New culture technique of human eliminable feeder-assisted target cell sheet production.

Cultured cell sheets for transplantation generally have been co-cultured with animal feeder cells, which carry risks because of different species and results in non-contact culture between the feeder and target cells. We developed a new technique to produce human eliminable feeder-assisted target cell sheets by novel human-derived genetically modified feeder cells. Three genes (human-derived telomerase reverse transcriptase gene, enhanced green fluorescent protein gene, and herpes simplex virus thymidine kinase gene) were transducted into human stromal cells, which enabled genetically modified feeder cells to be immortalized, labeled, and eliminated as needed. A target cell sheet was produced as one sheet by assisting the genetically modified feeder cells and successfully transplanted in vivo without their contamination. Genetically modified human eliminable feeder cells could be a promising tool for cultivated cell sheet transplantation.

A 27-gauge instrument system for transconjunctival sutureless microincision vitrectomy surgery.

OBJECTIVES: To evaluate the efficiency, preliminary safety, and feasibility of a 27-gauge instrument system for transconjunctival microincision vitrectomy surgery (MIVS) in a variety of vitreoretinal diseases. DESIGN: Experimental, interventional case series. PARTICIPANTS: Thirty-one eyes (31 patients) underwent a variety of vitreoretinal procedures using the 27-gauge transconjunctival MIVS system to treat epiretinal membrane (n = 10), idiopathic macular holes (n = 7), diabetic vitreous hemorrhage (n = 5), vitreous opacity with suspicion of intraocular lymphoma (n = 4), focal diabetic traction retinal detachment (n = 3), macular traction syndrome (n = 1), and macular edema secondary to central retinal vein occlusion (n = 1). METHODS: We developed a 27-gauge instrument system that includes an infusion line, a high-speed vitreous cutter, an illumination system, and a variety of vitreoretinal instruments, such as membrane forceps and sharp-tipped endophotocoagulation probes. The duty cycle of the 27- and 25-gauge cutters was measured for several cut rates using a high-speed imaging camera. Infusion and aspiration rates were measured using balanced saline solution (BSS) and porcine vitreous with different aspiration levels. Surgical outcomes, including anatomic success, visual outcomes, operating times, and intraoperative and postoperative complications, were evaluated. MAIN OUTCOME MEASURES: Duty cycle of cutters, infusion and aspiration rates, and surgical results of 27-gauge vitrectomy. RESULTS: Although the infusion and aspiration rates of the 27-gauge system measured in BSS were reduced to an average of 62% and 80%, respectively, compared with those of the 25-gauge system, the duty cycle of the 27-gauge cutter, 61% at 1000 cpm and 38% at 1500 cpm, was equal to or better than those of the 25-gauge cutter (62% and 28%, respectively). Analysis of the fluid dynamics showed that vented gas-forced infusion can be set to range from 20 to 30 mmHg to control intraocular pressure (IOP) during 27-gauge vitrectomy. Anatomic success was achieved in all study eyes (100%); 20 eyes (65%) had visual improvement of 3 lines or more. No eyes required conversion to larger gauge instrument. All sclerotomies self-sealed without hypotony (IOP < or = 7 mmHg) from 1 day postoperatively. CONCLUSIONS: Although the fluid dynamics and cutting efficiency of 27-gauge instruments are lower compared with 25-gauge MIVS, the 27-gauge system is feasible and may reduce concerns about wound sealing-related complications in selected cases. FINANCIAL DISCLOSURE(S): Proprietary or commercial disclosure may be found after the references.

Prevalence and features of keratitis with quantitative polymerase chain reaction positive for cytomegalovirus.

