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Biomedical subjects

D G Ahearn

Publications and source records attributed to D G Ahearn.

At least 19 recordsLinked to original sources

DNA relatedness, karyotyping and gene probing of Candida tropicalis, Candida albicans and its synonyms Candida stellatoidea and Candida claussenii.

Isolates of Candida albicans with varied phenotypes, including sucrose-negative variants (C. stellatoidea, serotypes A and B) and avirulent germ tube-negative forms (C. claussenii) showed significant (greater than 90%) DNA relatedness to classical C. albicans, but insignificant relatedness to C. tropicalis and sucrose-negative C. tropicalis. A transverse alternating-field gel electrophoresis procedure (TAFE) showed discrete karyotype patterns among the phenotypic variants of C. albicans including the sucrose-negative C. stellatoidea. The number of chromosome-sized DNA bands for C. tropicalis (7 bands) were within the range of bands observed for C. albicans (5 to 10 bands). The general DNA-migration pattern for C. albicans appeared distinct from that of C. tropicalis. An aspartyl proteinase (PrA) gene probe from C. albicans hybridized with chromosomal DNA from C. albicans, C. claussenii and C. stellatoidea but not with that from C. tropicalis.

Aspartic Acid Endopeptidases

Hemagglutination (fimbriae) and hydrophobicity in adherence of Serratia marcescens to urinary tract epithelium and contact lenses.

The capacity of 59 isolates of Serratia marcescens, obtained from urinary tract infections, wounds, and contact lenses or their paraphernalia, to agglutinate erythrocytes from different animal species was tested. Three main patterns were found: mannose-sensitive agglutination of guinea-pig, fowl or horse erythrocyte; mannose-resistant agglutination of chicken or pigeon erythrocytes alone or in combination with mannose-sensitive agglutination; and no agglutination. Hemagglutination capacity was associated with isolates from urinary tract infection, but not with isolates associated with contact lenses. Adherence to human urinary tract epithelium did not correlate with the hemagglutination patterns nor with the origin of the isolates. Some strains of different hemagglutination pattern were selected for the study of hydrophobicity and adherence to contact lens polymers. Hydrophobicity, as determined by degree of partition in hexadecane and water (BATH-values), correlated neither with degree of adherence to contact lens polymers nor with the hemagglutination pattern. For a representative strain there was an excellent correlation (r2 = 0.98) between adherence and the water content (hydrophobicity) of the lens polymers. These results suggest that, as with tissues, other factors interact with hydrophobicity in causing adherence to plastics.

Alkanes

Comparative efficacies of soft contact lens disinfectant solutions against microbial films in lens cases.

Biofilms of Pseudomonas aeruginosa, Serratia marcescens, Staphylococcus epidermidis, Streptococcus pyogenes, and Candida albicans, established in the wells of a polyethylene contact lens case, retained viability to certain soft contact lens disinfectant solutions after exposure for the manufacturer's minimum recommended disinfection times. The relative order of resistance of bacterial biofilms was as follows: S marcescens was greater than P aeruginosa, which was greater than S epidermidis, which was greater than S pyogenes. Air drying of biofilms for 10 hours increased the efficacy of the disinfectant solutions, but drying was not enough to decrease the incidence of recovery to 0% for all solutions. Hydrogen peroxide was more effective against biofilms than disinfectant solutions formulated with chlorhexidine gluconate or polyquaternium-1 or polyaminopropyl biguanide. We recommend that determination of efficacy of contact lens disinfectant solutions should include challenges against biofilms.

Bacteria

Adherence of Pseudomonas aeruginosa to rigid gas-permeable contact lenses.

We examined the adherence of a human corneal isolate of Pseudomonas aeruginosa to unused rigid gas-permeable and hydrogel contact lenses. Adherence to rigid gas-permeable lenses was greater than the adherence to hydrogels. The lower incidence of microbial keratitis associated with rigid gas-permeable lenses, as compared with hydrogel lenses, may be attributed to the more rigorous cleaning and disinfection possible for rigid gas-permeable lenses and/or patient noncompliance with complicated disinfection systems used with hydrogel lenses.

Bacterial Adhesion

Candida haemulonii from clinical specimens in the USA.

Classical yeast identification procedures and DNA relatedness studies confirmed the occurrence of Candida haemulonii among clinical specimens in the USA, particularly isolations from the foot. None of the six clinical isolates studied produced identical API 20C profile codes.

Base Composition

Microbial contamination of contact lens storage cases and solutions.

