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S M Fleiszig

Publications and source records attributed to S M Fleiszig.

27 records · Page 2Linked to original sources
PubMed1995

Rapid and sensitive method for evaluating Pseudomonas aeruginosa virulence factors during corneal infections in mice.

A murine corneal scratch model has been used extensively to study various aspects of the pathogenesis of Pseudomonas aeruginosa, a common etiologic agent of corneal infections. This model uses mild inhalation anesthetics which keep the animals immobile for a relatively short time and promote the interaction between the infecting organisms and the corneal wound. Under these circumstances, only a small number of P. aeruginosa isolates delivered at inocula of > 10(7) CFU are infectious. We determined that this model is useful for studying other P. aeruginosa strains given at lower doses if injectable anesthetics are administered prior to infection to keep the animals immobile for 15 to 30 min. Under these conditions, eight clinical isolates of P. aeruginosa tested at doses of 10(8) CFU per eye induced corneal perforation and/or phthisis in C3H/HeN mice. The 50% infective doses of several strains were between 3 x 10(2) and 1 x 10(5) CFU per mouse eye. When this modified anesthetic procedure was used to evaluate the roles of different P. aeruginosa virulence factors in eye infections, pathology was not observed when eyes were inoculated with 10(8) CFU of strains deficient in production of a complete lipopolysaccharide or the RpoN sigma factor. A strain with a point mutation in the fur gene, involved in production of iron-regulated factors, showed decreased virulence, while a mutant deficient in both hemolytic and nonhemolytic phospholipase C was fully virulent. By modifying the anesthesia procedure, the corneal scratch model allows rapid evaluations of the roles of P. aeruginosa virulence factors in corneal infections.

Animals↗
PubMed1994

Modulation of Pseudomonas aeruginosa adherence to the corneal surface by mucus.

To gain access to the corneal epithelium and cause infections keratitis, bacterial pathogens must first interact with ocular surface factors that could affect bacterial adherence. In this study, we demonstrated that the mucus layer, and, in particular, the mucin fraction of mucus, modulated adherence to intact corneal epithelium of Pseudomonas aeruginosa but not that of Staphylococcus aureus or Streptococcus pyogenes. Removal of endogenous mucus from rat or rabbit eyes increased the adherence of P. aeruginosa by 3- to 10-fold. Ocular mucus obtained from rat eyes, porcine stomach mucin, or bovine submaxillary gland mucin inhibited adherence of P. aeruginosa to uninjured corneal epithelium. The mucin fraction of ocular mucus, purified by ultracentrifugation, was found to contain the inhibitory activity, and inhibition was demonstrated at concentrations of mucin as low as 35 micrograms/ml. Ocular mucin was the only material tested that inhibited adherence of P. aeruginosa to an injured cornea. However, the binding of P. aeruginosa to immobilized substrates in vitro did not predict which fraction would possess antiadherence activity: bacteria bound well to whole ocular mucus, mucin, the nonmucin fraction of ocular mucus, and dilute human tears as well as to porcine stomach mucin and bovine submaxillary gland mucin. The effectiveness of the mucin fraction of ocular mucus at inhibiting the binding of P. aeruginosa to the cornea implies that this material is a barrier that protects the surface of the eye from P. aeruginosa adherence.

Animals↗
PubMed1994

Pseudomonas aeruginosa invades corneal epithelial cells during experimental infection.

Pseudomonas aeruginosa is considered an extracellular pathogen. Using assays to determine intracellular survival in the presence of gentamicin, we have demonstrated that some strains of P. aeruginosa are able to invade corneal cells during experimental bacterial keratitis in mice. Although intracellular bacteria were detectable 15 min after inoculation, the number of intracellular bacteria increased in a time-dependent manner over a 24-h period. Levels of invasion were similar when bacteria were grown as a biofilm on solid medium and when they were grown in suspension. Intracellular bacteria survived in vitro for at least 24 h, although only minimal bacterial multiplication within cells was observed. P. aeruginosa PAK and Escherichia coli HB101 did not cause disease in this model and were not isolated from corneas after 24 h even when an inoculum of 10(8) CFU was applied. Transmission electron microscopy of corneal epithelium from eyes infected for 8 h revealed that intracellular bacteria were present within membrane-bound vacuoles, which suggests that bacterial entry was an endocytic process. At 24 h, the observation of many bacteria free in the cytoplasm indicated that P. aeruginosa was able to escape the endocytic vacuole. The ability of some P. aeruginosa strains to invade corneal epithelial cells may contribute to the pathogenesis or to the progression of disease, since intracellular bacteria can evade host immune effectors and antibiotics commonly used to treat infection.

Animals↗
PubMed1993

The role of pili in the attachment of Pseudomonas aeruginosa to unworn hydrogel contact lenses.

Contamination of contact lenses is thought to increase the risk of infectious keratitis, yet factors promoting attachment of bacteria to contact lenses are not fully understood. It has been suggested that strains of Pseudomonas aeruginosa attach to mucosal surfaces via pili which are appendages found on some strains. This study investigated the role of pili and the effect of incubation time on the attachment of P. aeruginosa to 20 unworn hydrogel lenses representative of each of the four FDA categories. Ten lenses were incubated for 15 minutes and another ten for 180 minutes. Lenses were incubated with either PAK + P. aeruginosa which possessed pili or its isogenic mutant pair, PAK-, which was genetically similar except for the absence of pili. Bacteria were quantified, following homogenization of the contact lens, by viable counts. Non-piliated bacteria were significantly more likely to adhere to the lenses (p < 0.001). A significant interaction between lens type and incubation time was observed (p < 0.05); thus it is difficult to generalize about either of these effects in isolation. These results show that surface characteristics may confer an attachment advantage to bacteria.

