Choriocapillary Flow Correlation with Axial Length in High Myopia - An Angiography Study with Optical Coherence Tomography.

PURPOSE: To investigate parameters of retinal and choroidal microcirculation quantitatively with optical coherence tomography angiography (OCTA) in high myopic children, and to explore potential correlations with age, axial length (AL), spherical equivalent (SE), and central retinal thickness (CRT). METHODS: En face angiograms were generated with an OCTA device and evaluated with automated density and flow analyzer algorithms. Perfusion parameters were correlated with age, AL, SE, and CRT using Spearman's rank correlation analysis. Repeatability and reproducibility of perfusion parameter measurements were calculated in a high myopic cohort. RESULTS: Repeatability and reproducibility of OCTA measurements were good, ranging from 3.6 - 6.5%. Strong positive correlation was identified between age and CRT (rho = 0.673, p = 0.00) as well as between AL and SE (rho = 0.844, p = 0.00). There was a strong negative correlation between AL and choriocapillary flow density (CCFD) (rho = - 0.612, p = 0.00), and a moderate negative correlation between age and superficial parafoveal retinal vessel density (SPRVD) as well as CCFD (rho = - 0.497, p = 0.013 and rho = - 0.483, p = 0.023, respectively). CONCLUSION: OCTA appears to be a reliable tool for the quantitative investigation of retinal and choroidal microcirculation in a high myopic pediatric cohort. CCFD reduction was associated with increasing AL in this cohort.

Results with Posterior Scleral Reinforcement for Progressive Highly Myopic Children in Hungary.

PURPOSE: We have been performing posterior scleral reinforcement in our ophthalmological department since 1992 on progressive highly myopic eyes. Here, we report on our results with this technique in the foregoing 7 years in a retrospective comparative design. METHODS: Thirty-eight eyes of 32 patients, operated according to Snyder-Thompson's method, were enrolled in this study, and a control group of 9 age- and myopia-matched children's 14 eyes was built for comparison. Pre- and postoperative best-corrected visual acuity, subjective refractive error (spherical equivalent of spectacle dioptres), and axial length were recorded. Changes within groups were calculated, as well as baseline parameters and their changes during follow-up, and compared between the groups. Correlation analysis was performed to identify factors that could influence myopia progression. RESULTS: Myopic progression was significantly lower in the operated than in the nonoperated group, both in terms of mean annual axial length as well as refractive error changes (0.21 ± 0.08 mm versus 0.49 ± 0.19 mm and 0.18 ± 0.29 D versus 0.6 ± 0.33 D, respectively). Mean overall visual improvement was more explicit in operated eyes as compared to those left untreated (0.15 ± 0.09 versus 0.01 ± 0.1). No association of any factor with myopia progression could be identified. We encountered no serious or lasting complications. CONCLUSION: In our clinical practice, posterior scleral reinforcement according to Snyder-Thompson proved to be a safely applicable and effective surgical method to stop or significantly retard pathological increases in axial length and dioptres, and thus can help prevent the onset of myopic degenerative lesions, and irreversible visual impairment in the long run.

Cellular Factor XIII, a Transglutaminase in Human Corneal Keratocytes.

Cellular factor XIII (cFXIII, FXIII-A2), a transglutaminase, has been demonstrated in a few cell types. Its main function is to cross-link proteins by isopeptide bonds. Here, we investigated the presence of cFXIII in cells of human cornea. Tissue sections of the cornea were immunostained for FXIII-A in combination with staining for CD34 antigen or isopeptide cross-links. Isolated corneal keratocytes were also evaluated by immunofluorescent microscopy and flow cytometry. FXIII-A in the corneal stroma was quantified by Western blotting. FXIII-A mRNA was detected by RT-qPCR. The cornea of FXIII-A-deficient patients was evaluated by cornea topography. FXIII-A was detected in 68 ± 13% of CD34+ keratocytes. Their distribution in the corneal stroma was unequal; they were most abundant in the subepithelial tertile. cFXIII was of cytoplasmic localization. In the stroma, 3.64 ng cFXIII/mg protein was measured. The synthesis of cFXIII by keratocytes was confirmed by RT-qPCR. Isopeptide cross-links were detected above, but not within the corneal stroma. Slight abnormality of the cornea was detected in six out of nine FXIII-A-deficient patients. The presence of cFXIII in human keratocytes was established for the first time. cFXIII might be involved in maintaining the stability of the cornea and in the corneal wound healing process.

