Antibiotic resistance of bacterial pathogens isolated from the conjunctiva in the Antibiotic Resistance Monitoring in Ocular micRoorganisms (ARMOR) surveillance study (2009-2021).

Antibiotic resistance in bacterial ocular infections is of significant clinical concern and may affect treatment outcomes. We report on in vitro antibiotic susceptibility rates and trends among conjunctival-sourced isolates collected in the Antibiotic Resistance Monitoring in Ocular micRoorganisms (ARMOR) surveillance study. A total of 2214 conjunctival isolates (918 Staphylococcus aureus, 589 coagulase-negative staphylococci [CoNS], 194 Streptococcus pneumoniae, 171 Pseudomonas aeruginosa, and 342 Haemophilus influenzae) obtained between 2009-2021 were analyzed. Staphylococci were commonly resistant to azithromycin (≥54.8%) and oxacillin (≥29.3%). Resistance among S. pneumoniae isolates was notable for azithromycin (34.0%) and penicillin (28.9%), while P. aeruginosa and H. influenzae isolates were highly susceptible to most tested antibiotics. Methicillin-resistant staphylococci demonstrated greater concurrent resistance to other antibiotics than methicillin-susceptible isolates and exhibited high rates of multidrug resistance (≥74.0%). Among staphylococci, antibiotic resistance increased with patient age, and there were small decreases in resistance to several drugs over the 13-year period. These findings indicate that resistance to antibiotics routinely used in ophthalmic practice remains high among conjunctival isolates.

Antibiotic susceptibility of bacterial pathogens isolated from the aqueous and vitreous humour in the Antibiotic Resistance Monitoring in Ocular micRoorganisms (ARMOR) Surveillance Study: 2009-2020 update.

OBJECTIVES: We evaluated antibiotic resistance among intraocular isolates obtained from presumed endophthalmitis cases collected from 2009 through 2020 in the Antibiotic Resistance Monitoring in Ocular micRoorganisms (ARMOR) study, the only ongoing nationwide surveillance study tracking in vitro resistance in ocular pathogens. METHODS: Presumed endophthalmitis isolates obtained from the aqueous humour and vitreous humour were collected from participating centres, and minimum inhibitory concentrations were determined and interpreted per Clinical and Laboratory Standards Institute methods and available breakpoints. RESULTS: A total of 307 presumed endophthalmitis isolates (aqueous humour, n = 88; vitreous humour, n = 219) were obtained from 43 clinical sites, including 188 coagulase-negative staphylococci (CoNS), 61 Staphylococcus aureus, 31 Streptococcus pneumoniae, 14 Pseudomonas aeruginosa, and 13 Haemophilus influenzae isolates. Of the CoNS isolates, 47.9% (90/188) were methicillin resistant, 58.0% (109/188) were azithromycin resistant, and 46.3% (87/188) were ciprofloxacin resistant. Of the S. aureus isolates, 45.9% (28/61) were methicillin resistant, 57.4% (35/61) were azithromycin resistant, and 44.3% (27/61) were ciprofloxacin resistant. Multidrug resistance (MDR; i.e., resistance to ≥3 antibiotic classes) was prevalent among staphylococci, particularly methicillin-resistant strains, of which >70% exhibited MDR. Resistance among S. pneumoniae isolates was notable for azithromycin and penicillin, each 38.7% (12/31), and for polymyxin B among P. aeruginosa 100.0% (14/14), whereas no resistance was observed for H. influenzae isolates to the antibiotics tested. CONCLUSION: In vitro antibiotic resistance was common among presumed endophthalmitis isolates collected in the ARMOR surveillance study. These data could inform antibiotic selection for infection prophylaxis and/or treatment of intraocular infections.

In vitro time-kill experiments with besifloxacin, moxifloxacin and gatifloxacin in the absence and presence of benzalkonium chloride.

OBJECTIVES: To compare the bactericidal activity of besifloxacin, moxifloxacin and gatifloxacin and determine the contribution of the preservative benzalkonium chloride (BAK) to bactericidal activity. METHODS: Time-kill experiments were performed against four species (n=12) with besifloxacin, moxifloxacin and gatifloxacin, in the presence or absence of BAK, at t=0, 5, 15, 30, 45, 60, 120 and 360 min, according to standard CLSI methods. RESULTS: In the presence of BAK, bactericidal activity was observed within 5 min, regardless of the fluoroquinolone tested. The bactericidal activity of BAK was unaffected by the concurrent presence of besifloxacin and rapid killing (within 5 to 15 min) was not observed at BAK concentrations below 50 mg/L. However, when tested without BAK, besifloxacin was bactericidal in as little as 45 min, while moxifloxacin and gatifloxacin required at least 120 min; besifloxacin kill rates against fluoroquinolone-susceptible and -resistant strains were at least 2- to 4-fold faster than those of gatifloxacin or moxifloxacin. CONCLUSIONS: Besifloxacin was the most rapidly bactericidal fluoroquinolone tested, followed by gatifloxacin and moxifloxacin, both of which had similar activity. Our studies demonstrate that the previously reported rapid in vitro killing by gatifloxacin formulations was probably due to the concurrent presence of 50 mg/L BAK, which is much higher than the 3.2 mg/L BAK observed in human tears 1 min after instillation of ophthalmic gatifloxacin solutions [Friedlaender MH, Breshears D, Amoozgar B et al. The dilution of benzalkonium chloride (BAK) in the tear film. Adv Ther 2006; 23: 835-41].

