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1.
Biomed Opt Express ; 15(4): 2681-2696, 2024 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-38633089

RESUMO

We present proof of concept for a handheld contact-type system capable of simultaneous optical coherence tomography (OCT) imaging of the retina and wide-field digital fundus color photography. The study focuses on demonstrating the feasibility of the proposed approach, particularly for eventual use in pediatric patients during examination under anesthesia in the operating room and in the neonatal intensive care unit. Direct contact of the probe with the cornea allows the photographer to maintain a stable position during imaging, reducing motion artifacts in the OCT images. Additionally, it simplifies the alignment process and increases the field of view of the optics. By integrating OCT and fundus imaging into a single device, the proposed compact modular design eliminates the need for separate, space-consuming systems dedicated to each imaging modality.

2.
JAMA Ophthalmol ; 2024 Mar 21.
Artigo em Inglês | MEDLINE | ID: mdl-38512246

RESUMO

Importance: Evaluation of the microbiological diagnostic profile of multidrug-resistant Pseudomonas aeruginosa keratitis and potential management with rose bengal-photodynamic antimicrobial therapy (RB-PDAT) is important. Objective: To document the disease progression of carbapenemase-resistant P aeruginosa keratitis after an artificial tear contamination outbreak. Design, Setting, and Participants: This retrospective observation case series included 9 patients 40 years or older who presented at Bascom Palmer Eye Institute and had positive test results for multidrug-resistant P aeruginosa keratitis between January 1, 2022, and October 31, 2023. Main Outcomes and Measures: Evaluation of type III secretion phenotype, carbapenemase-resistance genes blaGES and blaVIM susceptibility to antibiotics, and in vitro and in vivo outcomes of RB-PDAT against multidrug-resistant P aeruginosa keratitis. Results: Among the 9 patients included in the analysis (5 women and 4 men; mean [SD] age, 73.4 [14.0] years), all samples tested positive for exoU and carbapenemase-resistant blaVIM and blaGES genes. Additionally, isolates were resistant to carbapenems as indicated by minimum inhibitory concentration testing. In vitro efficacy of RB-PDAT indicated its potential application for treating recalcitrant cases. These cases highlight the rapid progression and challenging management of multidrug-resistant P aeruginosa. Two patients were treated with RB-PDAT as an adjuvant to antibiotic therapy and had improved visual outcomes. Conclusions and Relevance: This case series highlights the concerning progression in resistance and virulence of P aeruginosa and emphasizes the need to explore alternative therapies like RB-PDAT that have broad coverage and no known antibiotic resistance. The findings support further investigation into the potential effects of RB-PDAT for other multidrug-resistant microbes.

3.
Artigo em Inglês | MEDLINE | ID: mdl-38465836

RESUMO

PURPOSE: To quantify the angular-dependence of monofocal intraocular lens (IOL) power. SETTING: Ophthalmic Biophysics Laboratory, Kallam Anji Reddy campus, L V Prasad Eye Institute, Hyderabad, India. DESIGN: Laboratory study. METHODS: Experiments were performed on IOLs from two different manufacturers (APPALENS 207, Appasamy Associates and SN60WF, Alcon Laboratories). IOL powers ranged from 17 to 25 D. The IOLs were mounted in a fluid-filled chamber and the on-axis and off-axis powers were measured using a laser ray tracing system over the central 3 mm zone with delivery angles ranging from -30° to +30° in 5° increments. The position of the best focus was calculated for each IOL at each angle. The angular dependence of IOL power was compared with theoretical predictions. RESULTS: Peripheral defocus increased significantly with increasing incidence angle and power. The peripheral defocus at ±30° increased from 5.8 D to 8.5 D when the power increased from 17.5 D to 24.5 D for APPALENS 207 and from 4.9 D to 7.4 D when the power increased from 17 D to 25 D for SN60WF. The mean difference between the measured and theoretical tangential power at ±30° was 0.50 ± 0.16 D for the APPALENS 207 and -0.40 ± 0.10 D for the SN60WF, independent of IOL power. CONCLUSIONS: IOLs introduce a significant amount of peripheral defocus which varies significantly with IOL power and design. Given that peripheral defocus is related to lens power, replacement of the natural lens (approximately 24 D) with an IOL will produce a significant difference in peripheral defocus profile after surgery.