The effect of ranibizumab and aflibercept treatment on the prevalence of outer retinal tubulation and its influence on retreatment in neovascular age-related macular degeneration.

BACKGROUND: We aimed to analyze the differences in the prevalence of outer retinal tubulation (ORT) in neovascular age-related macular degeneration (AMD) treated with anti-vascular endothelial growth factor (anti-VEGF) agents, either aflibercept or ranibizumab. Our further aim was to examine the changes in the frequency of injections of ranibizumab before and after ORT appearance. METHODS: Two hundred thirty six eyes of 230 patients were included in the study (184 eyes treated with ranibizumab by pro re nata regimen (PRN), 52 eyes with aflibercept bimonthly) and followed for 6-24 months. Using optical coherence tomography (OCT), the first appearance of ORT was documented, and fixed time point evaluations were also made every six months to determine the existence of ORT. The number of injections, the presence or absence of subretinal hyperreflective material (SHRM) at treatment initiation and visual acuity were also noted. RESULTS: The survival analysis with Cox proportional hazard model showed no significant difference between the ranibizumab and aflibercept groups in relation to the development of ORT (p = 0.79, hazard ratio 0.92). In the PRN treated ranibizumab group the number of injections showed significant decrease after ORT development (p = 0.004). When SHRM was present at treatment initiation the chance of developing ORT was 2.75 and 11.14 times higher in the ranibizumab and aflibercept groups, respectively. CONCLUSIONS: The prevalence of ORT increased over time independently from the chosen anti-VEGF drug. Our results suggest that upon the appearance of ORT a decrease in retreatments can be expected.

[Ophthalmological symptoms of measles and their treatment]. / A kanyaró szemészeti tünetei és kezelése.

Measles, caused by the Morbilli virus, is a highly (about 95 %) contagious disease affecting primarily children, but without proper immunisation, adults can also be infected. The leading symptoms of the disease are high fever that presents after an incubation period of 9-10 days and the red rash that begins several days after the fever starts. Beyond specific generalized symptoms, measles may have ocular symptoms. The most commonly occurring conjunctivitis, the so-called "red eye symptom", is not characteristic only for measles infection, however, by taking the generalized symptoms it can suggest the diagnosis at the beginning of the disease. Conjunctivitis of varying severity is noticed in the half of the cases without using ophthalmological instrumentation. Using ophthalmological instrumentation, the mild forms of conjunctivitis can be diagnosed, by meticulous ophthalmological examination, further eye diseases can be discovered. The viral conjunctivitis can progress to keratitis and bacterial superinfection can occur. If the infection presents in childhood it can affect the posterior segment. The fight against measles is very effective in Hungary since the vaccination has been introduced, and the lack of vaccination is also the primary cause of the risk to the disease. In the diagnosis, symptomatic treatment of the disease and the curbing of possible mass infections, the practicing physician (general practitioner) has a key role. The correct care of the infected patient in Hungary is provided by a methodological letter, professional information and legal guides. Orv Hetil. 2017; 158(39): 1523-1527.

[Experiences with CO2 laser-assisted sclerectomy surgery]. / CO2-lézerrel végzett mélysclerectomiával szerzett tapasztalatok.

