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Mutation Patterns of gyrA, gyrB, parC and parE Genes Related to Fluoroquinolone Resistance in Ureaplasma Species Isolated from Urogenital Specimens (비뇨생식기계 검체로부터 분리된 Ureaplasma 종의 Fluoroquinolone 내성과 관련된 gyrA, gyrB, parC, parE 유전자의 돌연변이 양상)

  • Cho, Eun-Jung;Hwang, Yu Yean;Koo, Bon-Kyeong;Park, Jesoep;Kim, Young Kwon;Kim, Sunghyun
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.2
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    • pp.74-81
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    • 2016
  • Ureaplasma species can normally colonize in the bodies of healthy individuals. Their colonization is associated with various diseases including non-gonococcal urethritis, chorioamnionitis, neonatal meningitis, and prematurity. In 2012, the sum of the resistant and intermediate resistant rates of Ureaplasma spp. to ofloxacin and ciprofloxacin was 66.08% and 92.69%, respectively. DNA point mutations in the genes encoding DNA gyrase (topoisomerase II) and topoisomerase IV are commonly responsible for fluoroquinolone resistance. Each enzyme is composed of two subunits encoded by gyrA and gyrB genes for DNA gyrase and parC and parE genes for topoisomerase IV. In the current study, these genes were sequenced in order to determine the role of amino acid substitutions in Ureaplasma spp. clinical isolates. From December 2012 to May 2013, we examined mutation patterns of the quinolone resistance-determining region (QRDR) in Ureaplasma spp. DNA sequences in the QRDR region of Ureaplasma clinical isolates were compared with those of reference strains including U. urealyticum serovar 8 (ATCC 27618) and U. parvum serovar 3 (ATCC 27815). Mutations were detected in all ofloxacin- and ciprofloxacin-resistant isolates, however no mutations were detected in drug-susceptible isolates. Most of the mutations related to fluoroquinolone resistance occurred in the parC gene, causing amino acid substitutions. Newly found amino acid substitutions in this study were Asn481Ser in GyrB; Phe149Leu, Asp150Met, Asp151Ile, and Ser152Val in ParC; and Pro446Ser and Arg448Lys in ParE. Continuous monitoring and accumulation of mutation data in fluoroquinolone-resistant Ureaplasma clinical isolates are essential to determining the tendency and to understanding the mechanisms underlying antimicrobial resistance.

Detection of Vancomycin-Resistant Enterococci and Related Genes Using VITEK 2 System and Multiplex Real-time PCR Assay (VITEK 2 시스템과 Multiplex Real-time PCR을 이용한 반코마이신 내성 장알균(VRE)과 내성관련 유전자 검출)

  • Jeong, Min-Kyung;Yu, Young-Bin;Kim, Sang-Ha;Kim, Sunghyun;Kim, Young-Kwon
    • Korean Journal of Clinical Laboratory Science
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    • v.49 no.4
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    • pp.401-406
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    • 2017
  • In this study, using the VITEK 2 system, 74 samples (22.6%) out of 327 specimens were identified by the growth of Enterococcosel media (EV6 agar) supplemented with $6{\mu}g/mL$ of vancomycin. Enterococcus faecium was identified as 55 strains (74.3%), Enterococcus casseliflavus as 2 strains (2.7%), Enterococcus avium as 1 strain (1.4%), and Enterococcus gallinarum as 16 strains (21.6%). Among the 55 phenotypes of Enterococcus faecium, 42 (76.4%), 9 (16.4%), and 4 strains (7.3%) showed the vanA, vanB, and vanC phenotype, respectively. The 16 strains of Enterococcus gallinarum and 2 strains of Enterococcus casseliflavus showed the vanC phenotype and the 1 strain of Enterococcus avium had the vanB phenotype. The one strain of Enterococcus faecium propagated only in EV4 and was susceptible to both vancomycin and teicoplanin according to the antimicrobial susceptibility test using the VITEK 2 system. The vancomycin resistance phenotype gene was not detected by PCR. A total of 327 specimens were cultured in Enterococcosel broth supplemented with $6{\mu}g/mL$ of vancomycin (EV6 broth), and 120 strains (36.7%) were isolated. These 120 strains were subjected to vancomycin resistant genotyping by a multiplex real-time polymerase chain reaction and 51 strains (42.5%) showed vanA; 5 strains (4.2%) showed vanA and vanC; and 18 strains (15%) showed vanC. Vancomycin resistance genotypes were not detected in the remaining 46 strains (38.3%).