We compared microbial contamination of contact lens storage cases of asymptomatic contact lens wearers (Group 1; No. = 118; sampled once) and of contact lens wearers with manufacturer's lens-care instructions reinforced (Group 2; No. = 62; sampled three, six, 12, and 20 weeks after initial advisement). A significantly higher incidence of contamination of contact lens storage cases and solutions was observed among samples from Group 1 (132 of 247 samples) as compared to samples from Group 2 (30 of 500 samples; P = .000). Contact lens storage cases of individuals in Group 2 who used hydrogen peroxide systems (four of 78) showed a significantly lower incidence of contamination as compared to individuals who used other chemical disinfection (11 of 62 soft lens users; 10 of 59 rigid gas-permeable lens users; P = .041). Biofilms, adhered microorganisms embedded in a glycocalyx, in contact lens storage cases were not always inactivated by the addition of fresh solutions. Cleaning and periodic replacement of contact lens storage cases is recommended.

Bacteria

Involvement of a cell wall receptor in the mode of action of an anti-Candida toxin of Pichia anomala.

Hanes-Woolf, Dixon, and Hill plots of growth rates of Candida albicans RC1 grown in various concentrations of glucose and a Pichia anomala WC65 toxin suggested the presence of toxin-binding sites. Indirect immunofluorescence microscopy with antitoxin antibodies demonstrated binding of the toxin to the cell wall. Resistance to the toxin of a mutant Saccharomyces cerevisiae deficient in cell wall beta-1-6-D-glucan suggests that the glucan either served as the receptor or influenced the number or composition of the receptor. Immunofluorescence that appeared to be associated with the cell membrane of toxin-treated spheroplasts of C. albicans was also observed. Spheroplasts of the resistant mutant of S. cerevisiae were sensitive to the toxin.

Candida albicans

Occurrence of Clavispora lusitaniae, the teleomorph of Candida lusitaniae, among clinical isolates.

Of 13 clinical isolates of Candida lusitaniae from diverse geographical regions, 7 represented the mating types (6 alpha, 1 a) of the ascomycete Clavispora lusitaniae. Selected nonfertile isolates showed significant DNA relatedness (greater than 90%) to representatives of both mating types. Phenotypic physiological characteristics, such as cellobiose fermentation and rhamnose assimilation, proved insufficient for separation of Clavispora lusitaniae and Clavispora opuntiae.

Candida

Electrophoretic karyotyping of typical and atypical Candida albicans.

Electrophoretic karyotypes of atypical isolates of Candida albicans, e.g., strains that were germ tube negative, failed to express proteinase activity, demonstrated low virulence for mice, formed hyperchlamydoconidia, produced hyperhyphae, or were sucrose negative (including the type strain of Candida stellatoidea), were compared with those of typical C. albicans. Karyotypes of whole-cell DNA of classical C. albicans examined with transverse alternating-field electrophoresis under specific conditions were composed of seven DNA bands with a specific migration pattern. Certain atypical strains and representatives of the three serotypes of C. stellatoidea produced discrete karyotypes with 5 to 10 bands. All isolates demonstrated a significant degree of DNA relatedness, suggesting their conspecificity. Densitometric tracings of DNA bands provided an objective and standardized method for comparing bands within the gels.

Candida albicans

Purification and characterization of the anti-Candida toxin of Pichia anomala WC 65.

Pichia anomala WC 65 secretes a toxin that is inhibitory to a variety of yeasts, including strains of the animal pathogen Candida albicans. The toxin was purified to homogeneity by ultrafiltration, ethanol precipitation, ion-exchange chromatography with a Mono Q column, and gel permeation chromatography with a Superose 12 column. The toxin had a molecular weight of 83,300 as determined by electrophoresis on sodium dodecyl sulfate-polyacrylamide gradient gels and a molecular weight of 85,290 as determined by gel permeation chromatography. The isoelectric point of the toxin was pH 5.0. The toxin was stable between pH 2.0 and 5.0. Chemical analysis of the purified toxin indicated that the toxin was a glycoprotein composed of about 86% protein and 14% carbohydrate. At high concentrations, the toxin showed a tendency to aggregate, with loss of biological activity against C. albicans, Pichia bimundalis, and Saccharomycodes ludwigii. Purified toxin expressed killing activity against C. albicans in contrast to the static activity of the crude toxin.

Candida albicans

In situ survival of plasmid-bearing and plasmidless Pseudomonas aeruginosa in pristine tropical waters.

Two rare wild-type strains of Pseudomonas aeruginosa were mixed in membrane diffusion chambers and then introduced into a natural freshwater environment for 72 h. The plasmid-containing strain (R serotype 15) and the plasmidless strain (H serotype 5) had initial bacterial densities of 2 x 10(5) cells per ml. Samples collected from the chambers were analyzed for viable and direct counts and for acquired-resistance frequencies. Suspected transconjugant-to-donor ratios ranged from 0.5 to 1.3; transfer percentages ranged from 13 to 70%. [3H]thymidine uptake indicated DNA synthesis in both strains as well as in transconjugants. These studies indicate that rare wild-type bacterial strains with large plasmid loads can survive as well as can bacteria with low plasmid loads when exposed to the in situ conditions of a tropical freshwater habitat. These results also suggest that genetic modification of indigenous microbiota through conjugation or transformation is feasible when rare wild-type strains or genetically engineered microorganisms are released in large numbers in tropical aquatic ecosystems.