INTRODUCTION: CO2 laser- assisted sclerectomy surgery (CLASS) can be used for the surgical treatment of open-angle glaucoma. AIM: To introduce our results with CLASS. METHOD: We performed 21 CLASS operations using OT-134-IOPtiMate (IOPtima Ltd, Ramat-Gan, Israel). Patients were examined on the 1st day, and in the 1st, 3rd, 6th, 9th and 12th months postoperatively. We evaluated intraocular pressure (IOP), antiglaucomatous medication-use, visual acuity, complications. RESULTS: Mean age was 65.6 yrs. Complete success (no hypotensive medication required to target IOP) was achieved in 61.1% (18 patients) at 6 months, whereas in 50% (10 patients) at 12 months. Qualified success (hypotensive medication required to target IOP) was achieved in 72.2% and in 70%, preoperative mean IOP was 29.2 ± 9.4 Hgmm, which falled to 17.7 ± 4.9 Hgmm and 17.3 ± 4.3 Hgmm, respectively. Antiglaucomatous medication use falled significantly from 2.90 ± 0.83 to 2.05 ± 1.46. Apart from 1 macroperforation, no serious complication occurred. CONCLUSIONS: With CLASS it is possible to effectively lower intraocular pressure in open-angle glaucoma. Orv Hetil. 2017; 158(18): 701-705.

[Case report on choroidal effusion after oral acetazolamide administration: an unusual manifestation of a well-known idiosyncratic effect?] / Acetazolamid orális alkalmazása mellett jelentkezo chorioidealeválás két esete: ismert idioszinkráziás hatás szokatlan megjelenési formája?

Sulpha drugs are widely employed in medicine for various diseases and disorders. During the last several decades, numerous papers had been published on supra ciliary and posterior choroidal effusion likely presenting as an idiosyncratic effect of these drugs especially of acetazolamide. In each publication, the effusion was associated with either an acute angle-closure glaucoma or transitory myopia or both of these as leading symptoms. In the current publication, authors report on two cases where the acetazolamide-induced choroidal effusion was an accidental finding without either a myopic shift in refraction or an acute elevation in intraocular pressure. To our best knowledge, ours is the first report in the literature describing this unusual, "silent" form of a sulpha drug-induced choroidal effusion. Since the choroidal involvement may vary in size and location, and is not necessarily associated with acute glaucoma and myopia, one can assume that a considerable amount of acetazolamide-related ocular side-effects will not be discovered. The above case report aims to draw the attention of other specialities to the need for ophthalmic examination for their patients taking sulpha drugs with acute visual deterioration. Orv Hetil. 2017; 158(50): 1998-2002.

Triamcinolone regulated apopto-phagocytic gene expression patterns in the clearance of dying retinal pigment epithelial cells. A key role of Mertk in the enhanced phagocytosis.

BACKGROUND: The apopto-phagocytic gene expression patterns during clearance of dying cells in the retina and the effect of triamcinolone (TC) upon these processes have relevance to development of age-related macular degeneration (AMD). METHODS: ARPE-19 cells and primary human retinal pigment epithelium (hRPE) were induced to undergo cell death by anoikis and the clearance of these cells by living hRPE/ARPE-19 or human monocyte-derived macrophages (HMDMs) in the presence or absence of TC was quantified by flow cytometry. TaqMan low-density gene expression array determining known markers of phagocytosis and loss-of-function studies on selected apopto-phagocytic genes was carried out in HMDM engulfing anoikic cells. RESULTS: The glucocorticoid TC had a profound phagocytosis-enhancing effect on HMDM engulfing anoikic ARPE-19 or hRPE cells, causing a selective upregulation of the Mer tyrosine kinase (MERTK) receptor, while decreasing the expression of the AXL receptor tyrosine kinase and thrombospondin-1 (THSB-1). The key role of the MERTK could be demonstrated in HMDM engulfing dying cells using gene silencing as well as blocking antibodies. Similar pathways were found upregulated in living ARPE-19 engulfing anoikic ARPE-19 cells. Gas6 treatment enhanced phagocytosis in TC-treated HMDMs. CONCLUSIONS: Specific agonists of the Mertk receptor may have a potential role as phagocytosis enhancers in the retina and serve as future targets for AMD therapy. GENERAL SIGNIFICANCE: The use of Gas6 as enhancer of retinal phagocytosis via the MerTK receptor, alone or in combination with other specific ligands of the tyrosine kinase receptors' family may have a potential role in AMD therapy.

