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Studies on T-OMP and the development of antimicrobial tolerance in Pseudomonas aeruginosa PAO1Winder, Catherine Louise January 2000 (has links)
<i>Pseudomonas aeruginosa</i> displays high levels of tolerance and resistance to many antimicrobial agents. Much of this tolerance is related to the nature of the Gram-negative cell envelope and in particular, the outer membrane. The outer membrane plays an important role in excluding harmful molecules from the cell, whilst being selectively permeable to other solutes via its implanted proteins (outer membrane proteins or OMPs). In order to exert their antibacterial action, antimicrobial agents must enter the cell and attain sufficiently high concentrations at their target site(s). The OMPs are highly sensitive to environmental changes and have a physiological ability to respond to such changes. It is thought that the altered cell envelope structure contributes to the accessibility of antimicrobial agents into the cell interior and resistance to such agents is related to over expression or loss of certain OMPs. Brozel and Cloete (1994) observed a gradual increase in tolerance to increasing concentrations of biocide upon exposure of <i>P. aeruginosa</i> to KathonTM, a commercial biocide containing 1.15% v/v 5-chloro-N-methylisothiazolone (CMIT) and 0.35% v/v N-methylisothiazolone (MIT). This adaptation was associated with the concurrent disappearance of a 35kDa OMP, designated T-OMP. Therefore, they concluded that the biocide entered the sensitive cells via the T-OMP and that the observed resistance was the result of the absence of this OMP. The aim of this investigation was to induce tolerance in cultures of <i>P. aeruginosa</i> PAOl towards the pure active forms of the three isothiazolone biocides 1,2-benzisothiazolone (BIT), MIT, CMIT and the thiol-interactive agent thiomersal (used as a positive control). An increase was observed in the minimum inhibitory concentrations (MIC) of all four biocides by at least 58% between the sensitive and resistant cultures. In some cases the percentage increase in MIC was in excess of 150%. However, when the tolerant cells were removed from the presence of the biocide, the MIC began to decrease, indicating a loss in tolerance. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDSPAGE) analysis of the OMP profiles from the tolerant-induced cultures illustrated the loss of T-OMP in all cases. Analysis of the sensitive and resistant cultures using twodimensional polyacrylamide gel electrophoresis (2D-PAGE) indicated that the T-OMP disappeared in the tolerant cultures. However, these observations also suggested that other outer membrane alterations occur concurrently in T-OMP depleted tolerant cells. Investigations into the cross-resistance of the resistant cultures towards the other test biocides, indicated that resistance was, to some extent, transferable, once it had been developed towards one member of the biocide group. Following routine passaging of the resistant cultures on gradient plates two distinct colonial morphologies were observed, mucoid and non-mucoid. An increase in the cell surface hydrophobicity was noted between the mucoid and non-mucoid cultures, which indicated a loss or reduction in the B-band OPolysaccharide. However, there were no observable differences in the lipopolysaccharide banding patterns between the mucoid and non-mucoid cells. These observations suggested that other alterations were occurring in the tolerant cells upon exposure to biocide, over and above the simple disappearance of T-OMP. Therefore, it is suggested that the observed tolerant development in biocide exposed cells, was not solely due to the loss of T-OMP. Investigations into Gram-negative bacteria isolated from contaminated industrial samples preserved with isothiazolone compounds exhibited higher MICs towards the preservative biocides than would normally be expected in the species of bacteria isolated and identified. However, there were no observable alterations in their OMP profiles.
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Gene expression in the genus DeinococcusPurvis, Ian James January 1984 (has links)
No description available.
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Mercury resistance genes in a natural Bacillus populationHart, Mark Christopher January 1998 (has links)
No description available.