PURPOSE: To assess corneal scrapings and aqueous humor samples analyzed by polymerase chain reaction (PCR) that were positive for cytomegalovirus (CMV) in patients with keratitis of unknown origin and to investigate their clinical manifestations. DESIGN: Retrospective, interventional case series. PARTICIPANTS: Seventy-eight patients with epithelial (n=37), stromal (n=12), or endothelial keratitis (n=29) of unknown origin examined at the Osaka University Medical Hospital. METHODS: Clinical examination and tears, corneal scrapings, and aqueous humor specimens were evaluated by real-time PCR for CMV. MAIN OUTCOME MEASURES: Quantification of CMV DNA at the diagnosis of each type of keratitis with unknown origin and monitoring during the therapeutic course for CMV-positive cases. RESULTS: No cases of epithelial or stromal keratitis had CMV DNA. Seven of 29 corneal endotheliitis cases (24.1%) were positive for CMV. Cytomegalovirus-positive cases of corneal endotheliitis characterized by localized corneal edema and keratic precipitates included 4 patients who had undergone penetrating keratoplasty and were refractory to the treatment for graft rejection and 3 patients with idiopathic endotheliitis. Cytomegalovirus DNA copy numbers were estimated and ranged from 6.3x10(4) to 3.6x10(6)/ml. In all positive cases, the numbers of CMV DNA copies decreased within weeks during treatment with systemic and topical ganciclovir (GCV) combined with a topical steroid. Five eyes (62.5%) had clinical improvement. In cases of endothelial keratitis, diabetes mellitus was significantly higher in patients positive for CMV (71.4%) than in patients negative for CMV (18.2%, P=0.016, chi-square test). CONCLUSIONS: A total of 24.1% of cases with corneal edema of unknown origin were CMV positive and should be included in the differential diagnosis of idiopathic corneal endotheliitis or graft edema after penetrating keratoplasty, especially for bullous keratopathy. Real-time PCR for CMV, based on the diagnosis and monitoring of the clinical course, may be useful. Cytomegalovirus corneal endotheliitis requires early appropriate treatment using GCV. Because clinical remission after GCV may depend on the area of normal endothelium, early diagnosis and therapy are important for CMV corneal endotheliitis.

Nuclear cataract caused by a lack of DNA degradation in the mouse eye lens.

The eye lens is composed of fibre cells, which develop from the epithelial cells on the anterior surface of the lens. Differentiation into a lens fibre cell is accompanied by changes in cell shape, the expression of crystallins and the degradation of cellular organelles. The loss of organelles is believed to ensure the transparency of the lens, but the molecular mechanism behind this process is not known. Here we show that DLAD ('DNase II-like acid DNase', also called DNase IIbeta) is expressed in human and murine lens cells, and that mice deficient in the DLAD gene are incapable of degrading DNA during lens cell differentiation--the undigested DNA accumulates in the fibre cells. The DLAD-/- mice develop cataracts of the nucleus lentis, and their response to light on electroretinograms is severely reduced. These results indicate that DLAD is responsible for the degradation of nuclear DNA during lens cell differentiation, and that if DNA is left undigested in the lens, it causes cataracts of the nucleus lentis, blocking the light path.

Spectral-domain optical coherence tomographic findings in myopic foveoschisis.

PURPOSE: The purpose of this study was to investigate the sensitivity of various scan modes of spectral-domain optical coherence tomography (SD-OCT) for detecting pathologies that may accompany myopic foveoschisis, which is important for making surgical decisions and planning surgical strategies for myopic foveoschisis. METHODS: Twenty-one eyes of 18 patients were diagnosed with myopic foveoschisis by SD-OCT. Patients were examined with SD-OCT using both a five-line raster scan and a three-dimensional scan at the same visit. The detection rates of pathologies such as macular hole, epiretinal membrane, retinal vascular microfolds, internal limiting membrane detachment, paravascular microhole, and photoreceptor inner and outer segments defect were compared between modes. RESULTS: The three-dimensional scanning mode of the SD-OCT tended to be superior to both the one-line and five-line raster scanning modes for detecting inner and outer segments, epiretinal membrane, macular hole, paravascular microhole, and internal limiting membrane. The three-dimensional mode had a significantly higher detection rate (71%) of retinal vascular microfolds than both the 1-line (19%, P < 0.01) and 5-line raster scanning modes (33%, P < 0.05). CONCLUSION: The three-dimensional scan mode of SD-OCT was more sensitive for detecting the pathologies that accompany myopic foveoschisis, providing important information for vitreous surgery.

