Search PubMed⌕ Search

Biomedical subjects

R J Doyle

Publications and source records attributed to R J Doyle.

At least 73 records · Page 4Linked to original sources
PubMed1990

Inhibition of the cooperative adhesion of Streptococcus sanguis to hydroxylapatite.

The adhesion of Streptococcus sanguis to hydroxylapatite is a process involving several adhesins and receptors. Binding isotherms and Scatchard plots of the adhesion suggest that cooperative interactions occur at low cell densities. It was found that sulfolane, a hydrophobic-bond diluent, was capable of inhibiting the cooperative adhesion of S. sanguis to saliva-coated hydroxylapatite beads. Sodium thiocyanate, a chaotropic agent, inhibited not only cooperative adhesion, but also the adhesion thought to result from noncooperative interactions. It is suggested that strong chaotropic agents may not only inhibit adhesin-receptor complexes, but also may influence the secondary/tertiary structures of interacting species.

Bacterial Adhesion↗
PubMed1990

Lectins and their application to clinical microbiology.

Lectins are generally associated with plant or animal components, selectively bind carbohydrates, and interact with procaryotic and eucaryotic cells. Lectins have various specificities that are associated with their ability to interact with acetylaminocarbohydrates, aminocarbohydrates, sialic acids, hexoses, pentoses, and as other carbohydrates. Microbial surfaces generally contain many of the sugar residues that react with lectins. Lectins are presently used in the clinical laboratory to type blood cells and are used in a wide spectrum of applications, including, in part, as carriers of chemotherapeutic agents, as mitogens, for fractionation of animal cells, and for investigations of cellular surfaces. Numerous studies have shown that lectins can be used to identify rapidly certain microorganisms isolated from a clinical specimen or directly in a clinical specimen. Lectins have been demonstrated to be important diagnostic reagents in the major realms of clinical microbiology. Thus, they have been applied in bacteriology, mycology, mycobacteriology, and virology for the identification and/or differentiation of various microorganisms. Lectins have been used successfully as epidemiologic as well as taxonomic markers of specific microorganisms. Lectins provide the clinical microbiologist with cost-effective and potential diagnostic reagents. This review describes the applications of lectins in clinical microbiology.

Animals↗
PubMed1990

Mechanism of enhancement of microbial cell hydrophobicity by cationic polymers.

Polycationic polymers have been noted for their effects in promoting cell adhesion to various surfaces, but previous studies have failed to describe a mechanism dealing with this type of adhesion. In the present study, three polycationic polymers (chitosan, poly-L-lysine, and lysozyme) were tested for their effects on microbial hydrophobicity, as determined by adhesion to hydrocarbon and polystyrene. Test strains (Escherichia coli, Candida albicans, and a nonhydrophobic mutant, MR-481, derived from Acinetobacter calcoaceticus RAG-1) were vortexed with hexadecane in the presence of the various polycations, and the extent of adhesion was measured turbidimetrically. Adhesion of all three test strains rose from near zero values to over 90% in the presence of low concentrations of chitosan (125 to 250 micrograms/ml). Adhesion occurred by adsorption of chitosan directly to the cell surface, since E. coli cells preincubated in the presence of the polymer were highly adherent, whereas hexadecane droplets pretreated with chitosan were subsequently unable to bind untreated cells. Inorganic cations (Na+, Mg2+) inhibited the chitosan-mediated adhesion of E. coli to hexadecane, presumably by interfering with the electrostatic interactions responsible for adsorption of the polymer to the bacterial surface. Chitosan similarly promoted E. coli adhesion to polystyrene at concentrations slightly higher than those which mediated adhesion to hexadecane. Poly-L-lysine also promoted microbial adhesion to hexadecane, although at concentrations somewhat higher than those observed for chitosan. In order to study the effect of the cationic protein lysozyme, adhesion was studied at 0 degree C (to prevent enzymatic activity), using n-octane as the test hydrocarbon. Adhesion of E. coli increased by 70% in the presence of 80 micrograms of lysozyme per ml. When the negatively charged carboxylate residues on the E. coli cell surface were substituted for positively charged ammonium groups, the resulting cells became highly hydrophobic, even in the absence of polycations. The observed "hydrophobicity" of the microbial cells in the presence of polycations is thus probably due to a loss of surface electronegativity. The data suggest that enhancement of hydrophobicity by polycationic polymers is a general phenomenon.

Acinetobacter↗
PubMed1989

Inhibition by maltose, isomaltose, and nigerose of the synthesis of high-molecular-weight D-glucans by the D-glucosyltransferases of Streptococcus sobrinus.

