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Biomedical subjects

C Robinson

Publications and source records attributed to C Robinson.

At least 19 recordsLinked to original sources

Plaque biofilms: the effect of chemical environment on natural human plaque biofilm architecture.

The architecture of microbial biofilms especially the outer regions have an important influence on the interaction between biofilm and local environment particularly on the flux of materials into and out of biofilm compartments and as a consequence, biofilm metabolic behaviour. In the case of dental plaque biofilms, architecture will determine access of nutrients including acidogenic substrates and therapeutic materials to the microbial biomass and to the underlying tooth surface. Manipulation of this architecture may offer a means of altering mass transfer into the whole biofilm and biomass and raises the possibility of improving access of therapeutics. Plaque biofilms formed in vivo on human enamel were subjected to a number of different chemical conditions while under observation by confocal laser scanning microscopy in reflection mode. In this way the outer 50-100 microm or so of the biofilms was examined. Density and distribution of biomass were recorded as degree of reflectance. The amount and density of biofilm biomass increased from the plaque saliva interface towards the interior. Plaque biofilms were robust and little affected by mechanical manipulation, high ionic strength or low pH (2.5). Detergent (SLS), however, often appeared to either remove biomass and/or dramatically reduce its density.

Biofilms↗

Effects of systemic fluoride and in vitro fluoride treatment on enamel crystals.

Systemically administered fluoride at a concentration of 75 ppm increases the surface roughness of developing enamel crystals in rats, which may be significant in advancing our understanding of the biological mechanism of fluorosis. Thus, the aim of this study was to investigate whether the increased surface roughness may be a result of surface restructuring by the direct action of fluoride at the crystal surface. We examined the fluoride dose-dependent roughening of enamel crystal surfaces in vivo, in the rat, and whether this roughening could be mimicked by the in vitro treatment of rat enamel crystals with neutral pH fluoride solutions. Our results showed that enamel crystal surface roughness increased after treatment with increasing fluoride ion concentrations, whether applied in vitro or administered systemically. This suggests a mechanism, alongside others, for the increased surface roughness of crystals in fluorotic enamel.

Administration, Oral↗

Model testing using data on 137Cs from Chernobyl fallout in the Iput River catchment area of Russia.

Data collected for 10 years following the Chernobyl accident in 1986 have provided a unique opportunity to test the reliability of computer models for contamination of terrestrial and aquatic environments. The Iput River scenario was used by the Dose Reconstruction Working Group of the BIOMASS (Biosphere Modelling and Assessment Methods) programme. The test area was one of the most highly contaminated areas in Russia following the accident, with an average contamination density of 137Cs of 800,000 Bq m-2 and localized contamination up to 1,500,000 Bq m-2, and a variety of countermeasures that were implemented in the test area had to be considered in the modelling exercise. Difficulties encountered during the exercise included averaging of data to account for uneven contamination of the test area, simulating the downward migration and changes in bioavailability of 137Cs in soil, and modelling the effectiveness of countermeasures. The accuracy of model predictions is dependent at least in part on the experience and judgment of the participant in interpretation of input information, selection of parameter values, and treatment of uncertainties.

Cesium Radioisotopes↗

Model testing using data on 131I released from Hanford.

The Hanford test scenario described an accidental release of 131I to the environment from the Hanford Purex Chemical Separations Plant in September 1963. Based on monitoring data collected after the release, this scenario was used by the Dose Reconstruction Working Group of BIOMASS to test models typically used in dose reconstructions. The primary exposure pathway in terms of contribution to human doses was ingestion of contaminated milk and vegetables. Predicted mean doses to the thyroid of reference individuals from ingestion of 131I ranged from 0.0001 to 0.8 mSv. For one location, predicted doses to the thyroids of two children with high milk consumption ranged from 0.006 to 2 mSv. The predicted deposition at any given location varied among participants by a factor of 5-80. The exercise provided an opportunity for comparison of assessment methods and conceptual approaches, testing model predictions against measurements, and identifying the most important contributors to uncertainty in the assessment result. Key factors affecting predictions included the approach to handling incomplete data, interpretation of input information, selection of parameter values, adjustment of models for site-specific conditions, and treatment of uncertainties.

Iodine Radioisotopes↗

Surface chemistry of enamel apatite during maturation in relation to pH: implications for protein removal and crystal growth.

