SubFoveal Choroidal Imaging in High Myopic Nepalese Cohort.

Purpose: Evidence suggests that choroid is thinner in myopes as compared to nonmyopes. However, choroidal thickness varies with the refractive error, age, axial length, and ethnicity. The purpose of this study was to determine the subfoveal choroidal thickness (SFCT) in high myopic Nepalese subjects and to investigate its association with the mean spherical equivalent refractive error (MSE), axial length, and age. Methods: Ninety-two eyes of 92 high myopic subjects (MSE ≤ -6 diopters) and 83 eyes of 83 emmetropic subjects (MSE: 0.00 Diopters) were included in the study. SFCT was assessed using spectral domain optical coherence tomography, and the axial length was measured using partial coherence interferometry. SFCT was measured manually using the inbuilt tool within the imaging software. Results: SFCT in the high myopic subjects was significantly thinner (mean ± SD: 224.17 ± 68.91 µm) as compared to the emmetropic subjects (353.24 ± 65.63 µm) (mean difference, 127.76 ± 130.80 µm, and p < 0.001). In high myopic subjects, there was a significant negative correlation of choroidal thickness with the axial length (rho = -0.75; p < 0.001) and MSE (rho = -0.404; p < 0.01). Regression analysis demonstrated a decrease of choroidal thickness by 40.32 µm (p < 0.001) for every 1 millimeter increase in the axial length and by 11.65 µm (p < 0.001) for every 1 diopter increase in the MSE. Conclusion: High myopic Nepalese subjects had significantly thinner choroid as compared to emmetropes. The MSE and axial length were inversely correlated with the SFCT. Age had no effect on SFCT in this study. These findings may have implications in interpreting choroidal thickness values in clinical and epidemiological studies in myopes, especially in the south Asian population.

Baseline metrics that may predict future myopia in young children.

PURPOSE: We used baseline data from the PICNIC longitudinal study to investigate structural, functional, behavioural and heritable metrics that may predict future myopia in young children. METHODS: Cycloplegic refractive error (M) and optical biometry were obtained in 97 young children with functional emmetropia. Children were classified as high risk (HR) or low risk (LR) for myopia based on parental myopia and M. Other metrics included axial length (AXL), axial length/corneal radius (AXL/CR) and refractive centile curves. RESULTS: Based on the PICNIC criteria, 46 children (26 female) were classified as HR (M = +0.62 ± 0.44 D, AXL = 22.80 ± 0.64 mm) and 51 (27 female) as LR (M = +1.26 ± 0.44 D, AXL = 22.77 ± 0.77 mm). Based on centiles, 49 children were HR, with moderate agreement compared with the PICNIC classification (k = 0.65, p < 0.01). ANCOVA with age as a covariate showed a significant effect for AXL (p < 0.01), with longer AXL and deeper anterior chamber depth (ACD) (p = 0.01) in those at HR (differences AXL = 0.16 mm, ACD = 0.13 mm). Linear regression models showed that central corneal thickness (CCT), ACD, posterior vitreous depth (PVD) (=AXL - CCT - ACD-lens thickness (LT)), corneal radius (CR) and age significantly predicted M (R = 0.64, p < 0.01). Each 1.00 D decrease in hyperopia was associated with a 0.97 mm elongation in PVD and 0.43 mm increase in CR. The ratio AXL/CR significantly predicted M (R = -0.45, p < 0.01), as did AXL (R = -0.25, p = 0.01), although to a lesser extent. CONCLUSIONS: Although M and AXL were highly correlated, the classification of pre-myopic children into HR or LR was significantly different when using each parameter, with AXL/CR being the most predictive metric. At the end of the longitudinal study, we will be able to assess the predictability of each metric.

Binocular vision findings in normally-sighted school aged children who used digital devices.