Clinical findings in unilateral acute idiopathic maculopathy: new findings in acute idiopathic maculopathy.

We report a case of unilateral acute idiopathic maculopathy (UAIM) with new clinical findings. A 34-year-old Japanese man had a neurosensory retinal detachment (approximately 5 disk diameters) with yellowish-white exudates at the macula in the left eye (visual acuity (VA) 0.4). Fluorescein angiography (FA) showed early hypofluorescent spots and late pooling in the subretinal space. Three weeks after onset, indocyanine green angiography (IA) showed numerous hypofluorescent spots at the lesion. Optical coherence tomography (OCT) showed subretinal fluids and an elevated choroidal lesion with low reflectivity, suggesting choroidal edema. The VA and fundus appearance spontaneously resolved without treatment three months after onset. The VA was 1.0 six months after onset. Irregular pigmentation remained at the macular lesion. The main UAIM pathology may be outer retinal layer and retinal pigment epithelial inflammation. FA, IA, and OCT suggested that choroidal inflammation may be involved in the pathogenesis of UAIM.

Foveal microstructure and visual acuity after retinal detachment repair: imaging analysis by Fourier-domain optical coherence tomography.

PURPOSE: To evaluate foveal microstructural changes in eyes with anatomically successful repair of rhegmatogenous retinal detachments (RRDs). DESIGN: Retrospective, consecutive, observational case series. PARTICIPANTS: Fifty-three eyes of 51 consecutive patients with macula-on RRDs (15 eyes) or macula-off RRDs (38 eyes) after anatomically successful surgical repair. METHODS: A microscopic fundus examination was conducted followed by Fourier-domain optical coherence tomography (FD-OCT) to assess the postoperative foveal microstructure. The correlation between the postoperative best-corrected visual acuity (BCVA) and microstructural findings at the fovea was evaluated. MAIN OUTCOME MEASURES: Images of the foveal microstructure obtained by FD-OCT and the BCVA measured on the same day. RESULTS: We obtained FD-OCT images a mean of 10.3+/-7.3 months (range, 1-25) postoperatively. Foveal anatomic abnormalities were detected in 33 eyes (62%); disruption of the junction between the photoreceptor inner and outer segments (IS/OS) in 23 eyes (43%), of which 9 eyes (39%) had a disrupted external limiting membrane (ELM); residual subretinal fluid in 6 eyes (11%), epiretinal membranes in 12 eyes (23%), and cystoid macular edema in 2 eyes (4%). Disruption of the photoreceptor IS/OS junction was observed only in macula-off eyes, whereas other microstructural abnormalities were observed in both macula-on and macula-off eyes. In preoperative macula-off eyes, the postoperative BCVA was significantly correlated with the integrity of the photoreceptor IS/OS and ELM signals detected by FD-OCT postoperatively (r=0.805; P<0.001). Of the 16 eyes followed by FD-OCT, the photoreceptor IS/OS junction was restored in 7 (64%) of the 11 eyes with a disrupted back-reflection line from the IS/OS junction, but without disrupted ELM signals at the initial examination. Of the 5 eyes with disrupted back-reflection lines from both IS/OS junction and ELM at the initial examination, the photoreceptor layer was not restored completely during the follow-up period in any eyes. CONCLUSIONS: After anatomically successful RRD repair, FD-OCT is a valuable, noninvasive tool for evaluating foveal microstructural changes. The integrity of the photoreceptor IS/OS junction and ELM signals detected by FD-OCT may account for visual restoration in patients with preoperative macula-off RRDs. Preservation of the ELM postoperatively may predict the subsequent restoration of the photoreceptor layer.

