Biomedical subjects

M D Willcox

Publications and source records attributed to M D Willcox.

At least 91 records · Page 5Linked to original sources

PubMed1995

Fibronectin binding by Streptococcus milleri group strains and partial characterisation of the fibronectin receptor of Streptococcus anginosus F4.

The Streptococcus milleri group were shown to bind fibronectin (Fn) to their cell-surface and this binding increased the adhesion of cells to hydroxyapatite. The binding of Fn to Streptococcus anginosus F4 was studied in more detail. Fn binding to bacterial cells increased the association of the bacteria with the polymorphonuclear leukocytes obtained from the peritoneal cavity of rats but did not increase killing of the bacteria. The cell-surface receptor was a protein of M(r) 14,000 which was released from cells after mutanolysin digestion. The binding was specific, with cells having a maximum number of binding sites per cell of 770. Electron microscopy, using gold-labelled Fn, localised the receptor to areas between daughter cells.

Animals↗

PubMed1995

Potential pathogenic properties of members of the "Streptococcus milleri" group in relation to the production of endocarditis and abscesses.

The "Streptococcus milleri" (SMG) group have been shown to possess factors in vitro that may be involved in pathogenesis. All SMG strains are able to bind fibronectin via a cell-surface protein; the binding ranged from 12 to 198 mol/cell. Strains also bound to platelet-fibrin or fibrin clots and fibrinogen, giving maximum adhesion values of 16.5%, 21.8% and 151 mol/cell respectively. Members of the species S. constellatus produced thrombin-like activity. Lancefield group C SMG aggregated rat platelets, a bacterial cell-surface protein acting as mediator in the reaction. Most of the in-vitro factors did not correlate with each other, an indication that SMG strains possess a wide variety of pathogenic properties that may be involved in the production of abscesses or endocarditis. However, there was a correlation between the binding of large amounts of fibrinogen ( > 100 mol/cell) and the ability to aggregate platelets. This suggests that fibrinogen binding may aid in platelet aggregation.

Abscess↗

PubMed1995

The effect of sodium hypochlorite on potential pathogenic traits of Candida albicans and other Candida species.

Strains of Candida albicans, Candida krusei, Candida kefyr, Candida tropicalis, Candida parapsilosis and Candida guilliermondii were grown in the presence or absence of concentrations of sodium hypochlorite below the minimal inhibitory concentration and tested for a range of characteristics that may be associated with pathogenicity. Sodium hypochlorite is used routinely in hospitals in Australia for disinfection procedures, and these experiments were designed to assess the efficacy of hypochlorite as a sterilizing agent for acrylic dentures. Candida showed varying abilities to adhere to surfaces that may be present in the oral cavity. Sodium hypochlorite reduced the adhesion of all C. albicans strains and most other Candida species to both polystyrene and buccal epithelial cells. A biofilm of Streptococcus gordonii reduced the adhesion of most C. albicans strains and most other Candida species to polystyrene. However, Candida species were able to coaggregate with S. gordonii in suspension, with one strain of C. albicans, GDH 2346, showing greater coaggregating ability than the other strains or species. Sodium hypochlorite increased coaggregation of all C. albicans strains and most other Candida species. Examination of cell wall proteins from strains of C. albicans and a strain of C. parapsilosis showed that growth in hypochlorite increased the amount of protein in some existing bands and, in one strain of C. albicans, increased the number of detectable protein bands ranging from 56 to 26 kDa. Only 4 strains of C. albicans were able to produce hyphae, and 3 of these same strains and C. parapsilosis were able to produce proteinase. Hypochlorite increased the rate of blastospore to hyphal transition but had no effect on proteinase production or activity. It is concluded that hypochlorite at a concentration below the minimal inhibitory concentration may function as an anti-adhesin for Candida species but may not affect their more pathogenic characteristics.

Acrylic Resins↗

PubMed1995

Degradative enzymes of oral streptococci.