PURPOSE: To determine the binocular vision status in normally-sighted school aged children who used digital devices. METHODS: A cross-sectional study was conducted at B.P. Koirala Lions Center for Ophthalmic Studies, Kathmandu, Nepal for a duration of one year. One hundred and eighty school aged children (71 female and 109 male) aged 7 to 17 years were included in the study. All the children underwent detailed ophthalmic and binocular vision examinations. The duration of the use of digital devices by the children were asked to either the parents or guardians present at the time of the study. The study participants were divided into two groups: children who used digital devices for the last six months (users group) and those who hadn't used digital devices for the last six months (non users group). The users group was again divided into two subgroups: children who used digital devices for less than 3 hours per day and a day per week (low digital device users subgroup) and children who used digital devices for more than 3 hours per day and all days in a week (high digital device users subgroup). RESULTS: Accommodative amplitudes, accommodative facility, and positive fusional vergence for both near and distance were significantly reduced in the high digital device users group than in the low digital device users subgroup (p <0.01). Stereo acuity, near point of convergence, and negative fusional vergences for both near and distance were not statistically significantly different between the two subgroups. Prevalence of accommodative and vergence anomalies (except convergence insufficiency) was more in the high digital device users subgroup than in the low digital device users subgroup (p<0.01). CONCLUSIONS: Children who used digital devices for a significantly greater amount of time had significantly reduced amplitudes of accommodation, accommodative facility, and positive fusional vergence both at near and distance.

Visual function in guinea pigs: behavior and electrophysiology.

CLINICAL RELEVANCE: Guinea pig visual function is characterised based on behavioural and electrophysiological measures and retinal ganglion cell density is examined to further develop the guinea pig as a model of human ocular conditions. BACKGROUND: Guinea pigs are an important model of human ocular conditions. Here, guinea pig spatial frequency discrimination, pattern and full-field photopic electroretinography (ERG), and retinal ganglion cell distribution were investigated. METHODS: Adult guinea pigs (n = 6) were included. Optomotor responses to square-wave gratings from 0.3 to 2.4 cycles per degree (cpd) were assessed. Pattern ERG responses were recorded using square-wave gratings from 0.025 to 0.25 cpd at 100% contrast, alternating at a temporal frequency of 1.05 Hz. Full-field ERG responses were recorded using a 10.0 cd.s/m2 flash. Ganglion cell density was determined histologically from retinal whole mounts. RESULTS: Maximum spatial frequency discrimination was 1.65 ± 0.49 cpd for stimuli rotating temporally to nasally and 0.75 ± 0.16 cpd for stimuli rotating nasally to temporally. For pattern ERG, a maximum amplitude of 3.50 ± 1.16 µV for the first negative to positive peak (N1P1) was elicited with a 0.025 cpd grating, and 2.5 ± 0.1 µV for the positive to second negative peak (P1N2) was elicited with a 0.05 cpd grating. For full-field ERG, a-wave amplitude was 19.2 ± 4.24 µV, b-wave amplitude was 33.6 ± 8.22 µV, and the PhNR was 24.0 ± 5.72 µV. Peak retinal ganglion cell density was 1621 ± 129 cells/mm2, located 1-2 mm superior to the optic nerve head. CONCLUSION: Guinea pigs show directional selectivity for stimuli moving in the temporal to the nasal visual field. Guinea pigs demonstrate a quantifiable PhNR in the full-field ERG and negative and positive waveforms in the pattern ERG. The visual streak is located in the superior retina.

Retinal ganglion cell ablation in guinea pigs.

