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ABSTRACT: PURPOSE: Bacterial adhesion to contact lenses is believed to be the initial step for the development of several adverse reactions that occur during lens wear such as microbial keratitis. This study examined the effect of combinations of proteins on the adhesion of bacteria to contact lenses. METHODS: Unworn balafilcon A and senofilcon A lenses were soaked in commercially available pure protein mixtures to achieve the same amount of various proteins as found ex vivo. These lenses were then exposed to Pseudomonas aeruginosa and Staphylococcus aureus. Following incubation, the numbers of P. aeruginosa or S. aureus that adhered to the lenses were measured. The possible effect of proteins on bacterial growth was investigated by incubating bacteria in medium containing protein. RESULTS: Although there was a significant (p < 0.003) increase in the total or viable counts of one strain of S. aureus (031) on balafilcon A lenses soaked in the lysozyme/lactoferrin combination, the protein adhered to lenses did not alter the adhesion of any other strains of P. aeruginosa or S. aureus (p > 0.05). Growth of S. aureus 031 (p < 0.0001) but not of P. aeruginosa 6294 was stimulated by addition of lysozyme/lactoferrin combination (2.8/0.5 mg/mL). Addition of lipocalin did not affect the growth of any strains tested (p > 0.05). CONCLUSIONS: Adsorption of amounts of lysozyme and lactoferrin or lipocalin equivalent to those extracted from worn contact lenses did not affect the adhesion of most strains of S. aureus or P. aeruginosa to lens surfaces.
Optometry and vision science: official publication of the American Academy of Optometry 05/2013; · 1.53 Impact Factor
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ABSTRACT: PURPOSE: To evaluate subjective responses, rates of solution-induced corneal staining (SICS), and corneal infiltrative events (CIEs) associated with a silicone hydrogel when worn under a daily disposable (DD) or daily wear (DW) modality. METHODS: Data were obtained from seven prospective clinical trials, involving 283 subjects wearing Senofilcon A contact lenses (Johnson and Johnson Vision Care, Jacksonville, FL). Four groups wore the contact lens on a DW basis using multipurpose solutions (DW-MPS, n = 160), two groups using hydrogen peroxide (DW-H2O2, n = 83), and one group as a DD (n = 40). Participants were followed for 3 months using the same protocol. Subjective ratings were compared between DD and DW groups using a linear mixed model. Rates of SICS and CIE were calculated as the percent frequency of first events within the 3-month study. Association of SICS and CIE with subjective responses was analyzed using a linear mixed model. RESULTS: End-of-day comfort and dryness ratings were significantly better for DD (8.5 ± 1.7 and 8.6 ± 1.7, respectively) compared with DW-H2O2 (7.7 ± 1.8, p = 0.01; 7.6 ± 1.9, p = 0.006, respectively) and DW-MPS (7.7 ± 1.7, p = 0.004; 7.6 ± 2.0, p = 0.003, respectively). Compared with DD, a higher incidence of CIE and SICS was found for DW-MPS (0.0 vs. 3.9%, p = 0.021; 0.9 vs. 8.6%, p = 0.002, respectively). Comfort at insertion and end of day and end-of-day dryness scores were significantly lower for participants who experienced SICS (8.2 ± 1.6, 7.0 ± 1.9, and 7.0 ± 2.2) than those who did not (8.8 ± 1.2, p = 0.004; 7.9 ± 1.7, p = 0.002; and 7.9 ± 1.8, p = 0.003, respectively). Comfort at insertion scores were significantly lower for participants who experienced CIEs than those who did not (8.2 ± 1.5 vs. 8.8 ± 1.4, p = 0.032). CONCLUSIONS: Senofilcon A clinical response is modulated by the lens care products. All lens care products tested reduced subjective responses relative to DD modality, whereas MPS increased the incidence of CIEs and SICS. Senofilcon A performs best when used on a DD basis.
Optometry and vision science: official publication of the American Academy of Optometry 03/2013; · 1.53 Impact Factor
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ABSTRACT: The deposition of biological material (biofouling) onto polymeric contact lenses is thought to be a major contributor to lens discomfort and hence discontinuation of wear. We describe a method to characterize lipid deposits directly from worn contact lenses utilizing liquid extraction surface analysis coupled to tandem mass spectrometry (LESA-MS/MS). This technique effected facile and reproducible extraction of lipids from the contact lens surfaces and identified lipid molecular species representing all major classes present in human tear film. Our data show that LESA-MS/MS is a rapid and comprehensive technique for the characterization of lipid-related biofouling on polymer surfaces.