Members of the Streptococcus sanguis group (SSG) and Streptococcus milleri group (SMG) were screened for their ability to produce glycosidase, arylamidase (peptidase), protease, dextranase and glycosyltransferase activities. Species within each group produced unique patterns of activity. The most commonly produced glycosidases were beta-D-glucosidase, beta-D-galactosidase, N-acetyl-beta-D-glucosaminidase and N-acetyl-beta-D-galactosaminidase and the least commonly produced glycosidase activity was beta-fucosidase with Streptococcus intermedius (SMG) being the only species capable of producing the activity. For arylamidase activity, the most commonly produced type was lysine-arylamidase. Glycosidase and arylamidase activities were localized to particular sub-cellular fractions. alpha-galactosidase was found only in culture supernatant fluids whereas N-acetyl-beta-D-glucosaminidase was found in all fractions; the culture supernatant, cell wall, cell membrane and cytoplasm. No arylamidase activity was seen in culture supernatants. Phe-arg-arylamidase was found only in cytoplasmic fractions whereas val-pro-argarylamidase was found in cell walls, cell membranes and cytoplasmic fraction. Protease activity was measured as the degradation of bovine serum albumin (BSA) and casein. Casein was degraded by a number of strains whereas no species/strains were able to degrade BSA. Streptococcus intermedius, Streptococcus constellatus (SMG), Streptococcus mitior and Streptococcus defectivus (SSG) were the only species that produced hyaluronidase and no species produced chondroitin sulphatase. The groups were also examined for their abilities to produce glycosyltransferase and dextranase. Strep. sanguis, Streptococcus gordonii, Streptococcus mitis and Streptococcus oralis produced glucosyltransferase and, with the exception of the latter species, fructosyltransferase. No species within the SMG was capable of producing either glycosyltransferase.(ABSTRACT TRUNCATED AT 250 WORDS)

Aminopeptidases↗

PubMed1995

Changes to the ocular biota with time in extended- and daily-wear disposable contact lens use.

Gram-negative bacteria may play a role in the etiology of certain soft contact lens (SCL)-related diseases. Contact lens (CL) wear may modify the normal ocular biota, providing a more favorable environment for potential pathogens. This study reports temporal changes in ocular biota in daily-wear (DW) and extended-wear (EW) disposable SCL use in experienced and neophyte wearers. Lid margin and bulbar conjunctival biota were sampled prior to CL fitting in 26 previous DW SCL users, 18 previous EW SCL users, and 26 neophytes. Wearers were fitted with an etafilcon A CL in one eye and a polymacon CL in the fellow eye. Lenses were worn on a daily basis by the 26 previous DW SCL wearers and on an EW basis by the remaining 44 subjects. The ocular biota was further sampled after 1, 3, 6, 9, and 12 months of wear. The ocular biota consisted of coagulase-negative staphylococci, Corynebacterium spp., Micrococcus spp., and Propionibacterium spp. Potential pathogens were rarely isolated at baseline. No significant trend of increasing ocular colonization was shown for extended CL wear. Lid and conjunctival colonization increased with DW SCL use (P < 0.001), although this increase occurred for nonpathogenic species only. Fewer potential pathogens were isolated from DW SCL than from EW SCL users (P < 0.05). The lid margin consistently showed greater colonization than the conjunctiva and may be a source of potential pathogens during CL wear. Hydrogel CL wear appears to modify the ocular biota. An increased number of commensal organisms were present in DW SCL use. EW SCL use altered the spectrum of organisms isolated. These alterations may suppress the normal ocular defense mechanisms and may be relevant in the pathogenesis of CL-related disease.

Adult↗

PubMed1994

Lancefield group C Streptococcus milleri group strains aggregate human platelets.

Lancefield group C Streptococcus milleri group (SMG) strains are the only SMG types that are able to aggregate human platelets. Complete aggregation occurred within 10 min of mixing bacterial cells and platelets together in the ratio 8:1. Substances which (i) chelated cations; (ii) inhibited the cycloxygenase pathway in platelets; (iii) reduced the availability of ADP and disrupted platelet membrane stability; (iv) reduced bacterial aggregation of platelets. The platelet-interacting substance on the surface of the SMG appeared to be proteinacious as digestion of bacterial cells with protease inhibited aggregation whereas treatment with lipase, periodate or antisera to Lancefield group C polysaccharide had no effect.

Abscess↗

PubMed1994

Surface-associated properties of Actinomyces strains and their potential relation to pathogenesis.

Twenty-nine strains from the Actinomyces species were tested for a range of surface properties. Results show considerable heterogeneity both between different species and within some of the species, especially Actinomyces naeslundii. Two commonly used A. naeslundii strains, T14V and ATCC 12104, fell within the low (salivary aggregation and collagen binding by T14V), moderate (surface charge and haemagglutination) or high range of values (hydrophobicity, saliva-coated hydroxyapatite adhesion, polystyrene binding by T14V, fibrinogen binding by T14V and collagen binding by A. naeslundii ATCC 12104). Both strains adhered well to saliva-coated hydroxyapatite; T14V bound the highest amount of fibrinogen, ATCC 12104 had the highest number of cells bound to collagen and T14V was not bound at all. The heterogeneity of these characteristics highlights the need to include a range of strains of Actinomyces in studies on their pathogenicity. Statistical correlations were found between a number of properties, for example saliva-coated hydroxyapatite adhesion and hydrophobicity, and between haemagglutination and hydrophobicity.