Microincision vitrectomy surgery and intravitreal bevacizumab as a surgical adjunct to treat diabetic traction retinal detachment.

PURPOSE: To investigate the feasibility and efficacy of microincision vitrectomy surgery (MIVS) combined with intravitreal bevacizumab (IVB) as a surgical adjunct for treating traction retinal detachment (TRD) secondary to severe proliferative diabetic retinopathy (PDR). DESIGN: Retrospective, comparative, consecutive, interventional case series. PARTICIPANTS: Seventy-one eyes of 59 consecutive patients who underwent primary vitrectomy for diabetic TRD and were followed up for more than 6 months after surgery. METHODS: Eyes that received IVB (1 mg) as a preoperative adjunct followed by MIVS (IVB/MIVS group) from November 2005 through December 2007 were compared with eyes that underwent conventional 20-gauge pars plana vitrectomy (20-g PPV group) from September 2003 through October 2005. MAIN OUTCOME MEASURES: Primary and ultimate anatomic success, intraoperative and postoperative complications, and final visual success with at least 6 months of follow-up. RESULTS: This series included 38 eyes (33 patients) in the IVB/MIVS group and 33 eyes (26 patients) in the 20-g PPV group. The primary and ultimate anatomic success rates (95% vs. 91% and 100% in both groups, respectively) and the mean visual acuity changes did not differ significantly between groups; the surgical time and intraoperative bleeding in the IVB/MIVS group decreased significantly compared with the 20-g PPV group (P<0.001). The rate of visual improvement of 3 lines or more at the 6-month follow-up was 68% in the IVB/MIVS group and 49% in the 20-g PPV group, respectively. Progression of the preexisting TRD after IVB occurred in 7 eyes (18%). Absence of previous laser photocoagulation (P = 0.025) and the presence of a ring-shaped fibrovascular membrane (P = 0.013) were relevant findings in eyes with these IVB-induced complications. CONCLUSIONS: Intravitreal bevacizumab plus MIVS offers comparable anatomic success compared with conventional 20-gauge PPV in patients with TRD resulting from severe PDR. This technique shortens the surgical time with fewer intraoperative complications and favorable visual recovery. However, caution should be taken because of rapid progression of the preexisting TRD after IVB in some patients. FINANCIAL DISCLOSURE(S): The author(s) have no proprietary or commercial interest in any materials discussed in this article.

Retardation of retinal vascular development in apelin-deficient mice.

OBJECTIVE: Apelin is an endogenous ligand for the G protein-coupled receptor, APJ, and participates in multiple physiological processes. To identify the roles of endogenous apelin, we investigated the phenotype of apelin-deficient (apelin-KO) mice. METHODS AND RESULTS: Apelin-KO mice showed impaired retinal vascularization and ocular development, which were analyzed by histology, immunohistochemistry, real-time polymerase chain reaction, and the mouse corneal micropocket assay. Apelin-KO mice showed significantly impaired retinal vascularization in the early postnatal period. Retinal apelin/APJ mRNAs were transiently upregulated during the first 2 postnatal weeks but were undetectable in adults. There were no differences in VEGF or FGF2 mRNA expression, or in the morphology and localization of GFAP-positive astrocytes, in the apelin-KO retinas at P5. The corneal pocket assay showed that angiogenic responses to VEGF and FGF2 were remarkably decreased in apelin-KO mice. The reduced responses to VEGF and FGF2 in apelin-KO mice were partially restored by apelin, but apelin alone did not induce angiogenesis. CONCLUSIONS: Our results suggest that spatiotemporally regulated apelin/APJ signaling participates in retinal vascularization in a cooperative manner with VEGF or FGF2, and contributes to normal ocular development.

Tissue plasminogen activator-assisted vitrectomy: surgical drainage of submacular hemorrhage.