The Analyst 03/2013; 138(5):1316-20. · 4.23 Impact Factor
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ABSTRACT: The aim of this study was to quantify specific proteins deposited on daily wear silicone hydrogel lenses used in combination with multipurpose disinfecting solutions (MPDSs) by applying multiple-reaction-monitoring mass spectrometry (MRM-MS).
Balafilcon A or senofilcon A contact lenses used with different MPDSs on a daily wear schedule were collected. Each worn lens was extracted and then digested with trypsin. MRM-MS was applied to quantify the amounts of lysozyme, lactoferrin, lipocalin-1, proline-rich protein-4, and keratin-1 in the extracts.
The amount of protein extracted from the contact lenses was affected by the individual wearers, lens material, and type of care system used. Higher amounts of proteins were extracted from lenses after wear when they were used with an MPDS containing polyhexamethylene biguanide (PHMB) and poloxamer 407 compared with MPDSs containing polyquaternium-1 (PQ-1)/alexidine dihydrochloride with Tetronic 904 or PQ-1/ PHMB with poloxamine and sulfobetaine (p<0.05). There was a correlation between the amount of lipocalin-1 or keratin-1 extracted from lenses and symptoms of ocular dryness.
The MRM-MS technique is a promising approach that could be used to reveal associations of individual proteins deposited on lenses with performance of contact lenses during wear.
Molecular vision 01/2013; 19:390-9. · 2.20 Impact Factor
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ABSTRACT: PURPOSE: To develop a stable antimicrobial contact lens which is effective against the International Organization for Standardization (ISO) panel microorganisms, Acanthamoeba castellanii and drug resistant strains of Pseudomonas aeruginosa and Staphylococcus aureus. Methods: Melimine was covalently incorporated into Etafilcon A lenses. The amount of peptide present on the lens surface was quantified using amino acid analysis. After coating, the heat stability (121oC), lens surface hydrophobicity and in vitro cytotoxicity to mouse L929 cells of the lenses were investigated. Antimicrobial activity against the microorganisms was evaluated by viable plate count and fluorescence microscopy; measuring the proportion of cell death compared to control lenses with no melimine Results: The most effective concentration was determined to be 152±44 µg lens-1 melimine on the lens surface. After coating, lenses were relatively hydrophilic and were non toxic to mammalian cells. The activity remained high after autoclaving (e.g. 3.1, 3.9, 1.2 and 1.0 log inhibition against P. aeruginosa, S. aureus, A. castellanii and Fusarium solani respectively). Fluorescence microscopy confirmed significantly reduced (p <0.001) adhesion of viable bacteria to melimine contact lenses. Viable count confirmed that lenses were active against all the bacteria and fungi from the ISO panel, Acanthamoeba and gave at least 2 log inhibition against all the multidrug resistant S. aureus and P. aeruginosa strains. Conclusions: Melimine may offer excellent potential for development as a broad spectrum antimicrobial coating for contact lenses, showing activity against all the bacterial and fungal ISO panel microorganisms, Acanthamoeba and antibiotic resistant strains of P. aeruginosa and S. aureus.