Two D-glucosyltransferases are produced by Streptococcus sobrinus C211. One (GTF-S) catalyzes the conversion of sucrose into soluble alpha-(1----6)-linked alpha-(1----3)-branched D-glucans, and the other (GTF-I), of sucrose into alpha-(1----3)-linked alpha-(1----6)-branched D-glucans. These enzymes were studied by using maltose, isomaltose, and nigerose as inhibitors. Maltose and isomaltose were found to be competitive inhibitors of GTF-S, whereas nigerose has no effect on GTF-S activity. The Ki values for maltose and isomaltose were determined to be 11 and 15mM, respectively. Maltose, isomaltose, and nigerose competitively inhibit GTF-I. The Ki values for these inhibitors were found to be approximately 0.8, 2.5, and 15mM, respectively. The inhibitory properties of each disaccharide are interpreted in terms of conformational comparisons with sucrose.

Carbohydrate Conformation↗
PubMed1989

Glucosyltransferases of Streptococcus sobrinus C211 are both stimulated and inhibited by hydrogen peroxide.

There are 2 glucosyltransferases (GTF) produced by Streptococcus sobrinus C211. One enzyme, GTF-S, produces a water-soluble glucan that is a-1,6-linked, with short a-1,3 branches, and the other enzyme, GTF-I, produces a water-insoluble glucan that is a-1,3-linked with a-1,6 branches. Hydrogen peroxide was found not only to be a potent inhibitor of GTF activity, but also a stimulator of GTF activity when employed at relatively low concentrations. At 0.88 M, H2O2 completely inhibited insoluble glucan synthesis, whereas at a 0.29 M concentration, H2O2 enhanced synthesis of the same glucan. Soluble glucan synthesis was also inhibited by H2O2 at 1.47 M. Low concentrations of hydrogen peroxide with GTF-S, however, caused the enzyme to convert from soluble glucan production to insoluble glucan production. 13C-Nuclear magnetic resonance spectra of glucans produced by peroxide-treated GTF confirmed that the production of a-1,3 linked glucans was increased with H2O2-treated GTF-S.

Glucans↗
PubMed1988

Glucan-binding factor in saliva.

High-molecular-weight polymers of alpha-1,6-linked D-glucans are insoluble in alcohol solutions. Whole, but not parotid, saliva prevented the precipitation of D-glucans by 80% (vol/vol) ethanol, showing that the whole saliva contained a factor which complexed with the glucan to render it alcohol soluble. The glucan-binding factor was retained on a column of Sephacryl S-200 which had been preequilibrated with 80% ethanol. The factor was then eluted with water. Passive hemagglutination assays revealed that the glucan-binding factor could sensitize erythrocytes to agglutination with anti-poly(glycerolphosphate), suggesting that the active glucan-binding component with lipoteichoic acid. The glucan-solubilizing factor was resistant to heat (100 degrees C), proteases, sialidase, lysozyme, lactoperoxidase, trichloroacetic acid, and Triton X-100. When sucrose was added to saliva, a suspension of Streptococcus cricetus AHT, or a suspension of Streptococcus sanguis 10556, relatively large amounts of glucan-binding factor were released in a soluble form. In addition, penicillin G caused the release of the glucan-solubilizing component from a suspension of S. cricetus AHT. It is suggested that whole saliva contains a component, tentatively identified as lipoteichoic acid, which can complex with glucans in a relatively hydrophobic solvent. This type of complex formation may be important in the adhesion of oral streptococci to saliva-coated surfaces.

Glucans↗
PubMed1988

Specificity of the glucan-binding lectin of Streptococcus cricetus.

The specificity of the glucan-binding lectin (GBL) of Streptococcus cricetus AHT was determined. Examination of the kinetics of aggregation of cell suspensions with glucans containing various percentages of alpha-1,6, alpha-1,4, alpha-1,3, and alpha-1,2 anomeric linkages revealed that only glucans with at least 80% alpha-1,6 linkages promoted strong aggregation. Moreover, only linear glucans with molecular weights greater than 5 X 10(5) were capable of causing rapid aggregation of the bacteria. The lectin was observed to be present on S. cricetus strains, on Streptococcus sobrinus, and on several Streptococcus mutants strains. Preincubation of suspensions of S. cricetus AHT with glucan T10 (molecular weight of 10,000) before the addition of high-molecular-weight glucan resulted in competitive inhibition in a concentration-dependent manner. Inhibition was achieved also with isomaltopentaose, isomaltohexaose, and isomaltooctaose, but at higher concentrations than glucan T10. In contrast, no inhibition was observed with maltoheptaose, providing additional evidence for the specificity of GBL. Treatment of suspensions of S. cricetus AHT with trypsin before and after aggregation with high-molecular-weight glucan revealed a substantial level of protection of GBL when in a bound state. Collectively, these results indicated that GBL has an absolute affinity for glucans rich in alpha-1,6 linkages and possesses an active site which recognizes internal sequences and accommodates isomaltosaccharides of at least nine residues. This unusual specificity may contribute to the colonization of S. cricetus, S. sobrinus, and S. mutans in glucan-containing plaque in the oral cavity.