Topoisomerase mutations that are associated with high-level resistance to earlier fluoroquinolones in Staphylococcus aureus have less effect on the antibacterial activity of besifloxacin.

BACKGROUND: The impact of mutations in DNA gyrase and topoisomerase IV on minimum inhibitory concentrations (MICs) was investigated to better understand why besifloxacin has a higher potency against Staphylococcus aureus when compared to other fluoroquinolones, which was especially pronounced against ciprofloxacin-resistant isolates. METHODS: MICs were determined for 52 clinical isolates against besifloxacin, moxifloxacin, gatifloxacin, ciprofloxacin, and levofloxacin. The genes encoding GyrA, GyrB, ParC, and ParE were sequenced and the potential impact of mutations assessed in light of recent structural data. RESULTS: For all fluoroquinolones tested, the MICs increased with the number of mutations in the quinolone resistance-determining regions. However, this increase was the smallest for besifloxacin and the largest for ciprofloxacin and levofloxacin. In addition to the commonly observed mutations in ParC and GyrA, more unusual mutations in ParE, such as Asp-432→His or Pro-585→Ser, were also detected. CONCLUSIONS: Compared to earlier fluoroquinolones, the higher potency of besifloxacin suggests that the drug's unique combination of a 7-azepinyl ring and an 8-chloro-substituent results in unique interactions with DNA gyrase and topoisomerase IV.

Characterization of Polybacterial versus Monobacterial Conjunctivitis Infections in Pediatric Subjects Across Multiple Studies and Microbiological Outcomes with Besifloxacin Ophthalmic Suspension 0.6.

INTRODUCTION: The choice of empiric therapy for bacterial conjunctivitis should be guided by an awareness of typical causative pathogen distributions. Bacterial conjunctivitis can be polybacterial, although pediatric-specific data are lacking. METHODS: This was a post-hoc analysis of data in pediatric subjects (1-17 years) from five bacterial conjunctivitis trials evaluating besifloxacin ophthalmic solution 0.6%. RESULTS: Of the 730 pediatric subjects with culture-confirmed conjunctivitis, nearly one-fourth (23.6%) had polybacterial infections and three-fourths (76.4%) had monobacterial infections at baseline. In both polybacterial and monobacterial infections, the most prevalent organisms were Haemophilus influenzae, Streptococcus pneumoniae, Staphylococcus aureus, and Streptococcus mitis/S. mitis group. In polybacterial versus monobacterial infections, S. mitis/S. mitis group (8.7% vs 4.3%; P=0.032) and Moraxella catarrhalis (4.7% vs 0.5%; P<0.001) were identified more frequently, whereas S. pneumoniae (14.0% vs 28.1%; P<0.001) was identified less frequently, as the dominant infecting species. MICs for individual species were similar for tested antibiotics regardless of polybacterial or monobacterial infection, except Staphylococcus epidermidis for which fluoroquinolone MICs were ≥3 dilutions higher for isolates of this species sourced from polybacterial compared to monobacterial infections. Treatment with besifloxacin resulted in microbial eradication in 79.1% of polybacterial and 92.3% of monobacterial infections (P≤0.005 vs vehicle). DISCUSSION: One in four pediatric bacterial conjunctivitis infections is polybacterial, highlighting the need for a broad-spectrum antibiotic when choosing empiric therapy.

Bactericidal activity of besifloxacin against staphylococci, Streptococcus pneumoniae and Haemophilus influenzae.

OBJECTIVES: Besifloxacin is a novel fluoroquinolone that was recently approved for topical treatment of bacterial conjunctivitis. The compound was shown to be active in vitro against a broad spectrum of bacteria, including isolates resistant to other antibacterials. Here, the bactericidal activity of besifloxacin was evaluated against the most common bacterial conjunctivitis pathogens. METHODS: MIC, MBC and time-kill experiments with besifloxacin and comparators were performed according to CLSI guidelines. Quinolone resistance-determining regions (QRDRs) were sequenced using standard PCR-based techniques. RESULTS: MIC and MBC data indicated that besifloxacin was the most potent fluoroquinolone tested against Staphylococcus aureus (n = 30), Staphylococcus epidermidis (n = 15) and Streptococcus pneumoniae (n = 35), while all fluoroquinolones were highly active against Haemophilus influenzae (n = 40). Besifloxacin MBC:MIC ratios were < or = 4 for 97.5% of all isolates tested (n = 120). All fluoroquinolones tested, as well as tobramycin, were bactericidal, while azithromycin was bactericidal against S. pneumoniae and H. influenzae, but bacteriostatic against the staphylococci. Time-kill assays with all four species showed that besifloxacin caused > or = 1000-fold killing within 2 h for 11 of 12 isolates. Only one isolate treated with moxifloxacin and three ciprofloxacin-treated isolates achieved the same level of bactericidal activity under the same conditions. Unlike the comparator fluoroquinolones, besifloxacin maintained a high potency and bactericidal activity even against strains that contained multiple mutations in the genes encoding DNA gyrase and topoisomerase IV. CONCLUSIONS: Overall, besifloxacin demonstrated rapid bactericidal activity against the four major human pathogens tested here, including isolates that showed in vitro resistance to other fluoroquinolones, beta-lactams, macrolides or aminoglycosides.