A thick subretinal hemorrhage involving the macula is currently treated with pneumatic displacement or surgical removal. We evaluated the long-term effects of tPA-assisted surgical drainage of a submacular hemorrhage by reviewing the medical records of 12 eyes with submacular hemorrhage followed for a mean follow-up time of 6.9 years. The final best corrected visual acuity (BCVA) improved in 10 eyes (83%), was unchanged in 2 (17%), and was reduced in 0 (0%). The mean preoperative BCVA was 6/200, and the best postoperative BCVA was 20/47, and the final BCVA was 20/143. The late postoperative complications, including recurrent submacular hemorrhage, enlargement of the CNV, and retinal degeneration, reduced the final BCVA. These results indicate that tPA-assisted surgical removal of a subretinal hemorrhage can lead to improved long-term vision. Controlling the choroidal neovascularization may be important for retaining the best postoperative vision.

Macular autofluorescence in eyes with cystoid macula edema, detected with 488 nm-excitation but not with 580 nm-excitation.

BACKGROUND: Fundus autofluorescence (AF) derives from lipofuscin in the retinal pigment epithelium (RPE). Because lipofuscin is a by-product of phagocytosis of photoreceptors by RPE, AF imaging is expected to describe some functional aspect of the retina. In this study we report distribution of AF in patients showing macular edema. METHODS: Three eyes with diabetic macular edema (DME) and 11 with retinal vein occlusion (RVO), associated with macular edema (ME) were examined. ME was determined by standard fundus examination, fluorescein angiography (FA) and optical coherence tomography (OCT). AF was recorded using a Heidelberg confocal scanning laser ophthalmoscope (cSLO) with 488 nm laser exciter (488 nm-AF), and a conventional Topcon fundus camera with halogen lamp exciter and 580 nm band-pass filter (580 nm-AF). Color fundus picture, FA image and these two AF images were analyzed by superimposing all images. RESULTS: All subjects presented cystoid macular edema (CME) with petaloid pattern hyperfluorescence in FA. In 488 nm-AF, all eyes (100%) showed macular autofluorescence of a similar shape to that of the CME in FA. In contrast, in 580 nm-AF only one eye (7%) presented this corresponding petaloid-shaped autofluorescence. In all cases, peripheral retinal edemas did not show autofluorescence corresponding to the leakage in FA. CONCLUSIONS: In eyes with CME, analogous hyperautofluorescence to the CME was always observed in 488 nm-AF, while it was rarely observed in 580 nm-AF. Moreover, this CME hyperautofluorescence was only seen in the macular area. We hypothesize that autofluorescence from CME may be considered as a "pseudo" or "relative" autofluorescence, due to macular stretching following CME that may result in lateral displacement of macular pigments (MPs) and subsequent reduction of MPs density, as MPs block 488 nm-AF more intensely than 580 nm-AF. Although this phenomenon may not directly indicate change of RPE function, it may be used as a method to assess or track CME non-invasively.

One-year results of combined photodynamic therapy and intravitreal bevacizumab injection for retinal pigment epithelial detachment secondary to age-related macular degeneration.

BACKGROUND: To evaluate the efficacy of combined photodynamic therapy (PDT) and intravitreal bevacizumab injection in eyes with a serous pigment epithelial detachment (PED) associated with age-related macular degeneration (AMD). METHODS: Twenty-two eyes with a serous PED exceeding two disc areas associated with AMD with choroidal vascular abnormalities [choroidal neovascularization (n = 10), polypoidal choroidal vasculopathy (n = 9), and retinal angiomatous proliferation (n = 3)] received combined PDT and intravitreal bevacizumab, and were followed about every 6 weeks for more than 1 year. Additional treatments were given for residual or recurrent lesions. The main outcome measures were changes in the PED height measured by optical coherence tomography, and the best-corrected visual acuity. RESULTS: After one treatment, the PED resolved in 12 eyes (55%) and the PED decreased in ten eyes (45%). There was no recurrence in eight (36%) eyes; however, PED recurred in 14 eyes. At 1 year, the average PED height decreased to 413 microns from the baseline 751 microns (p < 0.001). Twenty eyes (91%) had improved or stabilized vision; two eyes had decreased vision due to a retinal pigment epithelial tear and subretinal hemorrhage. CONCLUSIONS: Combined PDT and intravitreal bevacizumab may decrease the PED height and stabilize visual acuity at 1 year.