An Evaluation of Vitek MS System for Rapid Identification of Bacterial Species in Positive Blood Culture (혈액배양 양성검체에서 패혈증 원인균 신속동정을 위한 Vitek MS 시스템의 유용성 평가)

  • Park, Kang-Gyun;Kim, Sang-Ha;Choi, Jong-Tae;Kim, Sunghyun;Kim, Young-Kwon;Yu, Young-Bin
    • Korean Journal of Clinical Laboratory Science
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    • v.49 no.4
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    • pp.407-412
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    • 2017
  • The aim of this study was to shorten the time required for subculture and bacterial identification and obtain a simple and rapid identification method for new test methods for bloodstream infections. The following results were obtained using a mass spectrometer. In Vitek 2, 208 (81.8%) cases were well-identified and 45 isolates were not identified in blood cultures. Among 208 cases, 146 (57.5%) were Gram positive bacteria and 108 (42.5%) were Gram negative bacteria. In total, 233 were identified to the species level and 21 were identified to the genus level. The identification error was found to be Propionibacterium acnes as Clostridium bifermentans. The accuracy of Enterobacteriaceae, glucose non-fermentative bacilli (GNFB), and staphylococci were 81/83 (97.6%), 12/15 (80.0%), and 72/85 (84.7%), respectively. The concordance rate of Vitek 2 and Vitek MS by the direct method was 81.8% and 45 isolates were not identified. Most of the unidentified bacteria were Gram positive bacteria (N=37). The Gram positive bacteria were streptococci (14), coagulase-negative staphylococci (CNS) (11), enterococci (3), Staphylococcus aureus (2), Micrococcus spp. (2), Bacillus spp. (2) and Actinomyces odontolyticus, Finegoldia magna, and Peptostreptococcus spp. The results reporting time was reduced to 24~72 hours compared to the conventional method. The rate of identification of the aerobic and anaerobic cultures was similar, but the use of an anaerobic culture did not require a dissolution process, which could shorten the sample preparation time. These results suggest that the method of direct identification in blood cultures is very useful for the treatment of patients. In further studies, it might be necessary to further improve the method for identifying streptococci and CNS, which were lacking in accuracy in this study.

Vertical and horizontal distribution of squids in relation to oceanographic structure in the North Pacific Ocean (북태평양 오징어류의 연직 및 수평분포)

  • 김영승;이주희;박영철;황선재;김두남
    • Journal of the Korean Society of Fisheries and Ocean Technology
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    • v.35 no.2
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    • pp.102-117
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    • 1999
  • Vertical distribution of squids in relation to oceanographic structure was analyzed on the basis of experimental squid hand jigging fishing by R/V Pusan 851 in the area of 34°∼47° N, 150° E∼170°W in the North Pacific in summer from 1987 through 1993 with exception of 1991. The 6 species of squids showed different patterns of vertical and horizontal distribution as following; Boreopacific gonate squid (Gonatopsis borealis) were mainly caught in the layer of 71-80m fishing depth of the Subarctic Domain with water temperature of 6∼11℃ and salinity of 32.2∼33.6‰ and distributed in the latitudes of 41°∼43° N. Boreal clubhook squid (Onychoteuthis borealijaponica) were mainly caught in the layer of 11∼20m fishing depth of the Subarctic Domain with water temperature of 10∼12℃ and salinity of 32.9∼33.6‰ and distributed in the latitudes of 41°∼42N°. Tapanese flying squid (Todarodes pacificus) were mainly caught in the layer of 11∼20m fishing depth of the Transition Zone and the Subtropical Domain with water temperature of 15∼18℃ and salinity of 33.6∼34.0‰ and distributed in the latitudes of 40°∼42°N. Neon flying squid (Ommastrephes bartrami) were mainly caught in the layer of surface∼10m fishing depth of the Subarctic Convergence Zone and the Transition Zone with water temperature of 16∼17℃ and salinity of 33.7∼34.4‰ and distributed in the latitudes of 39 °∼41°N. Luminous flying squid (Symplectoteuthis luminosa) were mainly caught in the layer of 11 20m fishing depth of the Transition Zone and the Subtropical Domain with water temperature of 18∼20℃ and salinity of 33.8∼34.6‰ and distributed in the latitudes of 37°∼39°N. Purpleback flying squid (Symplectoteuthis oualaniensis) were mainly caught in the layer of surface∼10m fishing depth of the Subtropical Domain with water temperature of 24∼25℃ and salinity of 34.2∼34.4‰ and distributed in the latitude of 36°∼37°N.