Actinomyces↗

PubMed1993

Albumin-binding proteins on the surface of the Streptococcus milleri group and characterization of the albumin receptor of Streptococcus intermedius C5.

Members of the Streptococcus milleri group (SMG) that react with Lancefield group C antisera were shown to bind large amounts of albumin although there was no direct relation between these two properties as polyclonal antisera to Lancefield group C antigen did not prevent the binding of albumin. There was a specificity for albumin binding, with albumin from man, monkeys, cat, dog and mouse being bound to a greater degree than albumin from cow, horse, goat or rabbit. Gold-labelled albumin was shown to be located close to the surface of strains by transmission electron microscopy. A cell-surface protein of M(r) 24,000, which was liberated by lysozyme treatment of cells, was shown to be the cell-surface receptor on Streptococcus intermedius C5. The receptor was physically dissimilar from protein G, an albumin- and IgG-binding protein of 'large-colony' Lancefield group C and G streptococci.

Animals↗

PubMed1993

Biochemical properties of Streptococcus sobrinus reisolates from the gastrointestinal tract of a gnotobiotic rat.

Streptococcus sobrinus strain 6715-13-201 was inoculated into the oral cavity of a gnotobiotic rat and then reisolated from different portions of the gastrointestinal tract. Fourteen isolates, selected on the basis of their colonial morphology, were then screened for their ability to adhere to saliva-coated hydroxyapatite (SHA) in vitro, and their ability to produce extracellular polysaccharide from sucrose, and low pH in glucose broth. Certain isolates were also tested for their cariogenic potential as monoinfectants in gnotobiotic rats. All isolates differed in their abilities to adhere to SHA, with most showing an increased level of adhesion in the presence of sucrose, but this did not correlate with their ability to be aggregated by dextran. Most isolates were capable of producing glucosyltransferases (with only one exception) and dextranases (also one exception). There was more variability in the production of dextranase inhibitor. No isolate was capable of producing dextranase inhibitor in the absence of dextranase production. There were no correlations between the ability of isolates to adhere in vitro or produce/utilize polysaccharides and their ability to produce caries in vivo. Due to the differences between strains in their abilities to adhere, produce polysaccharides, utilize polysaccharides or produce a low pH and the lack of correlation between any of these parameters and cariogenicity, the results suggest that the ability of strains to colonize and produce caries depends on a number of different characteristics, no one of which is essential.

Animals↗

PubMed1993

Coaggregation of oral lactobacilli with streptococci from the oral cavity.

The ability of oral lactobacilli to coaggregate with streptococci and actinomycetes was investigated. Of the 7 species of lactobacilli studied, only two were capable of coaggregation and the coaggregation was restricted to streptococci. Lactobacillus salivarius strains (2/4) coaggregated with Streptococcus salivarius, Streptococcus gordonii, Streptococcus crista and tufted Streptococcus sanguis II strains. Lactobacillus fermentum (2/3) coaggregated with S. gordonii and S. sanguis. The coaggregation between L. salivarius and S. salivarius, S. gordonii or tufted S. sanguis II strains was mediated by a protein on the surface of the lactobacilli and was not inhibited by lactose. The coaggregation between L. fermentum and the streptococci was mediated by protein on the surface of the streptococci and was inhibited by lactose.

Actinomyces↗

PubMed1991

An examination of strains of the bacterium Streptococcus vestibularis for relative cariogenicity in gnotobiotic rats and adhesion in vitro.

The cariogenicity and adhesion of six strains of Streptococcus vestibularis were compared with those of strains of Strep. salivarius. All strains of Strep. vestibularis produced low levels of caries, confined to the fissures, whereas the two strains of Strep. salivarius produced high levels of caries, with one strain producing approximal as well as fissure caries. The values for adhesion to saliva-coated hydroxyapatite of Strep. vestibularis in the absence (median 4.74%) and the presence (median 4.67%) of sucrose were not statistically different, nor did they differ significantly from those of Strep. salivarius. Strains of Strep. vestibularis were able to adhere to buccal epithelial cells (median 1.19%) as well as could Strep. salivarius strain HB (1.65%). Neither sucrose nor saliva greatly aggregated the strains of either species. Strep. vestibularis did not adhere to hexadecane (median 18.5%) to the same extent as did Strep. salivarius strains (median 69%). There was a significant correlation between the adhesion in the presence and absence of sucrose (p less than 0.01). Strep. vestibularis strains could not coaggregate with either actinomycetes or Veillonella spp. whereas Strep. salivarius strains were able to coaggregate with Veillonella spp.

