Purpose. To investigate the relationship between the susceptibility of bacteria to topical antimicrobials and clinical outcome in microbial keratitis. Methods. Clinical outcome data were collected from patients with microbial keratitis from whom a bacterium had been isolated during the period 2003 to 2006. The minimum inhibitory concentration (MIC) was determined for the isolates against 10 antimicrobials. The determinants of the primary clinical outcome, the ratio of healing time (closure of epithelial defect) to ulcer size (HT/UA), was analyzed in a general linear model. Results. Complete clinical outcome and MIC data were available for 421 patients. Sixteen (4%) patients required enucleation and 23 (5%) surgical treatment; in 382 (91%) the ulcer healed with intensive topical antimicrobial therapy. There were significant correlations between HT/UA and organism type (P = 0.001), nearest distance of the ulcer to the limbus (0.02), and MIC of the first antimicrobial used or lowest MIC of combined therapy (P = 0.006). In a model including patients who received monotherapy with a fluoroquinolone who had no subsequent change in their treatment and whose ulcers healed without surgical intervention, there were significant linear associations between clinical outcome and MIC for Pseudomonas spp. (P = 0.047), Staphylococcus aureus (P = 0.04), and Enterobacteriaceae (P = 0.045), but not for Streptococcus spp. (P = 0.85) and coagulase-negative staphylococci (CNS) (P = 0.88). Conclusion. With fluoroquinolone monotherapy, there was significant association between the MIC of the antimicrobial prescribed and the clinical outcome with all bacteria except CNS and Streptococcus spp. The approach used in this study, if used prospectively, could allow topical breakpoint susceptibility concentrations to be determined for individual antimicrobial and bacterial combinations.

The opportunistic pathogen Pseudomonas aeruginosa employs the type III secretion system (T3SS) and four effector proteins, ExoS, ExoT, ExoU, and ExoY, to disrupt cellular physiology and subvert the host’s innate immune response. Of the effector proteins delivered by the T3SS, ExoU is the most toxic. In P. aeruginosa infections, where the ExoU gene is expressed, disease severity is increased with poorer prognoses. This is considered to be due to the rapid and irreversible damage exerted by the phospholipase activity of ExoU, which cannot be halted before conventional antibiotics can successfully eliminate the pathogen. This review will discuss what is currently known about ExoU and explore its potential as a therapeutic target, highlighting some of the small molecule ExoU inhibitors that have been discovered from screening approaches.

Pseudomonas aeruginosa is a common opportunistic bacterial pathogen that causes a variety of infections in humans. Populations of P. aeruginosa are dominated by common clones that can be isolated from diverse clinical and environmental sources. To determine whether specific clones are associated with corneal infection, we used a portable genotyping microarray system to analyze a set of 63 P. aeruginosa isolates from patients with corneal ulcers (keratitis). We then used population analysis to compare the keratitis isolates to a wider collection of P. aeruginosa from various nonocular sources. We identified various markers in a subpopulation of P. aeruginosa associated with keratitis that were in strong disequilibrium with the wider P. aeruginosa population, including oriC , exoU , katN , unmodified flagellin, and the carriage of common genomic islands. The genome sequencing of a keratitis isolate (39016; representing the dominant serotype O11), which was associated with a prolonged clinical healing time, revealed several genomic islands and prophages within the accessory genome. The PCR amplification screening of all 63 keratitis isolates, however, provided little evidence for the shared carriage of specific prophages or genomic islands between serotypes. P. aeruginosa twitching motility, due to type IV pili, is implicated in corneal virulence. We demonstrated that 46% of the O11 keratitis isolates, including 39016, carry a distinctive pilA , encoding the pilin of type IV pili. Thus, the keratitis isolates were associated with specific characteristics, indicating that a subpopulation of P. aeruginosa is adapted to cause corneal infection.

In vascularized organ transplants, gender mismatches have higher rates of immunological rejection. We investigated the influence of gender incompatibility, including H-Y incompatibility, on corneal transplant graft rejection and failure. Patients were included who had undergone a first corneal transplant for keratoconus (KC), Fuchs endothelial dystrophy (FED), pseudophakic bullous keratopathy (PBK), infection and other indications. A Cox regression model was fitted for each indication to determine factors affecting graft failure and rejection at 5 years. The impact of gender, including H-Y, matching was analyzed after accounting for other factors, including known risk factors. Of 18 171 patients, 4314 had undergone a transplant for FED, 4783 for KC, 3669 for PBK, 1903 for infection and 3502 for other disorders. H-Y mismatched (male [M]→female [F]) corneas were at greater risk of graft failure or rejection. For FED, F→F were 40% less likely to fail (p < 0.0001) and 30% less likely to reject (p = 0.01); M→M were 20% less likely to fail (p = 0.04) and 30% less likely to reject (p = 0.01). For KC, M→M matched corneas were 30% less likely to fail (p = 0.05) and 20% less likely to reject (p = 0.01) compared with H-Y mismatches. H-Y antigen mismatched (M→F) patients were at greater risk of rejection or graft failure.