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REDUCING BACTERIAL RESISTANCE THROUGH BETTER ANTIBIOTIC PRESCRIPTION PRACTICESOuma, Christine 01 January 2008 (has links)
The objective of this study was to find a regression procedure that can better explain the relationship between patterns of antibiotic use and proportions of bacterial resistance. The sample for the study is comprised of 44 University Health System Consortium (UHC) member hospitals, and the data for antibiotic use and proportions of resistance are from the years 2002 to 2005. The hospitals are spread across the Northeast, South, Southwest, Midwest, and Northwest regions of the USA. Based on statistical analysis, MRSA continues to have the highest proportion of resistance among the bacteria examined and has increased significantly since 2002. The antibiotic use in the study was measured in indices called diversity indices. There were six such measures in the study. The study, first using ordinary least squares regression, did not find one single diversity index that adequately predicted the proportion of resistance. There were also concerns that the diversity indices could be measuring the same thing, and therefore all should not be used in the model. The correlations between the three general diversity indices were strong, positive, and linear. Likewise, the three Gram-negative indices were also positively correlated with one another. Multicollinearity diagnostics also showed that there were serious dependencies among general diversity indices. Given the multicollinearity results and the correlation coefficients for the indices, it can be concluded that all six indices should not be in the same model together. Logistic regression and weighted least squares regression using the logit transformation were also performed, and just like the ordinary least squares results, there was no one single diversity index or a combination of diversity indices that adequately predicted the proportion of resistance.
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Synthesis and Biological Activity of Aminoglycosides and 1,4-Naphthoquinone DerivativesYatchang, Marina Fosso 01 December 2012 (has links)
The research described in this dissertation is at the interface of organic chemistry and biology, and it aimed at designing and synthesizing biologically active molecules for the possible development of therapeutic agents. Spinal muscular atrophy is an incurable disease that affects 1 in every 6000 babies, making it the leading genetic cause of infant mortality. While no treatment is available, efforts are being taken to solve this issue. Part of the work outlined in this dissertation was carried out in collaboration with researchers from the University of Missouri to investigate a potential therapeutic for this disease. In addition, the continuous outbreak of diseases caused by bacteria demands for new and improved antibiotics that could help eradicate those pathogens. My research thus allowed me to discover molecules with interesting activity against bacteria for the possible development of potential antibacterial agents. Finally, my research also allowed me to develop potential agro fungicides, which are still very much needed nowadays. Many crop diseases are due to fungal infections,which globally cause enormous economic losses. The use of fungicides is still the main strategy to control these diseases. However, current agro fungicides show some limitations. This is illustrated with Fusarium head blight (FHB), a destructive and costly disease of wheat, barley and other small grains, whose economic losses in the Central United States alone were estimated to $2.7 billion.
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Increased expression of <i>ompA, ompX, dedA</i>, and <i>gutS</i> genes in <i>Enterobacter</i> sp. YSU in the presence of seleniteAl-Akash, Ahmed M. 11 December 2020 (has links)
No description available.
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Antimicrobial peptides : structure, function and resistanceVargues, Thomas January 2009 (has links)
Higher eukaryotes produce a vast range of antimicrobial peptides (AMPs) that play important roles in their defence against microbial infection. Beta defensins are small (3-5 kDa), cationic peptides that display broad, potent antimicrobial activity against a range of microbes and also act as chemoattractants of important immunomodulatory cells. To generate highly pure peptides for structural and functional studies, we developed a method to prepare recombinant human beta defensin-2 (HBD2). The HBD2 gene was synthesised by recursive PCR with codons optimised for expression in Escherichia coli. HBD2 was expressed as an insoluble fusion to a His-tagged ketosteroid isomerase. After cleavage from the fusion with cyanogen bromide, 1H NMR spectroscopy and mass spectrometry confirmed that the oxidised HBD2 was folded and possessed the correct b-defensin disulfide bond topology. The recombinant HBD2 was active against E. coli, P. aeruginosa, S. aureus and C. albicans and was also a chemoattractant against HEK293 cells expressing the chemokine receptor CCR6. 15N-labelled HBD2 was also prepared and was highly suitable for future structural studies. Since defensins are thought to interact with bacterial membranes we also tested the recombinant HBD2 in biophysical studies (surface plasmon resonance, SPR, Biacore). We observed different binding to artificial model membranes containing either E. coli Kdo2-lipid A or phospholipids. Bacterial resistance to AMPs has been linked to the covalent modification of the outer membrane lipid A by 4-amino-4-deoxy-L-arabinose (L-Ara4N). This neutralises the charge of the LPS, thereby decreasing the electrostatic attraction of cationic peptides to the bacterial membrane. The pathogen Burkholderia cenocepacia displays extremely high resistance to AMPs and other antibiotics and the Ara4N pathway appears to be essential. To explore this further we expressed recombinant forms of two enzymes (ArnB and ArnG) from the B. cenocepacia Ara4N pathway. Purified ArnB is a pyridoxal 5’-phosphate (PLP)-dependent transaminase and we tested its ability to bind amino acid substrates. We investigated the binding of inhibitors L- and D-cycloserine to ArnB and tested their antibiotic activity against Burkholderia strains. We also studied the B. cenocepacia ArnG – a proposed membrane protein undecaprenyl-L-Ara4N flippase – and showed that the protein behaved as a dimer by non-denaturing gel analysis. The B. cenocepacia ArnG failed to complement E. coli knock-out strains encoding the equivalent flippase proteins ArnE and ArnF, suggesting that ArnG is a Burkholderia-specific protein.
