• Title/Summary/Keyword: virulence expression

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Panton-Valentine Leukocidin Positive Methicillin-Susceptible Staphylococcus aureus: A Case Report of Two Pediatric Patients with Thrombotic Complications

  • Ng, David Chun Ern;Anand, L. Alexis;Khiu, Fu Lung;Tan, Kah Kee
    • Pediatric Infection and Vaccine
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    • v.25 no.1
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    • pp.50-53
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    • 2018
  • Staphylococcus aureus is a well-recognized human pathogen that causes a wide range of infections as a result of its extensive virulence factors. One of these factors is Panton-Valentine leukocidin (PVL), a potent pore-forming cytotoxin that has been linked to invasive S. aureus infections. PVL is one of the important virulence factors for S. aureus and has been largely recognized as one of the markers for community-acquired methicillin-resistant S. aureus. However, the presence of PVL in methicillin-susceptible S. aureus infections is not widely reported in the literature. Thrombotic sequelae of S. aureus infections associated with PVL expression are uncommon in children. We hereby report two children with thrombotic complications associated with PVL-producing methicillin-susceptible S. aureus. Both patients responded well to antibiotic and anticoagulant therapies, and survived without any long-term sequelae.

Putative response regulator two-component gene, CaSKN7, regulate differentiation and virulence in Candida albicans

  • Lee, Jung-Shin;Minyoung Lim;Yim, Hyung-Soon;Kang, Sa-Ouk
    • Proceedings of the Korean Biophysical Society Conference
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    • 2003.06a
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    • pp.50-50
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    • 2003
  • We have identified and analysed a putative response regulator two-component gene (CaSKN7) from Candida albicans and its encoding protein (CaSkn7). CaSKN7 has an open reading frame of 1677bp. CaSKN7 encodes a 559 amino acid protein (CaSkn7) with an estimated molecular mass of 61.1 kDa. CaSKN7 is a homologue of a Saccharomyces cerevisiae SKN7 that is the regulator involved in the oxidative stress response. To study the role of CaSKN7, we constructed a CAI4-derived mutant strain carrying a homozygous deletion of the CaSKN7 gene. In the caskn7 disruptant cells, the formation of germ tube require shorter time than that in the congenic wild-type strain but the growth of mycelium delayed in liquid media. In contrast, the caskn7 disruptant cells attenuate the differentiation in solid media and the virulence in mouse model system. Expression level of hypha-specific and virulence genes - HYR1, ECE1, HWP1, and ALS1 - in the caskn7 disruptant cells increased as compared with that in the congenic wild-type strain in 10% serum YPD. Skn7 in 5. cerevisiae was found to bind the HSE element from the SSA promoter, Also, CaSkn7 contains heat shock factor DNA-binding domain and the promoters of these genes have HSE-like sties. Therefore these results show that CaSKN7 regulate the differentiation and virulence of C. albicans.

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Arctigenin from Burdock Root Exhibits Potent Antibacterial and Anti-Virulence Properties against Pseudomonas aeruginosa

  • Abdulrahman E. Koshak;Mahmoud A. Elfaky;Hossam M. Abdallah;Dina A. I. Albadawi;Gamal A. Mohamed;Sabrin R. M. Ibrahim;Abdulrahim A. Alzain;El-Sayed Khafagy;Azza A. H. Rajab;Wael A. H. Hegazy
    • Journal of Microbiology and Biotechnology
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    • v.34 no.8
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    • pp.1642-1652
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    • 2024
  • Arctium lappa (Burdock) root is used in various culinary applications especially in Asian Cuisine. Arctigenin (ARC) is a polyphenolic compound abundant in the roots of the burdock plant from which it derives its name. The emergence of bacterial resistance is a growing global worry, specifically due to the declining availability of new antibiotics. Screening for the antibacterial candidates among the safe natural products is a promising approach. The present study was aimed to assess the antibacterial activity of ARC against Pseudomonas aeruginosa exploring its effect on the bacterial cell membrane. Furthermore, the anti-virulence activities and anti-quorum sensing (QS) activities of ARC were in vitro, in vivo and in silico assessed against P. aeruginosa. The current results showed the ARC antibacterial activity was owed to its disruption effect of the cell membrane. ARC at sub-MIC significantly decreased the formation of biofilm, motility, production of extracellular enzymes and in vivo protected mice against P. aeruginosa. These anti-virulence activities of ARC are owed to its interference with bacterial QS and its expression. Furthermore, ARC showed mild effect on mammalian erythrocytes, low probability to induce resistance and synergistically combined with antibiotics. In summary, the promising anti-virulence properties of ARC indicate its potential as an effective supplement to conventional antibiotics for treating severe P. aeruginosa infections.