Comparison of surgical treatments for central retinal vein occlusion; RON vs. cannulation of tissue plasminogen activator into the retinal vein.

PURPOSE: To compare the surgical outcomes and evaluate the effectiveness of two treatments for central retinal vein occlusion: radial optic neurotomy (RON) and cannulation of tissue plasminogen activator (tPA) into the retinal vein (tPA). METHODS: This study consisted of 22 eyes. The inclusion criterion was a best-corrected visual acuity of < or =20/60 due to central retinal vein occlusion. The exclusion criteria were previous treatment and the presence of ocular neovascularization. Patients were randomized into RON or tPA groups (n = 11 each). Best-corrected visual acuity, macular thickness, and complications were recorded. RESULTS: The mean best-corrected visual acuity changed from 16/200 at baseline to 20/167 at 12 months in RON (P = 0.217) and from 10/200 to 20/200 in tPA (P = 0.051). The preoperative macular thicknesses decreased from 1,059 microm to 406 microm 12 months postoperatively in RON (P < 0.001) and from 1,121 microm to 271 microm (P < 0.001) in tPA. Neovascular glaucoma developed in 2 eyes (18%) in RON and in 4 eyes (40%) in tPA. Visual field defects associated with surgery were seen in 2 eyes (18%) in RON. CONCLUSION: There was no significant difference in surgical outcomes between the two procedures. Although best-corrected visual acuity and macular edema improved, the incidence of neovascular glaucoma was high. It is, therefore, still uncertain whether these treatments are effective.

Presumed vascular traction-associated recurrence of retinal detachment in patients with myopic macular hole.

Optical coherence tomography (OCT) and OCT-ophthalmoscopic examinations were consecutively performed in two patients with focal retinal redetachment despite previously successful repair of the myopic macular hole-related retinal detachment by vitrectomy with removal of the internal limiting membrane. A microfold developed on the redetached retinal surface in both eyes soon after surgery. C-scan analysis by OCT-ophthalmoscope showed that the retinal microfolds coincided with a retinal artery adjacent to the preexisting macular hole. OCT revealed the focal retinal detachment was most prominent beneath the retinal microfold in both study eyes and did not reach the macular hole in one eye. The retinal microfold was most marked at the initiation of the recurrence and then attenuated with the elevation of the detached retina. Inward vascular tractional force appearing as a retinal microfold can be a critical trigger of recurrent retinal detachment in patients with myopic macular hole.

Artificial vision by direct optic nerve electrode (AV-DONE) implantation in a blind patient with retinitis pigmentosa.

The purpose of this study was to evaluate the efficacy and safety of artificial vision by using a direct optic nerve electrode (AV-DONE) in a blind patient with retinitis pigmentosa (RP). This device, comprising three wire electrodes (0.05 mm in diameter), was implanted into the optic disc of a patient with RP with no light perception vision and the device was left implanted. Six months later, visual sensations were elicited by electrical stimulation through each electrode and the thresholds for the phosphene perception elicited by pulses of 0.25-ms duration/phase and a pulse frequency of 320 Hz were 30, 5, and 70 microA for each electrode. The phosphenes, which ranged in size from that of a match head to an apple, were round, oval, or linear, primarily yellow, and focally distributed. The area of the phosphenes changed when the electrical stimulation was supplied from different electrodes. No complications arose during the follow-up period. Localized visual sensations were produced in a blind patient with advanced RP, suggesting that our system could lead to the development of a useful visual prosthesis system.

