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      Microbial interactions and differential protein expression in Staphylococcus aureusCandida albicans dual-species biofilms

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          Abstract

          The fungal species Candida albicans and the bacterial species Staphylococcus aureus are responsible for a majority of hospital-acquired infections and often coinfect critically ill patients as complicating polymicrobial biofilms. To investigate biofilm structure during polymicrobial growth, dual-species biofilms were imaged with confocal scanning laser microscopy. Analyses revealed a unique biofilm architecture where S. aureus commonly associated with the hyphal elements of C. albicans. This physical interaction may provide staphylococci with an invasion strategy because candidal hyphae can penetrate through epithelial layers. To further understand the molecular mechanisms possibly responsible for previously demonstrated amplified virulence during coinfection, protein expression studies were undertaken. Differential in-gel electrophoresis identified a total of 27 proteins to be significantly differentially produced by these organisms during coculture biofilm growth. Among the upregulated staphylococcal proteins was l-lactate dehydrogenase 1, which confers resistance to host-derived oxidative stressors. Among the downregulated proteins was the global transcriptional repressor of virulence factors, CodY. These findings demonstrate that the hyphae-mediated enhanced pathogenesis of S. aureus may not only be due to physical interactions but can also be attributed to the differential regulation of specific virulence factors induced during polymicrobial growth. Further characterization of the intricate interaction between these pathogens at the molecular level is warranted, as it may aid in the design of novel therapeutic strategies aimed at combating fungal–bacterial polymicrobial infection.

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          Invasive methicillin-resistant Staphylococcus aureus infections in the United States.

          As the epidemiology of infections with methicillin-resistant Staphylococcus aureus (MRSA) changes, accurate information on the scope and magnitude of MRSA infections in the US population is needed. To describe the incidence and distribution of invasive MRSA disease in 9 US communities and to estimate the burden of invasive MRSA infections in the United States in 2005. Active, population-based surveillance for invasive MRSA in 9 sites participating in the Active Bacterial Core surveillance (ABCs)/Emerging Infections Program Network from July 2004 through December 2005. Reports of MRSA were investigated and classified as either health care-associated (either hospital-onset or community-onset) or community-associated (patients without established health care risk factors for MRSA). Incidence rates and estimated number of invasive MRSA infections and in-hospital deaths among patients with MRSA in the United States in 2005; interval estimates of incidence excluding 1 site that appeared to be an outlier with the highest incidence; molecular characterization of infecting strains. There were 8987 observed cases of invasive MRSA reported during the surveillance period. Most MRSA infections were health care-associated: 5250 (58.4%) were community-onset infections, 2389 (26.6%) were hospital-onset infections; 1234 (13.7%) were community-associated infections, and 114 (1.3%) could not be classified. In 2005, the standardized incidence rate of invasive MRSA was 31.8 per 100,000 (interval estimate, 24.4-35.2). Incidence rates were highest among persons 65 years and older (127.7 per 100,000; interval estimate, 92.6-156.9), blacks (66.5 per 100,000; interval estimate, 43.5-63.1), and males (37.5 per 100,000; interval estimate, 26.8-39.5). There were 1598 in-hospital deaths among patients with MRSA infection during the surveillance period. In 2005, the standardized mortality rate was 6.3 per 100,000 (interval estimate, 3.3-7.5). Molecular testing identified strains historically associated with community-associated disease outbreaks recovered from cultures in both hospital-onset and community-onset health care-associated infections in all surveillance areas. Invasive MRSA infection affects certain populations disproportionately. It is a major public health problem primarily related to health care but no longer confined to intensive care units, acute care hospitals, or any health care institution.
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            High resolution two-dimensional electrophoresis of proteins.