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Studies on a Nematode, Anguillicola crassa Parasitic in the Air Bladder of the Eel (뱀장어 부레에 기생(寄生)하는 선충(線虫), Anguillicola crassa에 관(關)한 연구(硏究))

  • Kim, Young-Gill;Kim, Eul-Bae;Kim, Jong-Yeon;Chun, Seh-Kyu
    • Journal of fish pathology
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    • v.2 no.1
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    • pp.1-18
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    • 1989
  • In Korea, studies on a Nematode, Anguillicola crassa parasitic in the air bladder of eel are not yet reported. This reason led the author to study the parasitic species, state and life history of the A. crassa parasitized in the air bladder of eel in order to take effective control measures against its damage. The size of fully developed eggs was 80 to $92(86.7){\times}62$ to $71(67.4)\;{\mu}m$, larva was 210 to $240(225){\times}18$ to $23(20.6)\;{\mu}m$. The intermediate host of A. crassa was Thermocyclops hyalinus, it was capable for parasitizing the eel after 4 days of invasion and then the size of larva was 360 to $420(390){\times}28$ to $35(31)\;{\mu}m$. Fifty days after eel had ingested the Thermocyclops hyalinus infected with larva of A. crassa, the larvae matured into adult worms in the air bladder of eel. The size of detected adult worms was 7.3 to $31.0(16.5){\times}0.5$ to 2.2(1.2) mm, 4.9 to $13.3(8.3){\times}0.3$ to 0.9(0.4) mm. Investigating the morphology of the worms, they were identified as A. crassa. Monthly the parasitic rate of the worms in the eel was high in June, September and December, but low in January to March. After the investigation on the significance between non-parasitic fish and parasitic fish, it was not significant, therefore it can be considered that there is no effect of infection in the growth of eel. Any abnormality of eels air bladder tissue was not seen by the infection of A. crassa. At 25.0 to $26.7^{\circ}C$ of water temperature the death time of Thermocyclops hyalinus by masoten treatment was 14 hours in 0.5 ppm, 20 hours in 0.4 ppm, 22 hours in 0.3 ppm, 30 hours in 0.2 ppm and 42 hours in 0.1 ppm.

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Study on Marteilioides chungmuensis Comps et al., 1986 Parasite of the Pacific Oyster, Crassostrea gigas Thunberg (참굴의 난(卵)에 기생(寄生)하는 Marteilioids chungmuensis Comps et al., 1986에 관하여)

  • Park, Mi-Seon;Chun, Seh-Kyu
    • Journal of fish pathology
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    • v.2 no.2
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    • pp.53-70
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    • 1989
  • An ovarian parasite, Marteilioides chungmuensis of the Pacific oyster, Crassostrea gigas has been observed on several occasions in the Pacific sector of production of this oyster species(Matsuzato et al., 1977 ; Chun, 1979). This study was carried out on the specimens collected at Hwado, Och'$\check{o}$n, and Sinchang respectively located the southern, western, and eastern coasts of Korean Peninsula from 1986 through 1988 to investigate M. chungmuensis to the Pacific oyster. Uitrastructural studies were also carried out on infected oysters, to allow detailed examination of the structure and consepuently the systematic position of this parasite. Infection rates of M. chungmuensis at Hwado and Och'$\check{o}$n oyster farms were 5.3% and 4.2% each in 1986, 6.7% and 2.8% each in 1987, but they were not found at Sinchang oyster habitat. M,. chungmuensis-infected oysters were found from June to November at Hwado and from June to October at Och'$\check{o}$n. Twenty five of three hundred oysters transplanted from Sinchang to Hwado were found infected with M. chungmuensis. Some abnormal eggs infected with M. chungmuensis are liberated through the gill together with normal mature eggs on the spawning and the rest remain necrotized after spawning season. The earliest known stages consist of a stem cell or primary cell, including a secondary cell in which ovoid haplosporosomes are found. During sporulation, 2 or 3 secondary are produced by exogenous budding from the first secondary cell and, each secondary cell evolves into a sporont upon the tertiary cell differentiation (enodogenous budding) ; then, haplosporosomes are formed in the young sporont. Internal cleavages involve the differentiation of one tricellular spore per sporont. The outermost spore cell contains membrane-bounded osmiophilic bodies : the middle and the inner, most spore cells contain high density cytoplasmic ribosomes. The mechanism of spore formation from the stem cell of M. chungmuensis is the simplest of the class Paramyxea known up to now.