4.
Lasers Med Sci ; 39(1): 72, 2024 Feb 21.
Artigo em Inglês | MEDLINE | ID: mdl-38379056

RESUMO

The purpose is to assess the efficacy of rose bengal photodynamic antimicrobial therapy (PDAT) using different irradiation energy levels and photosensitizer concentrations for the inhibition of fungal keratitis isolates. Seven different fungi (Aspergillus fumigatus, Candida albicans, Curvularia lunata, Fusarium keratoplasticum, Fusarium solani, Paecilomyces variotii, and Pseudallescheria boydii) were isolated from patients with confirmed infectious keratitis. Experiments were performed in triplicate with suspensions of each fungus exposed to different PDAT parameters including a control, green light exposure of 5.4 J/cm2, 2.7 J/cm2 (continuous and pulsed), and 1.8 J/cm2 and rose bengal concentrations of 0.1%, 0.05%, and 0.01%. Plates were photographed 72 h after experimentation, and analysis was performed to assess fungal growth inhibition. PDAT using 5.4 J/cm2 of irradiation and 0.1% rose bengal completely inhibited growth of five of the seven fungal species. Candida albicans and Fusarium keratoplasticum were the most susceptible organisms, with growth inhibited with the lowest fluence and minimum rose bengal concentration. Fusarium solani, Pseudallescheria boydii, and Paecilomyces variotii were inhibited by lower light exposures and photosensitizer concentrations. Aspergillus fumigatus and Curvularia lunata were not inhibited by any PDAT parameters tested. Continuous and pulsed irradiation using 2.7 J/cm2 produced similar results. Rose bengal PDAT successfully inhibits the in vitro growth of five fungi known to cause infectious keratitis. Differences in growth inhibition of the various fungi to multiple PDAT parameters suggest that susceptibilities to PDAT are unique among fungal species. These findings support modifying PDAT parameters based on the infectious etiology.


Assuntos
Anti-Infecciosos , Byssochlamys , Curvularia , Fusarium , Ceratite , Scedosporium , Humanos , Rosa Bengala/farmacologia , Fármacos Fotossensibilizantes/farmacologia , Ceratite/tratamento farmacológico , Ceratite/radioterapia , Ceratite/microbiologia
5.
Transl Vis Sci Technol ; 12(8): 9, 2023 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-37561510

RESUMO

Purpose: To report the biological stability and postoperative outcomes of a second-generation, single-piece, flexible synthetic cornea in a rabbit model. Methods: Device materials and design were amended to enhance biointegration. Optic skirt design devices were made from compact perfluoroalkoxy alkane with porous expanded polytetrafluoroethylene ingrowth surface overlying the skirt and optic wall. Sixteen devices were implanted into intrastromal pocket in rabbit eyes. Rabbits were randomly assigned to 6- and 12-month follow-up cohorts (n = 8 in each) postoperatively. Monthly examinations and optical coherence tomography assessed cornea-device integration, iridocorneal angle, optic nerve, and retina. Results: There were no intraoperative complications. All devices were in situ at exit, with clear optics. No retroprosthetic membrane, glaucoma, cataract formation, or retinal detachment was observed. Two rabbits in the 6-month group had mild, focal anterior lamella thinning without retraction adjacent to the optic near tight sutures. Three postoperative complications occurred in the 12-month group. One rabbit diagnosed with endophthalmitis was euthanized on day 228. Mild sterile focal retraction of anterior lamella occurred in two rabbits, which were terminated on days 225 and 315. Light microscopic examination of enucleated globes demonstrated fibroplasia with new collagen deposition into the porous scaffold without significant inflammation, encapsulation, or granuloma formation. Conclusions: Clinical evaluations, imaging, and histopathological findings indicate favorable outcomes of this synthetic corneal device in a rabbit model. Early feasibility studies in humans are being planned. Translational Relevance: Favorable 12-month results of the device in rabbits demonstrate vision-restoring potential in corneally blind individuals at high risk of failure with donor keratoplasty.


Assuntos
Córnea , Próteses e Implantes , Animais , Coelhos , Colágeno , Córnea/cirurgia , Próteses e Implantes/efeitos adversos , Retina/diagnóstico por imagem , Retina/cirurgia
6.
Invest Ophthalmol Vis Sci ; 64(2): 12, 2023 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-36753168