Guinea pigs are a common model of human ocular conditions; however, their visual function has not been fully characterized. The purpose of this study was to determine the contributions of retinal ganglion cells to structural and functional measures in guinea pigs. Healthy adult guinea pigs (n = 12) underwent unilateral optic nerve crush. Retinal structure was assessed with spectral domain optical coherence tomography (OCT), and thickness of the ganglion cell/nerve fiber layer (GC/NFL) was determined. Visual function was assessed with optomotor tracking of a drifting grating and light adapted electroretinograms (ERGs). From flash ERGs, a-wave, b-wave, oscillatory potentials (OPs), and photopic negative response (PhNR) were analyzed. From pattern ERGs, N1P1 and P1N2 were analyzed. Histological studies were done at various time points for ganglion cell quantification. Optomotor tracking was absent in optic nerve crush eyes following optic nerve crush. Significant thinning of the GC/NFL was evident four weeks following the crush. Flash ERGs revealed a significant reduction in the OP1 amplitude two weeks following crush (P < 0.01) and in the PhNR amplitude six weeks following crush (P < 0.01). There were no significant changes in a-wave, b-wave, or pattern ERG responses (P > 0.05 for all). In vivo OCT imaging showed progressive thinning of inner retinal layers. Ganglion cell density, quantified histologically, was significantly reduced by 75% in the optic nerve crush eye compared to the control eye at four weeks following crush. These findings indicate that retinal ganglion cells contribute to the PhNR and OP1 components of the full field flash ERG, but not significantly to the pattern ERG in guinea pigs. This study demonstrates that OCT imaging and full field flash ERGs are valuable in assessing retinal ganglion cell loss in vivo in guinea pigs and will help to further establish the guinea pig as a model of human ocular pathologies.

In vivo assessment of foveal geometry and cone photoreceptor density and spacing in children.

The fovea undergoes significant developmental changes from birth into adolescence. However, there is limited data examining cone photoreceptor density, foveal pit shape, and foveal avascular zone (FAZ) size in children. The purpose of this study was to determine whether overall foveal structure differs as a function of age and refractive status in children. Forty-eight healthy children (ages 5.8 to 15.8 years) underwent optical coherence tomography imaging to quantify foveal point thickness and foveal pit diameter, depth, and slope. Adaptive optics scanning laser ophthalmoscope (AOSLO) images of foveal capillaries and cone photoreceptors were acquired in a subset of children to quantify FAZ metrics and cone densities at 0.2, 0.3, and 0.5 mm eccentricities. Results show that foveal pit and FAZ metrics were not related to age, axial length, or refractive status. However, linear cone density was lower in myopic versus non-myopic children at eccentricities of 0.2 mm (mean ± SD = 50,022 ± 5,878 cones/mm2 vs 58,989 ± 4,822 cones/mm2, P < 0.001) and 0.3 mm (43,944 ± 5,547 cones/mm2 vs 48,622 ± 3,538 cones/mm2, P < 0.001). These results suggest FAZ and foveal pit metrics do not systematically differ with age in children, while myopic eyes have decreased linear cone density near the foveal center. Significance Statement: The development of the fovea begins prior to birth and continues through the early teenage years until it reaches adult-like properties. Although the majority of changes during childhood are related to the maturation and migration of cone photoreceptors, in vivo data describing cone packing in children is limited. We assessed overall foveal structure in children as young as 5.8 years old by quantifying cone density and spacing, foveal avascular zone size, and foveal pit morphometry to investigate potential structural differences as a function of age and refractive status. While foveal avascular zone and foveal pit metrics did not significantly differ with age, results indicate that myopic children have lower linear cone densities close to the foveal center compared to non-myopic children.

The optic nerve head, lamina cribrosa, and nerve fiber layer in non-myopic and myopic children.

