Yuki Muraoka

Association Between Hyperreflective Foci in the Outer Retina, Status of Photoreceptor Layer, and Visual Acuity in Diabetic Macular Edema

PURPOSE To determine if hyperreflective foci in the outer retina are associated with photoreceptor integrity and the logarithm of minimal angle of resolution (logMAR) visual acuity (VA) in patients with diabetic macular edema (DME). DESIGN Retrospective, observational, cross-sectional study. METHODS Patients (n=76; 108 eyes) with clinically relevant macular edema and no serous retinal detachment were analyzed retrospectively. Spectral-domain optical coherence tomography (SD-OCT) images were obtained for all patients. We investigated the relationship between the hyperreflective foci in the outer retinal layers of the external limiting membrane (ELM) at the fovea and the photoreceptor integrity and VA. RESULTS SD-OCT showed that 58 eyes (53.7%) had hyperreflective foci in the outer retinal layers, and 107 eyes (99.1%) had hyperreflective foci in the inner retinal layers. The logMAR VA was significantly (P<.0001) worse in eyes with hyperreflective foci in the outer retinal layers than in eyes without them (0.463±0.382 vs 0.127±0.206, respectively). Disruption of the ELM line on OCT was significantly (P<.0001, for both comparisons) associated with both hyperreflective foci in the outer retinal layers and poor logMAR VA. Disruption of the junction of the inner and outer segment line (IS/OS) also was related to hyperreflective foci in the outer retinal layers and poor logMAR VA (P<.0001 for both comparisons). CONCLUSIONS The presence of hyperreflective foci in the outer retina is closely associated with a disrupted ELM and IS/OS line on SD-OCT images and decreased VA in DME.

Optical Coherence Tomographic Reflectivity of Photoreceptors beneath Cystoid Spaces in Diabetic Macular Edema

PURPOSE To investigate the relationship between the cystoid spaces in the outer plexiform layer (OPL) and the characteristics of the photoreceptors beneath the cystoid spaces in patients with diabetic macular edema (DME). METHODS In this observational cross-sectional study, 123 eyes of 96 consecutive patients with clinically significant macular edema were retrospectively reviewed. The characteristics of the photoreceptors on optical coherence tomography (OCT) images represented by the external limiting membrane (ELM) and the junction between inner and outer segments (IS/OS), and their association with the overlying cystoid spaces were investigated. RESULTS The areas beneath the cystoid spaces in the OPL had a longer transverse length of disrupted or faint IS/OS and disrupted ELM lines than those without cystoid spaces (P < 0.001, P < 0.001, and P = 0.009). The IS/OS lines beneath the cystoid spaces had higher reflectivity than those in areas without cystoid spaces (P < 0.001). Enlarged cystoid spaces extending from the inner nuclear layer to the OPL were associated with disrupted IS/OS or ELM but not faint IS/OS (P < 0.001, P < 0.001, and P = 0.467). The transverse length of disrupted IS/OS at the fovea was correlated with the logarithm of the minimum angle of resolution (logMAR) more than the association between foveal thickness and logMAR (r = 0.49, P < 0.001 vs. r = 0.28, P = 0.002). The ELM descended to the RPE more frequently in eyes with single-lobulated fluorescein pooling in the foveal avascular zone than those with multi-lobulated pooling (P < 0.001). CONCLUSIONS OCT showed that the cystoid spaces in the OPL were accompanied by photoreceptor damage beneath the cystoid spaces in DME.

Longitudinal and Simultaneous Imaging of Retinal Ganglion Cells and Inner Retinal Layers in a Mouse Model of Glaucoma Induced by N-Methyl-D-Aspartate

PURPOSE To investigate the longitudinal profile of N-methyl-D-aspartate (NMDA) injection-induced damage in retinal ganglion cells (RGCs) by imaging retinal Thy 1-cyan fluorescent protein (CFP) expression and inner retinal layers using a custom-made imaging device containing short-wavelength confocal scanning laser ophthalmoscope (scSLO) and speckle noise-reduced spectral-domain optical coherence tomography (SD-OCT). METHODS Simultaneous scSLO and SD-OCT examinations were performed in Thy 1-CFP mice injected with NMDA (1-20 nanomoles). CFP-expressing RGCs were counted using scSLO images. Ganglion cell complex (GCC: retinal nerve fiber layer, ganglion cell layer, and inner plexiform layer) thickness around the optic disc was measured in SD-OCT images. RESULTS The RGCs rapidly decreased 1 day after NMDA injection in a dose-dependent manner (65.3%, 71.7%, 49.5%, and 27.1% of the preinjection level, 2, 5, 10, and 20 nanomoles, respectively) and continued to decrease slightly (to 53.7%, 44.1%, 28.3%, and 20.2% of the preinjection level on days 14, 2, 5, 10, and 20 nanomoles, respectively). In contrast, dose-dependent reduction of GCC thickness was first detected 4 days after injection. The thickness further decreased to 84.6%, 75.7%, 76.5%, and 71.4% of the preinjection level on day 14 (2, 5, 10, and 20 nanomoles, respectively). CONCLUSIONS NMDA-induced RGC damage is characterized by rapid RGCs loss followed by gradual reduction in GCC thickness. Simultaneous imaging of CFP expression in the RGCs and inner retinal layers provides a sensitive, reliable, and new method for longitudinal evaluation of progressive RGC damage in experimental models of glaucoma.