RESUMO

Purpose: To determine whether lens mechanical dynamics change with age and with accommodative demands. Methods: Lens thickness microfluctuations were measured using a high-speed custom-built spectral domain optical coherence tomography system in five young adults (20 to 25 years old) at 0 diopters (D), 2 D, 4 D, and maximum accommodative demand and in five prepresbyopes (38 to 45 years old) under relaxed and maximal accommodation. For each state, the measurements were repeated four times during the same session. Images of the central 2-mm zone of the lens comprising 170 A-lines/frame were acquired for 10 seconds, and axial lens thickness change was measured. Lens thickness microfluctuations (µm²/Hz) were assessed by integrating the power spectrum of lens thickness microfluctuations between 0 and 4 Hz. Results: The amplitude of lens microfluctuations was higher in the accommodated states than in the relaxed state in both age groups. Lens microfluctuations were higher in young adult participants than in prepresbyopes, with a significant difference in relaxed and maximally accommodated states (P = 0.04 and P = 0.04). In the young participants, the amplitude of microfluctuations reached a plateau at maximum accommodation. Conclusions: Lens mechanical dynamics are both age and accommodation dependent. The decrease in lens thickness microfluctuations with age is consistent with an age-related increase in lens stiffness or decrease of the ciliary muscle displacement. The lens does not contribute to the high-frequency component of ocular dioptric microfluctuations.


Assuntos
Acomodação Ocular , Cristalino , Adulto Jovem , Humanos , Adulto , Pessoa de Meia-Idade , Cristalino/diagnóstico por imagem , Cristalino/fisiologia , Corpo Ciliar/diagnóstico por imagem
7.
Am J Ophthalmol ; 243: 125-134, 2022 11.
Artigo em Inglês | MEDLINE | ID: mdl-35952754

RESUMO

PURPOSE: To evaluate the intermediate-term clinical outcomes of Rose Bengal Photodynamic Antimicrobial Therapy (RB-PDAT) for infectious keratitis; secondarily, to evaluate the surgical outcomes of individuals who underwent optical keratoplasty after RB-PDAT. DESIGN: Retrospective cohort study. METHOD: A retrospective chart review was performed of 31 eyes from 30 consecutive individuals with infectious keratitis refractory to standard medical therapy who underwent RB-PDAT at the Bascom Palmer Eye Institute between January 2016 and July 2020. Data collected included demographics, risk factors for infectious keratitis, microbiological diagnosis, best spectacle-corrected visual acuity (BCVA), clinical outcomes after RB-PDAT, and complication rates post-keratoplasty. RB-PDAT was performed as described in previous studies. Graft survival was evaluated using Kaplan-Meier curves with log-ranks in individuals who underwent keratoplasty after RB-PDAT. RESULTS: The mean age of the study population was 53 ± 18.0 years. In all, 70% were female; 53.3% self-identified as non-Hispanic White and 43.3% as Hispanic. Mean follow-up time was 28.0 ± 14.4 months. Risk factors included contact lens use (80.6%), history of infectious keratitis (19.3%), and ocular surface disease (16.1%). Cultures were positive for Acanthamoeba (51.6%), Fusarium (12.9%), and Pseudomonas (6.5%). Of the individuals with Acanthamoeba infection, 22.5% were treated with concomitant Miltefosine. Clinical resolution was achieved in 77.4% of patients on average 2.72 ± 1.85 months after RB-PDAT, with 22.5% requiring therapeutic penetrating keratoplasties and 54.8% subsequently requiring optical penetrating keratoplasties. At 2 years, the overall probability of graft survival was 78.7%, and the graft failure rate was 21.3%. CONCLUSION: RB-PDAT is a potential adjunct therapy for infectious keratitis that may reduce the need for a therapeutic penetrating keratoplasty. Patients who undergo keratoplasty after RB-PDAT may have a higher probability of graft survival at 1 year postoperatively.


Assuntos
Anti-Infecciosos , Ceratite , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Antibacterianos/uso terapêutico , Anti-Infecciosos/uso terapêutico , Ceratite/tratamento farmacológico , Ceratoplastia Penetrante , Estudos Retrospectivos , Rosa Bengala/uso terapêutico , Resultado do Tratamento , Acuidade Visual
8.
J Cataract Refract Surg ; 48(9): 1016-1022, 2022 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-35297814

RESUMO

PURPOSE: To determine whether patient-customized paraxial eye models that do not rely on exact ray tracing and do not consider aberrations can accurately predict pseudophakic refraction. SETTING: Bascom Palmer Eye Institute, Miami, Florida. DESIGN: Prospective study. METHODS: Cataract surgery patients with and without a history of refractive surgery were included. Manifest refraction, corneal biometry, and extended-depth optical coherence tomography (OCT) imaging were performed at least 1 month postoperatively. Corneal and OCT biometry were used to create paraxial eye models. The pseudophakic refraction simulated using the eye model was compared with measured refraction to calculate prediction error. RESULTS: 49 eyes of 33 patients were analyzed, of which 12 eyes from 9 patients had previous refractive surgery. In eyes without a history of refractive surgery, the mean prediction error was 0.08 ± 0.33 diopters (D), ranging from -0.56 to 0.79 D, and the mean absolute error was 0.27 ± 0.21 D. 31 eyes were within ±0.5 D, and 36 eyes were within ±0.75 D. In eyes with previous refractive surgery, the mean prediction error was -0.44 ± 0.58 D, ranging from -1.42 to 0.32 D, and the mean absolute error was 0.56 ± 0.46 D. 7 of 12 eyes were within ±0.5 D, 8 within ±0.75 D, and 10 within ±1 D. All eyes were within ±1.5 D. CONCLUSIONS: Accurate calculation of refraction in postcataract surgery patients can be performed using paraxial optics. Measurement uncertainties in ocular biometry are a primary source of residual prediction error.


