Mariela C Aguilar

Assessment of rose bengal versus riboflavin photodynamic therapy for inhibition of fungal keratitis isolates.

PURPOSE To compare the in vitro effect of rose bengal and riboflavin as photosensitizing agents for photodynamic therapy (PDT) on fungal isolates that are common causes of fungal keratitis. DESIGN Experimental study. METHODS Three isolates (Fusarium solani, Aspergillus fumigatus, Candida albicans) recovered from patients with confirmed fungal keratitis were used in the experiments. Isolates were grown on Sabouraud-Dextrose agar, swabbed, and prepared in suspension, and 1 mL aliquots were inoculated onto test plates in triplicate. Test plates were separated into 5 groups: Group 1, no treatment; Group 2, 0.1% rose bengal alone; Group 3, 518 nm irradiation alone; Group 4, riboflavin PDT (riboflavin + 375 nm irradiation); and Group 5, rose bengal PDT (rose bengal + 518 nm irradiation). Irradiation was performed over a circular area using either a green light-emitting diode (LED) array (peak wavelength: 518 nm) or an ultraviolet-A LED array (peak wavelength: 375 nm). Test plates were irradiated with an energy density of 5.4 J/cm(2). Later, plates were placed in a 30 C incubator and observed for growth. RESULTS Rose bengal-mediated PDT successfully inhibited the growth of all 3 fungal isolates in the irradiated area. All other groups exhibited unrestricted growth throughout the plate. CONCLUSIONS Rose bengal-mediated PDT successfully inhibited the growth of 3 types of fungi. No other experimental groups, including riboflavin-mediated PDT, had any inhibitory effect on the isolates. The results might be useful for the treatment of patients suffering from corneal infection.

New Methods for Quantification of Visual Photosensitivity Threshold and Symptoms

Purpose Visual photosensitivity is a common symptom difficult to measure and diagnose, and is found in many ocular and neurological disorders. We developed two novel reproducible quantitative assessments of visual photosensitivity. Methods We designed and built the ocular photosensitivity analyzer (OPA), an automated instrument to determine light intensity visual photosensitivity threshold (VPT), and developed the Visual Light Sensitivity Questionnaire-8 (VLSQ-8), an eight-question survey to assess the presence and severity of photosensitivity symptoms. We evaluated the test–retest variability and obtained normative values of these two approaches in 35 healthy normal subjects, distributed evenly over five age groups from eight to 60 years. Each subject underwent two test sessions, each with VLSQ-8, eye examination, and OPA, four weeks apart, between April 2015 and June 2016. Results Log-transformed VPTs (log10lux) and VLSQ-8 results were highly reproducible between the two sessions (VPT intraclass correlation coefficient [ICC] = 0.86; 95% confidence interval [CI] = 0.71–0.93; binocular testing, VLSQ-items ICC range = 0.53–0.87). No consistent significant differences in VPTs were found with monocular (P = 0.053, session 1) or binocular (P = 0.26) testing. Subjects in age group >30 to 40 years had significantly higher VPTs than those in other age groups (P ≤ 0.011) except the >40 to 50 years age group (P = 0.11). Photosensitivity symptoms assessed by the VLSQ-8 generally were low and highly reproducible with ≥88% of responses between the 2 sessions being within one category of each other. Conclusions Our results provide reliability data and normative results toward validation of two novel approaches to quantify visual photosensitivity and provide support for their potential use in ocular and neurologic conditions as well as in clinical trials. Translational Relevance The new quantitative photosensitivity approaches are potential measures to characterize disease severity, monitor disease progression, and evaluate treatment efficacy.

Rose Bengal Photodynamic Antimicrobial Therapy: A Novel Treatment for Resistant Fusarium Keratitis

Purpose: To evaluate the efficacy of rose bengal PDAT for the management of a patient with multidrug-resistant Fusarium keratoplasticum keratitis unresponsive to standard clinical treatment. Methods: This case report presents a clinical case of F. keratoplasticum keratitis not responsive to standard medical care. In vitro studies from patients culture isolated responded to rose bengal PDAT. Patient received two treatments with rose bengal 0.1% and exposure to green light with a total energy of 2.7 J/cm2. Results: In vitro results demonstrated the efficacy of rose bengal PDAT a multidrug-resistant F. keratoplasticum species. There was complete fungal inhibition in our irradiation zone on the agar plates. In the clinical case, the patient was successfully treated with 2 sessions of rose bengal PDAT, and at 8-month follow-up, there was neither recurrence of infection nor adverse effects to report. Conclusions: Rose bengal PDAT is a novel treatment that may be considered in cases of aggressive infectious keratitis. Further studies are needed to understand the mechanisms of PDAT in vivo.