Expression of Helicobacter pylori urease in plants to use as an edible vaccine

  • Gang, Gwi-Hyeon;Han, So-Cheon;Gang, Tae-Jin;Yang, Mun-Sik
    • 한국생물공학회:학술대회논문집
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    • 2003.10a
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    • pp.186-189
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    • 2003
  • Helicobacter pylori is the etiologic agent of human gastritis and peptic ulceration and produces urease as the major protein component on its surface. H. pylori urease is known to serve as a potent immunogen as well as major virulence factor. In order to express the recombinant urease in tobacco plants, a DNA fragment containing the minimal H. pylori urease gene cluster was subcloned into a plant expression vector. The recombinant vector was transformed to tobacco plants. The integration of the recombinant plasmids into tobacco chromosomal genome was verified by genomic PCR. Expression to mRNA was confirmed by Northern blot analysis, and expression to recombinant urease protein was observed by Western blot analysis. These results showed that the recombinant urease can be produced in tobacco plants and will be tested for immune response to use as an edible vaccine.

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Gene Expression Profiles Following High-Dose Exposure to Gamma Radiation in Salmonella enterica serovar Typhimurium

  • Lim, Sangyong;Jung, Sunwook;Joe, Minho;Kim, Dongho
    • Journal of Radiation Industry
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    • v.2 no.3
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    • pp.111-119
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    • 2008
  • Microarrays can measure the expression of thousands of genes to identify the changes in expression between different biological states. To survey the change of whole Salmonella genes after a relatively high dose of gamma radiation (1 kGy), transcriptome dynamics were examined in the cells by using DNA microarrays. At least 75 genes were induced and 89 genes were reduced two-fold or more after irradiation. Several genes located in pSLT plasmid, cyo operon, and Gifsy prophage were induced along with many genes encoding uncharacterized proteins.While, the expression of genes involved in the virulence of Salmonella as well as metabolic functions were decreased. Although the radiation response as a whole could not be illustrated by using DNA microarrays, the data suggest that the response to high dose of irradiation might be more complex than the SOS response.

Cloning, Sequencing and Expression of apxIA, IIA, IIIA of Actinobacillus pleuropneumoniae Isolated in Korea (국내 분리 흉막폐렴균의 apxIA, IIA, IIIA 유전자 Cloning, 염기서열 분석 및 단백질 발현)

  • Shin, Sung-jae;Cho, Young-wook;Yoo, Han-sang
    • Korean Journal of Veterinary Research
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    • v.43 no.2
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    • pp.247-253
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    • 2003
  • Actinobacillus pleuropneumoniae causes a highly contagious pleuropneumoniae in swine. The bacterium produces several virulence factors such as exotoxin, LPS, capsular polysaccharide, etc. Among them, the exotoxin, called Apx, has been focused as the major virulence factor, and the toxin consists of 4 gene cluster. apx CABD. apxA is the structural gene of toxin and has four different types, I, II, III, and IV. As the first step of development of a new subunit vaccine, the three different types of apxA gene were amplified from A. pleuropneumoniae isolated from Korea by PCR with primer designed based on the N- and C-terminal of the toxin. The sizes of apxIA, IIA and IIIA were 3,073, 2,971 and 3,159bps, respectively. The comparison of whole DNA sequences of apxIA, IIA and IIIA genes with those of the reference strain demonstrated 98%, 99% and 98% homology, respectively. In addition, the phylogenetic analysis was performed based on the amino acid sequences compared with 12 different RTX toxin family using the neighbor-joining method. ApxA proteins of Korean isolates were identical with reference strains in this study. All ApxA proteins were expressed in E. coli with pQE expression vector and identified using Western blot with polyclonal antibodies against culture supernatants of A. pleuropneumoniae serotype 2 or 5. The sizes of each expressed ApxA protein were about 120, 110, 125 kDa (M.W.), respectively. The results obtained in this study could be used for the future study to develop a new vaccine to porcine pleuropneumoniae.

The Transcription Cofactor Swi6 of the Fusarium graminearum Is Involved in Fusarium Graminearum Virus 1 Infection-Induced Phenotypic Alterations

  • Son, Moonil;Lee, Yoonseung;Kim, Kook-Hyung
    • The Plant Pathology Journal
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    • v.32 no.4
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    • pp.281-289
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    • 2016
  • The transcription cofactor Swi6 plays important roles in regulating vegetative growth and meiosis in Saccharomyces cerevisiae. Functions of Swi6 ortholog were also characterized in Fusarium graminearum which is one of the devastating plant pathogenic fungi. Here, we report possible role of FgSwi6 in the interaction between F. graminearum and Fusarium graminearum virus 1 (FgV1) strain DK21. FgV1 perturbs biological characteristics of host fungi such as vegetative growth, sporulation, pigmentation, and reduction of the virulence (hypovirulence) of its fungal host. To characterize function(s) of FgSWI6 gene during FgV1 infection, targeted deletion, over-expression, and complementation mutants were generated and further infected successfully with FgV1. Deletion of FgSwi6 led to severe reduction of vegetative growth even aerial mycelia while over-expression did not affect any remarkable alteration of phenotype in virus-free isolates. Virus-infected (VI) FgSWI6 deletion isolate exhibited completely delayed vegetative growth. However, VI FgSWI6 over-expression mutant grew faster than any other VI isolates. To verify whether these different growth patterns in VI isolates, viral RNA quantification was carried out using qRT-PCR. Surprisingly, viral RNA accumulations in VI isolates were similar regardless of introduced mutations. These results provide evidence that FgSWI6 might play important role(s) in FgV1 induced phenotype alteration such as delayed vegetative growth.