Assuntos
Lentes Intraoculares , Facoemulsificação , Biometria , Humanos , Implante de Lente Intraocular , Óptica e Fotônica , Estudos Prospectivos , Refração Ocular , Estudos Retrospectivos , Acuidade Visual
9.
J Ophthalmic Inflamm Infect ; 12(1): 11, 2022 Mar 05.
Artigo em Inglês | MEDLINE | ID: mdl-35247126

RESUMO

PURPOSE: To report the increasing trends in Nocardia keratitis species diversity and in vitro antibiotic susceptibility, to demonstrate contact lens wear as a risk factor, and to report visual acuity outcomes after treatment. METHODS: A retrospective clinical case series was performed at a single academic referral center which identified 26 patients with culture-confirmed Nocardia keratitis between 2014 and 2021. A combination of conventional microbiology and molecular techniques were used to identify isolates. Antibiotic susceptibilities were determined using both commercial and in-house laboratory methods. Microbiology and electronic medical records were used to characterize patients' clinical profiles. RESULTS: Patients' median age was 32.5 years with a 2:1 male to female ratio. Eighty-four percent (n = 21/25) of patients were diagnosed within two weeks of symptom onset. Nocardia amikacinitolerans (n = 11/26) was the most recovered Nocardia isolate among study patients. Sixty-four percent (n = 16/25) of all isolates, including all 11 N. amikacinitolerans isolates, were resistant to amikacin. All isolates were susceptible to trimethoprim sulfamethoxazole. Contact lens wear was the leading identified risk factor (n = 23/26) in this population. Median time to resolution was 44 days (n = 23, range: 3-190 days). Seventy-one percent of patients (n = 15/21) had a final visual acuity of 20/40 or better. CONCLUSION: Amikacin resistant Nocardia isolates were the majority in the current study. Trimethoprim sulfamethoxazole may be the preferred alternative antibiotic treatment based on in vitro susceptibilities. Contact lens wear was the major risk factor for Nocardia keratitis in South Florida. Overall visual acuity treatment outcomes of patients were favorable.

10.
Exp Eye Res ; 216: 108951, 2022 03.
Artigo em Inglês | MEDLINE | ID: mdl-35051430

RESUMO

PURPOSE: To determine the effect of temperature on the accommodative response of non-human primate crystalline lenses during simulated accommodation. METHODS: Eight lenses from 7 cynomolgus monkeys (Macaca fascicularis, ages: 4.5-7.3 years; post-mortem time: 17.0 ± 16.4 h) were mounted in a lens stretcher. Stretching experiments were performed on each lens at 24 °C (room temperature), then the tissue was warmed to 35 °C (intraocular temperature) and the stretching experiments were repeated. The lens diameter, thickness, anterior and posterior surface radii of curvature, optical power, and the stretching force (load) were measured at each stretch position and the linear optomechanical relationships were quantified: load-lens diameter, load-thickness, power-load, load-anterior radius, and load-posterior radius. The rate of change for each parameter was quantified by performing a linear regression. The slopes of the linear regressions were compared at the two temperatures using a paired sample t-test. RESULTS: The average changes in the lens with stretching at 24 °C and 35 °C were: 3.07 ± 0.17 and 2.58 ± 0.15 for load-lens diameter (g/mm), -2.38 ± 0.20 and -2.00 ± 0.32 for load-thickness (g/mm), -13.35 ± 1.21 and -13.75 ± 1.26 for power-load (D/g), 0.41 ± 0.10 and 0.34 ± 0.05 for load-anterior radius of curvature (g/mm), and 1.35 ± 0.24 and 1.31 ± 0.35 for load-posterior radius of curvature (g/mm), respectively. The changes in load-diameter and load-thickness with lens stretching were significantly different for the two temperatures. CONCLUSIONS: Temperature influences the change in lens shape observed during simulated accommodation in non-human primate lenses. These results suggest that lens stretching experiments and other optomechanical measurement techniques on ex vivo crystalline lenses be conducted at 35 °C and that the temperature of the tissue sample be documented and maintained constant to ensure repeatability.


