Mary Mowrey-mckee - Academia.edu (original) (raw)
Papers by Mary Mowrey-mckee
PubMed, Jul 1, 2002
Purpose: To determine the cytotoxicity potential of soft contact lens disinfection solutions. Met... more Purpose: To determine the cytotoxicity potential of soft contact lens disinfection solutions. Methods: Three modifications of the United States Pharmacopeia (USP) elution test were conducted: trypan blue uptake test; regrowth of cells after exposure; and quantitation of viable cells after exposure test. Cycled lenses were also tested according to the USP direct-contact test. We compared the cytotoxicity profile of neutralized AOSept (CIBA Vision, Duluth, GA) disinfectant, SOLO-care Soft (CIBA Vision, Duluth, GA) brand multipurpose solution, OPTI-FREE Express (Alcon, Ft. Worth, TX) multipurpose disinfecting solution (with ALDOX), ReNu (Bausch & Lomb, Rochester, NY) multipurpose solution, ReNu MultiPlus (Bausch & Lomb, Rochester, NY) multipurpose solution, and COMPLETE Comfort PLUS (Allergan, Irvine, CA) multipurpose solution. Appropriate positive and negative controls were used for each test. Results: Neutralized AOSept, SOLO-care soft, and COMPLETE Comfort PLUS solutions were noncytotoxic by all four test methods. ReNu MPS and ReNu MultiPlus both were noncytotoxic by the USP direct contact test and the USP elution-based trypan blue uptake and cell regrowth tests, but both yielded less than 50% of viable cells. In the three USP Elution test methods, OPTI-FREE Express (with ALDOX) exhibited cytotoxicity. Conclusions: These solutions have shown widely varying cytotoxicity potential. Neutralized AOSept, SOLO-Care Soft, and COMPLETE Comfort Plus were noncytotoxic by all four tests. ReNu MultiPlus and ReNu MPS inhibited the growth of cells after exposure. OPTI-FREE Express (with ALDOX) may have a higher potential for ocular irritation correlating to severe cytotoxicity in vitro.
Optometry and Vision Science, Mar 1, 1993
Previous studies have demonstrated the presence of microorganisms on hydrogel contact lenses unde... more Previous studies have demonstrated the presence of microorganisms on hydrogel contact lenses under various usage conditions. We conducted this study to quantify and identify viable bacteria and fungi associated with hydrogel contact lenses while on the eye. We removed the lenses from patients' eyes using aseptic techniques and cultured them to identify loosely adherent, as well as lens bound, microorganisms. Lenses were vortexed in a transfer medium (thereafter called the lens extract) and the lenses were then incubated in an agar sandwich separately from the transfer medium. We cultured 108 lenses (82 daily wear and 26 extended wear) from 49 patients. Bacteria were cultured from 38% (41) of the lenses; for 31 of these 41 lenses bacteria were isolated only from the lens extracts (made by vortexing lenses in a transfer medium), suggesting a transient association with the lenses. No fungi were isolated. Counts of less than 10 colony forming units (CFU)/lens were observed on 89% of the lenses. Staphylococcus epidermidis were the most frequently isolated bacteria. A statistically significant relation was observed between increased CFU/lens and increased lens age for extended wear lenses (p = 0.028).