[Ocular symptoms and treatment of Ebola virus disease]. / Az Ebola-vírusos betegség szemészeti tünetei és kezelése.

Ocular signs and symptoms of Ebola infection initially suggest banal conjunctivitis, but in advanced cases severe haemorrhagic conjunctivitis appears and, in the final stage of the disease, retinal and chorioidal haemorrhages may occur which can cause even blindness. Although the viral infection accompanied by ocular symptoms of a non-specific conjunctivitis, the high fever present from the onset of the disease should raise the suspicion of Ebola infection. There is no causal therapy know so far, and the only adjunctive treatment may be delivered by an ophthalmologist. Because the virus can be detected in the tear, it can theoretically be the mediator of the infection and, therefore, ophthalmological examinations should be carried out with the highest caution. In case of suspected Ebola infection the nearest competent healthcare authority should be immediately alerted in order to take further actions.

Characterization of ex vivo cultured neuronal- and glial- like cells from human idiopathic epiretinal membranes.

BACKGROUND: Characterization of the neuro-glial profile of cells growing out of human idiopathic epiretinal membranes (iERMs) and testing their proliferative and pluripotent properties ex vivo is needed to better understand the pathogenesis of their formation. METHODS: iERMs obtained during uneventful vitrectomies were cultivated ex vivo under adherent conditions and assessed by standard morphological and immunocytochemical methods. The intracellular calcium dynamics of the outgrowing cells was assessed by fluorescent dye Fura-2 in response to acetylcholine (ACh)- or mechano- stimulation. RESULTS: The cells from the iERMs formed sphere-like structures when cultured ex vivo. The diameter of the spheres increased by 5% at day 6 and kept an increasing tendency over a month time. The outgrowing cells from the iERM spheres had mainly glial- and some neuronal- like morphology. ACh- or mechano- stimulation of these cells induced intracellular calcium propagation in both cell types; in the neuronal-like cells resembling action potential from the soma to the dendrites. Immunocytochemistry confirmed presence of glial- and neuronal cell phenotype (GFAP and Nestin-1 positivity, respectively) in the iERMs, as well as presence of pluripotency marker (Sox2). CONCLUSION: iERMs contain cells of neuronal- and glial- like origin which have proliferative and pluripotent potential, show functionality reflected through calcium dynamics upon ACh and mechano- stimulation, and a corresponding molecular phenotype.

Indapamide-induced transient myopia with supraciliary effusion: case report.

BACKGROUND: Ingestion of sulphonamide-derived drugs has been reported to possibly have ocular side-effects. Authors aimed to present a rare case of indapamide-induced transient myopia with ciliary body edema and supraciliary effusion. CASE PRESENTATION: A 39 years old caucasian female patient presented at the Department of Neurology with headache and sudden bilateral loss of distant vision. Neurological assessment and cranial CT scans were unremarkable. For her hypertension, twice a day bisoprolol 2.5 mg and once a day indapamide 1.5 mg tablets were prescribed several days before. At her presenting, ophthalmic findings were as follows: visual acuity 0.08-7.25Dsph = 1.0 and 0.06-7.25Dsph = 1.0; IOP 25 mmHg and 24 mmHg, anterior chamber depth (ACD) 2.32 mm and 2.49 mm, lens thickness (L) 4.02 mm and 4.09 mm in the right and the left eye, respectively. By means of ultrasound biomicroscopy (UBM), thickened (720 / 700 micron) and detached ciliary body, its forward movement (ciliary body-cornea angle 108' / 114') and forward rotated ciliary processes were seen. Angle opening distance (AOD500) were 300 / 314 microns. By the following days, the myopia gradually diminished, and a week after her first symptoms, her uncorrected visual acuity was 1.0 in both eyes, IOP 13 mmHg and 17 mmHg, ACD 3.68 mm and 3.66 mm, L 3.78 mm and 3.81 mm in the right and the left eye, respectively. Ciliary body edema and detachment disappeared (ciliary body thickness 225 / 230 micron), both of the ciliary body-cornea angle 134' / 140' and the AOD500 (650 / 640 microns) increased. At this point, the patient admitted that she had stopped taking indapamide two days before. CONCLUSIONS: Our case report is the third one in the literature to present indapamide-induced transient myopia, and the first to employ UBM for describing the characteristics of this rare condition. According to the findings, authors suggest that both ciliary muscle contraction and ciliary body edema may play role in the pathomechanism. UBM seems to be a useful tool in the differential diagnosis of acute myopia. Further, authors wish to draw attention to one of the potential adverse effects of this drug which was not listed by its package insert.