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Genetic methods for Rapid Detection of Medically Important Nosocomial BacteraThomas, Lee January 2007 (has links)
Master of Science / The role of the microbiology laboratory is (1) to provide infection control information, so that highly transmissible isolates may be identified and appropriate control measures instigated as rapidly as possible and (2) to provide adequate information to the clinician enabling correct antibiotic choices to be made, particularly in the critically ill. Microbiological data is by definition slow as it is culture dependent: this study focused on the development of genetic, culture-independent methods for detection of resistance in nosocomial pathogens that could be introduced into the routine microbiology department and would fit into the routine workflow with a consequent reduction in time to result. Initially a duplex real-time polymerase chain reaction was developed for the rapid identification and detection of S. aureus and methicillin-resistance. This was optimised for immediate as-needs testing of positive blood cultures signalling with “Gram positive cocci, possibly staphylococcus” evident on Gram stain, on a random access real-time PCR platform. This technology, allowing early identification of S. aureus and its susceptibility to methicillin, by simple automated methodology, may soon become the standard for all microbiology laboratories servicing the critically ill. The second part of the study involved the development of a selective broth and multiplex PCR for detection of three important nosocomial isolates at this institution, methicillin-resistant S. aureus (MRSA), carbapenem-resistant Enterobacteriaceae, and multi-resistant Acinetobacter baumannii (MRAB). A multiplex PCR using four primer sets was designed to detect low colonisation levels of these isolates after overnight incubation in selective broth, significantly reducing the time to result and associated costs. This potentially useful epidemiological screening tool is practical, reproducible and sensitive with the potential of moving to an automated test (using real-time PCR, for example) in the future. The availability of early negative results judged by simple visual scanning (or by densitometry), means that the result is less operator-dependent, potentially reducing error rate. The last part of the study dealt with an important resistance phenotype, aminoglycoside resistance. There had been no recent comprehensive local surveys performed to determine the frequency of aminoglycoside resistance amongst the Enterobacteriaceae, or to identify the genetic determinants and their transmissibility. The isolates collected for the study were all resistant to at least one of gentamicin, tobramycin or amikacin. Identification of integron cassette arrays and use of specific internal primers identified at least one genetic determinant for gentamicin and tobramycin resistance in 22 of 23 isolates. Three isolates had two aminoglycoside resistance genes, and three isolates had three aminoglycoside resistance genes identified (Table 6.1). Transferable gentamicin-resistant plasmids were predominant amongst Klebsiella spp., but less so amongst Enterobacter spp. and E. coli. Gentamicin-resistant Klebsiella spp. were often ESBL positive, the genetic determinants of which were typically co-transferred on a conjugative plasmid. The importance of screening at a local level was demonstrated by the unexpected predominance of aac(6')-IIc amongst Enterobacter spp. and the detection of a new gene (aac(6')-LT). This part of the study has provided an understanding of the primary aminoglycoside resistance genes present in the local setting and their association with other resistances. This knowledge will allow development of assays for patient screening (clinical isolates and colonising flora), to better understand the epidemiology of aminoglycoside resistance and to allow better choice of antibiotic therapy related to presence or absence of these genes.
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Cross resistance amongst coliphages / Robert E.W. HancockHancock, Robert Ernest William January 1974 (has links)
x, 153, xxviii leaves : ill. ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Thesis (Ph.D.)--University of Adelaide, Dept. of Microbiology, 1975
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Cross resistance amongst coliphages / Robert E.W. HancockHancock, Robert Ernest William January 1974 (has links)
x, 153, xxviii leaves : ill. ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Thesis (Ph.D.)--University of Adelaide, Dept. of Microbiology, 1975
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