Investigative ophthalmology & visual science 12/2012; · 3.43 Impact Factor
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ABSTRACT: PURPOSE: Compared with daily disposable wear schedule, continuous wear (CW) or extended wear of contact lenses has been associated with an increased risk of developing an ocular infection. Proof-of-principle studies were conducted to investigate the impact of daily replacement of lenses on the rate of contact lens-related ocular adverse events (AEs) during 30-night CW. METHODS: A total of 215 subjects were dispensed with silicone hydrogel lenses on a 30-night CW schedule but replaced lenses daily either each night before sleeping (n = 178 eyes) or each morning after waking (n = 252 eyes). Scheduled clinic visits were conducted at 1 week and 1 month. Neophytes were required to complete 1 week of daily wear before commencing CW. A historical control (n = 191 eyes) using the same site, subject demographics, and visit schedule but monthly lens replacement was used for AE rates. RESULTS: Logistic regression analysis showed a significant reduction in mechanical AEs (0.8 vs 5.2%, p = 0.01) and overall AEs (inflammatory and mechanical events) (4.0 vs 8.9%, p = 0.04) when lenses were replaced each morning compared with being replaced monthly. Estimation of handling-related lens contamination of unworn lenses in a subgroup of subjects showed isolation of Staphylococcus aureus from the lenses of 35% of subjects, and 65% of subjects had more than 1000 colony-forming units per lens of gram-positive bacterial contamination. CONCLUSIONS: Morning lens replacement during CW reduced mechanical and overall ocular AEs. Replacing lenses at night had no beneficial effects perhaps because the benefit of a fresh lens at night might be partially negated by contamination of the contact lens caused by lens handling before overnight eye closure. Contact lens wearers on an extended wear or CW schedule should be advised to minimize lens handling before sleep to reduce the risk of complications.
Optometry and vision science: official publication of the American Academy of Optometry 11/2012; · 1.53 Impact Factor
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ABSTRACT: PURPOSE.: We sought to determine whether human corneal limbal epithelial cells (HCLE) differ in their physiological response and production of inflammatory mediators during exposure to two multipurpose disinfecting solutions that differ in only four excipients. METHODS.: HCLE were exposed to dilutions (1%-20%) of OPTI-FREE Express and OPTI-FREE RepleniSH for 2, 6, or 18 h. Cell numbers were measured using CyQuant and metabolic activity assays. Morphology, viability, and apoptotic changes were examined by confocal microscopy after staining with Calcein AM/propidium iodide or Hoechst 33,342/propidiun iodide/YO-PRO-1. Cytokine responses and arachidonic acid metabolites were examined by enzyme-linked immunosorbent assay. RESULTS.: OPTI-FREE Express showed greater reductions in corneal cell metabolic activity (up to 3-fold) than OPTI-FREE RepleniSH over 18 h. Cells exposed to OPTI-FREE Express were highly vacuolated, whereas those exposed to OPTI-FREE RepleniSH had morphology consistent with a presumed apoptotic response. OPTI-FREE Express elicited higher levels of interleukin (IL)-6 (maximum 4225 ± 300 pg/mL) and IL-8 (maximum 1094 ± 250 pg/mL) from cells than OPTI-FREE RepleniSH (maximum of 1717 ± 225 pg/mL and 930 ± 300 pg/mL, respectively) at all concentrations tested after 18 h. HCLE did not produce leukotriene B4 or prostaglandin E2 on stimulation, and IL-1β was produced in low levels, but its production was not different between multipurpose disinfecting solution types. CONCLUSIONS.: The findings reported here are the first to describe the pattern of cytokine production produced in corneal epithelial cells in response to contact lens solutions. Alteration of only four excipients in the formulations had such effects on corneal epithelia; however, these differences do not explain differences in the incidences of corneal infiltrative events between these solutions. This might indicate that changes in the rates of infiltrative events result from interactions of the solutions with the contact lens surface, or directly with the corneal immune system, or as the result of gram-negative contamination of lens cases.
Optometry and vision science: official publication of the American Academy of Optometry 10/2012; 89(10):1460-7. · 1.53 Impact Factor
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ABSTRACT: PURPOSE:: To examine the effectiveness of heating contact lens cases after disinfection on reducing microbial contamination. METHODS:: One strain each of Pseudomonas aeruginosa (071) and Staphylococcus aureus (31) were used to set up robust biofilms in polypropylene contact lens cases. The effect of dilutions (from 1:10 to 1:1000) of trypticase soy broth (TSB) in phosphate-buffered saline and incubation time (24 to 48h) on the ability of strains to from biofilms with high levels of bacteria were first examined. Then the effect of increasing the temperature of incubation (from 14°C to 60°C) of biofilms during drying was examined. In the final set of experiments, biofilms of strains were subjected to heating in a warming device set to deliver 60°C for 3 hours, and the effect of this temperature after disinfection with a multipurpose disinfecting solution (MPDS; containing polyquat and Aldox) was examined by culturing the number of viable bacterial cells remaining. RESULTS:: A dilution of 1:100 TSB for S. aureus 31 and 1:1000 TSB for P. aeruginosa 071 together with an incubation time of 24 hours gave high numbers of viable cells of these 2 strains adhered to the contact lens cases. Having established the biofilms of bacteria, heating these to 60°C for 3 hours resulted in significant reductions in the number of viable cells that could be cultured (1 log reduction for S. aureus 31, P=0.0003; 3.5 log reduction for P. aeruginosa 071, P=0.002). Exposing the biofilms of cells to a disinfection cycle (6h at ambient temperature) in the presence of the MPDS and air drying at ambient temperature resulted in 2441±1237 colony-forming units/lens well for S. aureus 31 and 7401±4374 colony-forming units/lens well for P. aeruginosa 071. Increasing the drying temperature to 60°C resulted in zero viable cells (i.e., ≥4log reduction) for either bacterial type. CONCLUSIONS:: Using a warming device for contact lens cases after a disinfection cycle with an MPDS during drying for 3 hours results in substantial kill of biofilms of P. aeruginosa and S. aureus that have been formed in the wells of the cases.