The Effect of Transformation on the Virulence of Streptococcus pneumoniae

  • Zhang Xue-Mei;Yin Yi-Bing;Zhu Dan;Chen Bao-De;Luo Jin-Yong;Deng Vi-Ping;Liu Ming-Fang;Chen Shu-Hui;Meng Jiang-Ping;Lan Kai;Huang Yuan-Shuai;Kang Ge-Fei
    • Journal of Microbiology
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    • v.43 no.4
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    • pp.337-344
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    • 2005
  • Although pneumococcus is one of the most frequently encountered opportunistic pathogen in the world, the mechanisms responsible for its infectiveness have not yet been fully understood. In this paper, we have attempted to characterize the effects of pneumococcal transformation on the pathogenesis of the organism. We constructed three transformation-deficient pneumococcal strains, which were designated as Nos. 1d, 2d, and 22d. The construction of these altered strains was achieved via the insertion of the inactivated gene, comE, to strains 1, 2 and 22. We then conducted a comparison between the virulence of the transformation-deficient strains and that of the wild-type strains, via an evaluation of the ability of each strain to adhere to endothelial cells, and also assessed psaA mRNA expression, and the survival of hosts after bacterial challenge. Compared to what was observed with the wild-type strains, our results indicated that the ability of all of the transformation-deficient strains to adhere to the ECV304 cells had been significantly reduced (p < 0.05), the expression of psaA mRNA was reduced significantly (p < 0.05) in strains 2d and 22d, and the median survival time of mice infected with strains Id and 2d was increased significantly after intraperitoneal bacterial challenge (p < 0.05). The results of our study also clearly indicated that transformation exerts significant effects on the virulence characteristics of S. pneumoniae, although the degree to which this effect is noted appears to depend primarily on the genetic background of the bacteria.

Mass expression of Apx I and Apx II of Actinobacillus pleuropneumoniae in Escherichia coli (대장균에서 흉막폐렴균 독소 Apx I과 Apx II의 대량발현)

  • Kim, Tae-Jung;Lee, Bong-Joo;Lee, Jae-Il
    • Korean Journal of Veterinary Research
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    • v.45 no.2
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    • pp.185-189
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    • 2005
  • Actinobacillus pleuropneumoniae is the causative agent of a porcine contagious pleuropneumonia. Among several virulence factors including exotoxin (Apx toxins), LPS, transferrin-binding proteins, OMPs, and some proteases, Apx toxins have been major targets for the protection study. In this study, cloning and expression of A. pleuropneumoniae Apx I and Apx II toxin, which are produced by all highly virulent strains, were performed by Escherichia coli expression system. Genes coding Apx I and II toxin were amplified from the A. pleuropneumoniae serotype 5 genomic DNA using polymerase chain reaction and cloned to a prokaryotic expression vector, pRSET. Expression of the Apx I and Apx II coding sequences in E. coli resulted in the formation of insoluble inclusion bodies purified according to a denaturing purification protocol, which employs the use of guanidium. Recombinant proteins were purified using $Ni^{2+}$-charged resin affinity purification. This expression and purification system made it possible to produce Apx I and Apx II in large amounts for further immunologic studies.

Comparative Analysis of Growth-Phase-Dependent Gene Expression in Virulent and Avirulent Streptococcus pneumoniae Using a High-Density DNA Microarray

  • Ko, Kwan Soo;Park, Sulhee;Oh, Won Sup;Suh, Ji-Yoeun;Oh, TaeJeong;Ahn, Sungwhan;Chun, Jongsik;Song, Jae-Hoon
    • Molecules and Cells
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    • v.21 no.1
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    • pp.82-88
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    • 2006
  • The global pattern of growth-dependent gene expression in Streptococcus pneumoniae strains was evaluated using a high-density DNA microarray. Total RNAs obtained from an avirulent S. pneumoniae strain R6 and a virulent strain AMC96-6 were used to compare the expression patterns at seven time points (2.5, 3.5, 4.5, 5.5, 6.0, 6.5, and 8.0 h). The expression profile of strain R6 changed between log and stationary growth (the Log-Stat switch). There were clear differences between the growth-dependent gene expression profiles of the virulent and avirulent pneumococcal strains in 367 of 1,112 genes. Transcripts of genes associated with bacterial competence and capsular polysaccharide formation, as well as clpP and cbpA, were higher in the virulent strain. Our data suggest that late log or early stationary phase may be the most virulent phase of S. pneumoniae.