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A study on the concentration of trace metal elements in whole blood of the women in reproductive ages in urban and rural area (도시 및 농촌지역 가임연령 여성들의 혈중 미량금속원소의 함량에 관한 조사)

  • Jun, Jin-Ho
    • Journal of Preventive Medicine and Public Health
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    • v.17 no.1
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    • pp.95-106
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    • 1984
  • To acquire the essential basic data to the establishment of control measure for the hazardous health effect that could be caused by harmful metals, the author measured the concentrations of trace metals in whole blood of women of $20{\sim}39$ years old living in urban and rural area using atomic absorption spectrophotometer. The summarized results were as follows; 1. The mean concentration of zinc in whole blood was $10.69{\pm}8.07{\mu}g/ml$ in rural area. The frequency distribution by zinc concentration level was nearly L-type and the cumulative frequency distribution was showed bimodal type in both area. 2. The mean iron concentration in whole blood was $323.09{\pm}87.15{\mu}g/ml$ and $322.07{\pm}104.74{\mu}g/ml$ in urban and rural area, respectively. The frequency distribution was similar to normal distribution type in both area, but the cumulative distribution was unimodal type in urban area and bimodal type in rural area. 3. The mean magnesium concentration was $41.08{\pm}19.58{\mu}g/ml$ and $40.28{\pm}16.82{\mu}g/ml$ in the area, respectively. The frequency distribution type had skewness to the right and the cumulative frequency distribution was unimodal type in both area. 4. The mean copper concentration was $1.417{\pm}0.761{\mu}g/ml$ and $1.375{\pm}0.743{\mu}g/ml$ in the area, respectively. The frequency distribution type had skewness to the right and the cumulative frequency distribution was bimodal type in both area. 5. The mean manganese concentration was $0.079{\pm}0.039{\mu}g/ml$ and $0.07{\pm}0.058{\mu}g/ml$ in the area, respectively. The frequency distribution type had skewness to the right in both area but slight irregular in rural area and the cumulative distribution was unimodal and bimodal type in urban and rural area, respectively. 6. The mean cadmium concentration in whole blood was $0.031{\pm}0.026{\mu}g/ml$ in urban and $0.028{\pm}0.023{\mu}g/ml$ in rural area. The frequency distribution type had skewness to the right and cumulative frequency distribution was bimodal type in both area.

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Epidemiology of Psychosocial Distress in Korean Employees (우리나라 직장인 스트레스의 역학적 특성)

  • Chang, Sei-Jin;Kang, Myung-Gun;Cha, Bong-Suk;Park, Jong-Ku;Hyun, Sook-Jung;Park, Jun-Ho;Kim, Seong-Ah;Kang, Dong-Mug;Chang, Seong-Sil;Lee, Kyung-Jae;Ha, Eun-Hee;Ha, Mi-Na;Koh, Sang-Baek
    • Journal of Preventive Medicine and Public Health
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    • v.38 no.1
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    • pp.25-37
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    • 2005
  • Objective : To estimate the magnitude of psychosocial distress and examine eligible factors associated with the development of psychosocial distress in Korean employees, using a nationwide sample. Methods : A total of 6,977 workers were recruited from 245 companies. A structured questionnaire was used to assess sociodemographics, health-related behaviors, job characteristics, social support at work, personality traits (locus of control, type A behavior pattern), self-esteem, and psychosocial distress. Results : The results showed that 23 % of workers were categorized as high stress, 73% as moderate, and 5% as normal. Hierarchical multiple regression analysis showed that psychosocial distress was more common in younger workers, both male and female. Regular exercise was negatively associated with increase of psychosocial distress. In job characteristics, as expected, low decision latitude, high job insecurity, and low social support at work were related to high psychosocial distress. Personality traits such as locus of control and type A behavior pattern, and self-esteem were more powerful predictors of psychosocial distress than general characteristics, health-related behavior, and job characteristics. There were some gender differences. While men who are less educated and single (unmarried, divorced, and separated) experienced higher levels of psychosocial distress than those who are educated and married, women who feel high job demand experienced higher levels of psychosocial distress than those who feel low job demand. Conclusions : The proportion of the high stress group was higher than expected, and psychosocial factors like social support and personality characteristics (e. g. locus of control, type A behavior pattern and self-esteem) were more significant factors for psychosocial distress than other variables. This finding suggests that some psychosocial factors, especially inadequate social support, low self-esteem and lack of internal locus of control for the development of psychosocial distress, will also operate as an intervention strategy in the worksite stress reduction program. It is strongly required that worksite stress reduction programs should be established in at both occupational and level as well as in individual levels.

