Phenotype Variability in Czech Patients Carrying PAX6 Disease-Causing Variants.

The aim of this study was to report PAX6 disease-causing variants in six Czech families, to describe the associated phenotypes, and to perform functional assessment of the splice site variants. Detailed ophthalmic examination was performed. The PAX6 coding region was directly sequenced in three probands. Two probands were analysed by exome sequencing and one by genome sequencing. The effect of two variants on pre-mRNA splicing was evaluated using an exon trapping assay. Six different heterozygous PAX6 variants were identified, with c.111_120del and c.1183+1G˃T being novel. Both c.1183+1G˃T and c.1032+1G>A were proved to cause aberrant splicing with exon skipping and subsequent frameshift. The phenotypic features were variable between and within families. One individual, aged 31 years, presented with mild unilateral ptosis accompanied by aniridia in the right eye, partial aniridia in the left eye, and bilateral congenital cataracts, without marked foveal hypoplasia. Bilateral microcornea, partial aniridia, congenital cataracts, and a large posterior segment coloboma were found in another proband, aged 32 years. One child, aged 8 years, had bilateral high myopia, optic nerve colobomas, anterior polar cataracts, but no iris defects. Another individual, aged 46 years, had bilateral congenital ptosis, iris hypoplasia, keratopathy with marked fibrovascular pannus, anterior polar cataract, and foveal hypoplasia combined with impaired glucose tolerance. However, his daughter, aged 11 years, showed classical features of aniridia. Our study extends the genetic spectrum of PAX6 disease-causing variants and confirms that the associated phenotypic features may be very broad and different to the 'classical' aniridia.

[Ocular Cicatricial Pemphigoid - a Retrospective Study]. / Ocní jizevnatý pemfigoid - retrospektivní studie.

INTRODUCTION: Ocular cicatricial pemphigoid (OCP) is rare, severe, sight threatening autoimmune disease of the conjunctiva, which affects elderly patients, more often women. AIM: To evaluate the success rate of stabilisation of ocular findings in patients with OCP. METHODS: Retrospective study of patients from Centre of Conjunctival and Corneal Diseases at Department of Ophthalmology, General University Hospital and 1st Medical Faculty of Charles University in Prague in 1992-2013 was performed. Frequency of OCP clinical stages, visual acuity (VA), disease activity and ocular complications of referred patients were monitored. Moreover, type of immunosuppressive (IS) therapy, the number of relapses of the disease and progress of OCP clinical stages were evaluated. Especially, we evaluated effects and side effects of mycophenolate mofetil (MM) therapy. In addition to that, type and frequency of ocular surgery that was carried out to the patients before and after the referral were recorded. Furthermore, we evaluated percentage of patients with mucous membranous pemphigoid (MMP). Also, the positive yield of diagnostic methods was assessed. RESULTS: The OCP was diagnosed and monitored in 51 patients (21 men and 30 women) during 21 years, the average age on the day of diagnosis was 68,4 years, the average period of observation was 57 months. 55 % of eyes were referred to our department at clinical stage 3, 27 % at stage 4. VA was maintained in 76 % of eyes, improved in 5 % of eyes and in 19 % of eyes deteriorated. Activity of OCP was detected during the first examination in 96 % of patients, the most common complications at that time was corneal ulcer or perforation. Patients were treated by immunosuppressive therapy, most often in combination: corticosteroids (47 patients), azathioprine (28 patients), cyclophosphamide (25 patients), MM (16 patients), sulphasalazine (5 patients), dapsone (5 patients). We ascertained relapses in 40 % of patients. The progression to the next stage of OCP were found in 7 eyes (6,9 %) and 95 eyes (93,1 %) remained stable. Activity of disease was well controlled in 11 patients out of 16 (69 %) by MM, IS therapy of remain 5 patients (31 %) had to be changed. Side effects of MM such as lymphopenia were present in 1 patient. Before OCP was diagnosed, patients underwent cataract surgery with the intraocular lens implantation, cryoepilation of eyelashes and eyelid plastic surgery, especially entropion. The most common indicated surgery in our clinic was amniotic membrane transplantation and retro position of muscular cutaneous leaf. 31 % of patients were diagnosed with MMP. Positive results of conjunctival biopsy were detected in 48 % from 42 examined samples and 22 % from 32 examined samples had positive results of indirect immunofluorescence (anti-desmosoms). CONCLUSION: OCP diagnosis is established on the basis of patient´s ophthalmic history and clinical findings. Positive results of direct and indirect immunofluorescence support the diagnosis. Activity of the disease and progression of OCP is effectively suppressed by systemic immunosuppressive therapy (for example MM), mainly if started at early stage of the disease. KEY WORDS: ocular cicatricial pemphigoid, immunosuppressive therapy, direct and indirect immunofluorescence, mycophenolate mofetil.