Investigative Ophthalmology & Visual Science, May 1, 2004
Purpose: The cytotoxicity potential of poly-hexa-methyl-biguanide (PHMB) was determined using col... more Purpose: The cytotoxicity potential of poly-hexa-methyl-biguanide (PHMB) was determined using colorimetric in vitro assays with murine fibroblast cells (L929) and an immortalized human corneal epithelial cell line (HCE-T). These colorimetric assays are useful for the quantitative factor-induced cytotoxicity within a 24 to 72 hour period of cell culture. Soft contact lens care products containing PHMB corresponding soft contact lens care product excipient solutions, without PHMB, and PHMB in PBS were tested. Methods: Lens care solutions with and without PHMB were diluted 1:3 with growth medium. PHMB in PBS was diluted at 10, 5, 2.5, 1.25 and 0.625 ppm in growth medium. Tests used were the cell viability assays, MTS/PES (MTS/PES) and Alamar Blue (AB), and the cell membrane integrity assay using neutral red uptake and release, (NRUR). The endpoint was spectrophotometric measurement using a microplate reader for MTS/PES at 490 nm and NRUR at 540 nm and a cytofluorometer for AB at 530/580 excitation/emission. The data were expressed as the ratio of the test solution's absorbance value to the absorbance value of the negative control and the final percentage calculated. These percentage values were evaluated by their IC50 (inhibition of control by 50%) and compared using ANOVA/Tukey HSD test and t-test for statistical significance at a p<0.05 to the negative control and between samples. Results: Based on these studies, the following lens care solutions were noncytotoxic at 48 hours exposure using the MTS/PES, AB and NRUR: SOLOcare ® PLUS (CIBA Vision) and the corresponding excipient solution without PHMB and AQuify™ MPS (SOLOcare ® AQUA) (CIBA Vision) and the corresponding excipient solution without PHMB, and PHMB solutions of 5, 2.5, 1.25 and 0.625 ppm. The following solutions were cytotoxic using the same assays: ReNu MultiPlus ® (Bausch & Lomb), ReNu MultiPlus ® (Bausch & Lomb) excipient solution without PHMB and PHMB at 10 ppm. Discussion: The PHMB concentration of the marketed contact lens solutions is approximately 1 ppm. PHMB at 1ppm was noncytotoxic using the three in vitro assays, MTS/PES, AB and NRUR with L929 and HCE-T cells. The presence of PHMB in the ReNu solution did not increase the cytotoxicity indicating that the excipients are responsible for the cytotoxicity observed with ReNu MultiPlus ®. SOLOcare ® PLUS and AQuify™ MPS were both noncytotoxic. These results suggest that not all PHMB-containing solutions are equal. Materials and Methods Plate HCE-T cells by adding 100 µl cell suspension (1.5 X 10 5 cells/ml) to each well of a 96 well flat bottomed plate. One column of 3-4 replicates was allowed for each sample of PHMB or control to be tested. The plates were incubated overnight or for 36 hours at 37°C, 5 % CO 2 in a humidified incubator. Confluency of the wells was checked before proceeding. The medium was aspirated and replaced with a fresh 100 µl of media. Neat solutions of the six soft contact lenses solutions and PHMB dissolved in was Dulbecco's Phosphate Buffered Saline (DPBS) were serially diluted on the 96 well plates. Solutions of the six soft contact lens/media were tested at 25%. Positive control of PHMB/media was tested at final concentrations of 10 ppm (0.001%), 5 ppm (0.0005%), 2.5 ppm (0.00025%), 1.25 ppm (0.000125%) and 0.625ppm (0.0000625%), by serial dilution on 96 well plates. The negative control for these experiments was Dulbecco's Phosphate Buffered Saline (DPBS) at the same dilution as used for the soft contact lens products. HCE-T cells were grown in UltraCulture™ media without antibiotics.
Investigative Ophthalmology & Visual Science, Apr 28, 2009
Investigative Ophthalmology & Visual Science, Nov 1, 2004
Purpose.-To investigate the effect of hypoxia-induced molecular responses of corneal epithelial c... more Purpose.-To investigate the effect of hypoxia-induced molecular responses of corneal epithelial cells on the surface of rabbit and human corneas and corneal cells in culture on interactions with Pseudomonas aeruginosa that may underlie increased susceptibility to keratitis. Methods.-Organ cultures of rabbit and human corneal tissue, primary rabbit and human corneal cells, and transformed human corneal cells from a patient with cystic fibrosis and the same cell line corrected for expression of wild-type cystic fibrosis transmembrane conductance regulator (CFTR), the cellular receptor for P. aeruginosa, were exposed to hypoxic conditions for 24 to 72 hours. Changes in binding and internalization of P. aeruginosa were measured using cellular association and gentamicin-exclusion assays, and expression of CFTR and activation of NF-κB in response to hypoxia were determined by confocal laser microscopy and quantitative measurements of NF-κB activation. Results.-Hypoxia induced in a time-and oxygen-concentration-dependent manner increased association and internalization of clinical isolates of P. aeruginosa in all cells tested. Hypoxia increased CFTR expression and NF-κB nuclear translocation in rabbit and human cells with wildtype CFTR. Corneal cells lacking CFTR had reduced NF-κB activation in response to hypoxia. Hypoxia did not affect the increase in corneal cell CFTR levels or NF-κB activation after P. aeruginosa infection. Conclusions.-Hypoxic conditions on the cornea exacerbate the binding and internalization of P. aeruginosa due to increased levels of CFTR expression and also induce basal NF-κB activation. Both of these responses probably exacerbate the effects of P. aeruginosa infection by allowing lower infectious doses of bacteria to induce disease and promote destructive inflammatory responses.