Eye & contact lens 08/2012;
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ABSTRACT: The objective of this study was to determine the bacterial adhesion to various silicone hydrogel lens materials and to determine whether lens wear modulated adhesion.
Bacterial adhesion (total and viable cells) of Staphylococcus aureus (31, 38, and ATCC 6538) and Pseudomonas aeruginosa (6294, 6206, and GSU-3) to 10 commercially available different unworn and worn silicone hydrogel lenses was measured. Results of adhesion were correlated to polymer and surface properties of contact lenses.
S. aureus adhesion to unworn lenses ranged from 2.8 × 10 to 4.4 × 10 colony forming units per lens. The highest adhesion was to lotrafilcon A lenses, and the lowest adhesion was to asmofilcon A lenses. P. aeruginosa adhesion to unworn lenses ranged from 8.9 × 10 to 3.2 × 10 colony forming units per lens. The highest adhesion was to comfilcon A lenses, and the lowest adhesion was to asmofilcon A and balafilcon A lenses. Lens wear altered bacterial adhesion, but the effect was specific to lens and strain type. Adhesion of bacteria, regardless of genera/species or lens wear, was generally correlated with the hydrophobicity of the lens; the less hydrophobic the lens surface, the greater the adhesion.
P. aeruginosa adhered in higher numbers to lenses in comparison with S. aureus strains, regardless of the lens type or lens wear. The effect of lens wear was specific to strain and lens. Hydrophobicity of the silicone hydrogel lens surface influenced the adhesion of bacterial cells.
Optometry and vision science: official publication of the American Academy of Optometry 07/2012; 89(8):1095-106. · 1.53 Impact Factor
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ABSTRACT: To determine the association of single nucleotide polymorphisms (SNPs) of defensin 1B and toll-like receptor 4 with contact lens keratitis susceptibility and severity, and to understand the factors that influence study participation.
Retrospective, case-control study.
Ninety cases of keratitis and 185 controls recruited from studies conducted at Moorfields Eye Hospital and throughout Australia from 2003 to 2005 were analyzed for genetic associations. The reasons for participation of a subset of 146 participants from 1 site were also investigated.
Buccal swab samples were collected on Whatman FTA cards and mailed by post for analysis. DEFB1 (rs1799946, -52, rs1800972, -44, and rs11362, -20) and TLR4 (rs4986790, D299G) SNPs were screened by pyrosequencing and analyzed using a regression model for susceptibility (sterile, microbial keratitis [MK], controls) and severity. Study participation was investigated for age, gender, condition, and phone follow-up also using regression analysis.
Relative risk of developing contact lens-related keratitis and more severe forms of the disease based on genetic profiles.
Carriers of risk alleles of DEFB1 -52 and -20 showed a trend toward increased susceptibility to keratitis (-52: odds ratio [OR], 1.45; 95% confidence interval [CI], 0.99-2.11; P = 0.051; -20: OR, 1.37; 95% CI, 0.95-1.98; P = 0.088). A DEFB1 promoter haplotype (G-G-A) had a tendency toward decreased susceptibility of MK (OR, 0.68; 95% CI, 0.45-1.03; P = 0.062) and reduced severity (OR, 0.56; 95% CI, 0.30-1.07; P = 0.066). The TLR4 D299G was not associated with type and severity of keratitis. Older age (OR, 1.07; 95% CI, 1.05-1.08) and follow-up phone call (OR, 2.0; 95% CI, 1.2-3.5) were independent predictors of study participation.