The purpose of this study was to evaluate the optic nerve head, lamina cribrosa, retina, and choroid in school age children using spectral domain optical coherence tomography (SD-OCT) and to assess these structural parameters in relation to age, axial length, and refractive error. Healthy children, ages 11.15 ± 2.62 years (range 6-15 years, n = 53), underwent cycloplegic autorefraction, biometry, and SD-OCT imaging in both eyes. Images were analyzed using custom written programs in MATLAB, after adjustment for lateral magnification. Peripapillary retinal nerve fiber layer (RNFL) thickness, retinal and choroidal thicknesses, Bruch's membrane opening (BMO) area, minimum rim width (MRW), and anterior lamina cribrosa surface depth (ALCSD) were determined and analyzed with age, axial length, and refraction. Results show that axial length increased and refractive error became more myopic with increasing age (R2 = 0.25, ß = 0.18, P < 0.0001 and R2 = 0.27, ß = -0.37, P < 0.0001, respectively). Minimum foveal thickness and central 1 mm retinal thickness increased with increasing age (R2 = 0.15, ß = 2.38, P < 0.01 and R2 = 0.11, ß = 3.16, P < 0.05, respectively). Age-adjusted raw values for peripapillary RNFL thickness decreased with increasing axial length (R2 = 0.11, ß = -3.18, P < 0.05); however, this relationship was not present when image magnification was corrected (R2 = 0.07, ß = 2.72, P = 0.09). BMO area increased with myopic refractive error (R2 = 0.16, ß = -0.10, P < 0.01). Age-adjusted vertical cup-to-disc ratio decreased with increasing axial length and myopic refractive error (R2 = 0.12, ß = -0.05, P < 0.05 and R2 = 0.11, ß = 0.03, P = 0.05, respectively). Mean MRW, mean ALCSD, and peripapillary choroidal thickness were not associated with age, axial length, or refraction. Mean MRW was significantly thinner in eyes with deeper ALCS (R2 = 0.41, ß = -0.83, P < 0.0001). These findings provide normal values for retinal and optic nerve head parameters in school age children, and also suggest that ocular remodeling occurs in some structures in school age children with normal eye growth and during early stages of myopia development.

Twenty-four hour ocular and systemic diurnal rhythms in children.

PURPOSE: Ocular diurnal rhythms have been implicated in myopia, glaucoma, diabetes, and other ocular pathologies. Ocular rhythms have been well described in adults; however, they have not yet been fully examined in children. The goal of this study was to investigate ocular and systemic diurnal rhythms over 24 h in children. METHODS: Subjects, ages 5 to 14 years (n = 18), wore a light, sleep, and activity monitor for one week to assess habitual sleep/wake patterns, then underwent diurnal measurements every 4 h for 24 h. Measurements included blood pressure, heart rate, body temperature, intraocular pressure (IOP), ocular biometry, and optical coherence tomography imaging. Saliva was collected for melatonin and cortisol analysis. Mean ocular perfusion pressure was calculated from IOP and blood pressure. Central corneal thickness, corneal power, anterior chamber depth, lens thickness, vitreous chamber depth, and axial length were determined from biometry. Total retinal thickness, retinal pigment epithelium (RPE) + photoreceptor outer segment thickness, photoreceptor inner segment thickness, and choroidal thickness were determined for a 1 mm diameter centred on the fovea. Subjects' amplitude and acrophase of diurnal variation for each parameter were determined using Fourier analysis, and mean acrophase was calculated using unit vector averaging. RESULTS: Repeated measures analysis of variance (ANOVA) showed that all parameters except anterior chamber depth exhibited significant variations over 24 h (p ≤ 0.005 for all). Axial length underwent diurnal variation of 45.25 ± 6.30 µm with an acrophase at 12.92 h, and choroidal thickness underwent diurnal variation of 26.25 ± 2.67 µm with an acrophase at 1.90 h. IOP was approximately in phase with axial length, with a diurnal variation of 4.19 ± 0.50 mmHg and acrophase at 11.37 h. Total retinal thickness underwent a significant diurnal variation of 4.09 ± 0.39 µm with an acrophase at 15.04 h. The RPE + outer segment layer was thickest at 3.25 h, while the inner segment layer was thickest at 14.95 h. Melatonin peaked during the dark period at 2.36 h, and cortisol peaked after light onset at 9.22 h. CONCLUSIONS: Ocular and systemic diurnal rhythms were robust in children and similar to those previously reported in adult populations. Axial length and IOP were approximately in phase with each other, and in antiphase to choroidal thickness. These findings may have important implications in myopia development in children.

Immunotoxin-Induced Ablation of the Intrinsically Photosensitive Retinal Ganglion Cells in Rhesus Monkeys.