Robotic remote controlled stereo slit lamp

Purpose Our purpose was to develop a robotic remotely operated stereo slit lamp system allowing three-dimensional stereo viewing and recording of the patients examination via local area network, Internet, and satellite. Methods A commercial slit lamp was modified to accept motors and servos to permit control of all optical and mechanical components of the device. The custom graphical user interface with dual high-resolution real-time stereoscopic imaging, control/position indicators, overview video, and audio were transmitted via local area network, Internet, and satellite. Under University of Miami Institutional Review Board authorization, Internet connectivity enabled multiple examiners to simultaneously view and control the slit lamp and to collaboratively discuss diagnosis and treatment options. The remote clinicians used a tablet, laptop, or desktop computer to view and control the slit lamp. Results The network, Internet, satellite-connected system was controllable from the United States, Europe, and Canada while acquiring high-resolution, real-time video in all subjects. Control of the slit lamp through Ethernet, WiFi, and 4G exhibited total system latencies of 464 ± 58, 483 ± 64, and 870 ± 66 milliseconds when transmitting within the continent, and Ethernet control exhibited a latency of 606 ± 130 milliseconds when transmitting between continents. High- and low-magnification images of healthy volunteers were acquired by a remote clinician. Conclusions The robotic remotely operated stereo slit lamp system allows three-dimensional stereo viewing and recording of the patients examination via local area network, Internet, and satellite. Translational Relevance The robotic remotely controlled stereo slit lamp system enables remote examination of human subjects.

Comparison of continuous versus pulsed photodynamic antimicrobial therapy for inhibition of fungal keratitis isolates in vitro (Conference Presentation)

Fungal keratitis can lead to pain and impaired vision. Current treatment options include antifungal agents and therapeutic penetrating keratoplasty. An emerging option for the management of keratitis is photodynamic antimicrobial therapy (PDAT) which uses a photosensitizer rose bengal activated with green light. Utilizing a pulsed irradiation, rather than the standard continuous irradiation may have a similar antimicrobial effect with less total energy. This study is to compare pulsed and continuous rose bengal mediated PDAT for inhibition of six fungal isolates on agar plates: Fusarium solani, Fusarium keratoplasticum, Aspergillus fumigatus, Candida albicans, Paecilomyces variotti, and Pseudoallescheria boydii. Isolates were mixed with 0.1% rose bengal and exposed to three irradiation conditions: (1) 30-minute continuous (10.8J/cm2), (2) 15-minute continuous (5.4J/cm2), (3) 30-minute pulsed (5.4J/cm2). Plates were photographed at 72 hours and analyzed with custom software. At 72 hours, 30-minute continuous rose bengal mediated PDAT inhibited all six fungal species. Fungal inhibition was analogous between 30-minute continuous and 30-minute pulsed test groups, with the exception of A. fumigatus. The 15-minute continuous irradiation was less effective when compared to both 30-minute continuous and 30-minute pulsed groups. These in vitro results demonstrate the potential strength of pulsed rose bengal mediated PDAT as an adjunct treatment modality for fungal keratitis.

Differential flow rate of commercially available triamcinolone with and without preservative through small-gauge needles.

OBJECTIVE To compare the flow rate through different gauge needles of triamcinolone acetonide with benzyl alcohol (TABA) versus preservative-free triamcinolone acetonide injectable suspension (TAIS). MATERIALS AND METHODS Experimental injections of 1 mL of TABA or TAIS were performed through a 27-, 30-, or 32-gauge needle. A piezoelectric pressure transducer connected to a personal computer was placed between the syringe and needle to record the force-time relationship and assess the flow rate (mL/s). Measurements were performed in triplicate. RESULTS Flow rates were similar between preparations using a 27-gauge needle. Using a 30-gauge needle, the flow rate was 0.058 mL/s for TABA and 0.178 mL/s for TAIS. Using a 32-gauge needle, no flow was sustained by TABA, whereas the flow rate of TAIS was 0.114 mL/s (P < .05). CONCLUSION TAIS can be injected through a 32-gauge needle without the risk of obstruction. Occlusions were seen with TABA using the 30- and 32-gauge needles.

Treatment of retinal detachment with an encircling band and buckle implant: A comparative pilot study between poly (styrene- b -isobutylene- b -styrene) (SIBS) and trimethyl terminated polydimethylsiloxane (PDMS)

Purpose: To demonstrate the biocompatibility of SIBS implants as compared to PDMS implants in the treatment of retinal detachment in New Zealand White (NZW) rabbit model.1,2 Introduction: Scleral encircling bands, fixation rings and buckles are utilized for closure of retinal breaks and retina reattachment. The FDA approved PDMS-implant is associated with several post-operative complications, involving thick-fibrotic encapsulations. SIBS, an elastomeric triblock copolymer, was recently FDA approved for use in a cardiovascular drug eluting stent (TAXUSTM, Boston Scientific Corp., MA) and showed excellent biocompatibility and slow drug release capability. Materials and Methods: SIBS (9-mol%-styrene) implants were fabricated (InnFocus LLC, USA) to match PDMS implants (Labtician, Inc, Canada) dimensions. 5 NZW rabbits received SIBS and 4, PDMS-implants. Post-operative exam sequence: day 1 and 2, week 1, 2, 3, 4 and 6, and monthly thereafter for up to 1 year. Anatomohistopathology exams sequence: one SIBS animal at 6 weeks and one animal of each treatment group at 3 and 6-months, and two at 12-months. Results: SIBS compared to PDMS animals exhibited less inflammation and a better buckling effect during the first 6 weeks. At POD 9 months, the conjunctival injection in the SIBS rabbit was none as opposed to the PDMS value and the buckling effect for both groups were equal. There were no visible signs of encapsulation with SIBS. There were no infections in the 9 animals and none of the implants extruded thus far (<10 months). Conclusion: SIBS encircling bands, sleeves, and buckle implants are well tolerated in the rabbit model.