Effect of contact lens wear on the release of tear mediators in keratoconus.

OBJECTIVES: The release of different cytokines and mediators in tears of patients with keratoconus (KC) wearing contact lenses (CLs) may contribute to the pathology of KC. METHODS: Cohort study was established in patients with KC wearing rigid gas permeable (RGP) CL (group I), patients with ametropia wearing silicone hydrogel (Si-Hi) CL (group II) and ametropic patients wearing RGP CL (group III). RESULTS: Our findings indicate that before CL wear, the release of epidermal growth factor (EGF) and tissue-type plasminogen activator (t-PA) was attenuated, whereas matrix metalloproteinase (MMP)-9, interleukin (IL)-6, chemokine (C-C motif) ligand 5 (CCL5), IL-13, and plasminogen activator inhibitor (PAI)-1 were enhanced in KC compared with ametropes. An increasing linear trend over time was found for MMP-9, EGF, and CXCL8 in KC and MMP-9, MMP-13, IL-6, and CXCL8 in group III. Significant differences were observed in the linear trend over time between groups I and III for MMP-13 and tissue inhibitor of metalloproteinases (TIMP)-1; between groups I and II for MMP-9 and CXCL8; and between groups III and II for MMP-9, CXCL8, and MMP-13. In KC, the release of MMP-9 at week 6 and nerve growth factor (NGF) at 10 min was higher, but NGF at week 2 was lower than that in group II. The release of MMP-13 and NGF at week 2 and 6 were lower in the KC group as compared with group III, and similarly, with IL-6 and CXCL8 at week 2 and PAI at all time points. CONCLUSIONS: Contact lens wear can influence the levels and dynamics of various mediators in the tears of patients with KC that might have an impact on the progression of the disease.

[Ophthalmologic manifestations of Bardet-Biedl syndrome]. / Tapasztalataink Bardet-Biedl-szindrómás gyermekek általános szemészeti és elektrofiziológiai vizsgálatával.

INTRODUCTION: Bardet-Biedl syndrome is characterised by retinal dystrophy, polydactily, obesity and slow mental development. AIM: The aim of the authors was to present ophthalmologic signs and symptoms of the syndrome. METHOD: Between 1980 and 2010, 4 children with Bardet-Biedl syndrome were evaluated at the Department of Ophthalmology, University of Szeged, Szeged, Hungary. Their age at the first visit was between 1 and 10 years. Basic ophthalmological and electrophysiological evaluation, as well as orthoptic examinations were performed. RESULTS: In two cases the electroretinographic curves were subnormal, and in two cases the electroretinographic curves showed no elevation. In the 4 children abnormal electroretinographic curves appeared at the ages of 1, 5, 10, and 18 years. Pigmentary changes on the periphery of the retina were detected in two cases. CONCLUSIONS: The different signs and symptoms of Bardet-Biedl syndrome may manifest at different ages. Electrophysiological changes failed to correlate with retinal alterations is these patients.