Eye & Contact Lens-science and Clinical Practice, Sep 1, 2007
PURPOSE: Acanthamoeba castellani, ATCC 30234, cysts, and trophozoites after a 6-hour exposure. ME... more PURPOSE: Acanthamoeba castellani, ATCC 30234, cysts, and trophozoites after a 6-hour exposure. METHODS: Trophozoite cultures were prepared at Bio-Concept Laboratories in vented tissue culture flasks containing peptone yeast glucose broth by incubation (35 degrees C +/-1 degrees C for 11 days). Cyst suspensions were prepared by incubation of trophozoites in phosphate-buffered saline plus heat-killed yeast on Page's saline agar plates (35 degrees C +/-1 degrees C for 14 days). The solutions were inoculated in triplicate in respective lens cases. After the 6-hour exposure, aliquots of challenged solutions were transferred to Dey-Engley neutralizing broth and further diluted in peptone yeast glucose broth in tissue culture plates to the-7 dilution. Flasks and plates were incubated for 14 days at 35 degrees C +/-1 degrees C and were examined with an inverted light microscope at day 14 for the presence of viable trophozoites. The most probable number method was used for approximate enumeration of the number of survivors. RESULTS: Mean log reductions for cysts were 1.8 for Clear Care/AOSEPT Plus, 2.0 for BLUE Vision/BLUE SEPT, 0.7 for Oxysept 1 Step, 0.5 for OPTI-FREE Express with Aldox, and 0.2 for easyvision one step+. Mean log reductions for trophozoites were 2.2 for Clear Care/AOSEPT Plus, 2.7 for BLUE Vision/BLUE SEPT, 2.5 for Oxysept 1 Step, 2.5 for OPTI-FREE Express with Aldox, and 1.8 for easyvision one step+. CONCLUSIONS: Only Clear Care/AOSEPT Plus and BLUE Vision/BLUE SEPT showed high levels of antimicrobial activity against the cyst form of A. castellani. Oxysept 1 Step showed mild activity against the cysts and easyvision one step+ and OPTI-FREE Express with Aldox showed virtually no antiacanthamoeba activity against the cyst form after 6 hours of exposure.
Optometry and Vision Science, Dec 1, 2001
Cutaneous and Ocular Toxicology, 2005
Determination of a D value for specific test organisms is a component of the efficacy evaluation ... more Determination of a D value for specific test organisms is a component of the efficacy evaluation of new contact lens disinfecting solutions. This parameter is commonly defined as the time required for the number of surviving microorganisms to decrease 1 logarithmic unit. The assumption made in establishing a D value is that the rate of kill exhibits first-order kinetics under the specified conditions. Such exponential kill rates are seen with thermal contact lens disinfection systems. A comparison of the death rate kinetics for a variety of chemical contact lens disinfecting solutions was undertaken to ascertain the suitability of D-value determination for these chemical disinfectants. The active agents of these different solutions included hydrogen peroxide, thimerosal, chlorhexidine, tris(2-hydroxyethyl)tallow ammonium chloride, thimerosal, polyaminopropyl biguanide, and polyquaternium-1. The solutions were challenged with 106 CFU of either Pseudomonas aeruginosa, Serratia marcescens, or Staphylococcus hominis per ml, and survival rate was determined. This study clearly demonstrates the nonlinear nature of the inactivation curves for most contact lens chemical disinfecting solutions for the challenge organisms. D-value determination is, therefore, an inappropriate method of reporting the biocidal activity of these solutions.
Clinical and Experimental Optometry, Mar 1, 1987
Formation of deposits on hydrophilic soft contact lenses causes patient discomfort and decreased ... more Formation of deposits on hydrophilic soft contact lenses causes patient discomfort and decreased lens performance. HPLC and SDS-polyacrylamide gel electrophoresis have been used to resolve, identify, and quantify the biological materials from lens deposits, using catalase as an internal standard. Within the limits of detection by silver staining methods (above 0.2pg), all proteins found in lens deposits were also found in human tear fluid (lactoferrin, albumin, tear-specific prealbumin, lysozyme, and an unidentified protein of Mr = 35,000); however, the proportions of proteins deposited on lenses differed markedly from those in tear fuid. Among individual patient lenses, the relative amounts of proteins in lens deposits also showed considerable variation. Specific staining of lenses and electrophoretic gels with Alcian Blue/Periodic Acid-Schiff s reagent indicated the presence of mucin components (glycoproteins and muco-polysaccharides) on heavily deposited lenses, but not on normal or lightly deposited lenses. Severity of lens deposits correlated positively with staining density by silver or AB/PAS on SDS-PAGE gels. Mucin caused loss of electrophoretic resolution (smearing) and formation of elemental silver "mirrors" on silver stained SDS-PAGE gels. This investigation provides (a) new quantitative micro-methods for protein analysis with individual soft contact lenses, and (b) chemical evidence that much is a major component of heavy deposits on hydrophilic lenses. New lens cleaning methods should therefore include dissolution of mucin.