[Towards clinical application of a visual prosthesis].

We have conducted studies to develop a visual prosthesis for blind patients with outer retinal disorders such as retinitis pigmentosa, using a suprachoroidal transretinal electrical stimulation (STS) system. The consortium of the Japan Artificial Vision Project brought together medical researchers and engineers in close cooperation. Various aspects of the STS system have been assessed such as safety, stability in animal eyes, measurements of spatial resolution, development of animal models of retinal degeneration, neuroprotective effects of transcorneal electric stimulation, development of a CMOS LSI-based flexible. A unique artificial vision system has been developed using suprachoroidal electrical stimulation. The system provides spatial resolution equivalent to counting fingers. Animal models of external retinal disorders have been developed. Transcorneal electrical stimulation proved successful in providing neuroprotection in these models. The commercially available visual prosthesis will be further developed using the STS system.

Enrichment of corneal epithelial stem/progenitor cells using cell surface markers, integrin alpha6 and CD71.

Corneal epithelial stem cells are believed to reside in the basal layer of the limbal epithelium, but no definitive cell surface markers have been identified. For keratinocytes, stem/progenitor cells are known to be enriched by cell surface markers, integrin alpha(6) and CD71, as a minor subpopulation which shows high integrin alpha(6) and low CD71 expressions (alpha(6)(bri)/CD71(dim)). In the present study, we investigated the possibility that corneal epithelial stem cells can be enriched by integrin alpha(6) and CD71. The alpha(6)(bri)/CD71(dim) cells were separated by fluorescence-activated cell sorting, as a minor subpopulation of the limbal epithelial cells. They were enriched for relatively small cells, showing a higher clonogenic capacity and expression of stem cell markers, but a lower expression of differentiation markers, compared to other cell populations. The cells were localized immunohistochemically in the basal region of the limbal epithelium. These results indicate that the alpha(6)(bri)/CD71(dim) subpopulation enriched corneal epithelial stem cells.

Induction of amyloid beta accumulation by ER calcium disruption and resultant upregulation of angiogenic factors in ARPE19 cells.

PURPOSE: To investigate the intracellular mechanisms that induce amyloid beta (Abeta) accumulation and angiogenesis in the human retinal pigment epithelial cell line ARPE19. METHODS: The authors used two endoplasmic reticulum (ER) stress-inducing reagents, thapsigargin (TG), which inhibits the sarcoplasmic/endoplasmic calcium (Ca)2+-ATPase, and tunicamycin (TM), which inhibits N-linked glycosylation. The expression pattern of Abeta-precursor protein (APP) splice variants was investigated by reverse transcription (RT)-PCR. Cellular expressions of both a series of Abeta metabolism-related factors and angiogenic factors were evaluated by real-time RT-PCR and Western blot (VEGF). Expression of caspase-4 was examined by real-time RT-PCR and Western blot to evaluate the effect of the ER stressor. Intracellular Ca elevation by TG was evaluated by Ca2+ imaging experiments. Dimethyl sulfoxide and staurosporine were used as a nonreagent control and as an apoptosis-inducing reagent through mitochondria not ER, respectively. RESULTS: TG-treated ARPE19 cells increased the mRNA expression of Abeta production-inducing APP splice variants and reduced that of neprilysin, a catabolic enzyme for Abeta. TG-treated ARPE19 cells produced increases in VEGF, TNF-alpha, TACE mRNA, and VEGF protein expressions and a decrease in PEDF mRNA expression. TG-treated ARPE19 cells induced the expression of active more than TM-treated casepase-4. The intracellular Ca concentration was elevated in only TG-treated ARPE19 cells. CONCLUSIONS: TG-treated ARPE19 cells showed both Abeta accumulation-inducible and angiogenic factor mRNA expression patterns. This study suggests the possibility that ER stress through ER calcium disruption may induce the expression not only of Abeta deposit-promoting factors but also angiogenic factors in the retinal pigment epithelium.