Corneal transplantation in Hungary (1946-2009).

BACKGROUND: To identify changing trends in indications for corneal transplantation in Debrecen, Hungary over the past 64 years. DESIGN: Retrospective study, at the Department of Ophthalmology, University of Debrecen, Hungary. PARTICIPANTS: Four thousand seven hundred and seventy-eight patients who underwent keratoplasty. METHODS: The analysis was based on medical charts, surgical descriptions and eye bank records. MAIN OUTCOME MEASURES: Keratoplasty indications between January 1946 and December 2009. For an easier overview, the 64-year interval was divided into seven time periods (1946-1955, I; 1956-1965, II; 1966-1975, III; 1976-1985, IV; 1986-1995, V; 1996-2005, VI; 2006-2009, VII). RESULTS: Over the 64 years, clinical indications for keratoplasty were corneal scarring (24.9%), regraft (18.6%), keratoconus (18.6%), pseudophakic/aphakic corneal oedema (12%), stromal corneal dystrophies (6%), non-infectious keratitis (4.7%), chemical injuries (3.3%), corneal degenerations (3%), mechanical trauma (1.7%), infectious keratitis (1.4%), endothelial corneal dystrophies (1.3%) and others (4.5%). During periods I-IV, corneal scarring was the most common indication for corneal transplantation. In period V, corneal ectasia became the leading clinical indication. Regraft was the most frequent indication in period VI. In the most recent years, an emerging tendency in pseudophakic/aphakic corneal oedema as the indication for keratoplasty was observed. CONCLUSIONS: In Hungary, the number of grafts has increased greatly in the past 64 years, the transplantation rates are similar to those of industrial countries. Indications for corneal transplantation have changed considerably over the last half decade from corneal scarring to corneal ectasia, regraft and pseudophakic/aphakic corneal oedema.

Myopia-26, the female-limited form of early-onset high myopia, occurring in a European family.

BACKGROUND: Female-limited early-onset high myopia, also called Myopia-26 is a rare monogenic disorder characterized by severe short sightedness starting in early childhood and progressing to blindness potentially by the middle ages. Despite the X-linked locus of the mutated ARR3 gene, the disease paradoxically affects females only, with males being asymptomatic carriers. Previously, this disease has only been observed in Asian families and has not gone through detailed investigation concerning collateral symptoms or pathogenesis. RESULTS: We found a large Hungarian family displaying female-limited early-onset high myopia. Whole exome sequencing of two individuals identified a novel nonsense mutation (c.214C>T, p.Arg72*) in the ARR3 gene. We carried out basic ophthalmological testing for 18 family members, as well as detailed ophthalmological examination (intraocular pressure, axial length, fundus appearance, optical coherence tomography, visual field- testing) as well as colour vision- and electrophysiology tests (standard and multifocal electroretinography, pattern electroretinography and visual evoked potentials) for eight individuals. Ophthalmological examinations did not reveal any signs of cone dystrophy as opposed to animal models. Electrophysiology and colour vision tests similarly did not evidence a general cone system alteration, rather a central macular dysfunction affecting both the inner and outer (postreceptoral and receptoral) retinal structures in all patients with ARR3 mutation. CONCLUSIONS: This is the first description of a Caucasian family displaying Myopia-26. We present two hypotheses that could potentially explain the pathomechanism of this disease.

Diabetic Retinopathy Screening in Patients with Diabetes Using a Handheld Fundus Camera: The Experience from the South-Eastern Region in Hungary.

