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Vibrio cholerae, the causative agent of Asiatic cholera, is a gram-negative motile bacterial species acquired via oral ingestion of contaminated food or water sources. Cholera has spread from the Indian subcontinent where it is endemic to involve nearly the whole world seven times during the past 185 years. V. cholerae serogroup O1, biotype El Tor, has moved from Asia to cause pandemic disease in Africa and South America during the past 35 years. A new serogroup, O139, appeared in south Asia in 1992, has become endemic there, and threatens to start the next pandemic. The facultative human pathogen V. cholerae represents a paradigm that evolved from environmental non-pathogenic strains by acquisition of virulence genes. The major virulence factors of V. cholerae, cholera toxin (CTX) encoded by the ctxAB genes residing in the genome of filamentous lysogenic bacteriophage (CTXɸ) and toxin coregulated pilus (TCP) encoded by vibrio pathogenicity island (VPI). CTX, a potent stimulator of adenylate cyclase, causes the intestine to secrete watery fluid rich in sodium, bicarbonate, and potassium, in volumes far exceeding the intestinal absorptive capacity, by ADP-ribosylation of the alpha subunit of the GTP-binding protein. Thus intestinal infection with V. cholerae results in the loss of large volumes of watery stool, leading to severe and rapidly progressing dehydration and shock. Without adequate and appropriate rehydration therapy, severe cholera kills about half of affected individuals. Today, cholera still remains a burden mainly for underdeveloped countries, which cannot afford to establish or to maintain necessary hygienic and medical facilities. During the last three decades, intensive research has been undertaken to unravel the virulence properties and to study the epidemiology of this significant human pathogen. This review provides an overview of the role of CTX in the occurrence of this disease in humans.
Two PCR-based methods for identification of emetic toxin producing Bacillus cereus strains were developed. The first set of primers cesBF and cesBR allowed for the amplification of cesBI 838 bp long fragment of cereulide biosynthesis operon for cereulide producing strains, or 421 bp long fragment of nonribosomal peptide synthetase (nrps) for emetic toxin non producing strains. Detection of both genes cesBI and nrps was possible in one PCR reaction. Among 24 strains of Cereus group tested, only one named 19W-cesB contained cesBI gene fragment. Bacillus cereus isolate 19W-cesB did not contain of any other genes of nonribosomal peptide synthetases responsible for the synthesis of other low molecular weight peptide toxins. The shdR and shdF second primer set allowed for specific amplification of the other 690 bp long fragment of cesBII gene. Only strain 19W-cesB allowed for the PCR synthesis of appropriate amplicon from all tested strains. Proposed methods may be fast and reliable techniques for detection of Bacillus cereus strains producing cereulide.
Altered tissue water homeostasis may contribute to edema formation during various stresses including bacterial infection. We observed induction of aquaporin-1 (AQP1) during Staphylococcus aureus infection of cultured cells indicating a potential mechanism underlying altered water homeostasis during infection. To investigate mechanisms of AQP1 induction, we examined the effects of the S. aureus alpha-hemolysin on AQP1 abundance in Balb/c fibroblasts. Fibroblasts incubated with 30 microg/ml hemolysin exhibited a 5-10 fold increase in AQP1 protein within 4-6 hours of exposure. The use of multiple signaling cascade inhibitors failed to affect hemolysin-mediated accumulation of AQP1. However, immunoprecipitation revealed an initial accumulation of ubiquitinated AQP1 followed by a decrease to baseline levels after 4 hours. Immunofluorescence indicated that following hemolysin exposure, AQP1 was no longer on the plasma membrane, but was found in a population of submembrane vacuoles. AQP1 redistribution was further indicated by surface biotinylation experiments suggesting diminished AQP1 abundance on the plasma membrane as well as redistribution out of lipid raft fractions. Live cell confocal microscopy revealed that the pattern of cell volume change observed following hemolysin exposure was altered in cells in which AQP1 was silenced. We conclude that alpha-toxin alters proteasomal processing and leads to intracellular accumulation of AQP1, which may likely contribute to disrupted cell volume homeostasis in infection.
Quorum sensing (QS), or cell-to-cell communication in bacteria, is achieved through the production and subsequent response to the accumulation of extracellular signal molecules called autoinductors. The main role of QS is regulation of production of virulence factors in bacteria. Bacterial pathogenicity is often manifested by the expression of various cell-associated and secreted virulence factors, such as exoenzymes, toxins and biofilm. In bacteria, the expression of virulence factors is controlled coordinately by the global regulatory QS systems, which includes the AI-1/LuxIR-, AI-2/LuxS-, AI-3/QsC-, AIP/Agr-based systems. The regulation of production of virulence factors is extremely complex and many components influence it.
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