Characterization of baseline polybacterial versus monobacterial infections in three randomized controlled bacterial conjunctivitis trials and microbial outcomes with besifloxacin ophthalmic suspension 0.6.

BACKGROUND/PURPOSE: To date, studies examining polymicrobial infections in ocular disease have mostly been limited to keratitis or endophthalmitis. We characterized polybacterial infections compared to monobacterial infections in prior clinical studies evaluating besifloxacin ophthalmic suspension 0.6% for the treatment of bacterial conjunctivitis and report on associated microbiological outcomes. METHODS: In this post-hoc analysis, microbiological data for subjects with conjunctivitis due to one or more than one bacterial species in three previous studies (two vehicle-, one active-controlled) of besifloxacin were extracted. Bacterial species identified at baseline were deemed causative if their colony count equaled or exceeded species-specific prespecified threshold criteria. In subjects with polybacterial infections, the fold-increase over threshold was used to rank order the contribution of individual species. Baseline pathogens and their minimum inhibitory concentrations (MICs) for common ophthalmic antibiotics were compared by infection type, as were microbial eradication rates following treatment with besifloxacin. RESULTS: Of 1041 subjects with culture-confirmed conjunctivitis, 17% had polybacterial and 83% had monobacterial conjunctivitis at baseline. In polybacterial compared to monobacterial infections, Haemophilus influenzae and Streptococcus pneumoniae were identified less frequently as the dominant infecting species (P = 0.042 and P<0.001, respectively), whereas Streptococcus mitis/S. mitis group was identified more frequently as dominant (P<0.001). Viral coinfection was also identified more frequently in polybacterial infections (P<0.001). Staphylococcus aureus was the most common coinfecting species in polybacterial infections and the second most common dominant species in such infections. With few exceptions, MICs for individual species were comparable regardless of infection type. Clinical microbial eradication rates with besifloxacin were high regardless of infection type (P≤0.016 vs vehicle at follow-up visits). CONCLUSIONS: Approximately one in five subjects with bacterial conjunctivitis are infected with more than one bacterial species underscoring the need for a broad-spectrum antibiotic for such infections. Besifloxacin treatment resulted in robust eradication rates of these infections comparable to monobacterial infections. TRIAL REGISTRATION: NCT000622908, NCT00347932, NCT00348348.

Trends in Antibiotic Resistance Among Ocular Microorganisms in the United States From 2009 to 2018.