PURPOSE: Diabetic retinopathy (DR) is the leading cause of vision loss among active adults in industrialized countries. We aimed to investigate the prevalence of diabetes mellitus (DM), DR and its different grades, in patients with DM in the Csongrád County, South-Eastern region, Hungary. Furthermore, we aimed to detect the risk factors for developing DR and the diabetology/ophthalmology screening patterns and frequencies, as well as the effect of socioeconomic status- (SES-) related factors on the health and behavior of DM patients. METHODS: A cross-sectional study was conducted on adults (>18 years) involving handheld fundus camera screening (Smartscope Pro Optomed, Finland) and image assessment using the Spectra DR software (Health Intelligence, England). Self-completed questionnaires on self-perceived health status (SPHS) and health behavior, as well as visual acuity, HbA1c level, type of DM, and attendance at healthcare services were also recorded. RESULTS: 787 participants with fundus camera images and full self-administered questionnaires were included in the study; 46.2% of the images were unassessable. T1D and T2D were present in 13.5% and 86.5% of the participants, respectively. Among the T1D and T2D patients, 25.0% and 33.5% had DR, respectively. The SES showed significant proportion differences in the T1D group. Lower education was associated with a lower DR rate compared to non-DR (7.7% vs. 40.5%), while bad/very bad perceived financial status was associated with significantly higher DR proportion compared to non-DR (63.6% vs. 22.2%). Neither the SPHS nor the health behavior showed a significant relationship with the disease for both DM groups. Mild nonproliferative retinopathy without maculopathy (R1M0) was detected in 6% and 23% of the T1D and T2D patients having DR, respectively; R1 with maculopathy (R1M1) was present in 82% and 66% of the T1D and T2D groups, respectively. Both moderate nonproliferative retinopathy with maculopathy (R2M1) and active proliferative retinopathy with maculopathy (R3M1) were detected in 6% and 7% of the T1D and T2D patients having DR, respectively. The level of HbA1c affected the attendance at the diabetology screening (HbA1c > 7% associated with >50% of all quarter-yearly attendance in DM patients, and with 10% of the diabetology screening nonattendance). CONCLUSION: The prevalence of DM and DR in the studied population in Hungary followed the country trend, with a slightly higher sight-threatening DR than the previously reported national average. SES appears to affect the DR rate, in particular, for T1D. Although DR screening using handheld cameras seems to be simple and dynamic, much training and experience, as well as overcoming the issue of decreased optic clarity is needed to achieve a proper level of image assessability, and in particular, for use in future telemedicine or artificial intelligence screening programs.

Involvement of p63 in the herpes simplex virus-1-induced demise of corneal cells.

BACKGROUND: The transcription factor p63 plays a pivotal role in the development and maintenance of epithelial tissues, including the ocular surface. In an effort to gain insight into the pathogenesis of keratitis caused by HSV-1, we determined the expression patterns of the p63 and Bax proteins in the Staatens Seruminstitute Rabbit Cornea cell line (SIRC). METHODS: SIRC cells were infected with HSV-1 at various multiplicities and maintained for different periods of time. Virus replication was measured by indirect immunofluorescence assay and Western blot analysis. Cell viability was determined by MTT assay. The apoptotic response of the infected cells was quantified by ELISA detecting the enrichment of nucleosomes in the cytoplasm. Western blot analysis was used to determine the levels of p63 and Bax proteins. RESULTS: Indirect immunofluorescence assays and Western blot analyses demonstrated the presence of HSV-1 glycoprotein D (gD) in the infected SIRC cell line, and the pattern of gD expression was consistent with efficient viral replication. The results of MTT and ELISA assays showed that HSV-1 elicited a strong cytopathic effect, and apoptosis played an important role in the demise of the infected cells. Mock-infected SIRC cells displayed the constitutive expression of DeltaNp63alpha. The expressions of the Bax-beta and TAp63gamma isoforms were considerably increased, whereas the level of DeltaNp63alpha was decreased in the HSV-1-infected SIRC cells. Experiments involving the use of acyclovir showed that viral DNA replication was necessary for the accumulation of TAp63gamma. CONCLUSION: These data suggest that a direct, virus-mediated cytopathic effect may play an important role in the pathogenic mechanism of herpetic keratitis. By disturbing the delicate balance between the pro-survival DeltaN and the pro-apoptotic TA isoforms, HSV-1 may cause profound alterations in the viability of the ocular cells and in the tissue homeostasis of the ocular surface.