Actinomyces↗

PubMed1991

Enumeration of oral streptococci on media containing different concentrations of sodium and potassium ions.

Laboratory "type" strains of oral streptococci were screened for their ability to grow on mitis-salivarius agar (MSA) in the presence of increasing concentrations of either Na+ or K+ up to 500 mmol/L. Strains were generally better able to withstand increasing concentrations of Na+ than K+, although low numbers of colony-forming units (cfus) were seen with the highest concentration of either cation. Two strains of Streptococcus mutans, Ingbritt 162 and Ingbritt 175, behaved differently when the concentration of cation was increased from 50 to 200 mmol/L; the latter showed a marked increase in the number of cfus when the Na+ concentration was increased from 50 to 200 mmol/L, whereas there was a decrease with strain Ingbritt 162. Strains of oral streptococci from the saliva of adults and children were isolated on modified MSA containing known concentrations of Na+ and K+ and further examined if they showed "mutans-like" colony morphology. The number of cfus generally dropped as the concentration of Na+ or K+ was increased from 200 to 350 or 500 mmol/L. Greater numbers of streptococci were tolerant to Na+ than to K+. Half of the isolates were members of the Streptococcus sanguis group (SSG), either Streptococcus mitis or S. sanguis II, and these were more tolerant to high concentrations of Na+ or K+ than other isolates that were identified as Streptococcus morbillorum, Streptococcus acidominimus, and Streptococcus milleri.

Adult↗

PubMed1990

In vivo dental plaque-forming ability and cariogenicity of the bacterium Streptococcus bovis in gnotobiotic rats.

Germ-free Fisher rats, fed on a high sucrose diet, were monoinfected with Streptococcus bovis strains. High levels of fissure caries were formed by three strains; one strain produced moderate levels; another produced only very low levels. No strain could produce approximal or buccal/lingual lesions. All strains could colonize the fissures of teeth in the lower jaw, as shown by scanning electron microscopy. The most cariogenic strains were associated with caries scores similar to those of Strep. salivarius and members of the mutants streptococci.

Animals↗

PubMed1990

Surface-associated properties of Streptococcus milleri group strains and their potential relation to pathogenesis.

Thirty strains from the Streptococcus milleri (anginosus) group (SMG) obtained from various sources were tested for a range of characters that could be associated with pathogenicity and the results were compared with those for type strains of S. sanguis, S. mutans and S. pyogenes. The SMG strains were heterogeneous in all tests. Most (18) belonged to one of the Lancefield groups with group F predominating. Adherence of strains isolated from abscesses to buccal epithelial cells was greater than that of other strains (p = 0.033). Compared with strains of S. sanguis, SMG strains were generally not aggregated by human saliva. They differed from the type strain of S. pyogenes in their relative ability to bind fibrinogen and fibronectin; they were less effective in binding fibrinogen (0.33-4.28% cf. 22% for S. pyogenes) and generally more effective in binding fibronectin (0.49-12.37% cf. 0.95%). Strains isolated from infections were statistically better at binding fibronectin than other strains (p less than 0.001). The ability of strains to adhere to saliva-coated hydroxyapatite (SHA) varied 10-fold, from 0.16-16.35%. The amount of fibronectin bound by SMG strains correlated with their ability to adhere to SHA (p less than 0.001). The hydrophobicity of the strains, as measured in the hexadecane partition assay, ranged from 0.0% to 99.0%. Some strains carried both positive and negative cell-surface charges and some strains with a highly hydrophobic cell surface also possessed a relatively high cell-surface charge. A minority of strains possessed a net positive cell-surface charge. Neither hydrophobicity nor cell-surface charge was related to the capacity of strains to adhere to SHA. Strains of SMG co-aggregated weakly with strains of Veillonella parvula, V. dispar, Actinomyces viscosus and A. naeslundii.

Bacterial Adhesion↗

PubMed1990

Cohesion between oral streptococci and Neisseria pharyngis on saliva-coated glass, in the presence and absence of sucrose.