Purpose: Intrinsically photosensitive retinal ganglion cells (ipRGCs) contain the photopigment melanopsin, and are primarily involved in non-image forming functions, such as the pupillary light reflex and circadian rhythm entrainment. The goal of this study was to develop and validate a targeted ipRGC immunotoxin to ultimately examine the role of ipRGCs in macaque monkeys. Methods: An immunotoxin for the macaque melanopsin gene (OPN4), consisting of a saporin-conjugated antibody directed at the N-terminus, was prepared in solutions of 0.316, 1, 3.16, 10, and 50 µg in vehicle, and delivered intravitreally to the right eye of six rhesus monkeys, respectively. Left eyes were injected with vehicle only. The pupillary light reflex (PLR), the ipRGC-driven post illumination pupil response (PIPR), and electroretinograms (ERGs) were recorded before and after injection. For pupil measurements, 1 and 5 s pulses of light were presented to the dilated right eye while the left pupil was imaged. Stimulation included 651 nm (133 cd/m2), and 4 intensities of 456 nm (16-500 cd/m2) light. Maximum pupil constriction and the 6 s PIPR were calculated. Retinal imaging was performed with optical coherence tomography (OCT), and eyes underwent OPN4 immunohistochemistry to evaluate immunotoxin specificity and ipRGC loss. Results: Before injection, animals showed robust pupil responses to 1 and 5 s blue light. After injection, baseline pupil size increased 12 ± 17%, maximum pupil constriction decreased, and the PIPR, a marker of ipRGC activity, was eliminated in all but the lowest immunotoxin concentration. For the highest concentrations, some inflammation and structural changes were observed with OCT, while eyes injected with lower concentrations appeared normal. ERG responses showed better preserved retinal function with lower concentrations. Immunohistochemistry showed 80-100% ipRGC elimination with the higher doses being more effective; however this could be partly due to inflammation that occurred at the higher concentrations. Conclusion: Findings demonstrated that the OPN4 macaque immunotoxin was specific for ipRGCs, and induced a graded reduction in the PLR, as well as, in ipRGC-driven pupil response with concentration. Further investigation of the effects of ipRGC ablation on ocular and systemic circadian rhythms and the pupil in rhesus monkeys will provide a better understanding of the role of ipRGCs in primates.

In Vivo Imaging of the Retina, Choroid, and Optic Nerve Head in Guinea Pigs.

PURPOSE: Guinea pigs are increasingly being used as a model of myopia, and may also represent a novel model of glaucoma. Here, optical coherence tomography (OCT) imaging was performed in guinea pigs. In vivo measurements of retinal, choroidal, and optic nerve head parameters were compared with histology, and repeatability and interocular variations were assessed. METHODS: OCT imaging and histology were performed on adult guinea pigs (n = 9). Using a custom program in MATLAB, total retina, ganglion cell/nerve fiber layer (GC/NFL), outer retina, and choroid thicknesses were determined. Additionally, Bruch's membrane opening (BMO) area and diameter, and minimum rim width were calculated. Intraobserver, interocular, and intersession coefficients of variation (CV) and intraclass correlation coefficients (ICC) were assessed. RESULTS: Retina, GC/NFL, outer retina and choroid thicknesses from in vivo OCT imaging were 147.7 ± 5.8 µm, 59.2 ± 4.5 µm, 72.4 ± 2.4 µm, and 64.8 ± 11.6 µm, respectively. Interocular CV ranged from 1.8% to 11% (paired t-test, p = 0.16 to 0.81), and intersession CV ranged from 1.1% to 5.6% (p = 0.12 to 0.82), with the choroid showing the greatest variability. BMO area was 0.192 ± 0.023 mm2, and diameter was 493.79 ± 31.89 µm, with intersession CV of 3.3% and 1.7%, respectively. Hyper reflective retinal layers in OCT correlated with plexiform and RPE layers in histology. CONCLUSION: In vivo OCT imaging and quantification of guinea pig retina and optic nerve head parameters were repeatable and similar between eyes of the same animal. In vivo visibility of retinal cell layers correlated well with histological images. ABBREVIATIONS: optic nerve head (ONH), retinal ganglion cell (RGC), spectral domain optical coherence tomography (SD-OCT), enhanced depth imaging (EDI), minimum rim width (MRW), hematoxylin and eosin (H & E).