Assuntos
Acomodação Ocular/fisiologia , Temperatura Corporal/fisiologia , Cristalino/fisiologia , Estresse Mecânico , Temperatura , Envelhecimento/fisiologia , Animais , Fenômenos Biomecânicos/fisiologia , Cristalino/diagnóstico por imagem , Macaca fascicularis , Tomografia de Coerência Óptica
11.
Transl Vis Sci Technol ; 11(1): 29, 2022 01 03.
Artigo em Inglês | MEDLINE | ID: mdl-35044443

RESUMO

Purpose: To examine and compare the efficacy of in vitro growth inhibition using rose bengal and riboflavin photodynamic antimicrobial therapy (PDAT) for Nocardia keratitis isolates. Methods: Nocardia asteroides complex, Nocardia amikacinitolerans, and Nocardia farcinica species were isolated from patients with confirmed Nocardia keratitis. Isolates were tested against three experimental groups: (1) no photosensitizer/no irradiation, (2) photosensitizer/no irradiation, and (3) photosensitizer/irradiation. Each isolate was prepared in suspension to a concentration of 1.5 × 108 CFU/mL. Bacterial suspensions were mixed with water or prepared 0.1% photosensitizer solution for a final bacterial concentration of 1.5 × 107 CFU/mL. Aliquots of 1 mL were plated on 5% sheep blood agar. Rose bengal and riboflavin PDAT plates were irradiated for 15 minutes with a 525- or 375-nm custom 6-mW/cm2 powered light source for a total fluence of 5.4 J/cm2. All experimental groups were repeated in triplicate. Plates were incubated in a 35°C non-CO2 incubator for 96 hours and photographed. Percent inhibition was evaluated using LabVIEW-based software. Results: All strains of Nocardia tested with 0.1% rose bengal and irradiated for 15 minutes demonstrated statistically significant inhibition of growth (P < 0.05). No other experimental groups displayed any bacterial inhibition. Conclusions: Rose bengal is superior to riboflavin PDAT against selected Nocardia isolates. In vivo testing is warranted to investigate the utility of rose bengal PDAT for severe Nocardia keratitis. Translational Relevance: In vitro results for three clinical strains of Nocardia support the possible use of rose bengal PDAT as a complementary treatment of Nocardia keratitis.


Assuntos
Anti-Infecciosos , Ceratite , Nocardia , Animais , Florida , Humanos , Ceratite/tratamento farmacológico , Riboflavina/farmacologia , Riboflavina/uso terapêutico , Rosa Bengala/farmacologia , Ovinos
12.
Ophthalmic Physiol Opt ; 42(2): 311-318, 2022 03.
Artigo em Inglês | MEDLINE | ID: mdl-34846070

RESUMO

PURPOSE: To evaluate the visual photosensitivity threshold and objective photosensitivity luminance in healthy eyes, thereby providing a normative dataset that will lead to a better understanding of diseases causing photophobia. METHODS: This was a prospective cross-sectional study. Emmetropes whose visual acuity was better than 0.18 logMAR (6/9) with no other ocular abnormality were included. Headache Impact Test-6 and visual light sensitivity questionnaires were administered. Visual photosensitivity threshold was measured subjectively using the ocular photosensitivity analyser. Objective photosensitivity luminance was assessed manually by evaluating videos recorded using an infrared camera and noting the intensity of light at the first squeezing reflex. RESULTS: Seventy five normal subjects (age range, 7-71 years) were included. Median age was 32.7 years (inter-quartile range, 20.3-47.9 years). Forty (53.3%) were males. Median Headache Impact Test score was 38 (inter-quartile range, 36-42) and visual light sensitivity questionnaire score was 11 (inter-quartile range, 8-15). Mean (standard deviation) right eye, left eye and binocular visual photosensitivity threshold was 3.34 (0.78), 3.33 (0.81) and 3.37 (0.78) loglux, respectively. There was a significant negative correlation of visual light sensitivity questionnaire scores with right eye, left eye and binocular visual photosensitivity thresholds, and positive correlation of age with binocular visual photosensitivity thresholds. Mean (standard deviation) right eye, left eye and binocular objective photosensitivity luminance was 3.25 (0.55), 3.35 (0.47) and 3.15 (0.52) loglux, respectively. Age was only positively correlated with binocular objective photosensitivity luminance, and there was no correlation between age and right eye or left eye objective photosensitivity luminance. CONCLUSIONS: The study characterised, for the first time, objective photosensitivity luminance and established normative data for both visual photosensitivity threshold and objective photosensitivity luminance. The data will help in understanding the pathophysiology of diseases causing photophobia, monitoring the disease progression and evaluating treatment modalities.