Age- and hypertension-dependent changes in retinal vessel diameter and wall thickness: an optical coherence tomography study.

PURPOSE To validate and evaluate the reliability of retinal vessel diameter measurements by optical coherence tomography (OCT). The effects of age and hypertension on vessel diameter were also examined. DESIGN Prospective, cross-sectional study. METHODS Two hundred thirty-eight eyes (238 subjects) with no ocular disease were included. Hypertension was present in 106 subjects and absent in 132 subjects. Spectralis HRA+OCT was used to scan a circular region around the optic disc. Outer and inner diameters of the 4 largest retinal arteries and veins were measured using OCT vascular wall reflections, and vessel wall thickness was calculated. RESULTS Intervisit, interexaminer, and interevaluator intraclass correlation coefficients of randomly selected vessel measurements were all greater than 0.90. Mean inner arterial and venous diameters were 87.8 ± 9.4 μm and 113.7 ± 12.5 μm, respectively. The OCT-measured mean inner arterial and venous diameters were significantly correlated to fundus photography caliber measurements (P = .005 and P = .001, respectively). Arterial and venous wall thicknesses were 17.4 ± 2.4 μm and 13.7 ± 2.1 μm, respectively, both of which were highly correlated with subject age (arterial: r = 0.612, P < .001, venous: r = 0.455, P < .001). Additionally, both mean arterial and venous wall thicknesses were significantly greater in subjects with hypertension than in age-matched subjects without hypertension (P = .020 and P = .015, respectively). CONCLUSIONS Retinal vessel diameter measurements obtained with OCT were highly reproducible and vessel wall thicknesses, calculated using outer and inner diameter measurements, were significantly thickened by both aging and systemic hypertension.

Morphologic and Functional Changes in Retinal Vessels Associated with Branch Retinal Vein Occlusion

OBJECTIVE To study the morphologic and functional changes in retinal veins of eyes affected with branch retinal vein occlusion (BRVO) by thin sectioning with optical coherence tomography (OCT). DESIGN Prospective, observational, cross-sectional study. PARTICIPANTS Twenty-five consecutive patients (25 eyes) with acute BRVO. METHODS Major retinal veins, arteries, and arteriovenous (A/V) crossing were examined by sequential thin sectioning by Spectralis HRA+OCT (Heidelberg Engineering, Heidelberg, Germany). The retinal blood flow was mimicked in vitro and scanned with Spectralis HRA+OCT. MAIN OUTCOME MEASURES Morphologic characteristics of normal and BRVO-affected retinal vessels seen in OCT sections. RESULTS Cross-sectional OCT images revealed physiologic retinal vessels as oval configurations with 4 distinctive hyperreflectivities in a line. The vessel walls showed the innermost and outermost hyperreflectivity, and the blood flow showed internal paired hyperreflectivities with an hourglass shape. No eye with disturbed blood flow due to BRVO showed this internal hyperreflectivity pattern. In vitro, OCT sections of the blood within the glass tube without flow showed homogeneous reflectivities. Increased blood flow velocity resulted in the development of heterogeneous internal reflectivity and hourglass-shaped hyperreflectivities. In all eyes with acute BRVO, sequential sectioning with OCT visualized 3-dimensional vascular architecture and the intravascular conditions at the A/V crossing. At the affected A/V crossing, arterial overcrossing was seen in 17 eyes and venous overcrossing was seen in 8 eyes. In eyes with arterial overcrossing, the retinal vein seemed to run deep under the artery at the A/V crossing, and the venous lumen often appeared to be preserved even at the A/V crossing. In all eyes with venous overcrossing, the retinal vein appeared to be compressed and choked between the internal limiting membrane and the arterial wall at the A/V crossing. Optical coherence tomography sectioning showed intravenous thrombi in 21 eyes, and the thrombi were detected downstream of the A/V crossing in all the cases. The detection of thrombus was significantly associated with ischemic pattern in BRVO (P=0.036). CONCLUSIONS In eyes with BRVO, sequential thin sections with OCT visualized 3-dimensional retinal vasculature. The present OCT findings suggest that BRVO may occur by 2 different mechanisms, depending on the relative anatomic positions of the crossing vessels. FINANCIAL DISCLOSURE(S) The author(s) have no proprietary or commercial interest in any materials discussed in this article.