Bacterial Adhesion↗
PubMed1993

Lipopolysaccharide in adherence of Pseudomonas aeruginosa to the cornea and contact lenses.

PURPOSE: To determine the role of smooth or rough lipopolysaccharide on adherence of Pseudomonas aeruginosa bacteria to the rat cornea in vitro and on contact lenses of differing types. METHODS: Adherence of a smooth (AK957) and isogenic rough strain (AK1012) of P. aeruginosa bacteria to rat corneas that were either normal, traumatized using a 20-gauge needle or treated for 15 min with 0.1N sodium hydrochloric acid was assessed by homogenization and viable counting. Adherence of these organisms to 43 unworn contact lenses representing the four Food and Drug Administration lens groups was also assessed using viable counts. RESULTS: Attachment to contact lenses was greater for the smooth strain for all four lens types (P < 0.001). No variation in adherence to the different lens types was observed. Smooth bacteria also adhered to the cornea to a greater extent than the rough strain, regardless of trauma type (P < 0.001). Adherence to traumatized corneas was greater than to nontraumatized corneas for both strains of P. aeruginosa bacteria (P < 0.01). Measurement of surface hydrophobicity of the two bacterial strains revealed that the smooth strain was more hydrophobic than the rough strain (P < 0.001), perhaps accounting for the adherence pattern. CONCLUSIONS: These results indicate that bacterial surface characteristics may be important determinants of adherence and could explain the propensity of certain bacterial strains to infect the cornea.

Animals↗
PubMed1992

Conjunctival flora in extended wear of rigid gas permeable contact lenses.

A longitudinal study was performed to examine the effect of rigid gas permeable (RGP) contact lenses (Boston Equalens II and Quantum II) on the conjunctival flora of 45 young healthy subjects. Microbial flora were determined before delivery of lenses. Subjects wore lenses on an extended wear basis, removing them every 7 days for cleaning and disinfection. Cultures were repeated after 2 months of lens wear and the microbial flora were found to be significantly altered compared to the prelens wear results (0.05 greater than p greater than 0.02). Changes to conjunctival flora included an increase in the number of eyes from which potentially pathogenic microorganisms were isolated, an increase in the number of eyes that were culture-negative, and a decrease in the number of eyes harboring only normal conjunctival flora. The increase in potentially pathogenic flora was not specific for Gram-negative bacteria, which are most often associated with infectious keratitis during contact lens wear.

Adolescent↗
PubMed1992

Microbial flora in eyes of current and former contact lens wearers.

Microbial flora from the right eye conjunctival sac of 84 consecutively presenting contact lens patients were compared with cultures from both surfaces of their lens after aseptic removal and with the flora of their storage cases. Similar results were obtained from contact lens and conjunctival cultures of each individual; however, there was no correlation between storage case isolates and lens or conjunctival flora, suggesting that in uncomplicated lens wear, the eye is highly efficient in eradicating microorganisms introduced via handling. Conjunctival flora during daily contact lens wear was similar to the conjunctival flora of a matched control group of non-lens wearers. However, bacteria that are considered to be part of the normal ocular flora were isolated significantly more often from former contact lens wearers. The data also indicated that the use of nonperoxide chemical lens disinfection was associated with a higher proportion of positive cultures for pathogenic microorganisms than the use of other forms of disinfection, for both current and former contact lens wearers. The isolation of potential pathogens was particularly common among elderly subjects using thick contact lenses for extended wear. These changes to conjunctival flora may contribute to the increased risk of ocular infection associated with contact lens wear.

Adult↗
PubMed1992

Extended contact lens wear enhances Pseudomonas aeruginosa adherence to human corneal epithelium.

Extended wear of soft contact lenses is associated with an increased risk of Pseudomonas aeruginosa infection of the cornea. To assess the role of bacterial adherence in the pathogenesis of these infections, superficial corneal epithelial cells and leukocytes from ten patients who use extended-wear soft lenses and ten control eyes were compared for their propensity to attach P. aeruginosa in vitro. Cells were washed from the cornea by saline irrigation, incubated with a 10-ml solution containing 10(7) colony-forming units/ml of bacteria at 35 degrees C for 30 min, collected on a filter, and prepared using a modified acridine orange staining method. Fluorescence microscopy showed bacterial adherence to corneal epithelial cells, leukocytes, and ocular mucus. The mean number of bacteria adhering to epithelial cells was 2.6 for control eyes and 6.6 for the lens-wearing eyes (P = 0.002). The percentage of epithelial cells attaching greater than or equal to four bacteria was higher for lens-wearing eyes than control eyes (57.4% versus 26.0%, P = 0.0005). There was no significant difference between contact lens-wearing eyes and control eyes in the number of leukocytes collected or in the number of bacteria attached to these cells. These results show that P. aeruginosa adherence to epithelial cells is enhanced in those who use extended-wear soft contact lenses, and this may contribute to the increased incidence of P. aeruginosa keratitis for this population.

Acridine Orange↗