Lettuces and Raw Milk (상추와 원유에서 분리한 황색 포도상구균의 유전형 및 표현형 특징)

  • Jung, Hye-Jin;Cho, Joon-Il;Park, Sung-Hee;Ha, Sang-Do;Lee, Kyu-Ho;Kim, Cheol-Ho;Song, Eun-Seop;Chung, Duck-Hwa;Kim, Min-Gon;Kim, Kwang-Yup;Kim, Keun-Sung
    • Korean Journal of Food Science and Technology
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    • v.37 no.1
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    • pp.134-141
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    • 2005
  • To characterize genotypic and phenotypic traits of Staphylococcus aureus isolates (n = 86) from lettuces and raw milk, major virulence-associated genes and antibiotic susceptibility were detected using PCR-based methods and disk diffusion method, respectively. All isolates possessed coagulase gene and showed five polymorphism types [500 bp (2.4%), 580 bp (17.4%), 660 bp (61.6%), 740 bp (17.4%), and 820 bp (1.2%)] due to variable numbers of tandem repeats present within the gene. Two or three different loci of hemolysin gene family were dominant in isolates, 47 of which (55%) possessed combination of hla/hld/hlg-2 genes as the most prevalent types. Among enterotoxin-encoding genes, sea was detected from 32 isolates (37%), sed from 1 isolate (1%), and sea and sed genes were co-detected from 4 isolates (5%), whereas seb, sec, and tsst-1 genes were not detected. All isolates were susceptible to ciprofloxacin, trimethoprim/sulfamethoxazole, oxacillin, and vancomycin, 85 isolates (99%) to penicillin G, 54 isolates (63%) to chloramphenicol, 51 isolates (59%) to erythromycin, and 7 isolates (8%) to clindamycin. Among resistant isolates, seven displayed multiantibiotic-resistance against two different antibiotics.

Analysis of rpoB Gene in Rifampin-Resistant M. Tuberculosis by Direct Sequencing and Line Probe Assay (염기서열결정과 Line Probe 분석법에 의한 Rifampin내성 결핵균의 rpoB 유전자 분석)

  • Lee, Min-Ki;Kim, Yun-Seong;Lee, Hyo-Jin;Cheon, Du-Su;Yun, Sang-Myung;Park, Sam-Seok;Kim, Cheol-Min;Park, Soon-Kew
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.2
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    • pp.251-263
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    • 1997
  • Background : The emergence of multidrug-resistant strains of Mycobacterium tuberculosis presents a significant challange to the treatment and control of tuberculosis, and there is an urgent need to understand the mechanisms by which strains acquire multidrug resistance. Recent advances in molecular methods for the detection of M. tuberculosis genetic targets have approached the sensitivity of culture. Furthermore the prospect of determining resistance in mycobacteria at the nucleic acid level particulary to first-line drugs like rifampin, isoniazid has provided a glimps of the next generation of sensitivity test for M. tuberculosis. Previous studies in RMP resistant M. tuberculosis have shown that mutation in $\beta$subunit of RNA polymerase is main mechanism of resistance. Method : In this study, rpoB gene for the $\beta$subunit of RNA polymerase from M. tuberculosis of 42 cultured samples (32 were RMP resistant and 10 were sensitive cases) were isolated and characterised the mutations. Direct sequencing data were compared with the results of INNO-LiPA Line Probe Assay (LiPA, Innogenetics, Belgium), commercial RMP resistance detecting kit using reverse hybridization method. Results : All of the RMP resistant samples were revealed the presence of mutation by LiPA. In 22 samples (68.8%) out of 32 RMP resistant cases, the mutation types were confirmed by the positive signal at one of 4 mutation bands in the strip. The most frequent type was R5 (S531L) which were 17 cases (77.3%). Results of direct sequencing were identified the exact characteristics of 8 mutations which were not confirmed by LiPA. S522W type point mutation and 9 base pair deletion at codon 513~515 were new identified mutations for the first time. Conclusion : Mutations in rpoB gene is the main mechanism of RMP resistance in M. tuberculosis and LiPA is a very useful diagnostic tool for the early diagnosis of RMP resistance in M. tuberculosis.