Genetic variation in DEFB1 that may lead to decreased protein expression of hBD-1 exhibits a tendency toward increased susceptibility and severity of contact lens-related keratitis. Investigation of these and other hBD genes that play important roles in animal models in a larger sample size is warranted. The approach of requesting samples from retrospective case series was generally feasible, although significant resources, including repeat phone calls, are required. More targeted strategies to recruit younger individuals to participate in genetic studies may be useful.
The author(s) have no proprietary or commercial interest in any materials discussed in this article.
Ophthalmology 07/2012; 119(10):1997-2002. · 5.45 Impact Factor
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ABSTRACT: Reduced tear film stability is reported to contribute to dry eye. Rabbits are known to have a more stable tear film than humans. Thus, we sought to examine the tears of rabbits and humans for metal cations, and to test how they influence tear film stability.
Tears were collected from 10 healthy humans and 6 rabbits. Tear osmolality was measured by vapor pressure osmometer, and metals analyzed using inductively coupled plasma (ICP) mass spectrometry or ICP atomic emission spectroscopy. The influence of divalent cations on tears was analyzed by measuring surface tension using the Langmuir trough in vitro, using different concentrations of cations in the subphase, and grading the tear break-up in rabbits in vivo after instillation of chelating agents.
Rabbit tears had a higher osmolality compared to humans. Major metals did not differ between species; however, rabbits had higher levels of Mg(2+) (1.13 vs. 0.39 mM) and Ca(2+) (0.75 vs. 0.36 mM). In rabbit tears in vitro, diminishing divalent cations resulted in a decrease in the maximum surface pressure from 37 to 30 mN/m. In vivo, an increase in the amount of tear film that was broken-up was found. In contrast, when changing divalent cation concentrations in human tears, the maximum surface pressure remained at 26 mN/m.
The normal osmolality of rabbit tears is significantly higher than that in humans. While divalent cations had little influence on human tears, they appear to have an important role in maintaining tear film stability in rabbits.
Investigative ophthalmology & visual science 04/2012; 53(7):3280-5. · 3.43 Impact Factor
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ABSTRACT: To investigate whether single nucleotide polymorphisms (SNPs) in interleukin (IL)-1β, IL-6, and IL-12β are associated with the susceptibility and severity of contact lens-related keratitis.
Retrospective, case control study.
One hundred twelve cases of keratitis and 225 controls were recruited from studies conducted at Moorfields Eye Hospital and in Australia during 2003 through 2005.
Buccal swab samples were collected on Whatman FTA cards and were mailed by post for analysis. IL-1β (-31), IL-6 (-174, -572, -597), and IL-12B (3'+1158) genotypes were analyzed with pyrosequencing and analyzed using a regression model for susceptibility (sterile, microbial keratitis, controls) and severity. Statistical significance was set at 0.05.
The relative risk of developing contact lens-related keratitis and more severe forms of the disease based on allele, genotype, and haplotype associations.
Carriers of IL-6 SNPs were more likely to experience moderate and severe events compared with those with nonmutated genotypes (-174 heterozygous: odds ratio [OR], 3.1; 95% confidence interval [CI], 1.1-8.3; homozygous: OR, 6.4; 95% CI, 1.4-28.4; -174/-597: OR, 4.1; 95% CI, 1.6-11.0). More severe keratitis and microbial keratitis were less likely to occur in wearers with the nonmutated IL-6 haplotype (severity OR, 0.4 [95% CI, 0.2-0.7]; microbial OR, 0.6 [95% CI, 0.4-0.9]). Wearers carrying an IL-12B SNP had an increased risk of sterile keratitis (OR, 9.7; 95% CI, 1.2-76.9) compared with controls.
The IL-6 SNPs are known to reduce protein expression of this cytokine and thus ocular immune defense, and carriers of these SNPs were more likely to experience more severe and microbial keratitis, suggesting that IL-6 decreases the severity and susceptibility of contact lens-related keratitis. Carriers of a functional SNP of IL-12B that is known to increase IL-12 expression and stability are more likely to experience sterile keratitis, suggesting that this is associated with the intense inflammatory reaction that occurs in this condition.