Apatite crystal growth rate and morphology in mineralized tissues are considered to be controlled by surface interaction with extracellular matrix proteins. During enamel maturation where protein is finally removed from crystal surfaces to permit massive crystal growth, pH oscillates between approximately 5.8 and approximately 7.2. With this in mind, a study of enamel apatite surface chemistry in terms of local environmental pH was undertaken. Using atomic force microscopy adhesion force measurements were made between hydroxylated or carboxylated cantilever tips and maturation stage crystals between pH 2 and 10. Adhesion force increased from pH 10 to a maximum at pH 6.6 presumably due to increased hydrogen bonding due to replacement of surface cations (Na, Ca, Mg) with protons and/or protonation of phosphate per se. Below pH 6.6 adhesion force decreased and became very variable indicating that the surface had become unstable probably due to removal of fully protonated phosphate from the surface by adherence to the cantilever tip. Frictional force measurements also revealed 2-3, approximately 30 nm diameter high friction domains in bands across the crystal long axis. Their location mirrored the binding pattern of similarly sized amelogenin aggregates seen in vitro. The data suggests that specific protein binding sites may exist on crystal surfaces and may be released at lower pH by protonation which would lower cationic charge on both crystal surface and ionic charge on the protein. Instability of the crystal surface could also play a role.

Animals↗

The complexity of pathways for protein import into thylakoids: it's not easy being green.

Numerous proteins are transported into or across the chloroplast thylakoid membrane. To date, two major pathways have been identified for the transport of luminal proteins (the Sec- and Tat-dependent pathways) and it is now clear that these protein translocases use fundamentally different transport mechanisms. Integral membrane proteins are inserted by means of at least two further pathways. One involves the input of numerous targeting factors, including SRP (signal recognition particle), FtsY and Albino3. Surprisingly, the other pathway does not involve any of the known chloroplastic targeting factors, and insertion is energy-independent, raising the possibility of an unusual 'spontaneous' insertion mechanism.

Intracellular Membranes↗

Penetration of fluoride into natural plaque biofilms.

Caries occurs at inaccessible stagnation sites where plaque removal is difficult. Here, the penetration through plaque of protective components, such as fluoride, is likely to be crucial in caries inhibition. We hypothesized that topically applied fluoride would readily penetrate such plaque deposits. In this study, plaque biofilms generated in vivo on natural enamel surfaces were exposed to NaF (1000 ppm F-) for 30 or 120 sec (equivalent to toothbrushing) or for 30 min. Biofilms were then sectioned throughout their depth, and the fluoride content of each section was determined with the use of a fluoride electrode. Exposure to NaF for 30 or 120 sec increased plaque fluoride concentrations near the saliva interface, while concentrations near the enamel surface remained low. Fluoride penetration increased with duration of NaF exposure. Removal of exogenous fluoride resulted in fluoride loss and redistribution. Penetration of fluoride into plaque biofilms during brief topical exposure is restricted, which may limit anti-caries efficacy.

Biofilms↗

Difference and consensus of whole cell Salmonella enterica subsp. enterica serovars matrix-assisted laser desorption/ionization time-of-flight mass spectrometry spectra.

AIM: Application of MALDI-TOF MS for characterization of strains of Salmonella enterica subsp. enterica. METHODS AND RESULTS: Whole cells were analysed by MALDI-TOF MS. Spectra with a maximum of 500 mass peaks between (m/z) 0 and 25000 were examined for consensus peaks manually and by a computer software algorithm. Consensus peaks were observed by both methods for spectra of Salmonella enterica serovars Derby, Hadar, Virchow, Anatum, Typhimurium and Enteritidis. CONCLUSIONS: Differences in numbers of consensus peaks in spectra obtained by manual and computer comparison indicated that development of the software involving statistical analysis of peak accuracy is necessary. SIGNIFICANCE AND IMPACT OF THE STUDY: Development of an analysis system for peak profiles in whole cell MALDI-TOF MS spectra to enable intra and interlaboratory comparison.

Bacteriophage Typing↗

Minimizing prion risk without compromising the microbial composition of biofilms grown in vivo in a human plaque model.

AIMS: To determine whether the stringency of sterilization procedures for biological components of in vivo dental plaque-generating devices based on enamel can be increased to minimize prion risk without compromising natural biofilm composition. METHODS AND RESULTS: The composition of in vitro biofilms, grown on hypochlorite-treated and untreated autoclaved enamel surfaces, was determined using culture-based methods and checkerboard DNA: DNA hybridization analysis. No differences were found between biofilms recovered from either substrate. SIGNIFICANCE: Several in situ models allow generation of plaque in the oral cavity, followed by recovery of intact biofilms for experimentation. Approaches allowing plaque formation on natural tooth surfaces are most valuable, but present a possible infection risk to volunteers wearing plaque-collecting devices, particularly with respect to prions. Hypochlorite treatment of biological material, as an adjunct to autoclaving, reduces infection risk without compromising biofilm composition and should be adopted in all future studies using plaque-generating devices incorporating enamel, where there is a potential prion threat, and further investigated in other biological hard tissues.

