• 제목/요약/키워드: resistance determinant

검색결과 72건 처리시간 0.022초

Expression, Purification, Crystallization and Preliminary X-Ray Crystallographic Analysis of CnrX from Cupriavidus metallidurans CH34

  • Kim, Kook-Han;Jung, Eun-Jung;Im, Ha-Na;Lelie, Daniel Van Der;Kim, Eunice Eun-Kyeong
    • Journal of Microbiology and Biotechnology
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    • 제18권1호
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    • pp.43-47
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    • 2008
  • The nickel and cobalt resistance of Cupriavidus metallidurans CH34 is mediated by the CnrCBA efflux pump encoded by the cnrYHXCBAT metal resistance determinant. The products of the three genes cnrYXH transcriptionally regulate expression of cnr. CnrY and CnrX are membrane-bound proteins, probably functioning as anti-sigma factors, whereas CnrH is a cnr-specific extracytoplasmic functions (ECF) sigma factor. The periplasmic domain of CnrX (residues 29-148) was cloned as a N-terminal His-tagged protein, expressed in Escherichia coli, and purified using affinity chromatography and gel filtration. The molecular mass was estimated to be about 13.6kDa by size exclusion chromatography, corresponding to a monomer. The tetragonal bipyramid crystals were obtained by mixing an equal volume of protein in 50mM Tris-HCl, pH 7.5, 1% glycerol, 100mM NaCl, 1mM DTT, and the reservoir solution of 15% w/v PEG 2000, 100mM lithium chloride at 277K in 2-4 days using hanging drop vapor diffusion. The protein concentration was 24mg/ml. The crystal that diffracted to $2.42{\AA}$ resolution belongs to space group $P4_1\;or\;P4_3$ with unit cell parameters of $a=b=32.14{\AA},\;c=195.31{\AA},\;{\alpha}={\beta}={\gamma}=90^{\circ}$, with one molecule of CnrX in the asymmetric unit.

임상검체와 가축으로부터 분리된 대장균을 대상으로 Quinolone계 항균제 내성인자 분석 (Analysis of Quinolone Resistance Determinants in Escherichia coli Isolated from Clinical Specimens and Livestock Feces)

  • 성지연
    • 대한임상검사과학회지
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    • 제50권4호
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    • pp.422-430
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    • 2018
  • 퀴놀론 항균제가 사람과 동물에게 부적절하고 광범위하게 사용될 경우 항균제내성인자의 출현 및 확산이 가속화 될 수 있다. 본 연구에서는 돼지의 직장면봉 검체(N=40) 및 임상 검체로(N=25)부터 분리된 총 65균주의 nalidixic acid 내성 대장균을 대상으로 quinolone 내성 기전을 조사하였다. 항균제 감수성은 디스크 확산법에 의해 결정되었다. Quinolone 내성과 관련된 유전자와 돌연변이를 조사하기 위해 PCR 및 DNA sequencing이 수행되었다. 총 65균주의 nalidixic acid 내성 대장균 중 62균주가 gyrA, parC, parE 유전자에 돌연변이를 포함하고 있었는데, gyrA 유전자에 돌연변이를 포함하고 있는 균주는 62균주(95.4%)였고, 35균주(53.8%)가 parC 유전자에 돌연변이를 갖고 있었으며, 7균주(10.8%)가 parE 유전자에 돌연변이를 포함하고 있었다. 35균주는 gyrA 와 parC 유전자에 모두 돌연변이를 가지고 있는 것으로 나타났다. 총 65균주의 대장균을 대상으로 plasmid-mediated quinolone resistance (PMQR) determinants를 조사하였다. 65균주의 nalidixic acid 내성 대장균 중 13균주에서 qnrS 유전자가 검출되었으나 이 중 10균주는 gyrA, parC, parE 유전자에 돌연변이를 포함하고 있는 것을 나타났다. 본 연구에서는 사람과 돼지로부터 분리된 대장균이 quinolone 계열 항균제에 내성을 나타내는데 중요한 역할을 하는 기전이 gyrA, parC, parE 유전자에 염색체 돌연변이가 발생하는 경우임을 확인하였는데 이 돌연변이들은 치료목적 또는 동물의 성장촉진을 위한 항균제의 과다사용으로 유발될 수 있다.