Assuntos
Fotofobia , Visão Binocular , Adolescente , Adulto , Idoso , Criança , Estudos Transversais , Humanos , Masculino , Pessoa de Meia-Idade , Fotofobia/diagnóstico , Estudos Prospectivos , Visão Binocular/fisiologia , Visão Ocular , Adulto Jovem
13.
Biomed Opt Express ; 12(11): 6746-6761, 2021 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-34858678

RESUMO

We have combined an anterior segment (AS) optical coherence tomography (OCT) system and a wavefront-based aberrometer with an approach that senses ocular wavefront aberrations using the OCT beam. Temporal interlacing of the OCT and aberrometer channels allows for OCT images and refractive error measurements to be acquired continuously and in real-time. The system measures refractive error with accuracy and precision comparable to that of clinical autorefractors. The proposed approach provides a compact modular design that is suitable for integrating OCT and wavefront-based autorefraction within the optical head of the ophthalmic surgical microscope for guiding cataract surgery or table-top devices for simultaneous autorefraction and ocular biometry.

14.
Biomed Opt Express ; 12(8): 5089-5106, 2021 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-34513244

RESUMO

We acquired 1325 nm OCT images of the sclera and ciliary muscle of human subjects. The attenuation coefficients of the sclera and ciliary muscle were determined from a curve fit of the average intensity profile of about 100 A-lines in a region of interest after correction for the effect of beam geometry, using a single scattering model. The average scleral attenuation coefficient was 4.13 ± 1.42 mm-1 with an age-related decrease that was near the threshold for statistical significance (p = 0.053). The average ciliary muscle attenuation coefficient was 1.72 ± 0.88 mm-1, but this value may be an underestimation due to contributions from multiple scattering. Overall, the results suggest that inter-individual variations in scleral attenuation contribute to variability in the quality of transscleral OCT images of the ciliary muscle and the outcome of transscleral laser therapies.

15.
Exp Eye Res ; 212: 108768, 2021 11.
Artigo em Inglês | MEDLINE | ID: mdl-34534541

RESUMO

The mouse lens is frequently used both in vivo and ex vivo in ophthalmic research to model conditions affecting the human lens, such as presbyopia. The mouse lens has a delicate structure which is prone to damage and biomechanical changes both before and after extraction from the whole globe. When not properly controlled for, these changes can confound the biomechanical analysis of mouse lenses. In this study, atomic force microscopy microindentation was used to assess changes in the Young's Modulus of Elasticity of the mouse lens as a function of mouse age and postmortem time. Old mouse lenses measured immediately postmortem were significantly stiffer than young mouse lenses (p = 0.028). However, after 18 h of incubation, there was no measurable difference in lens stiffness between old and young mouse lenses (p = 0.997). This demonstrates the need for careful experimental control in experiments using the mouse lens, especially regarding postmortem time.


Assuntos
Envelhecimento , Cápsula do Cristalino/fisiologia , Cristalino/fisiologia , Microscopia de Força Atômica/métodos , Animais , Elasticidade , Feminino , Cápsula do Cristalino/citologia , Cristalino/citologia , Camundongos , Modelos Animais
16.
PLoS One ; 16(7): e0254519, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34320020

RESUMO

BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) and multidrug-resistant (MDR) S. aureus strains are well recognized as posing substantial problems in treating ocular infections. S. aureus has a vast array of virulence factors, including superantigens and enterotoxins. Their interactions and ability to signal antibiotics resistance have not been explored. OBJECTIVES: To predict the relationship between superantigens and methicillin and multidrug resistance among S. aureus ocular isolates. METHODS: We used a DNA microarray to characterize the enterotoxin and superantigen gene profiles of 98 S. aureus isolates collected from common ocular sources. The outcomes contained phenotypic and genotypic expressions of MRSA. We also included the MDR status as an outcome, categorized as resistance to three or more drugs, including oxacillin, penicillin, erythromycin, clindamycin, moxifloxacin, tetracycline, trimethoprim-sulfamethoxazole and gentamicin. We identified gene profiles that predicted each outcome through a classification analysis utilizing Random Forest machine learning techniques. FINDINGS: Our machine learning models predicted the outcomes accurately utilizing 67 enterotoxin and superantigen genes. Strong correlates predicting the genotypic expression of MRSA were enterotoxins A, D, J and R and superantigen-like proteins 1, 3, 7 and 10. Among these virulence factors, enterotoxin D and superantigen-like proteins 1, 5 and 10 were also significantly informative for predicting both MDR and MRSA in terms of phenotypic expression. Strong interactions were identified including enterotoxins A (entA) interacting with superantigen-like protein 1 (set6-var1_11), and enterotoxin D (entD) interacting with superantigen-like protein 5 (ssl05/set3_probe 1): MRSA and MDR S. aureus are associated with the presence of both entA and set6-var1_11, or both entD and ssl05/set3_probe 1, while the absence of these genes in pairs indicates non-multidrug-resistant and methicillin-susceptible S. aureus. CONCLUSIONS: MRSA and MDR S. aureus show a different spectrum of ocular pathology than their non-resistant counterparts. When assessing the role of enterotoxins in predicting antibiotics resistance, it is critical to consider both main effects and interactions.