Bacteria↗

Copper ions inhibit the demineralisation of human enamel.

Cu2+ is cariostatic in rats reportedly due to it bacteriocidal properties. Here, we report the use of a simple abiotic model system to investigate whether Cu2+ has any inhibitory effect on the acid dissolution of human enamel. Crowns were exposed to a sequence of seven 10 mmol/l acetic acid challenges. The mineral dissolved during each challenge was then determined. CuSO4 (10 mmol/l) was present during the fourth of these challenges. Loss of calcium and phosphate were reduced by 57 and 63%, respectively, (P<0.0001) in the presence of Cu2+. Losses were also significantly reduced during the next acidic challenge in the absence of Cu2+. The degree of protection was found to approach maximum at about 5 mmol/l Cu2+. The well-known cariostatic properties of Cu2+ may therefore be due not only to its ability to inhibit bacterial growth but also to its ability to directly inhibit acid dissolution of enamel.

Acetic Acid↗

Daily use of dentifrice with and without xylitol and fluoride: effect on glucose retention in humans in vivo.

The effect of daily use of three different dentifrices on glucose retention after glucose mouth rinsing was tested in this study regarding xylitol and fluoride. Six experimental groups used three different dentifrices produced by two different companies: xylitol- and fluoride-containing dentifrice (XF), non-xylitol- and fluoride-containing dentifrice (F), and non-xylitol- and non-fluoride-containing dentifrice (NonX-NonF). Subjects were divided at random and rinsed their mouths for 15s with 20ml of 0.5M glucose solution. Glucose and lactate retention were determined by collecting samples of saliva from the approximal areas of the maxillary and mandibular anterior teeth and using the enzyme membrane test. Samples were collected 0, 1 and 2 months after the start of regular dentifrice use. There were significant differences in glucose retention in relation to the dentifrice used, month of sampling, site of sampling, and time since start of rinsing. Their contribution ratios were 2.0, 4.4, 11.7 and 7.4%, respectively (P<0.01). There were significant differences observed between the XF and NonX-NonF groups, with the XF group presenting lower glucose retention than the NonX-NonF group. The XF group presented lower glucose retention than the F group. The F group showed lower glucose retention than the NonX-NonF group. There were significant differences in lactate retention in relation to the month and site of sampling, and their contribution ratios were 3.3 and 2.8%, respectively (P<0.01). There were, however, no significant differences in glucose and lactate retention in relation to the dentifrice manufacturer. It was concluded that the XF dentifrice was the most effective, and the F dentifrice was more effective in reducing glucose retention than the NonX-NonF dentifrice.

Adult↗

Maggots and wound healing: an investigation of the effects of secretions from Lucilia sericata larvae upon interactions between human dermal fibroblasts and extracellular matrix components.

BACKGROUND: Through clinical observation, Lucilia sericata (greenbottle fly) larvae are credited with exerting the following beneficial effects upon a chronic nonhealing wound: removal of necrotic tissue ('debridement'), disinfection of the wound and active promotion of granulation tissue formation. As a major cellular component of granulation tissue, fibroblasts play an extensive role in healing. The composition of extracellular matrix (ECM) located in the wound is another important factor, partaking in a dynamic feedback loop with the fibroblasts that produce it. Fibroblast-ECM interactions therefore exert considerable influence upon new tissue formation. OBJECTIVES: To investigate the effects of L. sericata larval excretory/secretory products (ES) upon the behaviour of fibroblasts, seeded upon ECM component surfaces. METHODS: ES were collected by washing freshly hatched larvae in phosphate-buffered saline. Human dermal neonatal fibroblast cells were seeded upon fibronectin- or collagen-coated surfaces, together with untreated (or 'native') ES, heat-treated ES, or no ES (ES blank). Following incubation, fibroblast adhesion was determined using an adenosine triphosphate assay and, for confirmation, a total nucleic acid content assay. Cell spreading was observed using microscopy. The effect of ES upon fibronectin structure was observed using sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Peptide sequencing of suspected fibronectin fragments was performed using an 'electrospray' type time of flight mass spectrometer and 'Peptident' database search. RESULTS: ES significantly reduced fibroblast adhesion to both fibronectin and, to a lesser extent, collagen. Cell spreading was also reduced, yet cells remained viable. For both spreading and adhesion, heat-treated ES exerted significantly less activity than native, untreated ES. However, they still exhibited significant activity when compared with the ES blank. ES appeared to modify fibroblast adhesion indirectly via proteolytic fragmentation of the fibronectin protein surface. CONCLUSIONS: L. sericata larval secretions modify fibroblast adhesion and spreading across ECM protein surfaces, while keeping cells viable. Proteolytic activity of the ES played a significant role. If transferred to the wound situation, such alteration of fibroblast-ECM interactions may enhance new tissue formation.