충청지역의 임상검체로부터 분리된 Acinetobacter calcoaceticus-baumannii Complex를 대상으로 항균제 내성 유전자 비교분석 (Distribution of Antimicrobial Resistant Genes in Acinetobacter calcoaceticus-baumannii Complex Isolated from Clinical Specimens in Chungcheong, Korea)

  • 성지연
    • 대한임상검사과학회지
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    • 제49권4호
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    • pp.427-434
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    • 2017
  • Acinetobacter calcoaceticus-baumannii (Acb) complex에 속한 종들은 빈번하게 병원감염 및 기회감염을 일으킨다. 또한 다제내성인 경우가 많아 이 균들의 감염증 치료를 위한 항균제 선택이 매우 제한적이다. 본 연구에서는 ciprofloxacin 내성 Acinetobacter species 53균주를 대상으로 fluoroquinolone 내성기전을 조사했다. 항균제 감수성 양상을 조사하기 위해 디스크확산법이 시행되었다. Fluoroquinolone 내성과 관련된 유전자 및 돌연변이 검출을 위해 PCR과 염기서열분석이 이루어졌다. 본 연구에서 수집된 53균주의 ciprofloxacin 내성 Acinetobacter 중 47균주가 gyrA 유전자의 83번째 serine 아미노산 잔기와 parC 유전자의 80번째 serine 아미노산 잔기가 leucine 잔기로 치환된 sense mutations 가지고 있는 것으로 나타났다. gyrA와 parC 유전자에 sense mutations을 가지고 있는 47균주 중 44균주가 A. baumannii 였고 3균주는 A. pittii였다. 본 연구에서 조사대상이 되었던 Acb complex 균주들 중 plasmid-mediated quinolone resistance (PMQR) determinants를 가지고 있는 균주는 한나도 없었다. 46 균주의 ciprofloxacin 내성 A. baumannii 는 A, B, 또는 F형의 banding pattern을 보였는데 이는 충청지역에 위치한 일개의 병원에 ciprofloxacin 내성 A. baumannii가 수평확산 되어 있음을 의미한다. Fluoroquinolone 내성 Acb complex 균주의 집락화 및 확산을 막기 위해서 다제내성 균주들을 대상으로 항균제 내성인자들을 지속적으로 조사하고 모니터링할 필요가 있을 것으로 사료된다.

Diversity in Betasatellites Associated with Cotton Leaf Curl Disease During Source-To-Sink Movement Through a Resistant Host

  • Khan, Iftikhar Ali;Akhtar, Khalid Pervaiz;Akbar, Fazal;Hassan, Ishtiaq;Amin, Imran;Saeed, Muhammad;Mansoor, Shahid
    • The Plant Pathology Journal
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    • 제32권1호
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    • pp.47-52
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    • 2016
  • Cotton leaf curl is devastating disease of cotton characterized by leaf curling, vein darkening and enations. The disease symptoms are induced by DNA satellite known as Cotton leaf curl Multan betasatellite (CLCuMuB), dominant betasatellite in cotton but another betasatellite known as Chili leaf curl betasatellite (ChLCB) is also found associated with the disease. Grafting experiment was performed to determine if host plant resistance is determinant of dominant population of betasatellite in cotton (several distinct strains of CLCuMuB are associated with the disease). Infected scion of Gossypium hirsutum collected from field (the source) was grafted on G. arboreum, a diploid cotton species, resistant to the disease. A healthy scion of G. hirsutum (sink) was grafted at the top of G. arboreum to determine the movement of virus/betasatellite to upper susceptible scion of G. hirsutum. Symptoms of disease appeared in the upper scion and presence of virus/betasatellite in the upper scion was confirmed via molecular techniques, showing that virus/betasatellite was able to move to upper scion through resistant G. arboreum. However, no symptoms appeared on G. arboreum. Betasatelites were cloned and sequenced from lower scion, upper scion and G. arboreum which show that the lower scion contained both CLCuMuB and ChLCB, however only ChLCB was found in G. arboreum. The upper scion contained CLCuMuB with a deletion of 78 nucleotides (nt) in the non-coding region between Arich sequence and ${\beta}C1$ gene and insertion of 27 nt in the middle of ${\beta}C1$ ORF. This study may help in investigating molecular basis of resistance in G. arboreum.