Relationship between Fluorescein Pooling and Optical Coherence Tomographic Reflectivity of Cystoid Spaces in Diabetic Macular Edema

OBJECTIVE To study the characteristics of the reflectivity of the cystoid spaces and serous retinal detachment (SRD) on spectral-domain optical coherence tomography (SD-OCT) and the correlation with fluorescein findings in diabetic macular edema (DME). DESIGN Retrospective, observational, cross-sectional study. PARTICIPANTS Consecutive 134 eyes of 114 patients with clinically significant macular edema for whom SD-OCT and fluorescein angiography (FA) were performed on the same day. METHODS Fluorescein angiography using Heidelberg Retina Angiograph 2 (Heidelberg Engineering, Heidelberg, Germany) and OCT images using Spectralis OCT (Heidelberg Engineering) were obtained. The reflectivity of the cystoid spaces and SRD on the OCT images was evaluated qualitatively and quantitatively and compared with the fluorescein pooling intensity on FA images. MAIN OUTCOME MEASURES The relationship between the fluorescein pooling and the reflectivity characteristics of the cystoid spaces on SD-OCT images. RESULTS A total of 141 cystoid spaces in 101 eyes were delineated on OCT images, and 138 spaces (97.9%) had fluorescein pooling. Fifty-five cystoid spaces (39.9%) with marked fluorescein pooling intensity had lower reflectivity on OCT images than those with modest pooling (12.1±10.4 vs. 22.0±15.4, P < 0.001). The heterogeneity of the reflectivity of the cystoid spaces on the OCT images was associated significantly (P < 0.001) with modest fluorescein pooling. The hyperreflective foci in the cystoid spaces were correlated significantly with modest fluorescein pooling and higher or heterogeneous reflectivity on OCT images (P < 0.001, P < 0.001, and P=0.005, respectively). In addition, the cystoid spaces with microaneurysms had higher or heterogeneous reflectivity on OCT images more frequently than those without microaneurysms (P < 0.001 and P=0.019, respectively). The reflectivity levels in the SRD were significantly (P=0.005) lower than in the cystoid spaces, and only 1 eye (3.3%) had heterogeneous reflectivity on OCT images. CONCLUSIONS The results provided a novel interpretation of fluorescein pooling and OCT characteristics of cystoid spaces and SRD in DME and suggested several mechanisms by which the blood-retinal barrier is disrupted and concomitant edematous changes develop.

Real-Time Imaging of Rabbit Retina with Retinal Degeneration by Using Spectral-Domain Optical Coherence Tomography

Background Recently, a transgenic rabbit with rhodopsin Pro 347 Leu mutation was generated as a model of retinitis pigmentosa (RP), which is characterized by a gradual loss of vision due to photoreceptor degeneration. The purpose of the current study is to noninvasively visualize and assess time-dependent changes in the retinal structures of a rabbit model of retinal degeneration by using speckle noise-reduced spectral-domain optical coherence tomography (SD-OCT). Methodology/Principal Findings Wild type (WT) and RP rabbits (aged 4–20 weeks) were investigated using SD-OCT. The total retinal thickness in RP rabbits decreased with age. The thickness of the outer nuclear layer (ONL) and between the external limiting membrane and Bruchs membrane (ELM–BM) were reduced in RP rabbits around the visual streak, compared to WT rabbits even at 4 weeks of age, and the differences increased with age. However, inner nuclear layer (INL) thickness in RP rabbits did not differ from that of WT during the observation period. The ganglion cell complex (GCC) thickness in RP rabbits increased near the optic nerve head but not around the visual streak in the later stages of the observation period. Hyper-reflective change was widely observed in the inner segments (IS) and outer segments (OS) of the photoreceptors in the OCT images of RP rabbits. Ultrastructural findings in RP retinas included the appearance of small rhodopsin-containing vesicles scattered in the extracellular space around the photoreceptors. Conclusions/Significance In the current study, SD-OCT provided the pattern of photoreceptor degeneration in RP rabbits and the longitudinal changes in each retinal layer through the evaluation of identical areas over time. The time-dependent changes in the retinal structure of RP rabbits showed regional and time-stage variations. In vivo imaging of RP rabbit retinas by using SD-OCT is a powerful method for characterizing disease dynamics and for assessing the therapeutic effects of experimental interventions.