Ophthalmology 04/2012; 119(7):1320-7. · 5.45 Impact Factor
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ABSTRACT: Bacterial contamination of the contact lens surface has been demonstrated to cause corneal infiltrative events. A reduction in the rate of bacterially driven corneal infiltrative events associated with lens wear is one of the major goals of the contact lens industry. There is a concern over the potential of any antimicrobial strategy that there will be unwanted changes to the ocular microbiota or the development of resistance to the antimicrobial. The aim of this study was to investigate the effect of prophylactic topical antibiotic instillation during continuous wear of silicone hydrogel lenses on the normal ocular microbiota, the throat microbiota, and the ocular physiology.
Forty-two male subjects were dispensed with lotrafilcon A silicone hydrogel contact lenses for a 3-month, 30 night continuous wear, monthly replacement trial. Subjects were randomized into either tobramycin 0.3% (test) or saline (control) drop group. Two drops were instilled into each eye on waking and before sleep. At monthly visits, lenses were collected aseptically, and ocular and throat swabs were performed, followed by standard microbial recovery and identifications. Any corneal infiltrative event at scheduled or unscheduled visits was recorded.
Numbers of microbes recovered from eye swabs from the tobramycin (test) group were significantly lower than the control (p = 0.01). Gram-positive cocci were recovered less frequently from the test group (p = 0.001). There were no significant differences in the numbers and types of microbes recovered from lens samples, or the contamination rate of the lenses between the two groups. There were no changes in the numbers of fungi or bacteria from throat swabs. There was no evidence of changes to resistance profile of microbes in the throat. More eye swabs from the test group (68.5%) were culture-negative than swabs from control (46.5%; p = 0.002). The test group had less corneal staining superiorly (0.0 ± 0.0 vs. 0.3 ± 0.4; p = 0.025) but increased bulbar redness (2.2 ± 0.5 vs. 1.5 ± 0.4; p < 0.001) at the 3-month visit only, compared with control group.
Overall, there appeared to be a minimal safety risk with 3-month's prophylactic antibiotic drop use during continuous wear of silicone hydrogel lenses. Clinically, antibiotic drop use induced a mild to moderate increase in bulbar redness by the 3-month time-point. Antibiotic use reduced microbiota on lids but did not affect the microbiota of the throat or change resistance to tobramycin.
Optometry and vision science: official publication of the American Academy of Optometry 03/2012; 89(3):326-35. · 1.53 Impact Factor
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ABSTRACT: The aim was to examine the interaction of lactoferrin with gram-negative bacteria isolated from ocular adverse responses.
Strains representing the species Pseudomonas aeruginosa (six strains), Aeromonas hydrophila, Escherichia coli, Stenotrophomonas maltophilia, and Haemophilus influenza were tested. A modified enzyme-linked immunosorbent assay was used to measure the binding of the strains to native and deglycosylated lactoferrin. The effect on the viability of strains was measured by incubating strains in media containing lactoferrin as the sole carbon and nitrogen source. Siderophore production by strains was measured using an established assay.
All the strains except the single strain of E. coli (Ecol8) were capable of binding to lactoferrin. The ocular isolate of H. influenzae showed strong affinity for lactoferrin. The P. aeruginosa strains and the strain of S. maltophilia showed significantly reduced (80%-100% reduction; P<0.05) binding to lactoferrin that had been enzymatically deglycosylated, whereas deglycosylation had no effect on the binding of other strains/species tested. Most strains were able to grow and produce siderophores in the presence of lactoferrin as the sole carbon and nitrogen source.
The ability to bind to and grow on lactoferrin can be important for gram-negative pathogens that colonize the ocular environment, because this could allow bacteria to survive and propagate in the presence of tear fluid.
Eye & contact lens 01/2012; 38(4):208-13.
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ABSTRACT: Protein and lipid deposits on contact lenses may contribute to clinical complications. This study examined the effect of phospholipids on the adhesion of bacteria to contact lenses.
Worn balafilcon A (n = 11) and senofilcon A (n = 11) were collected after daily wear and phospholipids were extracted in chloroform:methanol. The amount of phospholipid was measured by electrospray ionization mass spectrometry. Unworn lenses soaked in phospholipids were exposed to Pseudomonas aeruginosa and Staphylococcus aureus. After 18 h incubation, the numbers of P. aeruginosa or S. aureus that adhered to the lenses were measured. Phospholipid was tested for possible effects on bacterial growth.