병원하수로부터 분리한 Gentamicin 저항성 세균에서 Tn3에 의한 aac(3)II의 발현 증가 (Increased Expression of aac(3)II by Tn3 in Gentamicin - Resistant Bacteria Isolated from Hospital Sewage)

  • 한효심;이문숙;정재성
    • 미생물학회지
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    • 제40권1호
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    • pp.60-64
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    • 2004
  • Gentamicin에 저항성을 나타내는 세균을 병원 하수로부터 분리하여 aminoglycoside-(3)- N-acetyltransferase를 암호화하는 유전자인 aac(3)II의 존재여부를 dot-blot hybridization으로 조사하였다. aac(3)II유전자의 일부를 탐침으로 사용한 결과 gentamicin저항성 세균의 41% (39/95)가 이 유전자를 가지고 있었다. aac(3)II와 Tn3에서 각각 설계된 primer를 사용한 PCR 결과 aac(3)II를 가지고 있는 39개 균주 중 13개 균주가 TnS-aac(3)II구조를 가지고 있음을 알 수 있었다. aac(3)II의 상류에 Tn3를 가지고 있는 13개 균주의 gentamicin에 대한 최소억제농도는 가지고 있지 않은 균주에 비해 상대적으로 높았다. 13개 균주를 동정한 결과 5개는 Eschelichia coli, 3개는 Acinetobacter johnsonii, Enterobacter agglomerans와 Micrococcus luteus가 각각 2개, 그리고 1개의 Pseudomonas facilis로 동정되었다. 이러한 결과들은 Tn3-aac(3)II구조가 gentamicin 저항성 세균들에서 널리 분포하고 있음을 말해준다.

A noble RBC aggregometer with vibration-induced disaggregation mechanism

  • Shin S.;Jang J.H.;Park M.S.;Ku Y.H.;Suh J.S.
    • Korea-Australia Rheology Journal
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    • 제17권1호
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    • pp.9-13
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    • 2005
  • The aggregation of red blood cells (RBCs) is a major determinant of blood flow resistance passing through various veins. Available techniques for measuring RBC aggregation often require pretreating and washing after each measurement, which is not optimal for day-to-day clinical use. A laser reflection technique has been combined with a vibration-aided disaggregation mechanism, which shows significant advances in aggregometer design, operation and data analysis. The essential features of this design are in its simplicity and a disposable element that is in contact with the blood sample. Using extremely small quantities of blood, the RBCs subjected to vibrations can be quickly and completely disaggregated. This is followed by measuring the backscattered light intensity. The measurements with the present sensor were compared with those of a commercial aggregometer and a strong correlation was found between them. The newly-developed optical aggregometer can measure the RBC aggregability difference between young and old cell suspension with ease and accuracy.

강인성이 건강증진행위, 신체증상에 미치는 효과-여대생을 대상으로- (Hardiness as a Stress-resistance Resource and as a Determinant of Health -promoting Behavior: in a Sample of Nursing Students)

  • 이미라;소희영;송영신
    • 지역사회간호학회지
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    • 제9권1호
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    • pp.21-29
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    • 1998
  • This study was undertaken in order to examine the relationship of hardiness and health-promoting behavior and the effects of hardiness on stress-related physical symptoms. The subjects were 104 female nursing students of one college in Kongju. The instruments used for this study were a survey of general characteristics, hardiness(25 items), health-promoting behavior(44 items), and physical symptoms(35 items), Analysis of data was done by use of mean, Pearson correlation coefficient, stepwise regression and a hierarchical multiple regression with an SAS program. The results of this study are as follows. 1) Significant correlations between health -promoting behavior and subscales of hardiness, that is, control(r=-.35, P<.00l), commitment(r= -.29, P<.0l), and challenge(r= -.23, p<.05) were found. 2) Control was the highest factor predicting health -promoting behavior. 3) Main and buffering effects of hardiness on current physical symptoms were not found.