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Studies on the Raw Starch Saccharifying Enzyme from the Aspergillus niger and Its Mutants (Aspergillus niger 및 그 변이주(變異株)의 생전분당화효소(生澱粉糖化酵素)에 관(關)한 연구(硏究))

  • Sohn, Cheon Bae;Park, Yoon Joong
    • Korean Journal of Agricultural Science
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    • v.10 no.1
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    • pp.166-185
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    • 1983
  • Aspergillus niger IFO 8541 (NRRL 3112) was investigated through a series of UV rays and N-Methyl-N'-Nitro-N-Nitrosoguanidine (NTG) treatments to induce mutants that produce highly active raw starch saccharifying enzyme, and two mutants with strong enzymatic productivity were obtained. The mutants obtained were investigated for their fungal characters, condition of enzyme production, and other activities. Furthermore, the raw starch saccharifying enzyme was purified and the characteristics of purified enzyme were studied. The results obtained were summarized as follows; 1. The color of conidial head of UV-46 mutant obtained from UV rays treatment was changed to tan type and the gelatinated starch saccharifying enzyme productivity and the raw starch saccharifying enzyme productivity increased up to twice and 1.8 times compared to the productivities of original Aspergillus niger IFO 8541 cultured on the wheat bran, respectively. 2. The conidial head color of NG-41 mutant obtained from NTG treatment became lighter than that of parent strain. The gelatinated starch saccharifying enzyme productivity and raw starch saccharifying enzyme productivity increased about 1.8 times, and twice over the Aspergillus niger IFO 8541 parent strain cultured on wheat bran, respectively. The productivity of ${\alpha}$-amylase increased about 3 times more than the parent strain. 3. Two peaks of glucoanlylase and a peak of ${\alpha}$-amylase were obtained when enzyme solution of mutants and parent strain were passed through DEAE-Sephadex A-50 column chromatography. Glucoamylase I showed only gelatinated starch saccharifying enzyme activity. However, glucoamylase II (raw starch saccharifying enzyme) showed both raw starch saccharifying enzyme activity and gelatinated starch saccharifying enzyme activity. 4. Mutant, UV-46 was strengthened in glucoamylase II productivity and mutant NG-41 was strengthened in ${\alpha}$-amylase productivity. 5. Glucoamylase II of mutants and parent strain were appeared to have the same enzymatic properties. 6. Glucoamylase II of mutants and parent strain were recognized as simple enzyme through electrophoresis. 7. The glucoamylase II crystallized showed rhombic board type. 8. The molecular weight, isoelectric point, optimum pH, and optimum temperature of the glucoamylase II crystallized were estimated as 76,000, 3.4, 3.5 and $60^{\circ}C$, respectively.

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Comparison of the Results for Sentinel Lymph Node Mapping in the Breast Cancer Patients using $^{99m}Tc$-Antimony Trisulfide Colloid, $^{99m}Tc$-Tin Colloid, and $^{99m}Tc$-Human Serum Albumin (유방암 환자에서 $^{99m}Tc$-Antimony Trisulfide Colloid, $^{99m}Tc$-Tin Colloid, $^{99m}Tc$-Human Serum Albumin을 이용한 감시림프절 매핑 성적의 비교)

  • Jang, Sung-June;Moon, Seung-Hwan;Kim, Seok-Ki;Kim, Bom-Sahn;Kim, Seok-Won;Chung, Ki-Wook;Kang, Keon-Wook;Lee, Eun-Sook
    • Nuclear Medicine and Molecular Imaging
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    • v.41 no.6
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    • pp.546-552
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    • 2007
  • Purpose: In the breast cancer patient, lymphatic mapping and sentinel lymph node biopsy are the most important procedure for axillary lymph node staging. We aimed to compare the three radiocolloids [$^{99m}Tc$-antimony trisulfide colloid (ASC), $^{99m}Tc$-tin colloid (TC), and $^{99m}Tc$-human serum albumin (HSA)] for sentinel lymph node mapping. Subjects and Methods: Totally, 397 patients with clinically N0 stage were enrolled. $^{99m}Tc$-ASC was injected in 202 out of 397 patients, $^{99m}Tc$-TC was injected in 120 patients, and $^{99m}Tc$-HSA was injected in the remaining 75 patients. The sentinel lymph nodes were localized by lymphoscintigraphy and selected using intraoperative gamma probe. All sentinel lymph nodes were investigated by intraoperative pathologic consultation. The axillary lymph nodes which were harvested by the lymph node dissection were also investigated. Results: The patients of each group showed similar clinical characteristics. There were no significant differences (p>0.05) in the identification rate of sentinel lymph nodes (IR), false negative rate (FNR), and negative predictive value (NPV). The axillary lymphadenectomy revealed axillary lymph node metastases in those three groups (ASC-33.2%, TC-31.7%, HSA-22.7%). The IR, FNR, and NPV were not significantly different among those groups. Conclusion: Those three $^{99m}Tc$-labeled radiocolloids showed equivalent results in sentinel lymph node mapping of breast cancer.