[Preimplantation genetic diagnosis and monogenic inherited eye diseases]. / Preimplantacní genetická diagnostika a dedicná onemocnení oka.

OBJECTIVE: Preimplantation genetic diagnosis (PGD) is an established application of genetic testing in the context of in vitro fertilization. PGD is an alternative method to prenatal diagnosis which aims to prevent the transmission of an inherited disorder to the progeny by implanting only embryos that do not carry genetic predisposition for a particular disease. The aim of this study is to provide an overview of eye disorders for which PGD has been carried out. METHODS: The European literature search focused on best practices, ethical issues, risks and results of PGD for inherited eye disorders. RESULTS: PGD is performed for a number of ocular disorders; a prerequisite for its application is however, the knowledge of a disease-causing mutation(s). The main advantage of this method is that the couple is not exposed to a decision of whether or not to undergo an abortion. Qualified counselling must be provided prior to the PGD in order to completely understand the risk of disability in any child conceived, consequences of disease manifestation, and advantages as well as limitations of this method. In the group of non-syndromic eye diseases and diseases in which ocular findings dominate, PGD has been performed in European countries for aniridia, choroideremia, congenital fibrosis of extraocular muscles, Leber congenital amaurosis, ocular albinism, retinitis pigmentosa, X-linked retinoschisis, Stargardt disease, blepharophimosis-ptosis-inverse epicanthus syndrome and retinoblastoma. Sexing for X-linked or mitochondrial diseases has been carried out for blue cone monochromatism, choroideremia, familial exudative vitreoretinopathy, Leber hereditary optic neuropathy, macular dystrophy (not further specified), Norrie disease, X-linked congenital stationary night blindness, X-linked retinoschisis and nystagmus (not further specified). CONCLUSION: In recent years, there has been an increase in potential to use PGD. The spectrum of diseases for this method has widened to include severe inherited eye diseases.Key words: preimplantation genetic diagnosis; monogenic eye diseases; in vitro fertilization.

[Tear Osmolarity in Patients with Severe Dry Eye Syndrome Before and After Autologous Serum Treatment: a Comparison with Tear Osmolarity in Healthy Volunteers]. / Osmolarita slz u pacientu s tezkým syndromem suchého oka pred a po aplikaci autologního séra. Porovnání s hodnotami zdravých dobrovolníku.

INTRODUCTION: Dry eye syndrome (DES) is a multifactorial disease of the tears and ocular surface. Recently, treatment with autologous serum eye drops (AS-ed) has been frequently used in these patients. Significant improvement correlates well with clinical, laboratory and subjective findings. It is assumed that one of the key factors in the development of the disease is increased tear osmolarity. The aims of our study were to test tear osmolarity measurements in clinical practice, to examine if osmolarity values differ before and after a 3-month application of 20% AS-ed, and to determine if the values differ between patients with severe DES and healthy individuals. METHODS: The study included 35 patients with severe DES (Schirmer test<5 mm/5 min) and 23 healthy volunteers. Tear osmolarity values (TearLab Osmolarity System), the Schirmer test (ST1), vital ocular surface staining and subjective feelings (the OSDI questionnaire) were assessed in patients with DES before and after treatment with 20% AS-ed. Further, the tear osmolarity values were compared between healthy subjects and patients with DES before and after treatment with AS-ed. RESULTS: The values of OSDI and vital staining significantly decreased in patients with DES after the treatment (p<0.0001). ST1 and tear osmolarity did not change significantly after the treatment. ST1 values in healthy individuals were significantly higher (p<0.0001) and the OSDI values significantly lower (p<0.0001) than the results obtained in patients before and after treatment. Tear osmolarity was not statistically different between healthy subjects (306 mosmol/l) and patients with DES (302 and 301 mosmol/l before and after treatment respectively). CONCLUSION: We demonstrated a positive effect of AS treatment on the ocular surface in patients with DES. However, the osmolarity values did not differ before and after treatment with AS, and they also did not differ significantly between DES patients and healthy individuals. In accordance with other recent studies, our results raise questions concerning the value of the TearLab Osmolarity System for evaluating therapeutic effect and also as a tool for DES diagnosis.