Current Eye Research, 1996
Following the solubilization of protein from patient worn soft contact lenses and subsequent anal... more Following the solubilization of protein from patient worn soft contact lenses and subsequent analysis via SDS-PAGE, an unidentified 30 kDa protein deposit was commonly observed. The mysterious deposit was found to accumulate on a variety of soft contact lens material. Acuvue, Cibasoft, Excelens and Newvue soft contact lenses were worn by three asymptomatic patients using both daily-wear and extended wear regimens. To characterize the unknown deposit, human tear samples and lens eluted protein were subjected to SDS-PAGE, immunoblotting, enzymatic assays and protein sequencing. Results show that the 30 kDa protein deposit is the homologous dimer of tear lysozyme. Polymerized lysozyme was found on each of the three lens materials within one h of wear. However, the dimer was not present in the normal tear film. Therefore, this dimerization phenomenon is the result of an aggregation and interaction of lysozyme with various soft contact lens polymers.
Investigative Ophthalmology & Visual Science, Apr 22, 2011
Clinical Infectious Diseases, 1991
Purpose. To evaluate the disinfection properties of multipurpose contact lens disinfection soluti... more Purpose. To evaluate the disinfection properties of multipurpose contact lens disinfection solutions, based on the International Organization for Standardization (ISO) 14729 guidelines. Methods. ReNu with MoistureLoc MultiPurpose Solution, OPTI-FREE Express with Aldox MultiPurpose Solution, Betadine 5% sterile ophthalmic preparation solution (povidone iodine), and 0.9% normal saline solution were inoculated with strains of Staphylococcus aureus and Pseudomonas aeruginosa. Surviving bacteria were quantified at specified times. Results. ReNu with Mois-tureLoc, OPTI-FREE Express, and 5% ophthalmic povidone iodine were effective in achieving a 5-log reduction in bacterial count. Additionally, all three products maintained their effectivity at 72 hours. However, ReNu with MoistureLoc and povidone iodine resulted in the greatest reduction in bacterial colonization. Conclusions. ReNu with MoistureLoc, OPTI-FREE Express, and 5% ophthalmic povidone iodine meet the ISO 14729 guidelines for standalone contact lens solutions. However, ReNu with Moisture-Loc and 5% ophthalmic povidone iodine are most efficient in reducing and maintaining low bacterial count for a period of 72 hours.
Investigative Ophthalmology & Visual Science, 2002
Le procede decrit, qui sert a la desinfection de lentilles de contact, consiste a placer la lenti... more Le procede decrit, qui sert a la desinfection de lentilles de contact, consiste a placer la lentille en contact avec une solution aqueuse isotonique contenant 0,6 a 2 % en poids de tromethamine (de preference 0,8 a 1,5 %) pendant une periode suffisante pour obtenir la desinfection de la lentille. Dans d'autres aspects, ledit procede consiste a ajouter a la solution 0,01 a 1 % en poids d'un agent de chelation (tel que de preference de l'acide ethylene-diamino-tetraacetique (EDTA) de disodium) et/ou un agent microbicide additionnel.
Investigative Ophthalmology & Visual Science, 2008
The publication costs of this article were defrayed in part by page charge payment. This article ... more The publication costs of this article were defrayed in part by page charge payment. This article must therefore be marked “advertisement” in accordance with 18 U.S.C. §1734 solely to indicate this fact.
La presente invention concerne une solution aqueuse destinee a la sterilisation et au stockage de... more La presente invention concerne une solution aqueuse destinee a la sterilisation et au stockage de dispositifs ophtalmiques, de preference des verres de contact, faits en hydrogel, de preference en hydrogel contenant un poly(oxyalkylene). La solution comprend un ou plusieurs tampons organiques tels qu'un tampon de Good ou un bis-aminopolyol et un agent organique d'ajustement de la tonicite avec des groupes hydroxyle en quantite suffisante pour donner une osmolarite comprise entre 200 et 450 mosm/l environ. Le pH de la solution est compris entre 5,5 et 8,5 environ, a condition que la solution aqueuse comprenne un tampon phosphate a une concentration inferieure ou egale a 15 mM environ et environ 5000 ppm de chlorure de sodium. L'invention concerne egalement un procede de sterilisation et de stockage d'un dispositif ophtalmique a l'aide de la solution aqueuse de l'invention.
