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Biomedical subjects

Paul Webster

Publications and source records attributed to Paul Webster.

At least 19 recordsLinked to original sources
PubMed2006

Identification of biofilm proteins in non-typeable Haemophilus Influenzae.

BACKGROUND: Non-typeable Haemophilus influenzae biofilm formation is implicated in a number of chronic infections including otitis media, sinusitis and bronchitis. Biofilm structure includes cells and secreted extracellular matrix that is "slimy" and believed to contribute to the antibiotic resistant properties of biofilm bacteria. Components of biofilm extracellular matrix are largely unknown. In order to identify such biofilm proteins an ex-vivo biofilm of a non-typeable Haemophilus influenzae isolate, originally from an otitis media patent, was produced by on-filter growth. Extracellular matrix fraction was subjected to proteomic analysis via LC-MS/MS to identify proteins. RESULTS: 265 proteins were identified in the extracellular matrix sample. The identified proteins were analyzed for COG grouping and predicted cellular location via the TMHMM and SignalP predictive algorithms. The most over-represented COG groups identified compared to their frequency in the Haemophilus influenzae genome were cell motility and secretion (group N) followed by ribosomal proteins of group J. A number of hypothetical or un-characterized proteins were observed, as well as proteins previously implicated in biofilm function. CONCLUSION: This study represents an initial approach to identifying and cataloguing numerous proteins associated with biofilm structure. The approach can be applied to biofilms of other bacteria to look for commonalities of expression and obtained information on biofilm protein expression can be used in multidisciplinary approaches to further understand biofilm structure and function.

Bacterial Proteins↗
PubMed2006

Cytoplasmic bacteria and the autophagic pathway.

Cytoplasmic bacteria may assist in our study of the autophagic pathway. This review highlights the use of Listeria monocytogenes for examining the assembly of autophagic vacuoles in mammalian cells. Inhibiting protein synthesis of cytoplasmic L. monocytogenes results in their being sequestered into the autophagic pathway. Autophagic vacuoles form around the easily identified bacterial particles making the assembly process easy to study using morphological and biochemical methods. L. monocytogenes, which appears to be ideally adapted to life in the cell cytoplasm, does not normally become a target of autophagy. In model systems the bacteria thrive within host cell cytoplasm, indicating the importance of de novo protein synthesis in avoiding the autophagic pathway. This observation indicates an interesting opportunity for identifying the bacterial mechanisms that are mobilized to avoid the autophagic pathway.

Animals↗
PubMed2006

Imidazole groups on a linear, cyclodextrin-containing polycation produce enhanced gene delivery via multiple processes.

The linear, cyclodextrin-containing polycation (CDP) is one of many non-viral gene delivery vectors that show improved transfection efficiency when modified to have pH-buffering capacity. The buffering activity is presumed to confer enhanced ability to escape the endocytic pathway. Here, the differences in delivery behavior between CDP and its pH-buffering, imidazole-containing variant (CDPim) are investigated in order to elucidate the mechanism(s) by which these related materials exhibit differences in gene delivery. In cell-free assays that include dye exclusion and heparan sulfate displacement, CDP appears to have weaker binding strength with nucleic acids than CDPim. Numerous analyses involving transfected cells, however, indicate that CDPim more readily releases nucleic acids in the intracellular setting. Together, these data suggest that differences in transfection efficiency between CDP and CDPim result from factors beyond buffering activity and endosomal escape.

Buffers↗
PubMed2006

Distribution of bacterial proteins in biofilms formed by non-typeable Haemophilus influenzae.

The ability to preserve the fragile ultrastructural organization of bacterial biofilms using cryo-preparation methods for electron microscopy has enabled us to probe sections through non-typeable Haemophilus influenzae (NTHi) biofilms and determine the localization of NTHi-specific lipooligosaccharide (LOS) and proteins within these structures. Some of the proteins we examined are currently being considered as candidates for vaccine development, so it is important that their distribution and accessibility within the biofilms formed by NTHi be determined. We have localized LOS to the extracellular matrix (ECM) of the biofilm and the P6 outer membrane protein to the membrane of what appear to be viable bacteria within the biofilm. The Hap and HWM1/HMW2 adhesive proteins were associated with bacteria within the biofilm and were present in the biofilm ECM. The IgA1 protease is a secreted protein that was also associated with NTHi in the biofilm and was in the ECM, but was more concentrated in the top region of the biofilm, suggesting a role in protecting biofilm bacteria from antibody attack.

Adhesins, Bacterial↗
PubMed2006

Presbycusic neuritic degeneration within the osseous spiral lamina.

OBJECTIVE: To describe a neglected anatomic variant occurring with presbycusis. STUDY DESIGN: Retrospective temporal bone histopathology study. METHODS: Quantitative analysis of peripheral hair cells, neurites, neurons, and the stria vascularis in temporal bones from individuals who had presbycusis. Fifty-three patients aged 65 years or older and with a down-sloping audiogram and clinical diagnosis of presbycusis were reviewed. Nine cases had normal hair and ganglion cell populations but reduced peripheral processes (neuritic presbycusis). These were compared with five normal-hearing controls on measurements of anterior middle and basal turn fiber bundle diameter and the ratio of basal to middle diameters. RESULTS: Thresholds at 4 and 8 kHz were significantly poorer in the neuritic presbycusis group than in the control group (p<or=0.004 and 0.05, respectively), as was speech discrimination score (p<or=0.028). The ratio of basal to middle turn diameters was significantly smaller in the neuritic presbycusis group (p<or=0.003). This effect was quite marked in that there was no overlap in ratios between the groups, with the maximum neuritic presbycusis group ratio smaller than the minimum control group ratio. There was a moderate negative correlation between ratio and threshold at 4 kHz (sigma=-0.49, p<or=0.075). CONCLUSION: Loss of peripheral neurites in the anterior basal cochlear segment is found in conjunction with presbycusis in temporal bones that have no other morphologic abnormalities. These cases can be identified by a gradual down-sloping audiogram in contrast to sensory (hair cell) presbycusis, which is characterized by a precipitous high tone loss.

Aged↗
PubMed2006

Characterization of rat spiral ligament cell line immortalized by adenovirus 12-simian virus 40 hybrid virus.

OBJECTIVES: Spiral ligament fibrocytes play an important role in inner ear ion homeostasis and are classified into several subtypes according to expression of specific enzymes such as Na+, K+ -ATPase, Ca++ -ATPase, and carbonic anhydrase. Although our understanding of the cell and molecular biology of spiral ligament fibrocytes has increased over time, access to these cells still remains a significant hurdle hindering future studies. In this study, we aimed to establish a rat spiral ligament cell line with minimal disruption of the original characteristics. METHODS: The primary spiral ligament fibrocytes were exposed to adenovirus 12-simian virus 40 hybrid virus for immortalization. Karyotypic analysis was performed after stabilization of the infected cells, and the population doubling time was compared to that of the primary cell. The cell line was characterized by immunolabeling and electron microscopy. RESULTS: We describe the establishment and characterization of a line of type I spiral ligament fibrocytes immortalized with an adenovirus 12-simian virus 40 hybrid virus. CONCLUSIONS: This cell line can be a useful research tool for investigating the role of spiral ligament fibrocytes in homeostasis and inflammation of the inner ear.

Adenoviridae↗