[Clinical findings in members of a Czech family with retinitis pigmentosa caused by the c.2426_2427delAG mutation in RPGR]. / Klinické nálezy u clenu ceské rodiny s retinitis pigmentosa podmínenou mutací v ORF15 genu RPGR.

PURPOSE: To describe the phenotype of members of the first Czech retinitis pigmentosa family with an identified molecular genetic cause (c.2426_2427delAG in RPGR), followed for more than 13 years. METHODS: Medical records were reviewed and a detailed ophthalmic examination including spectral-domain optical coherence tomography and full-field and multifocal electroretinography (ERG) was performed in two affected males, three female carriers and one unaffected female. RESULTS: A 22-year-old male who denied suffering from nyctalopia had a best corrected visual acuity (BCVA) of 0.63 in both eyes. Moderate myopia and myopic astigmatism were present bilaterally. Color vision and contrast sensitivity were normal. There was an eccentric constriction of the visual fields that spared the central 20 degrees in both eyes. Fundus examination revealed bilateral pigmentary changes in the mid-periphery. Full-field ERG documented a 10% rod and 20% cone response. The phenotype of his cousin, also aged 22 years, was more severe. He complained of nyctalopia since 12 years of age. His BCVA was 0.3 in the right eye and 0.5 in the left eye. Myopia and astigmatism were present bilaterally. Contrast sensitivity and color vision were severely impaired. Full field ERG was extinct, but some activity on multifocal ERG was still detectable. The constriction of the visual fields reached 5 degrees in both eyes. Fundus examination showed the typical retinitis pigmentosa appearance. All carriers denied that they suffered from nyctalopia, but two of them had decreased BCVA in at least one eye. None exhibited typical bone spicules or a tapetal-like reflex. Significant refractive errors were present in all eyes of the carriers. CONCLUSION: The finding of moderate or high myopia and astigmatism in males with retinitis pigmentosa as well as refractive errors in female relatives indicates possible X-linked inheritance, which may be especially important in pedigrees where the transmission pattern can not be clearly established. Our study highlights the inter-individual variability in phenotype observed in similar aged patients with identical ORF15 RPGR mutations.

[Problematic issues related to screening for diabetic retinopathy]. / Problemy pri sledování pacientu s diabetickou retinopatií.

The aim of this paper is to highlight the importance of screening for diabetic retinopathy and the education of diabetic patients. The key prerequisite of an effective screening is close collaboration between health care professionals. Successful screening detects DR at early stages and helps to prevent severe visual loss. Two cases of proliferative DR exhibiting the most serious complications (vitreous haemorrhage, traction retinal detachment and combined traction and rhegmatogenous retinal detachment, neovascular glaucoma) are reported. The first comorbid patient lost her vision despite the adequate treatment. The second patient was successfully managed with pars plana vitrectomy with final visual acuity better to the preoperative value.

[Importance of pars plana vitrectomy in the diagnosis of ocular toxocariasis]. / Význam pars plana vitrektomie v diagnostice ocní toxokarózy.

A retrospective study evaluates the importance of pars plana vitrectomy (PPV) in four patients with ocular form of toxocarosis. The results of laboratory determination of the titres of antibodies against Toxocara canis (the ELISA method) in samples of vitreous body were compared with those in the serum of the patient. The individual cases exemplify the problems in interpretation of laboratory findings and document the authors' experience in considering the diagnosis of ocular toxocarosis in individual cases. The authors' experience indicates that PPV is of diagnostic as well as of therapeutic significance.

[Multiple extrarenal complications in Wegener granulomatosis]. / Mnohocetné extrarenální komplikace Wegenerovy granulomatózy.

This is a case report of a patient with Wegener's granulomatosis (WG), who initially presented with a corneal perforation. In addition to the eye involvement, the pauciimmune necrotizing glomerulonephritis with crescent formation, E.N.T. and pulmonary involvement were diagnosed. The patient also suffered from the acute myocardial infarction, most likely due to coronal arteritis. In addition to the coronal ischaemia she also had vasculitis of the aortic valve due to the WG. Another rare complication was the massive intestinal bleeding. The patient had also a skin vasculitis and non-specific symptoms such as artralgias and fever. The correct diagnosis was supported by positive cANCA. A clinical and laboratory remission of the disease was achieved by combined immunosuppressive therapy. Subsequently, she developed a subglotic stenosis possibly due to reparative changes.