Investigative Ophthalmology & Visual Science, 2008
Investigative Ophthalmology & Visual Science, 2011
PubMed, Jul 1, 2002
Purpose: To determine the cytotoxicity potential of soft contact lens disinfection solutions. Met... more Purpose: To determine the cytotoxicity potential of soft contact lens disinfection solutions. Methods: Three modifications of the United States Pharmacopeia (USP) elution test were conducted: trypan blue uptake test; regrowth of cells after exposure; and quantitation of viable cells after exposure test. Cycled lenses were also tested according to the USP direct-contact test. We compared the cytotoxicity profile of neutralized AOSept (CIBA Vision, Duluth, GA) disinfectant, SOLO-care Soft (CIBA Vision, Duluth, GA) brand multipurpose solution, OPTI-FREE Express (Alcon, Ft. Worth, TX) multipurpose disinfecting solution (with ALDOX), ReNu (Bausch & Lomb, Rochester, NY) multipurpose solution, ReNu MultiPlus (Bausch & Lomb, Rochester, NY) multipurpose solution, and COMPLETE Comfort PLUS (Allergan, Irvine, CA) multipurpose solution. Appropriate positive and negative controls were used for each test. Results: Neutralized AOSept, SOLO-care soft, and COMPLETE Comfort PLUS solutions were noncytotoxic by all four test methods. ReNu MPS and ReNu MultiPlus both were noncytotoxic by the USP direct contact test and the USP elution-based trypan blue uptake and cell regrowth tests, but both yielded less than 50% of viable cells. In the three USP Elution test methods, OPTI-FREE Express (with ALDOX) exhibited cytotoxicity. Conclusions: These solutions have shown widely varying cytotoxicity potential. Neutralized AOSept, SOLO-Care Soft, and COMPLETE Comfort Plus were noncytotoxic by all four tests. ReNu MultiPlus and ReNu MPS inhibited the growth of cells after exposure. OPTI-FREE Express (with ALDOX) may have a higher potential for ocular irritation correlating to severe cytotoxicity in vitro.
Optometry and Vision Science, Mar 1, 1993
Previous studies have demonstrated the presence of microorganisms on hydrogel contact lenses unde... more Previous studies have demonstrated the presence of microorganisms on hydrogel contact lenses under various usage conditions. We conducted this study to quantify and identify viable bacteria and fungi associated with hydrogel contact lenses while on the eye. We removed the lenses from patients' eyes using aseptic techniques and cultured them to identify loosely adherent, as well as lens bound, microorganisms. Lenses were vortexed in a transfer medium (thereafter called the lens extract) and the lenses were then incubated in an agar sandwich separately from the transfer medium. We cultured 108 lenses (82 daily wear and 26 extended wear) from 49 patients. Bacteria were cultured from 38% (41) of the lenses; for 31 of these 41 lenses bacteria were isolated only from the lens extracts (made by vortexing lenses in a transfer medium), suggesting a transient association with the lenses. No fungi were isolated. Counts of less than 10 colony forming units (CFU)/lens were observed on 89% of the lenses. Staphylococcus epidermidis were the most frequently isolated bacteria. A statistically significant relation was observed between increased CFU/lens and increased lens age for extended wear lenses (p = 0.028).
Investigative Ophthalmology & Visual Science, May 1, 2004
Purpose: The cytotoxicity potential of poly-hexa-methyl-biguanide (PHMB) was determined using col... more Purpose: The cytotoxicity potential of poly-hexa-methyl-biguanide (PHMB) was determined using colorimetric in vitro assays with murine fibroblast cells (L929) and an immortalized human corneal epithelial cell line (HCE-T). These colorimetric assays are useful for the quantitative factor-induced cytotoxicity within a 24 to 72 hour period of cell culture. Soft contact lens care products containing PHMB corresponding soft contact lens care product excipient solutions, without PHMB, and PHMB in PBS were tested. Methods: Lens care solutions with and without PHMB were diluted 1:3 with growth medium. PHMB in PBS was diluted at 10, 5, 2.5, 1.25 and 0.625 ppm in growth medium. Tests used were the cell viability assays, MTS/PES (MTS/PES) and Alamar Blue (AB), and the cell membrane integrity assay using neutral red uptake and release, (NRUR). The endpoint was spectrophotometric measurement using a microplate reader for MTS/PES at 490 nm and NRUR at 540 nm and a cytofluorometer for AB at 530/580 excitation/emission. The data were expressed as the ratio of the test solution's absorbance