            A technique has been developed for the separation of proteins by two-dimensional polyacrylamide gel electrophoresis. Due to its resolution and sensitivity, this technique is a powerful tool for the analysis and detection of proteins from complex biological sources. Proteins are separated according to isoelectric point by isoelectric focusing in the first dimension, and according to molecular weight by sodium dodecyl sulfate electrophoresis in the second dimension. Since these two parameters are unrelated, it is possible to obtain an almost uniform distribution of protein spots across a two-diminsional gel. This technique has resolved 1100 different components from Escherichia coli and should be capable of resolving a maximum of 5000 proteins. A protein containing as little as one disintegration per min of either 14C or 35S can be detected by autoradiography. A protein which constitutes 10 minus 4 to 10 minus 5% of the total protein can be detected and quantified by autoradiography. The reproducibility of the separation is sufficient to permit each spot on one separation to be matched with a spot on a different separation. This technique provides a method for estimation (at the described sensitivities) of the number of proteins made by any biological system. This system can resolve proteins differing in a single charge and consequently can be used in the analysis of in vivo modifications resulting in a change in charge. Proteins whose charge is changed by missense mutations can be identified. A detailed description of the methods as well as the characteristics of this system are presented.
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              The distinct morphogenic states of Candida albicans.

              The human fungal pathogen, Candida albicans can grow in at least three different morphologies: yeast, pseudohyphae and hyphae. Further morphological forms exist during colony switching, for example, opaque phase cells are oblong, rather than the oval shape of yeast cells. Pseudohyphae and hyphae are both elongated and sometimes there has been little attempt to distinguish between them, as both are "filamentous forms" of the fungus. We review here the differences between them that suggest that they are distinct morphological states. We argue that studies on "filamentous forms" should always include a formal analysis to determine whether the cells are hyphae or pseudohyphae and we suggest some simple experimental criteria that can be applied to achieve this.
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                Author and article information

                Journal
                FEMS Immunol Med Microbiol
                fim
                Fems Immunology and Medical Microbiology
                Blackwell Publishing Ltd
                0928-8244
                1574-695X
                August 2010
                01 July 2010
                : 59
                : 3
                : 493-503
                Affiliations
                [1 ]simpleGraduate Program in Life Sciences, Molecular Microbiology and Immunology Program, University of Maryland – Baltimore Baltimore, MD, USA
                [2 ]simpleDepartment of Microbial Pathogenesis, University of Maryland – Baltimore, Dental School Baltimore, MD, USA
                [3 ]simpleDepartment of Oncology and Diagnostic Sciences, Dental School, University of Maryland – Baltimore Baltimore, MD, USA
                [4 ]simpleDepartment of Pathology, School of Medicine, University of Maryland Baltimore, MD, USA
                [5 ]simpleDepartment of Biological Sciences, Northern Arizona University Flagstaff, AZ, USA
                [6 ]simpleDepartment of Orthopedic Surgery, Center for Genomic Sciences, Allegheny-Singer Research Institute, Allegheny General Hospital Pittsburgh, PA, USA
                [7 ]simpleDepartment of Microbiology and Immunology, School of Medicine, University of Maryland – Baltimore Baltimore, MD, USA
                Author notes
                Correspondence: Mark E. Shirtliff, Department of Biomedical Sciences, Dental School, University of Maryland – Baltimore, 650 W. Baltimore Street, 8th Floor South, Baltimore, MD 21201, USA. Tel.: +1 410 706 2263; fax: +1 410 706 0865; e-mail: mshirtliff@ 123456umaryland.edu

                Editor: Roger Bayston

                Article
                10.1111/j.1574-695X.2010.00710.x
                2936118
                20608978
                dd6869da-9be4-4912-ad3a-9bb6fe71a1a4
                © 2010 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved

                Re-use of this article is permitted in accordance with the Creative Commons Deed, Attribution 2.5, which does not permit commercial exploitation.

                History
                : 23 February 2010
                : 15 May 2010
                : 25 July 2010
                Categories
                Research Articles

                Microbiology & Virology
                candida albicans,proteome,staphylococcus aureus,polymicrobial,biofilm
                Microbiology & Virology
                candida albicans, proteome, staphylococcus aureus, polymicrobial, biofilm

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