Physicochemical Characteristic of the Silkworm Sericin Cocoon (세리신잠견의 이화학적 특성)

  • 김수연;손해룡;배도규;김정호
    • Journal of Sericultural and Entomological Science
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    • v.45 no.1
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    • pp.10-17
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    • 2003
  • This study was carried out to analyze physicochemical characteristics of sericin cocoon from silkworm, Bombyx mori. The degumming loss increased with increasing treatment time up to 2 hr, and temperature up to 130$^{\circ}C$. At 130$^{\circ}C$, degumming loss of Nd-s jam and Nd$\^$H/ jam were 100% while that of Baegok jam was 24%. Nd-s jam and Baegok jam ha high glycine content of 29.1∼46.3 mol% where as Nd$\^$H/ jam had high serine content of 32.6 mol%. Thermal denaturation temperatures were found at 218$^{\circ}C$ for Nd-s jam, 216$^{\circ}C$ for Nd$\^$H/ jam, and 218$^{\circ}C$ for Baegok jam. Before degumming, crystallinities obtained by FT-IR analysis were 44.3, 43.7, and 59.9% for Nd-s jam, Nd$\^$H/ jam, and Baegok jam respectively. After degumming, crystallinity increased to 61.8% for Baegok jam. Before degumming, crystallinitics obtained from XRD were 35.9, 33.5, and 47.2%, for Nd-s jam, Nd$\^$H/ jam, and Baegok jam. After degumming, crystallinity increased to 49.8% for Baegok jam. The molecular weight of Nd$\^$H/ jam were 9,417 in 1 hr, 3,744 in 2 hr, 4,944 in hr, and 3,910 in 6 hr.

Dry Etching of GaAs and AlGaAs in Diffuion Pump-Based Capacitively Coupled BCl3 Plasmas (확산펌프 기반의 BCl3 축전결합 플라즈마를 이용한 GaAs와 AlGaAs의 건식 식각)

  • Lee, S.H.;Park, J.H.;Noh, H.S.;Choi, K.H.;Song, H.J.;Cho, G.S.;Lee, J.W.
    • Journal of the Korean Vacuum Society
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    • v.18 no.4
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    • pp.288-295
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    • 2009
  • We report the etch characteristics of GaAs and AlGaAs in the diffusion pump-based capacitively coupled $BCl_3$ plasma. Process variables were chamber pressure ($50{\sim}180$ mTorr), CCP power ($50{\sim}200\;W$) and $BCl_3$ gas flow rate ($2.5{\sim}10$ sccm). Surface profilometry was used for etch rate and surface roughness measurement after etching. Scanning electron microscopy was used to analyze the etched sidewall and surface morphology. Optical emission spectroscopy was used in order to characterize the emission peaks of the $BCl_3$ plasma during etching. We have achieved $0.25{\mu}m$/min of GaAs etch rate with only 5 sccm $BCl_3$ flow rate when the chamber pressure was in the range of 50{\sim}130 mTorr. The etch rates of AlGaAs were a little lower than those of GaAs at the conditions. However, the etch rates of GaAs and AlGaAs decreased significantly when the chamber pressure increased to 180 mTorr. GaAs and AlGaAs were not etched with 50 W CCP power. With $100{\sim}200\;W$ CCP power, etch rates of the materials increased over $0.3{\mu}m$/min. It was found that the etch rates of GaAs and AlGaAs were not always proportional to the increase of CCP power. We also found the interesting result that AlGaAs did not etched at 2.5 sccm $BCl_3$ flow rate at 75 mTorr and 100 W CCP power even though it was etched fast like GaAs with more $BCl_3$ gas flow rates. By contrast, GaAs was etched at ${{\sim}}0.3{\mu}m$/min at the 2.5 sccm $BCl_3$ flow rate condition. A broad molecular peak was noticed in the range of $500{\sim}700\;mm$ wavelength during the $BCl_3$ plasma etching. SEM photos showed that 10 sccm $BCl_3$ plama produced more undercutting on GaAs sidewall than 5 sccm $BCl_3$ plasma.