Ten strains of oral streptococci, comprising nine species, were screened for their in vitro cohesive interactions with a freshly isolated strain of Neisseria pharyngis. Strains of mutans streptococci, Streptococcus salivarius, Streptococcus sanguis I, Streptococcus milleri and Streptococcus mitis were able to cohere with N. pharyngis cells attached to a salivary pellicle adsorbed to glass. The cohesion was either due to coaggregation between the different genera or modification of the salivary pellicle by N. pharyngis MW2. S. sanguis II and Streptococcus oralis strains could not cohere with the N. pharyngis strain. Extracellular polysaccharide formed from sucrose was found to mediate cohesion between certain strains of mutans streptococci or S. oralis and N. pharyngis MW2.

Bacterial Adhesion↗

PubMed1989

Comparative cariogenicity and dental plaque-forming ability in gnotobiotic rats of four species of mutans streptococci.

The four species, each represented by two strains, all produced high levels of fissure caries in the upper and lower jaws of gnotobiotic Fisher rats. Streptococcus rattus strains and Streptococcus mutans NCTC 10832 gave low levels of approximal caries, whereas Streptococcus cricetus, Streptococcus sobrinus and Strep. mutans NCTC 10449 gave high levels. The only strains to produce buccal surface lesions were Strep. sobrinus and Strep. cricetus. All strains were able to form dense plaque in the central fissure of the second molar. Strep. cricetus, Strep. rattus and Strep. sobrinus strains were capable of producing smooth-surface plaque. This difference in plaque-forming ability did not correlate precisely with differences in cariogenicity, and such differences occurred between strains of the same species.

Animals↗

PubMed1989

A comparison of the adhesive properties and surface ultrastructure of the fibrillar Streptococcus sanguis 12 and an adhesion deficient non-fibrillar mutant 12 na.

Streptococcus sanguis 12 and a naturally occurring mutant, 12 na, were screened for their ultrastructure and adhesive properties in vitro. Negative staining showed that Strep. sanguis 12 carried three types of surface structure. The majority of cells carried long fibrils that in different batches ranged in length from 80 to 207 nm, and shorter fibrils which were 51.0 +/- 15.7 nm long. Both types of fibrils were primarily located at the poles of the cells. Occasionally cells were seen that carried fimbriae, which are structurally distinct from fibrils, and were 3-4 nm wide and less than 1.0 micron long. Strain 12 na carried no detectable surface structures. Ruthenium red staining revealed that both strains carried a loose, amorphous, extracellular polysaccharide layer attached to the cell wall. Streptococcus sanguis 12 na was 83% less adhesive than strain 12 in a saliva-coated hydroxyapatite assay, and 50% less adhesive in a buccal epithelial cell adhesion assay. In contrast, strain 12 na was more sensitive to aggregation by parotid saliva than strain 12, and both strains were equally aggregated by whole saliva. The cell surface hydrophobicity of the two strains was similar. Extraction of surface proteins by sodium lauroyl-sarcosinate followed by sodium dodecylsulphate polyacrylamide gel electrophoresis demonstrated that Strep. sanguis 12 expressed more high mol.wt proteins on its surface than strain 12 na. Using immunogold labelling, the fibrils of strain 12 labelled well with antiserum directed against the long fibrils, but so did the cell surfaces of both Strep. sanguis 12 and 12 na. High molecular weight proteins and cell surface fibrils may be associated with adhesion in this strain.

Bacterial Adhesion↗

PubMed1989

Surface structures, co-aggregation and adherence phenomena of Streptococcus oralis and related species.

Seven strains of Streptococcus oralis were found to possess surface structures. Four strains possessed long fimbriae which ranged in length from 266-366 nm, while the remaining three strains possessed shorter peritrichously distributed fibrils which ranged in length from 80-197 nm. The fibrillar strains were morphologically similar to strains of Streptococcus sanguis I and II and Streptococcus mitis. No strain of S. oralis produced tufts of fibrils like certain strains of S. sanguis I. Strains of Streptococcus milleri produced peritrichously distributed fimbriae which were morphologically dissimilar to the fimbriae produced by S. oralis strains. S. oralis strains were moderately to highly hydrophobic, but hydrophobicity could not be related to adhesion parameters or type or length or density of surface structures. Furthermore, there appeared to be no correlation between type of surface structures and adhesion parameters. Two of the three fibrillar strains of S. oralis and the peritrichously fibrillar strains of S. sanguis, together with one strain of S. milleri, were able to co-aggregate with actinomycetes.

Actinomyces↗