Amino Acid Sequence↗

Apertureless near-field Raman spectroscopy.

We report on the tip-enhanced Raman spectra of C60 obtained on a custom-built apertureless scanning near-field optical microscope. A commercial atomic force microscope tip coated with 100 nm thickness of gold was used to enhance locally the Raman signal and permit topographic and spectral information to be acquired simultaneously. We present preliminary data which demonstrate the tip enhancement effect using C60 as a test sample.

Journal Article↗

Subunit structures in hydroxyapatite crystal development in enamel: implications for amelogenesis imperfecta.

Previous freeze-etching studies of developing enamel revealed collinear arrays of spherical structures (approximately 50 nM dia) of similar width to the crystals of mature tissue. Concomitant with matrix degradation/processing, spherical structures became less distinct until, coincident with massive matrix loss, only crystal outlines were seen. More recently, using Atomic force microscopy technology, early crystals exhibited topology reminiscent of these collinear spherical structures. After matrix loss these were replaced by similarly sized bands of positive charge density on the crystal surfaces. The data suggest enamel crystals may form from mineral-matrix spherical subunits. Matrix processing may generate mineral nuclei and lead to their fusion and transformation into long apatite crystals. Support for this view derives from the appearance of short crystal segments in amelogenesis imperfecta (hypoplastic AI) or abnormally large crystals alongside 50 nM diameter spherical mineral subunits (hypomaturation AI). Mutation of matrix or processing enzymes leading to defective processing may have impaired mineral initiation, fusion, and subsequent growth.

Amelogenesis Imperfecta↗

Binding of dentin noncollagenous matrix proteins to biological mineral crystals: an atomic force microscopy study.

Noncollagenous matrix proteins (NCPs) of dental hard tissues (dentin, cementum) are involved, both temporally and spatially, in the mineralization of their collagen matrices. Two of the NCPs thought to initiate mineral nucleation and control crystal growth in dentin, are dentin phosphoproteins (DPP) and dentin sialoprotein (DSP). Control of crystal growth would depend on the binding capacity of these two molecules, which may be related to the charge domains on the crystals and/or the phosphorylation of the protein. Phosphophoryn (a highly phosphorylated DPP) and DSP were isolated, purified, and characterized from the immature root apicies of human teeth. Dephosphorylation of phosphophoryn was carried out using bovine intestinal alkaline phosphatase. Enamel crystals were prepared from the maturation stage of developing rat incisor enamel. Protein-coated crystals were prepared for viewing in an atomic force microscope fluid cell using tapping mode. Desorption of the proteins was achieved using a phosphate buffer and surface roughness measurements were obtained from all specimens. Time-lapsed images of the crystals showed "nanospheres" of protein distributed along the crystals but only the phosphophoryn-coated crystals showed a distinctive banding pattern, which was still visible after the phosphate desorption experiments. The surface roughness measurements were statistically greater (P <0.01) when compared to the control for only the phosphophoryn-coated specimens. It is hypothesized that the phosphophoryn binding may be associated with charge arrays on the crystal surface and its phosphorylation. Also, based on its affinity for the crystalsurfaces, phosphophoryn seems the most likely candidate for controlling dentin crystal growth and morphology.

Crystallization↗

The osteodystrophy of mucolipidosis type III and the effects of intravenous pamidronate treatment.

Mucolipidosis type III (ML III; McKusick 252600) is a rare lysosomal storage disease in which skeletal involvement is prominent, in particular the destruction of vertebral bodies and the femoral heads. We describe studies in two siblings with ML III that suggest the presence of a distinct metabolic bone disorder. Biochemical indices of bone turnover were increased, and transiliac bone biopsy demonstrated both trabecular osteopenia and marked subperiosteal bone resorption. Intravenous pamidronate treatment given monthly for a year was well tolerated and produced dramatic clinical effects, with reduction in bone pain and improvements in mobility, despite incomplete suppression of bone resorption as assessed by biochemical, radiographic and histological criteria. Bisphosphonate therapy may have an important role in the management of bone pain in ML III, as it does in the related lysosomal disorder of Gaucher disease.

Absorptiometry, Photon↗