Analysis of Clonorchis sinensis antigens and diagnosis of clonorchiasis using monoclonal antibodies (단세포군 항체를 이용한 간흡충 항원의 분석 및 간흡충증의 진단)

  • Yong, Tae-Sun;Im, Gyeong-Il;Jeong, Pyeong-Rim
    • Parasites, Hosts and Diseases
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    • v.29 no.3
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    • pp.293-310
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    • 1991
  • Clonorchis sinensis is a common parasite of man in Korea. Researches on the specific antigens of C. sinensis would be valuable not only because those elucidate the molecular characteristics of this fluke but also because it is applicable to immunodiagnosis. Although many monoclonal antibodies have been used in the field of parasite immunology, few articles on monoclonal antibodies against C. sinensis have been published so far. The aim of this study was to analyse C. sinensis antigens recognized by monoclonal antibodies, and to set up ELISA-inhibition test using C. sinensis specific monoclonal antibodies for improved specificity of immunodiagnostic tests. By fusion between spleen cells of the mice immunized with C. sinensis water-soluble crude adult worm antigens and plasmacytoma cells of mouse origin, 29 hybridoma clones secreting anti-C. sinensis monoclonal antibodies were made, and 8 clones among those were found specific. After cell cloning, isotypes of 6 selected specific monoclonal anti- bodies were determined to be IgGl, IgG2b and IgA. Four exposed antigenic determinants of natural infection were recognized by different specific monoclonal antibodies. By enzyme-immunoelectrotransfer blot, 10 KD, 34 KD antigenic determinants were found to be reacted with CsHyb 0714-20, CsHyb 0605-10 monoclonal antibodies, respectively, The antigenic determinant recognized by CsHyb 0714-20 monoclonal antibody was revealed to be located at the surface and parenchyme of a parasite by indirect immunoauorescent antibody technique, and those reacted with CsHyb 0605-10, CsHyb 0714-25 monoclonal antibodies were found at the parenchyme and intestine. The antigenic determinant reacted with CsHyb 0605-23 monoclonal antibody was found mainly around the uterine eggs. Four antigenic determinants recognized by specific monoclonal antibodies were all found to be present in the early eluted fractions of C. sinensis antigens separated by Sephadex G-200 gel filtration. By conventional ELISA, 75% of clonorchiasis cases were found positive, but 7.1% of normal controls and 37.5% of paragonimiasis cases showed false positives. However, by ELISA-inhibition test using C. sinensis specific monoclonal antibody (CsHyb 0605-23), 77.1% of clonorchiasis cases were found positive, and there were no false positives in normal controls or paragonimiasis cases, indicating 100% specificity. The ELISA- inhibition test using monoclonal antibodies was found to have same sensitivity and definitely high specificity in comparison with conventional ELISA for serodiagnosis of human clonorchiasis.

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AGL gene mutation and clinical features in Korean patients with glycogen storage disease type III

  • Ko, Jung-Min;Kim, Gu-Hwan;Yoo, Han-Wook
    • Journal of Genetic Medicine
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    • v.4 no.1
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    • pp.72-79
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    • 2007
  • Purpose : Glycogen storage disease type III (GSD-III) is a rare autosomal recessive disorder of glycogen metabolism. The affected enzyme, amylo-1,6-glucosidase, 4-alpha-glucanotransferase (AGL, glycogen debranching enzyme), is responsible for the debranching of the glycogen molecule during catabolism. The disease shows clinical and biochemical heterogeneity, reflecting genotype-phenotype heterogeneity among different patients. In this study, we aim at analyzing mutations of the AGL gene in three unrelated Korean GSD-III patients, and characterizing their clinical and laboratory findings. Methods : We characterized the clinical features of three unrelated Korean GSD-III patients by biochemical, histological and imaging studies. The 35 exons and part of exon-intron boundaries of AGL were analyzed by direct sequencing using genomic DNA extracted from the peripheral leukocytes of patients. Results : Diverse clinical features were observed in these patients including hepatomegaly (all patients), seizures (patient 2), grow th failure (patients 1 and 2), hyperlipidemia (patients 1 and 3), raised transaminase and creatine kinase concentrations (all patients), and mild cardiomyopathy (patient 2). Liver transplantation w as performed in patient 2 due to progressive hepatic fibrosis. A dministration of uncooked corn starch maintained normoglycemia and improved biochemical and growth profiles. DNA sequence analysis revealed mutations in 5 out of 6 alleles. Patient 1 was a compound heterozygote of c.1282 G>A (p.R428K) and c.1306delA (p.S603PfsX6), patient 2 had c.1510_1511insT (p.Y 504L fsX 10), and patient 3 had c.3416 T >C (p.L 1139P) and c.1735+1 G>T (p.Y 538_R578delfsX 4) mutations. A part from the p.R428K mutation, the 4 other substitutions identified w ere nov el. Conclusion : GSD-III patients display variable phenotypic characteristics resembling those of GSD-Ia. Molecular defects in the AGL gene of Korean GSD-III patients are genetically heterogeneous.