Importance: Antibiotic resistance in ocular infections can affect treatment outcomes. Surveillance data on evolving antibacterial susceptibility patterns inform the treatment of such infections. Objective: To assess overall antibiotic resistance profiles and trends among bacterial isolates from ocular sources collected during 10 years. Design, Setting, and Participants: This cross-sectional study of longitudinal data from the ongoing, nationwide, prospective, laboratory-based surveillance study, the Antibiotic Resistance Monitoring in Ocular Microorganisms (ARMOR) study, included clinically relevant isolates of Staphylococcus aureus, coagulase-negative staphylococci (CoNS), Streptococcus pneumoniae, Pseudomonas aeruginosa, and Haemophilus influenzae cultured from patients with ocular infections at US centers from January 1, 2009, to December 31, 2018. Main Outcomes and Measures: Minimum inhibitory concentrations were determined for various combinations of antibiotics and species. Odds ratios (ORs) were determined for concurrent antibiotic resistance; analysis of variance and χ2 tests were used to evaluate resistance rates by patient age and geographic region; Cochran-Armitage tests identified changing antibiotic susceptibility trends over time. Results: A total of 6091 isolates (2189 S aureus, 1765 CoNS, 590 S pneumoniae, 767 P aeruginosa, and 780 H influenzae) from 6091 patients were submitted by 88 sites. Overall, 765 S aureus (34.9%) and 871 CoNS (49.3%) isolates were methicillin resistant and more likely to be concurrently resistant to macrolides (azithromycin: S aureus: OR, 18.34 [95% CI, 13.64-24.67]; CoNS: OR, 4.59 [95% CI, 3.72-5.66]), fluoroquinolones (ciprofloxacin: S aureus: OR, 22.61 [95% CI, 17.96-28.47]; CoNS: OR, 9.73 [95% CI, 7.63-12.40]), and aminoglycosides (tobramycin: S aureus: OR, 18.29 [95% CI, 13.21-25.32]; CoNS: OR, 6.28 [95% CI, 4.61-8.56]) compared with methicillin-susceptible isolates (P < .001 for all). Multidrug resistance was observed among methicillin-resistant S aureus (577 [75.4%]) and CoNS (642 [73.7%]) isolates. Antibiotic resistance among S pneumoniae isolates was highest for azithromycin (214 [36.3%]), whereas P aeruginosa and H influenzae isolates showed low resistance overall. Differences in antibiotic resistance were found among isolates by patient age (S aureus: F = 28.07, P < .001; CoNS: F = 11.46, P < .001) and geographic region (S aureus: F = 8.03, P < .001; CoNS: F = 4.79, P = .003; S pneumoniae: F = 8.14, P < .001; P aeruginosa: F = 4.32, P = .005). Small changes in antibiotic resistance were noted over time (≤2.5% per year), with decreases in resistance to oxacillin/methicillin (oxacillin: -2.16%; 95% CI, -3.91% to -0.41%; P < .001) and other antibiotics among S aureus isolates, a decrease in ciprofloxacin resistance among CoNS (-1.38%; 95% CI, -2.24% to -0.52%; P < .001), and an increase in tobramycin resistance among CoNS (0.71%; 95% CI, -0.29% to 1.71%; P = .03). Besifloxacin retained consistently low minimum inhibitory concentrations. Conclusions and Relevance: Antibiotic resistance may be prevalent among staphylococcal isolates, particularly among older patients. In this study, a few small differences in antibiotic resistance were observed by geographic region or longitudinally.

An Evaluation of Staphylococci from Ocular Surface Infections Treated Empirically with Topical Besifloxacin: Antibiotic Resistance, Molecular Characteristics, and Clinical Outcomes.

INTRODUCTION: Understanding antibiotic resistance and toxin profiles among staphylococcal isolates in ocular infections can aid in therapeutic management and infection prevention strategies. We evaluated in vitro antibiotic resistance patterns and molecular traits of staphylococci isolated from patients with ocular surface infections. We also report on clinical outcomes for these patients following empirical treatment with topical besifloxacin ophthalmic suspension 0.6%. METHODS: This was a small observational study. Participating investigators from three clinical sites collected an initial ocular culture from the affected eye of patients presenting with ocular surface infections with presumed staphylococcal etiology. Clinical outcome data for patients with confirmed staphylococcal infections were collated later through retrospective review of patient medical records. Staphylococcal species identification in ocular cultures, in vitro antibiotic susceptibility testing, and PCR-based determination of methicillin resistance cassettes and toxin genotypes were conducted at a central laboratory. Isolates were categorized as susceptible or resistant based on systemic breakpoints, where available. RESULTS: Cultures were collected from 43 patients, and staphylococcal infections were confirmed in 25 patients. Two isolates of Staphylococcus aureus and 27 isolates of Staphylococcus epidermidis were identified. Both S. aureus isolates were methicillin-susceptible, lacked the gene encoding Panton-Valentine leukocidin, and carried few enterotoxin genes. Eight (30%) S. epidermidis were methicillin-resistant (MRSE), and 10 (37%) were ciprofloxacin-resistant. All but two MRSE isolates demonstrated multidrug resistance (MDR), and the staphylococcal cassette chromosome mec (SCCmec) type IVa was detected in five of the eight MRSE isolates. Clinical resolution of the ocular surface infection was reported in all 25 patients following treatment with besifloxacin. CONCLUSIONS: In this study, S. aureus contained few toxins, while SCCmec IVa and MDR was predominant among MRSE from ocular surface infections. Despite significant in vitro fluoroquinolone resistance, there were no cases of treatment failure with topical besifloxacin ophthalmic suspension 0.6%. FUNDING: Bausch Health US, LLC.

Antibiotic Resistance Among Pediatric-Sourced Ocular Pathogens: 8-Year Findings From the Antibiotic Resistance Monitoring in Ocular Microorganisms (ARMOR) Surveillance Study.