Retinal microstructural changes in eyes with resolved branch retinal vein occlusion: an adaptive optics scanning laser ophthalmoscopy study.

PURPOSE To assess macular photoreceptor abnormalities in eyes with resolved branch retinal vein occlusion (BRVO) using adaptive optics scanning laser ophthalmoscopy (AO-SLO). DESIGN Prospective observational cross-sectional case series. METHODS After complete resolution of macular edema and retinal hemorrhage, 21 eyes (21 patients) with BRVO underwent full ophthalmologic examination and imaging with optical coherence tomography (OCT) and a prototype AO-SLO system. Cone density and spatial mosaic organization were assessed using AO-SLO images. RESULTS Regular parafoveal cone mosaic patterns were clearly visualized with the prototype AO-SLO imaging system in the BRVO-unaffected side. However, in the side of the retina previously affected by the BRVO, cone mosaic patterns were disorganized and dark regions missing wave-guiding cones were apparent. Additionally, retinal capillaries were dilated, no longer had a uniform caliber, and had less direct paths through the retina. In the affected side, parafoveal cone density was significantly decreased, compared with the corresponding retinal area on the unaffected side (P < .001). Furthermore, the hexagonal Voronoi domain ratio and the nearest-neighbor distances were significantly lower than in the unaffected side (P < .05). These parameters were also correlated with photoreceptor layer integrity in the parafovea. CONCLUSIONS After BRVO-associated retinal hemorrhage and macular edema resolved, affected parafoveal cone density decreases and the cone mosaic spatial arrangement is disrupted, becoming more irregular. These cone microstructural abnormalities may extend to parafovea in the BRVO-unaffected side.

Novel VCP modulators mitigate major pathologies of rd10, a mouse model of retinitis pigmentosa.

Neuroprotection may prevent or forestall the progression of incurable eye diseases, such as retinitis pigmentosa, one of the major causes of adult blindness. Decreased cellular ATP levels may contribute to the pathology of this eye disease and other neurodegenerative diseases. Here we describe small compounds (Kyoto University Substances, KUSs) that were developed to inhibit the ATPase activity of VCP (valosin-containing protein), the most abundant soluble ATPase in the cell. Surprisingly, KUSs did not significantly impair reported cellular functions of VCP but nonetheless suppressed the VCP-dependent decrease of cellular ATP levels. Moreover, KUSs, as well as exogenous ATP or ATP-producing compounds, e.g. methylpyruvate, suppressed endoplasmic reticulum stress, and demonstrably protected various types of cultured cells from death, including several types of retinal neuronal cells. We then examined their in vivo efficacies in rd10, a mouse model of retinitis pigmentosa. KUSs prevented photoreceptor cell death and preserved visual function. These results reveal an unexpected, crucial role of ATP consumption by VCP in determining cell fate in this pathological context, and point to a promising new neuroprotective strategy for currently incurable retinitis pigmentosa.

Descriptive epidemiology of spot urine sodium-to-potassium ratio clarified close relationship with blood pressure level: the Nagahama study.

Objectives: We undertook descriptive epidemiology of spot urine sodium-to-potassium ratio (Na/K) in a population sample to clarify the close relationship between Na/K and blood pressure level independently of potential confounding factors. Methods: Study participants consisted of 9144 apparently healthy citizens (aged 54 ± 13 years). All clinical parameters were obtained at baseline. Results: Na/K was significantly higher in hypertensive individuals irrespective of antihypertensive medication status (normotension, 3.12 ± 1.82; untreated hypertension 3.50 ± 1.96; treated hypertension, 3.72 ± 2.53). As urinary Na (&bgr; = 0.092, P < 0.001) and K (&bgr; = −0.050, P < 0.001) levels were inversely associated with BP, Na/K (&bgr; = 0.118, P < 0.001) was more closely associated with BP than Na or K alone, as well as daily salt intake estimated from urinary Na (&bgr; = 0.088, P < 0.001). Several factors were significantly associated with Na/K, namely age, sex, obesity, blood pressure, renal function, salt restriction status, serum phosphate and urinary creatinine level, and fasting period and season at urine sample collection. However, the association between Na/K and BP was independent of these factors (adjusted &bgr; = 0.112, P < 0.001). No direct association was observed between Na/K and large arterial remodeling assessed by pulse wave analysis (P = 0.496) or retinal arteriolar morphological change (P = 0.431). Further, a genome-wide association study failed to identify any particular genotype influencing urinary Na and K levels. Conclusions: Although we clarified several factors that might affect spot urine Na/K, these relationships were not substantial enough to confound the association between urinary Na/K and BP. A simple measure of Na/K might be more representative of salt loading obtained from spot urine samples than Na excretion alone.