value to the absorbance value of the negative control and the final percentage calculated. These percentage values were evaluated by their IC50 (inhibition of control by 50%) and compared using ANOVA/Tukey HSD test and t-test for statistical significance at a p<0.05 to the negative control and between samples. Results: Based on these studies, the following lens care solutions were noncytotoxic at 48 hours exposure using the MTS/PES, AB and NRUR: SOLOcare ® PLUS (CIBA Vision) and the corresponding excipient solution without PHMB and AQuify™ MPS (SOLOcare ® AQUA) (CIBA Vision) and the corresponding excipient solution without PHMB, and PHMB solutions of 5, 2.5, 1.25 and 0.625 ppm. The following solutions were cytotoxic using the same assays: ReNu MultiPlus ® (Bausch & Lomb), ReNu MultiPlus ® (Bausch & Lomb) excipient solution without PHMB and PHMB at 10 ppm. Discussion: The PHMB concentration of the marketed contact lens solutions is approximately 1 ppm. PHMB at 1ppm was noncytotoxic using the three in vitro assays, MTS/PES, AB and NRUR with L929 and HCE-T cells. The presence of PHMB in the ReNu solution did not increase the cytotoxicity indicating that the excipients are responsible for the cytotoxicity observed with ReNu MultiPlus ®. SOLOcare ® PLUS and AQuify™ MPS were both noncytotoxic. These results suggest that not all PHMB-containing solutions are equal. Materials and Methods Plate HCE-T cells by adding 100 µl cell suspension (1.5 X 10 5 cells/ml) to each well of a 96 well flat bottomed plate. One column of 3-4 replicates was allowed for each sample of PHMB or control to be tested. The plates were incubated overnight or for 36 hours at 37°C, 5 % CO 2 in a humidified incubator. Confluency of the wells was checked before proceeding. The medium was aspirated and replaced with a fresh 100 µl of media. Neat solutions of the six soft contact lenses solutions and PHMB dissolved in was Dulbecco's Phosphate Buffered Saline (DPBS) were serially diluted on the 96 well plates. Solutions of the six soft contact lens/media were tested at 25%. Positive control of PHMB/media was tested at final concentrations of 10 ppm (0.001%), 5 ppm (0.0005%), 2.5 ppm (0.00025%), 1.25 ppm (0.000125%) and 0.625ppm (0.0000625%), by serial dilution on 96 well plates. The negative control for these experiments was Dulbecco's Phosphate Buffered Saline (DPBS) at the same dilution as used for the soft contact lens products. HCE-T cells were grown in UltraCulture™ media without antibiotics.
Investigative Ophthalmology & Visual Science, Apr 28, 2009
Investigative Ophthalmology & Visual Science, Nov 1, 2004
Purpose.-To investigate the effect of hypoxia-induced molecular responses of corneal epithelial c... more Purpose.-To investigate the effect of hypoxia-induced molecular responses of corneal epithelial cells on the surface of rabbit and human corneas and corneal cells in culture on interactions with Pseudomonas aeruginosa that may underlie increased susceptibility to keratitis. Methods.-Organ cultures of rabbit and human corneal tissue, primary rabbit and human corneal cells, and transformed human corneal cells from a patient with cystic fibrosis and the same cell line corrected for expression of wild-type cystic fibrosis transmembrane conductance regulator (CFTR), the cellular receptor for P. aeruginosa, were exposed to hypoxic conditions for 24 to 72 hours. Changes in binding and internalization of P. aeruginosa were measured using cellular association and gentamicin-exclusion assays, and expression of CFTR and activation of NF-κB in response to hypoxia were determined by confocal laser microscopy and quantitative measurements of NF-κB activation. Results.-Hypoxia induced in a time-and oxygen-concentration-dependent manner increased association and internalization of clinical isolates of P. aeruginosa in all cells tested. Hypoxia increased CFTR expression and NF-κB nuclear translocation in rabbit and human cells with wildtype CFTR. Corneal cells lacking CFTR had reduced NF-κB activation in response to hypoxia. Hypoxia did not affect the increase in corneal cell CFTR levels or NF-κB activation after P. aeruginosa infection. Conclusions.-Hypoxic conditions on the cornea exacerbate the binding and internalization of P. aeruginosa due to increased levels of CFTR expression and also induce basal NF-κB activation. Both of these responses probably exacerbate the effects of P. aeruginosa infection by allowing lower infectious doses of bacteria to induce disease and promote destructive inflammatory responses.