BACKGROUND: The Antibiotic Resistance Monitoring in Ocular Microorganisms (ARMOR) study is a nationwide longitudinal antibiotic resistance surveillance program specific to bacterial pathogens commonly encountered in ocular infections. We evaluated in vitro resistance rates and trends among isolates obtained from pediatric patients (≤17 years of age). METHODS: Clinical centers across the United States were invited to submit ocular isolates of Staphylococcus aureus, coagulase-negative staphylococci (CoNS), Streptococcus pneumoniae, Haemophilus influenzae and Pseudomonas aeruginosa to a central laboratory. Minimum inhibitory concentrations for various antibiotic classes were determined by broth microdilution per Clinical and Laboratory Standards Institute guidelines and interpreted as susceptible, intermediate or resistant based on available breakpoints. Longitudinal trends were analyzed using a Cochran-Armitage test for linear trends in a proportion. RESULTS: Of 4829 isolates collected from January 2009 to December 2016, 995 isolates, sourced primarily from hospitals and referral centers, were obtained from pediatric patients (n = 286 H. influenzae, n = 284 S. aureus, n = 213 CoNS, n = 150 S. pneumoniae and n = 62 P. aeruginosa). With few exceptions, P. aeruginosa and H. influenzae were generally susceptible to the antibiotics tested. Of S. aureus and CoNS isolates, respectively, 56% and 72% were resistant to azithromycin and 24% and 47% were methicillin-resistant (MR); concurrent resistance to other drug classes and multidrug resistance (≥3 drug classes) were prevalent among MR staphylococci. Of S. pneumoniae isolates, 38% and 35% demonstrated resistance to azithromycin and penicillin, respectively. Besifloxacin had the lowest minimum inhibitory concentration against the Gram-positive isolates. CONCLUSIONS: These in vitro data suggest antibiotic resistance is common among staphylococcal and pneumococcal isolates collected from pediatric patients with ocular infections. Methicillin resistance was prevalent among staphylococci with many strains demonstrating multidrug resistance. These findings may not be representative of resistance trends in community-based practices.

Antibiotic Resistance Rates by Geographic Region Among Ocular Pathogens Collected During the ARMOR Surveillance Study.

INTRODUCTION: The Antibiotic Resistance Monitoring in Ocular micRoorganisms (ARMOR) study is an ongoing nationwide surveillance program that surveys in vitro antibiotic resistance rates and trends among ocular bacterial pathogens. We report resistance rates by geographic region for isolates collected from 2009 through 2016. METHODS: Staphylococcus aureus, coagulase-negative staphylococci (CoNS), Streptococcus pneumoniae, Haemophilus influenzae, and Pseudomonas aeruginosa isolates from ocular infections were collected at clinical centers across the US and categorized by geographic region based on state. Minimum inhibitory concentrations (MICs) for various antibiotics were determined at a central laboratory, and isolates were classified as susceptible or resistant based on established breakpoints. Geographic differences in methicillin resistance among staphylococci were evaluated by χ2 test with multiple comparisons, whereas geographic differences in mean percentage antibiotic resistance were evaluated by one-way analyses of variance and Tukey's test. RESULTS: Overall, 4829 isolates (Midwest, 1886; West, 1167; Northeast, 1143; South, 633) were evaluated. Across all regions, azithromycin resistance was high among S. aureus (49.4-67.8%), CoNS (61.0-62.8%), and S. pneumoniae (22.3-48.7%), whereas fluoroquinolone resistance ranged from 26.1% to 47.8% among S. aureus and CoNS. Across all regions, all staphylococci were susceptible to vancomycin; besifloxacin MICs were similar to those of vancomycin. Geographic differences were observed for overall mean resistance among S. aureus, S. pneumoniae, and P. aeruginosa isolates (p ≤ 0.005); no regional differences were found among CoNS and H. influenzae isolates. Methicillin resistance in particular was higher among S. aureus isolates from the South and CoNS isolates from the Midwest (p ≤ 0.006). CONCLUSION: This analysis of bacterial isolates from the ARMOR study demonstrated geographic variation in resistance rates among ocular isolates, with greater in vitro resistance apparent in the South and Midwest for some organisms. These data may inform clinicians in selecting appropriate treatment options for ocular infections. FUNDING: Bausch & Lomb, Inc.

Evaluation of Efficacy and Safety of Brimonidine Tartrate Ophthalmic Solution, 0.025% for Treatment of Ocular Redness.

Purpose/Aims: This study assessed the efficacy and safety of brimonidine tartrate ophthalmic solution, 0.025% for treating ocular redness in adult subjects. MATERIALS AND METHODS: This was a single-center, double-masked, randomized, vehicle-controlled, parallel-group study in subjects ≥40 years, with ocular redness. Subjects were randomized 2:1 to brimonidine or vehicle, instilled QID for four weeks. Subjects completed four visits, the last occurring one week after treatment discontinuation. The investigator assessed ocular redness on a scale of 0-4 pre-instillation and 5-240 minutes post-instillation on Day 0, pre-instillation and 5 minutes post-instillation on Days 14 and 28, and on Day35; subjects assessed redness in diaries throughout the 28-day treatment period and following treatment discontinuation. Safety assessments included adverse events (AEs), rebound redness on treatment discontinuation, comprehensive ophthalmic exams, and vital signs. Drop comfort was assessed upon instillation, and 30 seconds and 1 minute post-instillation at Day 0. RESULTS: Fifty-seven subjects (brimonidine, n = 38; vehicle, n = 19) were randomized. Investigator-assessed ocular redness was significantly reduced with brimonidine across the entire post-instillation time period (overall treatment difference: -1.37; P < 0.0001) and at all individual time points (P < 0.0001). Subject-assessed ocular redness was also significantly lower with brimonidine (P ≤ 0.0005). No tachyphylaxis was evident. There were few ocular AEs, all mild to moderate in severity, and no redness rebound was observed upon brimonidine discontinuation. There were no effects on any safety measures, and both brimonidine and its vehicle were reported to be very comfortable. CONCLUSIONS: Brimonidine 0.025% appeared safe, well tolerated, and reduced ocular redness for at least 4 hours. No tachyphylaxis or rebound redness upon treatment discontinuation was observed.