Agglutination↗
PubMed1988

Expression of the glucan-binding lectin of Streptococcus cricetus requires manganous ion.

Streptococcus cricetus AHT exhibited a requirement for manganese for growth and expression of the glucan-binding lectin. While low concentrations of manganese (0.1 to 10 microM) were able to support growth, higher concentrations (greater than 100 microM) were required for full expression of the glucan-binding lectin. The manganous-aquo ion may be important in cellular adhesion and accumulation processes in dental plaque.

Bacterial Adhesion↗
PubMed1988

Surface characteristics of Pseudomonas cepacia.

Two major surface characteristics of Pseudomonas cepacia were examined in this study: reactivity with lectins and hydrophobicity. The results indicated that the surfaces of P. cepacia strains are heterogeneous with regard to the distribution of lectin receptors. Only lima bean agglutinin was found to strongly agglutinate all strains. The strains were also heterogeneous with regard to hydrophobicity as determined by adhesion to hexadecane. The degree of hydrophobicity, however, was not significantly altered when selected strains were mixed with either fibronectin or bovine serum albumin. In addition, the strains exhibited no apparent affinities for buccal epithelial cells and gave no evidence for an ability to haemagglutinate human red cells.

Agglutination↗
PubMed1987

Role of sialic acid in the kinetics of Streptococcus sanguis adhesion to artificial pellicle.

Evaluation of the kinetics of adhesion of Streptococcus sanguis 10556 to saliva-coated hydroxylapatite revealed that sialic acid played a role in the formation of a stable cell-substratum complex. In a previous paper (M. M. Cowan, K. G. Taylor, and R. J. Doyle, J. Dent. Res. 65:1278-1283, 1986) the adhesion was found to take place in two distinct stages: a reversible equilibrium, probably governed by long-range forces, followed by a transition to higher-affinity binding. In the present study, artificial pellicle was treated with neuraminidase, and kinetic adsorption and desorption experiments with S. sanguis were conducted. The depletion of sialic acid from pellicle decreased the initial adsorption rate constant only slightly. The rate constant describing the initial desorption was unaffected. However, no transition to the second (high-affinity) association occurred. While S. sanguis desorption from control pellicles exhibited two sequential rates, with the second rate being approximately 10 times slower than the first, all desorption from sialo-deficient pellicles occurred at one rate that was equivalent to the initial rate constant for control desorption. The cells did not reach an equilibrium with the sialo-deficient pellicle, even after 6 h. Competing sialic acid did not decrease the rate or extent of adsorption, but desorption occurred to a greater extent when cells had adsorbed in the presence of sialic acid. These data suggest that sialic acid plays little role in the initial association of cell and pellicle but that it is necessary for the transition to high-affinity binding and the concomitant decreased propensity to desorb.

Bacterial Adhesion↗
PubMed1987

Energetics of the initial phase of adhesion of Streptococcus sanguis to hydroxylapatite.

The initial adhesion of Streptococcus sanguis 10556 to artificial salivary pellicle and to bare hydroxylapatite was studied at several temperatures between 18 and 37 degrees C. When the natural logarithms of rate constants for adsorption and desorption were plotted against reciprocal temperatures in Arrhenius plots, curved lines were obtained, indicating that the thermodynamic quantities of enthalpy and entropy of activation were temperature dependent. For the bare hydroxylapatite system, the heat capacity (delta Cp = dH/dT) was large and negative. delta Cp was also negative for adhesion to saliva-coated hydroxylapatite, although its value was lower. Negative heat capacities, when coupled with favorable entropy, are often indicative of either electrostatic or hydrophobic interactions. When electrolyte (100 mM ammonium sulfate) was added to the cell-hydroxylapatite bead mixture, the rate and extent of adhesion were decreased. Addition of nonpolar p-dioxane (10% [vol/vol], final concentration) to the mixture enhanced binding. This suggests that electrostatic linkages participate in the primary adhesion of streptococci to both substrata. The strongly positive entropy values and the lesser temperature dependence of the saliva-coated hydroxylapatite system suggest that another entropy-driven process is imposed on the electrostatic linkages. This supports a role for hydrophobicity, suggesting that a combination of electrostatic and hydrophobic forces mediate the initial adhesion of S. sanguis to the salivary pellicle.