A broad range of sphingomyelins (SM) and phosphatidylcholines (PC) were detected from both types of worn lenses. SM (16:0) (m/z 703) and PC (34:2) (m/z 758) were the major phospholipids detected in the lens extracts. Phospholipids did not alter the adhesion of any strain of P. aeruginosa or S. aureus (p > 0.05). Phospholipids (0.1 mg/mL) showed no effect on the growth of P. aeruginosa 6294 or S. aureus 031.
Phospholipids adsorb/absorb to contact lenses during wear, however, the major types of phospholipids adsorbed to lenses do not alter bacterial adhesion or growth.
Optometry and vision science: official publication of the American Academy of Optometry 01/2012; 89(1):52-61. · 1.53 Impact Factor
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ABSTRACT: Antibiotic-resistant Staphylococcus aureus is of great concern, as it causes a wide range of life-threatening infections. The current study demonstrates that dihydropyrrolone (DHP)-coated polyacrylamide substrates are effective in reducing the number of culturable clinical isolates of S. aureus in vitro in a dose-dependent manner and are able to reduce the pathogenic potential of staphylococcal infection in a subcutaneous infection model. Covalently bound DHPs therefore show great potential for use as an antimicrobial strategy in device-related applications.
Antimicrobial Agents and Chemotherapy 12/2011; 56(2):1138-41. · 4.84 Impact Factor
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ABSTRACT: Expression of protease IV by Pseudomonas aeruginosa during ocular infections contributes significantly to tissue damage. However, several P. aeruginosa strains isolated from ocular infections or inflammatory events produce very low levels of protease IV. The aim of the present study was to characterize, genetically and phenotypically, the presence and expression of the protease IV gene in a group of clinical isolates that cause adverse ocular events of varying degrees, and to elucidate the possible control mechanisms of expression associated with this virulence factor. Protease IV gene sequences from seven clinical isolates of P. aeruginosa were determined and compared to P. aeruginosa strains PAO1 and PA103-29. Production and enzyme activity of protease IV were measured in test strains and compared to that of quorum-sensing gene (lasRI) mutants and the expression of other virulence factors. Protease IV gene sequence similarities between the isolates were 97.5-99.5 %. The strains were classified into two distinct phylogenetic groups that correlated with the presence of exo-enzymes from type three secretion systems (TTSS). Protease IV concentrations produced by PAOΔlasRI mutants and the two clinical isolates with a lasRI gene deficiency were restored to levels comparable to strain PAO1 following complementation of the quorum-sensing gene deficiencies. The protease IV gene is highly conserved in P. aeruginosa clinical isolates that cause a range of adverse ocular events. Observed variations within the gene sequence appear to correlate with presence of specific TTSS genes. Protease IV expression was shown to be regulated by the Las quorum-sensing system.
Journal of Medical Microbiology 09/2011; 61(Pt 2):180-90. · 2.50 Impact Factor
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ABSTRACT: Lens case contamination is a risk factor for microbial keratitis. The effectiveness of manufacturers' lens case cleaning guidelines in limiting microbial contamination has not been evaluated in vivo. This study compared the effectiveness of manufacturers' guidelines and an alternative cleaning regimen.
A randomized cross-over clinical trial with two phases (n = 40) was performed. Participants used the lens types of their choice in conjunction with the provided multipurpose solution (containing polyhexamethylene biguanide) for daily wear. In the manufacturers' guideline phase, cases were rinsed with multipurpose solution and air dried. In the alternative regimen phase, cases were rubbed, rinsed with solution, tissue wiped, and air-dried face down. The duration of each phase was 1 month. Lens cases were collected at the end of each phase for microbiological investigation. The levels of microbial contamination were compared, and compliance to both regimens was assessed.
The case contamination rate was 82% (32/39) in the manufacturers' guideline group, compared with 72% (28/39) in the alternative regimen group. There were significantly fewer (p = 0.004) colony forming units (CFU) of bacteria from cases used by following the alternative regimen (CFU range of 0 to 10, and median of 12 CFU per well) compared with that of the manufacturer's guidelines (CFU range of 0 to 10, and median of 28 CFU per well). The compliance level between both guidelines was not significantly different (p > 0.05).