Assuntos
Proteínas de Bactérias/genética , Farmacorresistência Bacteriana Múltipla/genética , Enterotoxinas/genética , Staphylococcus aureus/metabolismo , Superantígenos/genética , Antibacterianos/farmacologia , Proteínas de Bactérias/metabolismo , Clindamicina/farmacologia , Enterotoxinas/metabolismo , Oftalmopatias/microbiologia , Oftalmopatias/patologia , Genótipo , Gentamicinas/farmacologia , Humanos , Aprendizado de Máquina , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Staphylococcus aureus Resistente à Meticilina/metabolismo , Testes de Sensibilidade Microbiana , Fenótipo , Infecções Estafilocócicas/microbiologia , Infecções Estafilocócicas/patologia , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/isolamento & purificação , Superantígenos/metabolismo , Fatores de Virulência/genética , Fatores de Virulência/metabolismo
17.
Cornea ; 40(8): 1036-1043, 2021 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-34190718

RESUMO

PURPOSE: To evaluate the in vivo corneal changes after Rose Bengal photodynamic antimicrobial therapy (RB-PDAT) treatment in New Zealand White rabbits. METHODS: Sixteen rabbits were divided into 5 groups. All groups underwent deepithelialization of an 8 mm diameter area in the central cornea. Group 1: balanced salt solution drops only, group 2: 0.2% RB only, group 3: green light exposure (525 nm, 5.4 J/cm2) only, group 4: 0.1% RB-PDAT, and group 5: 0.1% RB-PDAT. All rabbits were followed clinically. Group 5 rabbits were followed using anterior segment optical coherence tomography (AS-OCT) and clinically. On day 35 after initial treatment, 1 rabbit from group 5 was re-exposed to green light (5.4 J/cm2) to evaluate reactivation of the remaining RB dye, and terminal deoxynucleotyl transferase-mediated UTP-biotin-nick-end labeling assay was performed on corneal cryosections. RESULTS: Complete reepithelization was observed, and corneas remained clear after treatment in all groups. In group 5, AS-OCT revealed a cross-linking demarcation line. AS-OCT showed RB fluorescence and collagen cross-linking in all treated eyes of group 5 animals after 5 weeks of treatment. Photobleached RB retention in the corneal stroma was corroborated by fluorescence confocal microscopy on frozen sections. There was no evidence of a sustained cytotoxic effect through terminal deoxynucleotyl transferase-mediated UTP-biotin-nick-end labeling at 5 weeks. CONCLUSIONS: RB-PDAT with 0.1% RB is a safe procedure. There was no difference clinically and on histopathology compared with control groups. In eyes where RB dye is retained in the corneal stroma after 1 month of treatment, oxidative stress is not evidenced at long term.


Assuntos
Colágeno/uso terapêutico , Reagentes de Ligações Cruzadas/farmacologia , Infecções Oculares Bacterianas/tratamento farmacológico , Ceratite/tratamento farmacológico , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/uso terapêutico , Rosa Bengala/uso terapêutico , Animais , Modelos Animais de Doenças , Infecções Oculares Bacterianas/diagnóstico , Feminino , Corantes Fluorescentes/uso terapêutico , Ceratite/diagnóstico , Microscopia Confocal , Projetos Piloto , Coelhos , Tomografia de Coerência Óptica/métodos
18.
Exp Eye Res ; 209: 108653, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-34097905