Ammonium Sulfate↗
PubMed1987

The functions of autolysins in the growth and division of Bacillus subtilis.

Some bacteria, such as streptococci, exhibit growth from discrete and well-defined zones. In Streptococcus faecalis, growth zones can be observed in the electron microscope, and the position of the zone can be used as a marker for cell cycle events. Growth of the cell surface of Bacillus subtilis appears to be by a much different mechanism from that of streptococci. Cell elongation takes place by the insertion at many sites in the cell cylinder of peptidoglycan components. The insertion occurs on the inner face of the wall, and upon cross linking, the new wall material becomes stress bearing and older wall is pushed to the surface. When old wall reaches the surface, it becomes susceptible to excision by autolysins, resulting in wall turnover; cell elongation, due to the stretching of the cross-linked peptidoglycan, therefore, accompanies turnover and does not require a specialized growth zone.

Amidohydrolases↗
PubMed1986

Induced release of Bacillus spores from sporangia by sodium sulphate.

Incubation of sporulating cultures of Bacillus anthracis, B. cereus, B. subtilis and B. thuringiensis in 1.0 mol/l sodium sulphate markedly increased the release of free spores from sporangia. It is postulated that the release of spores is due to activation of latent autolysins which hydrolyse sporangial cell walls. Sodium sulphate-induced lysis of sporangia represents a novel and highly effective method for the recovery of spores from cultures of Bacillus species.

Bacillus↗
PubMed1986

Microbes, warfare, religion, and human institutions.

A significant number of practicing microbiologists are not aware of the historical impact of infectious agents on the development of human institutions. Microbes have played a profound role in warfare, religion, migration of populations, art, and in diplomacy. Boundaries of nations have changed as a result of microbial diseases. Infectious agents have terminated some kingdoms and elevated others. There is a need for microbiologists to have a historical perspective of some of the major ways in which a pathogen may influence civilized populations. Conditions may exist in contemporary society for a repeat of some of the kinds of plagues suffered by previous societies. The purpose of this paper is to review examples of situations where pathogenic microbes have forced societal modifications on centers of human population.

Animals↗
PubMed1986

Kinetic analysis of Streptococcus sanguis adhesion to artificial pellicle.

Studies of equilibria between Streptococcus sanguis and artificial pellicle have suggested that there are multiple binding sites for the organism. In the present study, adhesion of S. sanguis to saliva-coated hydroxylapatite was examined by means of kinetic methods. Cell-pellicle complex formation was measured from initiation of binding to equilibrium. Rate constants were calculated for forward reactions (adsorption) and reverse reactions (desorption). Initial binding obeyed reversible, first-order kinetics, whereas desorption of bound cells followed biphasic kinetics. Initial desorption proceeded approximately ten times faster than the slower second rate. The results are consistent with the mechanism C + P reversible CP* in equilibrium with CP in which CP* represents the reversible equilibrium that shifts at a discrete rate to the high-affinity CP state. Thus, the biphasic binding behavior that has been previously deduced from equilibrium studies may be attributed to a time-dependent shift from close apposition to pellicle, stabilized by low-specificity forces, to a higher-affinity binding.

Adhesiveness↗
PubMed1985

Inside-to-outside growth and turnover of the wall of gram-positive rods.

Gram-positive, rod-shaped bacteria, given a pulse of peptidoglycan precursors, first exhibit a lag before the second or turnover phase of peptidoglycan commences. This is because new material is inserted on the inner face of the wall and gradually displaced through the wall. Based on this experimental observation, a mathematical model was constructed and compared with experimental data obtained in several laboratories for the first and second phases of wall turnover of Bacillus subtilis. The model allows the parameters of the process to be estimated for experiments with any labeling time. According to the surface stress theory the wall which is layed down immediately outside the cytoplasmic layer is in an unextended conformation. As subsequent additions of murein occur, the wall moves outward, becomes stretched, and bears the stress due to hydrostatic pressure. Ultimately, peptide and glycosyl bonds become cleaved. At the end of the lag phase the cleavage becomes so extensive that wall fragments are liberated into the medium. This strategy permits rod-shaped growth. In some experimental situations the half-life of wall radioactivity in this second phase roughly equals the doubling time; consequently, the exponential release probably does not represent random turnover but instead is the result of expansion of the underlying wall that continues to create strain which favors autolysis action. The slower turnover of the third phase, where there is a much slower loss, is also included in the analysis.

Bacillus subtilis↗