Colony Count, Microbial

Anti-Candida albicans activity of Pichia anomala as determined by a growth rate reduction assay.

Killer toxin activity of Pichia anomala WC65 appeared fungicidal for P. bimundalis WC38 and fungistatic for Candida albicans RC1. Inhibitory activity against sensitive C. albicans showed a linear relationship between toxin concentrations and the inverse of the reduced growth rates. The plot of toxin concentrations against growth rates was hyperbolic, as is characteristic of saturation kinetics. Sensitivity of C. albicans to the toxin decreased with increased cell age. The measurement of growth rate reduction provided a simple and accurate method for quantitation of toxin.

Antifungal Agents

Resistance of some common fungi to gamma irradiation.

Ten species of fungi representing the genera Alternaria, Aspergillus, Caldosporium, Curvularia, Fusarium, and Penicillium were examined for their relative resistance to gamma irradiation from a 137Cs source. Inactivation doses for dematiaceous fungi in agar medium ranged from 0.6 to greater than 1.7 megarads, whereas those for moniliaceous fungi were less than 0.3 megarad. D10 values (the dose required to reduce the inoculum by 1 log) for Curvularia geniculata (greater than 0.29 megarad) exceeded those for control spores of Bacillus pumilus (0.15 megarad).

Alternaria

Effects of protein, mucin, and human tears on adherence of Pseudomonas aeruginosa to hydrophilic contact lenses.

Pseudomonas aeruginosa isolated from a corneal ulcer was examined for its ability to adhere to hydrophilic contact lenses with adsorbed organic films. Generally, adherence was enhanced by mucin, lactoferrin, lysozyme, immunoglobulin A, bovine serum albumin, and a mixture of these macromolecules. The water content of the lenses also influenced bacterial adherence and protein adsorption. Adherence to lenses coated with human tear films was more complex; both enhanced and inhibited adherence were observed on lenses obtained from different individuals.

Bacterial Adhesion

Incidence and persistence of Pseudomonas aeruginosa in whirlpools.

Pseudomonas aeruginosa was isolated from seven commercial and two residential whirlpools that were treated with halogens. None of the commercial whirlpools was constantly maintained at appropriate disinfection levels. Superchlorination or the draining, cleaning, disinfection, and refilling of whirlpools markedly reduced densities of P. aeruginosa in whirlpool water, but the bacterial populations were rapidly reestablished (less than 10(3) cells per ml) when disinfectant concentrations decreased below recommended levels (chlorine, 3.0 ppm [3.0 micrograms/ml]; bromine, 6.0 ppm). P. aeruginosa in the water was replenished from various sources, such as hoses used to fill the whirlpool and the biofilm in the filter and piping of the whirlpool systems. Daily monitoring and adjustment of chemical characteristics (regardless of bather load) were essential for controlling densities of P. aeruginosa.

Bromine

Physical and biological parameters that determine the fate of p-chlorophenol in laboratory test systems.

Shake-flask and microcosm studies were conducted to determine the fate of para-chlorophenol (p-CP) in water and sediment systems and the role of sediment and nonsediment surfaces in the biodegradation process. Biodegradation of p-CP in estuarine water samples in shake flasks was slow over incubation periods of 300 h. The addition of detrital sediment resulted in immediate and rapid degradation evidenced by the production of 14CO2 from [14C]p-CP. The addition of sterile sediment, glass beads, or sand resulted in approximately four to six times more CO2 evolution than observed in the water alone. Densities of p-CP-degrading bacteria associated with the detrital sediment were 100 times greater than those enumerated in water. Bacteria in the water and associated with the sediment after preexposure of both water and sediment of p-CP demonstrated enhanced biodegradation. In some microcosms, p-CP was degraded completely in the top 1.0 cm of intact sediment beds. Sediment reworking activities by benthic invertebrates from one site were sufficient to mix p-CP deep into the sediment bed faster than biodegradation or molecular diffusion. p-CP was persistent at lower depths of the sediment, possibly a result of reduced oxygen conditions preventing aerobic biodegradation.

Bacteria

Adherence of Pseudomonas aeruginosa to hydrophilic contact lenses and other substrata.

Nonmucoid Pseudomonas aeruginosa isolated from corneal ulcers and contact lens cases and solutions were examined for their ability to adhere to polystyrene, glass, and hydrophilic contact lenses of varying water content and polymer composition. Adherence to the various substrates was strain specific. Adherence also was influenced by hydrophobicity and chemical composition of the substratum, as well as pH and electrolyte concentration. An extracellular polymeric adhesive appeared to be involved in the firm attachment of cells to soft contact lenses.

Bacterial Adhesion