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Determinant of Arterial Stiffness in Young Adults

  • Jo Yoon-Kyung;Jeon Justin Y.;Kim Eun-Sung;Jekal Youn-Suk;Eom Yong-Bin;Im Jee-Aee
    • 대한의생명과학회지
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    • 제12권3호
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    • pp.191-196
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    • 2006
  • Cardiovascular disease (CVD) risk factors may be acting several decades before CVD becomes manifest. Data from young subjects may be valuable in further elucidating at this issue. We evaluated the association between baPWV (brachial-ankle pulse wave velocity) and cardiovascular risk factors in apparently healthy young adults. A total of 46 male and 91 female adolescents aged $18{\sim}25 years$ were studied. baPWV increased in a dose-responsive manner as the number of metabolic syndrome components. In both gender groups, baPWV was positively correlated with age. In males, waist, circumference total cholesterol, and LDL-cholesterol were positively correlated with baPWV, and in females, blood pressure (BP) was positively correlated with baPWV. Age, gender, mean BP, and Homeostasis model assessment insulin resistance (HOMA-IR) were found to be independent factors associated with baPWV levels. In conclusion, mean BP, age, gender, and HOMA-IR were associated with baPWV in young adults. This result suggests that multiple cardiovascular risk factors may be associated with an increased risk of arterial stiffness in young adults.

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Exploiting tumor cell senescence in anticancer therapy

  • Lee, Minyoung;Lee, Jae-Seon
    • BMB Reports
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    • 제47권2호
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    • pp.51-59
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    • 2014
  • Cellular senescence is a physiological process of irreversible cell-cycle arrest that contributes to various physiological and pathological processes of aging. Whereas replicative senescence is associated with telomere attrition after repeated cell division, stress-induced premature senescence occurs in response to aberrant oncogenic signaling, oxidative stress, and DNA damage which is independent of telomere dysfunction. Recent evidence indicates that cellular senescence provides a barrier to tumorigenesis and is a determinant of the outcome of cancer treatment. However, the senescence-associated secretory phenotype, which contributes to multiple facets of senescent cancer cells, may influence both cancer-inhibitory and cancer-promoting mechanisms of neighboring cells. Conventional treatments, such as chemo- and radiotherapies, preferentially induce premature senescence instead of apoptosis in the appropriate cellular context. In addition, treatment-induced premature senescence could compensate for resistance to apoptosis via alternative signaling pathways. Therefore, we believe that an intensive effort to understand cancer cell senescence could facilitate the development of novel therapeutic strategies for improving the efficacy of anticancer therapies. This review summarizes the current understanding of molecular mechanisms, functions, and clinical applications of cellular senescence for anticancer therapy.

Characteristics of Bacteriocin and Mucin Production Phenotypes in Lactobacillus plantarum 27

  • Kim, Wang-Jung;Ha, Duk-Mo;Ray, Bibek
    • Journal of Microbiology and Biotechnology
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    • 제1권2호
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    • pp.96-101
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    • 1991
  • Phenotypic changes after plasmid curing experiment suggested that the bacteriocin production phenotype ($Bac^{+}$) might be linked to a chromosomal DNA and the mucin production phenotype ($Muc^{+}$) might be linked to a 62.5 kilobase (kb) plasmid (pMUC62) in Lactobacillus plantarum 27 isolated from meat starter culture. The non-mucoid ($Muc^{-}$) variants were missing pMUC62 but they produced bacteriocin as the wild strain ($Bac^{+}$). There was no difference in antibiotic resistance and sugar fermentation patterns between the wild strain ($Bac^{+}$ $Muc^{+}$) and the nonmucoid ($Bac^{+}$ $Muc^{-}$) variants. Antimicrobial spectrum of bacteriocin produced by both wild strain and $Muc^{-}$ variant of Lb. plantarum 27 included strains of Pediococcus acidilactici (A, M, H), Pediococcus sp. isolated from meat, Lactobacillus sp. isolated from meat, Lb. plantarum NCDO 955 and Staphylococcus aureus 485. Neither of the tested Gram negative bacteria were inhibited by bacteriocin. Antimicrobial activity of crude bacteriocin was retained after autoclaving, DNase or catalase treatment and exposure from pHs 4 to 9 but was lost after treating with several proteolytic enzymes and exposure at pH 10.

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