RESUMO

It is well known that human crystalline lens shape, dimensions and optical properties change throughout life and influence whole eye refraction. However, it is not clear if lens properties are associated with other ocular parameters. The purpose of the present study was to investigate the relationship of corneal and external globe dimensions with adult lens diameter (LD), lens thickness (LT) and lens power (LP) in order to determine if external factors influence lens properties. Postmortem human eyes (n = 66, age = 20-78 years) were obtained from the Ramayamma International Eye Bank, Hyderabad, India. Globe antero-posterior length (GAPL) and mean (average of horizontal and vertical) diameters of cornea (MCD) and globe (MGD) were measured using digital calipers. Eyes were dissected to produce ocular structures that contain the lens maintained in its accommodating framework, including intact zonules, ciliary body and sections of sclera. Specimens were mounted in a mechanical lens stretching system. LD, LT and LP were measured using high magnification retro-illumination photography, slit illumination photography and Scheiner principle-based optical system respectively in the unstretched (accommodated) state. Relationships between external globe and corneal dimensions and LD, LT or LP were assessed by multiple regression analysis. Age (0.012 ± 0.003 mm/year; p<0.001) and GAPL (0.185 ± 0.045 mm/mm; p<0.001) were significant (p<0.0001) predictors of LD. After adjusting for age-related increases, LD appears to be positively correlated with GAPL. Age (0.010 ± 0.004 mm/year; p = 0.009) and GAPL (-0.143 ± 0.060 mm/mm; p = 0.02) were significant (p = 0.001) predictors of LT. After adjusting for the age-related increase, LT appears to be negatively correlated with GAPL. Only age was a significant predictor of LP (-0.26 ± 0.04 D/year; p<0.001). The results suggest that, apart from aging, lens diameter and thickness are dependent on the anteroposterior length of the eye globe. Lens power is not influenced by globe dimensions.


Assuntos
Acomodação Ocular/fisiologia , Envelhecimento/fisiologia , Biometria/métodos , Córnea/anatomia & histologia , Olho/anatomia & histologia , Cristalino/anatomia & histologia , Refração Ocular/fisiologia , Adulto , Idoso , Córnea/crescimento & desenvolvimento , Olho/crescimento & desenvolvimento , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Tamanho do Órgão , Adulto Jovem
19.
Invest Ophthalmol Vis Sci ; 62(3): 15, 2021 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-33688927

RESUMO

Purpose: To characterize the peripheral defocus of isolated human crystalline lenses and its age dependence. Methods: Data were acquired on 116 isolated lenses from 99 human eyes (age range, 0.03-61 years; postmortem time, 40.1 ± 21.4 hours). Lenses were placed in a custom-built combined laser ray tracing and optical coherence tomography system that measures the slopes of rays refracted through the lens for on-axis and off-axis incidence angles. Ray slopes were measured by recording spot patterns as a function of axial position with an imaging sensor mounted on a positioning stage below the tissue chamber. Delivery angles ranged from -30° to +30° in 5° increments using a 6 mm × 6 mm raster scan with 0.5-mm spacing. Lens power at each angle was calculated by finding the axial position that minimizes the root-mean-square size of the spot pattern formed by the 49 central rays, corresponding to a 3-mm zone on-axis. The age dependence of the on-axis and off-axis optical power and the relative peripheral defocus (difference between off-axis and on-axis power) of lenses were quantified. Results: At all angles, lens power decreased significantly with age. Lens power increased with increasing delivery angle for all lenses, corresponding to a shift toward myopic peripheral defocus. There was a statistically significant decrease in the lens peripheral defocus with age. Conclusions: The isolated human lens power increases with increasing field angle. The lens relative peripheral defocus decreases with age, which may contribute to the age-related changes of ocular peripheral defocus during refractive development.


Assuntos
Envelhecimento/fisiologia , Cristalino/fisiologia , Refração Ocular/fisiologia , Erros de Refração/fisiopatologia , Acomodação Ocular/fisiologia , Adolescente , Adulto , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Pessoa de Meia-Idade , Tomografia de Coerência Óptica , Adulto Jovem
20.
Biomed Opt Express ; 12(1): 272-287, 2021 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-33520385

RESUMO

Rose bengal photodynamic antimicrobial therapy (RB-PDAT) treats corneal infection by activating rose bengal (RB) with green light to produce singlet oxygen (1O2). Singlet oxygen dosimetry can help optimize treatment parameters. We present a 1O2 dosimeter for detection of 1O2 generated during experimental RB-PDAT. The system uses a 520 nm laser and an InGaAs photoreceiver with bandpass filters to detect 1O2 luminescence during irradiation. The system was validated in RB solutions and ex vivo in human donor eyes. The results demonstrate the feasibility of 1O2 dosimetry in an experimental model of RB-PDAT in the cornea.

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