Analysis of Image Processing Characteristics in Computed Radiography System by Virtual Digital Test Pattern Method (Virtual Digital Test Pattern Method를 이용한 CR 시스템의 영상처리 특성 분석)

  • Choi, In-Seok;Kim, Jung-Min;Oh, Hye-Kyong;Kim, You-Hyun;Lee, Ki-Sung;Jeong, Hoi-Woun;Choi, Seok-Yoon
    • Journal of radiological science and technology
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    • v.33 no.2
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    • pp.97-107
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    • 2010
  • The objectives of this study is to figure out the unknown image processing methods of commercial CR system. We have implemented the processing curve of each Look up table(LUT) in REGIUS 150 CR system by using virtual digital test pattern method. The characteristic of Dry Imager was measured also. First of all, we have generated the virtual digital test pattern file with binary file editor. This file was used as an input data of CR system (REGIUS 150 CR system, KONICA MINOLTA). The DICOM files which were automatically generated output files by the CR system, were used to figure out the processing curves of each LUT modes (THX, ST, STM, LUM, BONE, LIN). The gradation curves of Dry Imager were also measured to figure out the characteristics of hard copy image. According to the results of each parameters, we identified the characteristics of image processing parameter in CR system. The processing curves which were measured by this proposed method showed the characteristics of CR system. And we found the linearity of Dry Imager in the middle area of processing curves. With these results, we found that the relationships between the curves and each parameters. The G value is related to the slope and the S value is related to the shift in x-axis of processing curves. In conclusion, the image processing method of the each commercial CR systems are different, and they are concealed. This proposed method which uses virtual digital test pattern can measure the characteristics of parameters for the image processing patterns in the CR system. We expect that the proposed method is useful to analogize the image processing means not only for this CR system, but also for the other commercial CR systems.

A Novel Chenodeoxycholic Derivative HS-1200 Induces Apoptosis in Human HT-29 Colon Cancer Cells (인체 대장암 세포주(HT-29)에서 담즙산 합성유도체(HS-1200)의 세포 사망 기전)

  • Oh Sin Geun;Yang Kwang Mo;Hur Won Joo;Yoo Young Hyun;Suh Hong Suk;Lee Hyung Sik
    • Radiation Oncology Journal
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    • v.20 no.4
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    • pp.367-374
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    • 2002
  • Purpose : To investigate the growth inhibitory effects, and the underlying mechanism of human colon cancer cell (HT-29) death, induced by a new synthetic bile acid derivative (HS-1200). Materials and Methods : Human colon cancer cells (HT-29), in exponential growth phase, were treated with various concentrations of a new synthetic bile acid derivative (HS-1200). The growth inhibitory effects on HT-29 cells were examined using a frypan blue exclusion assay. The extent of apoptosis was determined using agarose gel electrophoresis, TUNEL assays and Hoechst staining. The apoptotic cell death was also confirmed by Western blotting of PARP, caspase-3 and DNA fragmentation factor (DFF) analysis. To investigate the involvement of mitochondria, we employed immunofluorescent staining of cytochrome c and mitochondrial membrane potential analyses. Results : The dose required for the half maximal inhibition $(IC_{50})$ of the HT-29 cell growth was $100\~150\;{\mu}M$ of HS-1200. Several changes, associated with the apoptosis of the HT-29 cells, were reveal by the agarose gel eletrophoresis, TUNEL assays and Hoechst staining, following their treatment with $100\;{\mu}M$ of HS-1200. HS-1200 treatment also induced caspase-3, PARP and DFF degradations, and the western blotting showed the processed caspase-3 p20, PARP p85 and DFF p30 and p11 cleaved products. Mitochondrial events were also demonstrated. The cytochrome c staining indicated that cytochrome c had been released from the mitochondria in the HS-1200 treated cells. The mitochondrial membrane potential $(\Delta\Psi_m)$ was also prominently decreased in the HS-1200 treated cells. Conclusion : These findings suggest that the HS-1200 - induced apoptosis of human colon cancer cells (HT-29) is mediated via caspase and mitochondrial pathways.