Eye & Contact Lens-science and Clinical Practice, Sep 1, 2007
PURPOSE: Acanthamoeba castellani, ATCC 30234, cysts, and trophozoites after a 6-hour exposure. ME... more PURPOSE: Acanthamoeba castellani, ATCC 30234, cysts, and trophozoites after a 6-hour exposure. METHODS: Trophozoite cultures were prepared at Bio-Concept Laboratories in vented tissue culture flasks containing peptone yeast glucose broth by incubation (35 degrees C +/-1 degrees C for 11 days). Cyst suspensions were prepared by incubation of trophozoites in phosphate-buffered saline plus heat-killed yeast on Page's saline agar plates (35 degrees C +/-1 degrees C for 14 days). The solutions were inoculated in triplicate in respective lens cases. After the 6-hour exposure, aliquots of challenged solutions were transferred to Dey-Engley neutralizing broth and further diluted in peptone yeast glucose broth in tissue culture plates to the-7 dilution. Flasks and plates were incubated for 14 days at 35 degrees C +/-1 degrees C and were examined with an inverted light microscope at day 14 for the presence of viable trophozoites. The most probable number method was used for approximate enumeration of the number of survivors. RESULTS: Mean log reductions for cysts were 1.8 for Clear Care/AOSEPT Plus, 2.0 for BLUE Vision/BLUE SEPT, 0.7 for Oxysept 1 Step, 0.5 for OPTI-FREE Express with Aldox, and 0.2 for easyvision one step+. Mean log reductions for trophozoites were 2.2 for Clear Care/AOSEPT Plus, 2.7 for BLUE Vision/BLUE SEPT, 2.5 for Oxysept 1 Step, 2.5 for OPTI-FREE Express with Aldox, and 1.8 for easyvision one step+. CONCLUSIONS: Only Clear Care/AOSEPT Plus and BLUE Vision/BLUE SEPT showed high levels of antimicrobial activity against the cyst form of A. castellani. Oxysept 1 Step showed mild activity against the cysts and easyvision one step+ and OPTI-FREE Express with Aldox showed virtually no antiacanthamoeba activity against the cyst form after 6 hours of exposure.
Optometry and Vision Science, Dec 1, 2001
Cutaneous and Ocular Toxicology, 2005
Determination of a D value for specific test organisms is a component of the efficacy evaluation ... more Determination of a D value for specific test organisms is a component of the efficacy evaluation of new contact lens disinfecting solutions. This parameter is commonly defined as the time required for the number of surviving microorganisms to decrease 1 logarithmic unit. The assumption made in establishing a D value is that the rate of kill exhibits first-order kinetics under the specified conditions. Such exponential kill rates are seen with thermal contact lens disinfection systems. A comparison of the death rate kinetics for a variety of chemical contact lens disinfecting solutions was undertaken to ascertain the suitability of D-value determination for these chemical disinfectants. The active agents of these different solutions included hydrogen peroxide, thimerosal, chlorhexidine, tris(2-hydroxyethyl)tallow ammonium chloride, thimerosal, polyaminopropyl biguanide, and polyquaternium-1. The solutions were challenged with 106 CFU of either Pseudomonas aeruginosa, Serratia marcescens, or Staphylococcus hominis per ml, and survival rate was determined. This study clearly demonstrates the nonlinear nature of the inactivation curves for most contact lens chemical disinfecting solutions for the challenge organisms. D-value determination is, therefore, an inappropriate method of reporting the biocidal activity of these solutions.
Clinical and Experimental Optometry, Mar 1, 1987
Formation of deposits on hydrophilic soft contact lenses causes patient discomfort and decreased ... more Formation of deposits on hydrophilic soft contact lenses causes patient discomfort and decreased lens performance. HPLC and SDS-polyacrylamide gel electrophoresis have been used to resolve, identify, and quantify the biological materials from lens deposits, using catalase as an internal standard. Within the limits of detection by silver staining methods (above 0.2pg), all proteins found in lens deposits were also found in human tear fluid (lactoferrin, albumin, tear-specific prealbumin, lysozyme, and an unidentified protein of Mr = 35,000); however, the proportions of proteins deposited on lenses differed markedly from those in tear fuid. Among individual patient lenses, the relative amounts of proteins in lens deposits also showed considerable variation. Specific staining of lenses and electrophoretic gels with Alcian Blue/Periodic Acid-Schiff s reagent indicated the presence of mucin components (glycoproteins and muco-polysaccharides) on heavily deposited lenses, but not on normal or lightly deposited lenses. Severity of lens deposits correlated positively with staining density by silver or AB/PAS on SDS-PAGE gels. Mucin caused loss of electrophoretic resolution (smearing) and formation of elemental silver "mirrors" on silver stained SDS-PAGE gels. This investigation provides (a) new quantitative micro-methods for protein analysis with individual soft contact lenses, and (b) chemical evidence that much is a major component of heavy deposits on hydrophilic lenses. New lens cleaning methods should therefore include dissolution of mucin.