Serial measurements of higher-order aberrations after blinking in patients with dry eye.

PURPOSE: To study the sequential postblink changes in ocular higher-order aberrations (HOAs) in patients with dry eye. METHODS: A wavefront sensor was used to measure HOAs sequentially for 30 seconds in 20 eyes of 20 patients with dry eye. The 20 eyes were classified into two groups, with or without superficial punctate keratopathy (SPK) in the central cornea. During the measurement, subjects were required to blink every 10 seconds. The aberration data were analyzed in the central 4-mm diameter for coma-like, spherical-like, and total HOAs up to sixth-order Zernike polynomials. Total HOAs, as well as fluctuation index (FI) and stability index (SI) of the total HOAs over time were compared between the two groups. The sequential changes in coma-like aberration, spherical-like aberration, and total HOAs were also investigated. RESULTS: The total ocular HOAs were significantly (P = 0.001) greater in dry eyes with central SPK than in dry eyes without central SPK. The sequential pattern of the total ocular HOAs had higher initial and consistently higher values in dry eyes with central SPK, whereas that of dry eyes without central SPK showed consistently lower total HOAs that were similar to the pattern of normal eyes. CONCLUSIONS: Increased HOAs in dry eye at least partially result from SPK above the optical zone. The low tear volume in dry eye may not cause sequential increases in HOAs after blinking. Sequential measurement of HOAs may be useful for evaluating the sequential changes in optical quality in patients with dry eye.

Morphologic changes in acute central serous chorioretinopathy evaluated by fourier-domain optical coherence tomography.

OBJECTIVE: To investigate morphologic alterations around fluorescein leakage sites using Fourier-domain optical coherence tomography (FD OCT) in acute central serous chorioretinopathy (CSC). DESIGN: Observational case series. PARTICIPANTS: Twenty-one eyes with acute CSC with subjective symptoms for under 3 months. METHODS: Patients underwent measurement of visual acuity, fundus observations, and FD OCT examinations at every visit with the intervals of 2 to 4 weeks until subretinal fluid (SRF) resolved. Fluorescein angiography was performed at baseline to confirm dye leakage sites. Horizontal and vertical OCT scans (B-scans and consecutive raster scans) of the fovea and fluorescein leakage sites were obtained. MAIN OUTCOME MEASURES: Morphologic changes in the retinal pigment epithelium (RPE), detached retina, and subretinal space around the leakage sites were evaluated repeatedly during follow-up. RESULTS: The mean period between baseline and the final examination was 108 days (mean no. of examinations, 3.9). Among 23 leakage sites in 21 eyes, FD OCT showed RPE abnormalities in 22 (96%) sites (14 sites [61%] with a pigment epithelial detachment [PED] and 8 [35%] with a protruding or irregular RPE layer). Fibrinous exudates in the subretinal space and sagging/dipping of the posterior layer of the neurosensory retina above the leakage sites were seen at 12 (52%) and 10 (43%) leakage points, respectively. An RPE defect at the edge of or within the PED was observed in 5 leakage sites (22%); in 2 of these, a defect was detectable after the SRF decreased. The posterior surface of the detached retina was smooth in 17 eyes (81%) and granulated in 4 eyes (19%) (mean duration of subjective symptoms, 10 days and 42 days, respectively). The smooth posterior detached retina became granulated in the presence of residual SRF. A PED remained at the 5 leakage sites in 5 eyes (22%) despite SRF resolution. CONCLUSIONS: Fourier-domain OCT examinations showed detailed morphologic changes in eyes with acute CSC including an RPE defect within the PED at a leakage site through which fluid might pass from the sub-RPE to the subretinal area. Fourier-domain OCT findings may offer new information to facilitate understanding of the mechanisms of acute CSC.