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Development of Efficient Screening Methods for Melon Plants Resistant to Fusarium oxysporum f. sp. melonis (멜론 덩굴쪼김병에 대한 효율적인 저항성 검정법 개발)

  • Lee, Won Jeong;Lee, Ji Hyun;Jang, Kyoung Soo;Choi, Yong Ho;Kim, Heung Tae;Choi, Gyung Ja
    • Horticultural Science & Technology
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    • v.33 no.1
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    • pp.70-82
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    • 2015
  • This study was conducted to establish an efficient screening system to identify melon resistant to Fusarium oxysporum f. sp. melonis. F. oyxsporum f. sp. melonis GR was isolated from infected melon plants collected at Goryeong and identified as F. oxysporum f. sp. melonis based on morphological characteristics, molecular analyses, and host-specificity tests on cucurbits including melon, oriental melon, cucumber, and watermelon. In addition, the GR isolate was determined as race 1 based on resistance responses of melon differentials to the fungus. To select optimized medium for mass production of inoculum of F. oxysporum f. sp. melonis GR, six media were tested. The fungus produced the most spores (microconidia) in V8-juice broth. Resistance degrees to the GR isolate of 22 commercial melon cultivars and 6 rootstocks for melon plants were investigated. All tested rootstocks showed no symptoms of Fusarium wilt. Among the tested melon cultivars, only three cultivars were susceptible and the other cultivars displayed moderate to high resistance to the GR isolate. For further study, six melon cultivars (Redqueen, Summercool, Superseji, Asiapapaya, Eolukpapaya, and Asiahwanggeum) showing different degrees of resistance to the fungus were selected. The development of Fusarium wilt on the cultivars was tested according to several conditions such as plant growth stage, root wounding, dipping period of roots in spore suspension, inoculum concentration, and incubation temperature to develop the disease. On the basis of the test results, we suggest that an efficient screening method for melon plants resistant to F. oxysporum f. sp. melonis is to remove soil from roots of seven-day-old melon seedlings, to dip the seedlings without cutting in s pore s uspension of $3{\times}10^5conidia/mL$ for 30 min, to transplant the inoculated seedlings to plastic pots with horticulture nursery media, and then to cultivate the plants in a growth room at 25 to $28^{\circ}C$ for about 3 weeks with 12-hour light per day.

The Development of Vulnerable Elements and Assessment of Vulnerability of Maeul-soop Ecosystem in Korea (한국 마을숲 생태계 취약요소 발굴 및 취약성 평가)

  • Lim, Jeong-Cheol;Ryu, Tae-Bok;Ahn, Kyeong-Hwan;Choi, Byoung-Ki
    • Journal of the Korean Institute of Traditional Landscape Architecture
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    • v.34 no.4
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    • pp.57-65
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    • 2016
  • Maeul-soop(Village forest) is a key element of Korean traditional village landscape historically and culturally. However, a number of Maeul-soops have been lost or declined due to various influences since the modern age. For this Maeul-soop that has a variety of conservation values including historical, cultural and ecological ones, attention and efforts for a systematic conservation and restoration of Maeul-soop are needed. The purpose of the present study is to provide information on ecological restoration and sustainable use and management of Maeul-soops based on component plant species, habitat and location characteristics of 499 Maeul-soops spread throughout Korea. Major six categories of threat factors to Maeul-soop ecosystem were identified and the influence of each factor was evaluated. For the evaluation of weight by threat factors for the influence on the vulnerability of Maeul-soop ecosystem, more three-dimensional analysis was conducted using Analytic Hierarchy Process (AHP) analysis method. In the results of evaluation using AHP analysis method, reduction of area, among six categories, was spotted as the biggest threat to existence of Maeul-soops. Next, changes in topography and soil environment were considered as a threat factor of qualitative changes in Maeul-soop ecosystem. Influence of vegetation structure and its qualitative changes on the loss or decline of Masul-soop was evaluated to be lower than that of changes in habitat. Based on weight of each factor, the figures were converted with 100 points being the highest score and the evaluation of vulnerability of Maeul-soop was conducted with the converted figures. In the result of evaluation of vulnerability of Maeul-soops, grade III showed the highest frequency and a normal distribution was formed from low grade to high grade. 38 Maeul-soops were evaluated as grade I which showed high naturality and 10 Maeul-soops were evaluated as grade V as their maintenance was threatened. Also in the results of evaluation of vulnerability of each Maeul-soop, restoration of Maeul-soop's own area was found as top priority to guarantee the sustainability of Maeul-soops. It was confirmed that there was a need to prepare a national level ecological response strategy for each vulnerability factor of Maeul-soop, which was important national ecological resources.