Current Eye Research, 1996
Following the solubilization of protein from patient worn soft contact lenses and subsequent anal... more Following the solubilization of protein from patient worn soft contact lenses and subsequent analysis via SDS-PAGE, an unidentified 30 kDa protein deposit was commonly observed. The mysterious deposit was found to accumulate on a variety of soft contact lens material. Acuvue, Cibasoft, Excelens and Newvue soft contact lenses were worn by three asymptomatic patients using both daily-wear and extended wear regimens. To characterize the unknown deposit, human tear samples and lens eluted protein were subjected to SDS-PAGE, immunoblotting, enzymatic assays and protein sequencing. Results show that the 30 kDa protein deposit is the homologous dimer of tear lysozyme. Polymerized lysozyme was found on each of the three lens materials within one h of wear. However, the dimer was not present in the normal tear film. Therefore, this dimerization phenomenon is the result of an aggregation and interaction of lysozyme with various soft contact lens polymers.
Investigative Ophthalmology & Visual Science, Apr 22, 2011
Clinical Infectious Diseases, 1991
Purpose. To evaluate the disinfection properties of multipurpose contact lens disinfection soluti... more Purpose. To evaluate the disinfection properties of multipurpose contact lens disinfection solutions, based on the International Organization for Standardization (ISO) 14729 guidelines. Methods. ReNu with MoistureLoc MultiPurpose Solution, OPTI-FREE Express with Aldox MultiPurpose Solution, Betadine 5% sterile ophthalmic preparation solution (povidone iodine), and 0.9% normal saline solution were inoculated with strains of Staphylococcus aureus and Pseudomonas aeruginosa. Surviving bacteria were quantified at specified times. Results. ReNu with Mois-tureLoc, OPTI-FREE Express, and 5% ophthalmic povidone iodine were effective in achieving a 5-log reduction in bacterial count. Additionally, all three products maintained their effectivity at 72 hours. However, ReNu with MoistureLoc and povidone iodine resulted in the greatest reduction in bacterial colonization. Conclusions. ReNu with MoistureLoc, OPTI-FREE Express, and 5% ophthalmic povidone iodine meet the ISO 14729 guidelines for standalone contact lens solutions. However, ReNu with Moisture-Loc and 5% ophthalmic povidone iodine are most efficient in reducing and maintaining low bacterial count for a period of 72 hours.
Investigative Ophthalmology & Visual Science, 2002
Le procede decrit, qui sert a la desinfection de lentilles de contact, consiste a placer la lenti... more Le procede decrit, qui sert a la desinfection de lentilles de contact, consiste a placer la lentille en contact avec une solution aqueuse isotonique contenant 0,6 a 2 % en poids de tromethamine (de preference 0,8 a 1,5 %) pendant une periode suffisante pour obtenir la desinfection de la lentille. Dans d'autres aspects, ledit procede consiste a ajouter a la solution 0,01 a 1 % en poids d'un agent de chelation (tel que de preference de l'acide ethylene-diamino-tetraacetique (EDTA) de disodium) et/ou un agent microbicide additionnel.
Investigative Ophthalmology & Visual Science, 2008
The publication costs of this article were defrayed in part by page charge payment. This article ... more The publication costs of this article were defrayed in part by page charge payment. This article must therefore be marked “advertisement” in accordance with 18 U.S.C. §1734 solely to indicate this fact.
La presente invention concerne une solution aqueuse destinee a la sterilisation et au stockage de... more La presente invention concerne une solution aqueuse destinee a la sterilisation et au stockage de dispositifs ophtalmiques, de preference des verres de contact, faits en hydrogel, de preference en hydrogel contenant un poly(oxyalkylene). La solution comprend un ou plusieurs tampons organiques tels qu'un tampon de Good ou un bis-aminopolyol et un agent organique d'ajustement de la tonicite avec des groupes hydroxyle en quantite suffisante pour donner une osmolarite comprise entre 200 et 450 mosm/l environ. Le pH de la solution est compris entre 5,5 et 8,5 environ, a condition que la solution aqueuse comprenne un tampon phosphate a une concentration inferieure ou egale a 15 mM environ et environ 5000 ppm de chlorure de sodium. L'invention concerne egalement un procede de sterilisation et de stockage d'un dispositif ophtalmique a l'aide de la solution aqueuse de l'invention.
Investigative Ophthalmology & Visual Science, 2008
Investigative Ophthalmology & Visual Science, 2011