Infectivity of herpes simplex virus type-1 (HSV-1) amplicon vectors in dendritic cells is determined by the helper virus strain used for packaging.

Herpes simplex virus type-1 (HSV-1) amplicon vectors are being explored for a wide range of potential applications, including vaccine delivery and immunotherapy of cancer. While extensive effort has been directed towards the improvement of the amplicon "payload" in these vectors, relatively little attention has been paid to the effect of the packaging HSV-1 strains on the biological properties of co-packaged amplicon vectors. We therefore compared the biological properties of amplicon stocks prepared using a panel of primary HSV-1 isolates, a molecularly cloned strain used to package helper-free amplicons (designated here as F5), and two laboratory isolates (KOS and strain 17, which is the parent of the F5 clone). This analysis revealed considerable inter-strain variability in the ability of amplicon stocks packaged by different primary HSV-1 isolates to efficiently transduce established cell lines and primary human dendritic cells (DC). Amplicons packaged by both the F5 molecularly cloned virus and its laboratory-adapted parent (strain 17) were very inefficient at transducing DC, when compared to amplicons packaged by KOS or by several of the primary virus isolates. These finding have important implications for the future development of improved amplicon-based vaccine delivery systems and suggest that DC tropism may be an instrinsic property of some HSV-1 strains, independent of passage history or molecular cloning.

Besifloxacin Ophthalmic Suspension 0.6% Compared with Gatifloxacin Ophthalmic Solution 0.3% for the Treatment of Bacterial Conjunctivitis in Neonates.

PURPOSE: The aim of this study was to evaluate the safety and efficacy of topical besifloxacin ophthalmic suspension 0.6% compared with gatifloxacin ophthalmic solution 0.3% in the treatment of bacterial conjunctivitis in neonates. METHODS: This was a multicenter, randomized, double-masked, parallel group study. Subjects ≤31 days of age with severity grade ≥1 (scale 0-3) for both conjunctival discharge and conjunctival hyperemia were randomized to besifloxacin or gatifloxacin instilled three times daily for 7 days, and completed five study visits (three clinic visits and two phone calls). Primary endpoints included clinical resolution (absence of both conjunctival discharge and conjunctival hyperemia) at visit 5 (day 8 or 9) and ocular and non-ocular treatment-emergent adverse events (AEs). Bacterial eradication was a secondary endpoint. RESULTS: Thirty-three subjects were included in the intent-to-treat (ITT) population. All were aged <28 days, with a mean (standard deviation) age of 15.5 days (6.0), and 57.6% were female. Twenty-two subjects had culture-confirmed conjunctivitis in at least one eye (modified ITT [mITT] population), most often with Gram-positive bacteria. Visit 5 clinical resolution and bacterial eradication rates were comparable among besifloxacin- and gatifloxacin-treated study eyes (clinical resolution: 12/16 [75.0%] vs. 12/17 [70.6%] for the ITT population, and 11/13 [84.6%] vs. 7/9 [77.8%] for the mITT population; bacterial eradication: 12/13 [92.3%] vs. 8/9 [88.9%] for the mITT population, respectively). No AEs were reported in the besifloxacin treatment group, and AEs reported in the gatifloxacin group were considered not treatment-related. CONCLUSIONS: In this small study in neonates, both besifloxacin and gatifloxacin appeared effective and safe in the treatment of bacterial conjunctivitis. Larger studies are warranted.

Besifloxacin: Efficacy and Safety in Treatment and Prevention of Ocular Bacterial Infections.