The alternative guidelines are more effective in eliminating microbial contamination from lens cases than that of the current manufacturer's guideline. Simply incorporating rubbing and tissue-wiping steps in daily case hygiene reduces viable organism contamination.
Optometry and vision science: official publication of the American Academy of Optometry 07/2011; 88(10):E1180-7. · 1.53 Impact Factor
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ABSTRACT: The introduction of contact lens multipurpose disinfection solution (MPDS) that can be used in conjunction with a "no-rub" regimen has simplified lens care requirements. Once adhered to a surface, microorganisms can become less susceptible to disinfection. The aim of the study was to evaluate the effect of various regimen steps on the efficacy of MPDS when used with silicone hydrogel and conventional lenses.
Commercially available MPDSs containing polyquad or polyhexamethylene biguanide were used in conjunction with two types of silicone hydrogel (lotrafilcon B and galyfilcon A) and one type of conventional soft contact lenses (etafilcon A). Challenge microorganisms included Staphylococcus aureus ATCC 6538, Pseudomonas aeruginosa ATCC 9027, Serratia marcescens ATCC 13880, Fusarium solani ATCC 36031, Candida albicans ATCC 10231, or Acanthamoeba polyphaga Ros. The effect of regimen steps "rub and rinse," "rinse-only," or "no rub and no rinse" on the disinfection efficacy of test MPDSs was examined using the ISO 14729 Regimen Test procedure.
Overall, the greatest efficacy of MPDSs was observed when "rub and rinse" was performed before disinfection with each of the microorganisms tested, regardless of lens type. "No rub and no rinse" steps resulted in a greater load of microorganisms remaining on lenses compared with the other regimens (p < 0.05). When "rinse-only" was performed before disinfection, the MPDS containing polyquad performed generally better (p < 0.05) than MPDSs containing polyhexamethylene biguanide against bacteria. Significantly, less microorganisms were recovered from galyfilcon A than from other lenses (p < 0.05) when MPDSs were used with "rinse-only" step.
This study has demonstrated that "rub and rinse" is the most effective regimen and should be recommended in conjunction with all multipurpose lens care solutions and all contact lens types, particularly with silicone hydrogel lenses.
Optometry and vision science: official publication of the American Academy of Optometry 05/2011; 88(8):967-72. · 1.53 Impact Factor
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ABSTRACT: The aim of the study is to determine the adhesion of Gram positive and Gram negative bacteria onto conventional hydrogel (CH) and silicone hydrogel (SH) contact lens materials with and without lysozyme, lactoferrin, and albumin coating.
Four lens types (three SH-balafilcon A, lotrafilcon B, and senofilcon A; one CH-etafilcon A) were coated with lysozyme, lactoferrin, or albumin (uncoated lenses acted as controls) and then incubated in Staphylococcus aureus (Saur 31) or either of two strains of Pseudomonas aeruginosa (Paer 6294 and 6206) for 24 h at 37 °C. The total counts of the adhered bacteria were determined using the H-thymidine method and viable counts by counting the number of colony-forming units on agar media.
All three strains adhered significantly lower to uncoated etafilcon A lenses compared with uncoated SH lenses (p < 0.05). Lysozyme coating on all four lens types increased binding (total and viable counts) of Saur 31 (p < 0.05). However, lysozyme coating did not influence P. aeruginosa adhesion (p > 0.05). Lactoferrin coating on lenses increased binding (total and viable counts) of Saur 31 (p < 0.05). Lactoferrin-coated lenses showed significantly higher total counts (p < 0.05) but significantly lower viable counts (p < 0.05) of adhered P. aeruginosa strains. There was a significant difference between the total and viable counts (p < 0.05) that were bound to lactoferrin-coated lenses. Albumin coating of lenses increased binding (total and viable counts) of all three strains (p < 0.05).
Lysozyme deposited on contact lenses does not possess antibacterial activity against certain bacterial strains, whereas lactoferrin possess an antibacterial effect against strains of P. aeruginosa.
Optometry and vision science: official publication of the American Academy of Optometry 05/2011; 88(8):959-66. · 1.53 Impact Factor