Comparison of hydrophilic ophthalmic media on silicone oil emulsification.

The aim of this study was to investigate whether and how the biological media which are in contact with silicone oil play a role in the silicone emulsification process. Commercially available Oxane 1300 silicone oil and potential hydrophilic phases of the emulsions in the eye (porcine aqueous humor, porcine vitreous and balanced salt solution) were investigated separately and in a mixture or emulsions by means of surface tension, rheological, zeta potential measurements and microscopic investigation. The surface tension of biological media (vitreous and aqueous humor) was significantly lower than that of non-biological media, especially in the case of aqueous humor, which indicates a remarkable emulsification tendency with these phases. The biological media are able to form both oil-in-water and water-in-oil emulsions, which can be observed in the clinical practice as well. It was established that the vitreous has a more expressed emulsification ability compared with the aqueous humor because smaller and more stable droplets can form with silicon oil when the vitreous is still there. It can be concluded that the vitreous has a higher impact on emulsification than the aqueous medium, which can predict that the vitreous remaining after vitrectomy has a key role in emulsion formation in the eye with silicone oil endotamponade.

Keratoplasty in Hungary.

The first keratoplasty was performed in Hungary in 1909, but the transplantations were started in large numbers after the Second World War in Debrecen. Since 1992 a National Keratoplasty Register is functioning at the Department of Ophthalmology Debrecen. The present paper gives an overview on the current status of keratoplasty in Hungary based on the data of this Register. The number of corneal transplantations more than doubled in the past two decades. In the past 3 years (2005 to 2008) the number of transplantation seemed to stabilize around a bit more than 700. The ratio of preserved corneas disclosed a constant increase. The turning point was 1997, few years after the establishment of the Debrecen Eye Bank in 1994, when more preserved corneas were transplanted than fresh ones. The law regulating the procurement and the transplantation of corneas in our country is based on presumed consent and explicit objection which is the better type of regulation for the performance of keratoplasties in large numbers. The number of corneal transplantations in Hungary in the past decades was similar to the figures of industrial countries.

Ex vivo 3D human corneal stroma model for Schnyder corneal dystrophy - role of autophagy in its pathogenesis and resolution.

INTRODUCTION: Multilamellar bodies (MLBs) are concentric cytoplasmic membranes which form through an autophagy-dependent mechanism. In the cornea, the presence of MLBs is associated with Schnyder corneal dystrophy (SCD). Ex vivo 3D modelling of the corneal stroma and SCD can help study pathogenesis and resolution of the disorder. METHODS: Corneal stroma explants were isolated from cadavers and cultivated long-term for more than 3 months to achieve spontaneous 3D outgrowth of corneal stroma-derived mesenchymal stem-like cells (CSMSCs). The 3D tissues were then examined by transmission electron microscopy (TEM) for presence of MLBs, and by immunofluorescent labelling against markers for autophagy (p62, LC3). Autophagy was induced by classical serum starvation or rapamycin (RAP) treatment (50 nM), and inhibited by the autophagy inhibitor 3-methyladenine (3-MA, 10 mM) for 24 hours. RESULTS: CSMSCs can form spontaneously 3D outgrowths over a 3-4 weeks period, depositing their own extracellular matrix containing collagen I. TEM confirmed the presence of MLBs in the long-term (>3 months) 3D cultures, which became more abundant under starvation and RAP treatment, and decreased in number under autophagy inhibition with 3-MA. The presence of autophagy and its disappearance could be confirmed by an inversely related increase and decrease in the expression of LC3 and p62, respectively. CONCLUSIONS: MLB formation in long-standing CSMSC cultures could serve as a potential ex vivo model for studying corneal stroma diseases, including SCD. Inhibition of autophagy can decrease the formation of MLBs, which may lead to a novel treatment of the disease in the future.