This comprehensive review summarizes the mechanism of action, pharmacokinetics, efficacy, and safety of besifloxacin ophthalmic suspension, 0.6% and examines its role in the treatment of ocular surface bacterial infections. Besifloxacin possesses balanced activity against bacterial topoisomerase II (also called DNA gyrase) and topoisomerase IV. It has shown a low potential to select for bacterial resistance in vitro and demonstrated strong in vitro activity against many Gram-positive, Gram-negative, and anaerobic organisms, including methicillin-resistant Staphylococcus aureus and Staphylococcus epidermidis (MRSA and MRSE, respectively). Ocular pharmacokinetic studies have shown that besifloxacin achieves high, sustained concentrations in the tear fluid and conjunctiva following topical administration, with negligible systemic exposure. Large randomized, controlled clinical trials have established the efficacy and safety of besifloxacin administered three times daily for 5 days for treatment of acute bacterial conjunctivitis in both adults and children, with high rates of clinical resolution (up to more than 70% by day 5) and bacterial eradication (more than 90% by day 5), and a low incidence of adverse events. Additionally, besifloxacin applied twice daily for 3 days demonstrated greater efficacy than vehicle in treating bacterial conjunctivitis. Case reports, a large retrospective chart review, and animal studies have provided supporting evidence for the efficacy of besifloxacin in the management of acute bacterial keratitis. There is some evidence to suggest that besifloxacin may provide an advantage over other current-generation fluoroquinolones in antimicrobial prophylaxis for ocular surgery. Besifloxacin is an appropriate option for treatment of bacterial conjunctivitis, and its use in the treatment of bacterial keratitis and lid disorders, as well as for surgical prophylaxis, appears promising and warrants further evaluation.

Surveillance of the Activity of Aminoglycosides and Fluoroquinolones Against Ophthalmic Pathogens from Europe in 2010-2011.

PURPOSE/AIM: Bacterial infections of the ocular surface are commonly treated empirically with broad spectrum antibiotics. Due to concerns over increasing antibiotic resistance, we evaluated current susceptibility patterns of the ocular bacterial pathogens in Europe. MATERIALS AND METHODS: Non-consecutive ocular isolates of Staphylococcus aureus, coagulase-negative staphylococci (CoNS), Streptococcus pneumoniae, Haemophilus influenzae, and Pseudomonas aeruginosa were collected in 2011 from centers in France, Germany, Italy, Poland, Slovak Republic, Spain, and the United Kingdom. Centers were asked to provide similar numbers of methicillin-susceptible and -resistant staphylococcal isolates. Minimum inhibitory concentrations were determined for fluoroquinolones (besifloxacin, ciprofloxacin, moxifloxacin), aminoglycosides (tobramycin, gentamicin, netilmicin), oxacillin, chloramphenicol and erythromycin. Isolates were categorized as susceptible, intermediate, or resistant according to European Committee on Antimicrobial Susceptibility Testing (EUCAST) criteria. RESULTS: A total of 741 ocular isolates were obtained. Antibiotic resistance rates depended not only on the antibiotic and species, but also varied greatly by the country of origin. Resistance to ciprofloxacin, tobramycin, erythromycin, and to a lesser extent, chloramphenicol, was a concern for all staphylococci. Multidrug resistance was common among methicillin-resistant S. aureus (MRSA) and MRCoNS and isolates of S. pneumoniae, H. influenzae, and P. aeruginosa were frequently non-susceptible to erythromycin, beta-lactams, and ciprofloxacin/tobramycin, respectively. Resistance rates showed substantial differences among the seven countries tested. Fluoroquinolones and aminoglycosides showed differences in antibacterial potency and resilience toward the antibiotic resistance mechanisms. CONCLUSIONS: Methicillin-resistant staphylococcal isolates were frequently non-susceptible to a multitude of other antibiotics, making MRSA and MRCoNS a potentially significant concern. The broad range of resistance rates observed across Europe in this study confirms the importance of considering current local resistance patterns when antibacterial agents are chosen for empiric management of ocular infections.

Antibiotic Resistance Among Ocular Pathogens in the United States: Five-Year Results From the Antibiotic Resistance Monitoring in Ocular Microorganisms (ARMOR) Surveillance Study.

IMPORTANCE: The Antibiotic Resistance Monitoring in Ocular Microorganisms (ARMOR) study is the only ongoing nationwide antibiotic resistance surveillance program specific to ocular pathogens. OBJECTIVE: To report resistance rates and trends among common ocular isolates collected during the first 5 years of the ARMOR study. DESIGN, SETTING, AND PARTICIPANTS: This antibiotic resistance surveillance study was performed at an independent central laboratory. Clinical centers across the United States were invited to submit ocular isolates of Staphylococcus aureus, coagulase-negative staphylococci (CoNS), Streptococcus pneumoniae, Haemophilus influenzae, and Pseudomonas aeruginosa. Isolates were collected from January 1, 2009, through December 31, 2013, and analyzed from January 16 to May 15, 2015. MAIN OUTCOMES AND MEASURES: Minimum inhibitory concentrations for various antibiotic classes were determined by broth microdilution according to the guidelines of the Clinical and Laboratory Standards Institute. Minimum inhibitory concentrations were interpreted as susceptible, intermediate, or resistant based on established break points. RESULTS: A total of 3237 ocular isolates (1169 S aureus, 992 CoNS, 330 S pneumoniae, 357 H influenzae, and 389 P aeruginosa) were collected from 72 centers. Methicillin resistance was found among 493 S aureus isolates (42.2%; 95% CI, 39.3%-45.1%) and 493 CoNS isolates (49.7%; 95% CI, 46.5%-52.9%), and methicillin-resistant (MR) isolates had a high probability of concurrent resistance to fluoroquinolones, aminoglycosides, or macrolides (P < .001). Multidrug resistance to at least 3 additional antibiotic classes was found in 428 MR S aureus isolates (86.8%) and 381 MRCoNS isolates (77.3%). All staphylococcal isolates were susceptible to vancomycin. Resistance among S pneumoniae isolates was highest for azithromycin (113 isolates [34.2%]) whereas resistance among P aeruginosa and H influenzae was low against the antibiotics tested. Staphylococcal isolates from elderly patients were more likely to be MR, as were S aureus isolates obtained from the southern United States (P < .001). Methicillin resistance among staphylococci did not increase during the 5-year study period (P ≤ .22), and small decreases in resistance to ciprofloxacin among CoNS and MRCoNS and to tobramycin among CoNS (P ≤ .03) were found. CONCLUSIONS AND RELEVANCE: Methicillin resistance was prevalent among staphylococcal isolates from ocular infections, with many strains demonstrating multidrug resistance. These findings are consistent with resistance trends reported for nonocular staphylococcal isolates. Overall ocular resistance did not increase during the 5-year study period. Continued surveillance of ocular isolates provides critical information to guide selection of topical antibacterials used for empirical management of ocular infections.

