Yulina Aliwarga

A novel cationic-peptide coating for the prevention of microbial colonization on contact lenses.

Aims:  To develop an antimicrobial peptide with broad spectrum activity against bacteria implicated in biomaterial infection of low toxicity to mammalian cells and retaining its antimicrobial activity when covalently bound to a biomaterial surface.

Care regimen and lens material influence on silicone hydrogel contact lens deposition

Purpose. To quantitatively detect proteins and cholesterol extracted from worn silicone hydrogel contact lenses and determine the effect of various lens care solutions on deposit accumulation. Methods. Contact lenses, made from different polymers and worn on a daily wear schedule with different lens care solutions, were collected. Lipid and protein deposits were extracted by methanol:chloroform (1:1, v/v) and protein extraction solution (containing urea and surfactant), respectively. Lipid extracts were separated and cholesterol quantified using thin layer chromatography. Protein extracts were quantified using standard techniques. Results. Among all lenses tested, Balafilcon A lenses exhibited greatest extracted cholesterol (4.1 to 8.2 μg/lens) and total protein (5.4 to 23.2 μg/lens). AQuify was the most effective solution in reducing extracted deposits, especially extracted protein, from Balafilcon A lenses. AQuify and Opti-Free RepleniSH solutions were most effective in reducing extracted cholesterol from Senofilcon A and Galyfilcon A lenses, respectively. Use of Opti-Free Express solution resulted in more extracted protein from Lotrafilcon B lenses than use of other solutions. Generally, Lotrafilcon B, Senofilcon A, and Galyfilcon A lenses accumulated relatively low amount of proteins. Lotrafilcon B lenses accumulated the least amount of cholesterol deposit among all lenses tested regardless of solution used. Conclusions. Lens polymer (possibly associated with surface characteristics) is a prominent factor affecting lipid and protein accumulation. Within a lens polymer type, lens care solutions exhibit varying effectiveness in reducing protein and lipid accumulation.

Type III Secretion System-Associated Toxins, Proteases, Serotypes, and Antibiotic Resistance of Pseudomonas aeruginosa Isolates Associated with Keratitis

The association between possession of toxin gene-related type III secretory system, protease profiles, O serotypes, and antibiotic resistance patterns was characterized genetically and phenotypically in 46 keratitis isolates of Pseudomonas aeruginosa. There was no significant difference in exoU or exoS prevalence among the keratitis strains. Distinct protease profiles were seen in isolates harboring either exoU or exoS genes. One hundred percent (13/13) of serotype E (O:11) strains contained type III secretion system–associated cytotoxin gene exoU. Multidrug resistance was identified in 4% of Australian and 29% of Indian isolates. None of the Australian isolates was resistant to ciprofloxacin. In general, the rate of multidrug resistance in the exoU positive cytotoxic and serotype E (O:11) strains was significantly higher than in exoS positive invasive strains (p < 0.01). The results suggest that multidrug resistance may be more commonly associated with the corneal isolates of P. aeruginosa having type III secretion system-associated cytotoxin gene exoU and belonging to serotype E (O:11) group.

Effects of α-Toxin-Deficient Staphylococcus aureus on the Production of Peripheral Corneal Ulceration in an Animal Model

Purpose: To examine the role of Staphylococcus aureus α-toxin in contact lens-induced peripheral ulceration (CLPU). Materials and Methods: Proteolytic enzyme, hyaluronidase, α-toxin, and β-toxin production by S. aureus 8325-4 and its α-toxin-deficient mutant (S. aureus DU1090) were examined. Using a rabbit model of CLPU, animals were fitted with hydrogel contact lenses colonized by either S. aureus 8325-4 or the mutant strain. The clinical presentation, bacterial cultures, and histology of the ulceration were examined. Results: Both strains of S. aureus produced similar levels of caseinase, gelatinase, elastase, hyaluronidase, and β-toxin. S. aureus DU1090 induced weaker haemolysis of rabbit blood cells than S. aureus 8325-4. Ulceration in the S. aureus DU1090 eye was less frequent and less severe than that caused by S. aureus 8325-4. Conclusions: The enzyme production profile of S. aureus DU 1090 was similar to the parent strain. S. aureus strains may produce CLPU-like lesions irrespective of α-toxin production, but severe infectious lesions are produced only in the presence of α-toxin.

Antimicrobial peptides: a potential role in ocular therapy

Bacterial pathogens are often involved in contact lens‐related adverse responses. This study aimed to find antimicrobial peptides and proteins that effectively eradicate or inhibit ocular bacteria. The antimicrobials were screened against Gram‐ negative and Gram‐positive bacteria originating from ocular sources. The viability of these ocular bacteria was measured after exposure to the peptides and proteins. Two conditions were used to grow bacteria, low nutrient phosphate‐buffered saline and high nutrient tryptone soya broth. Samples were taken at different times up to 48 h. In low nutrient conditions, protamine was found to be the most effective against all strains. Melittin was very effective against all strains except Serratia and one Pseudomonas isolate which were partially affected. In high nutrient condition, only melittin was effective in killing Staphylococcus aureus. Protamine and the combination of protamine and melittin had the greatest effect in eradicating the bacteria tested in low nutrient condition. Protamine alone and its combination with melittin may have potential therapeutic agents for ocular infections in an era of emerging antibiotic resistance.