Antibacterial efficacy of prophylactic besifloxacin 0.6% and moxifloxacin 0.5% in patients undergoing cataract surgery.

BACKGROUND: The purpose of this study was to investigate the ocular bacterial flora in patients scheduled to undergo cataract surgery and compare the antibacterial effects of besifloxacin ophthalmic suspension 0.6% and moxifloxacin ophthalmic solution 0.5% in these patients. METHODS: This was a prospective, randomized, laboratory-masked clinical trial. Patients received besifloxacin or moxifloxacin "quater in die" or QID (four times a day) for 3 days before cataract surgery in the surgical eye and 1 hour before surgery in the nonsurgical fellow eye. Conjunctival and eyelid swabs were obtained from both eyes at baseline and after treatment, on the day of surgery (Visit 2). Swabs were processed for bacterial colony counts (in terms of colony-forming units) and species identification. In vitro antibiotic susceptibilities of isolates were determined using Clinical and Laboratory Standards Institute breakpoints. RESULTS: Fifty-nine patients (n=28 besifloxacin, n=31 moxifloxacin) completed the study. The majority (73%) of conjunctival samples were culture negative at baseline. The most frequent isolates were coagulase-negative staphylococci (CoNS, 89%), specifically Staphylococcus epidermidis (72%). Both fluoroquinolones reduced the lid CFU values when administered QID for 3 days (P≤0.019), but only besifloxacin reduced the lid CFU estimate 1 hour following instillation of a single drop (P=0.039). Fewer besifloxacin-treated eyes had lids that were culture positive for CoNS at Visit 2 compared with moxifloxacin-treated eyes regardless of dosing regimen (P≤0.03). The minimum inhibitory concentration (MIC90) of besifloxacin against methicillin-resistant S. epidermidis (MRSE) was eightfold lower than that of moxifloxacin. CONCLUSION: Besifloxacin appeared more effective in reducing bacterial counts on eyelids of patients undergoing cataract surgery, with significant reductions as early as 1 hour postdose, compared with moxifloxacin. Besifloxacin was more active in vitro against MRSE.

The facts of death.

Cell death is a physiological process critical for organismal development and required for the removal of damaged cells. Apoptosis, the purposeful killing of cells, is highly regulated, and abnormalities in apoptotic control mechanisms contribute to an assortment of human diseases. Depending on the nature of the death signal, separate independent signaling pathways become activated that coordinately participate in the execution of apoptosis via a common death effector machinery. This article reviews the specific morphological and biochemical hallmarks of apoptosis, the major participating players, and the various signaling pathways involved in triggering this highly complex form of cell death.

Unencapsulated Streptococcus pneumoniae from conjunctivitis encode variant traits and belong to a distinct phylogenetic cluster.

Streptococcus pneumoniae, an inhabitant of the upper respiratory mucosa, causes respiratory and invasive infections as well as conjunctivitis. Strains that lack the capsule, a main virulence factor and the target of current vaccines, are often isolated from conjunctivitis cases. Here we perform a comparative genomic analysis of 271 strains of conjunctivitis-causing S. pneumoniae from 72 postal codes in the United States. We find that the vast majority of conjunctivitis strains are members of a distinct cluster of closely related unencapsulated strains. These strains possess divergent forms of pneumococcal virulence factors (such as CbpA and neuraminidases) that are not shared with other unencapsulated nasopharyngeal S. pneumoniae. They also possess putative adhesins that have not been described in encapsulated pneumococci. These findings suggest that the unencapsulated strains capable of causing conjunctivitis utilize a pathogenesis strategy substantially different from that described for S. pneumoniae at other infection sites.