The aim of this study was to identify RS1 pathogenic variants in Czech patients with X-linked retinoschisis (XLRS) and to describe the associated phenotypes, including natural history, in some cases. Twenty-one affected males from 17 families were included. The coding region of RS1 was directly sequenced and segregation of the identified mutations was performed in available family members. In total, 12 disease-causing variants within RS1 were identified; of these c.20del, c.275G>A, c.[375_379del; 386A>T], c.539C>A and c.575_576insT were novel, all predicted to be null alleles. The c.539C>A mutation occurred de novo. Three patients (aged 8, 11 and 19 years) were misdiagnosed as having intermediate uveitis and treated with systemic steroids. Repeat spectral domain optical coherence tomography examinations in four eyes documented the transition from cystoid macular lesions to macular atrophy in the fourth decade of life. Four individuals were treated with topical dorzolamide and in two of them, complete resolution of the cystic macular lesions bilaterally was achieved, while one patient was noncompliant. Rebound phenomenon after discontinuation of dorzolamide for 7 days was documented in one case. Misdiagnosis of XLRS for uveitis is not uncommon; therefore, identification of disease-causing variants is of considerable benefit to the affected individuals.

• MeSH
• antihypertenziva aplikace a dávkování   terapeutické užití   MeSH
• dítě MeSH
• frekvence genu MeSH
• kojenec MeSH
• lidé MeSH
• mladiství MeSH
• mutace MeSH
• oční proteiny genetika   MeSH
• optická koherentní tomografie MeSH
• předškolní dítě MeSH
• retinoschisis farmakoterapie   genetika   patologie   MeSH
• rodokmen MeSH
• sulfonamidy aplikace a dávkování   terapeutické užití   MeSH
• thiofeny aplikace a dávkování   terapeutické užití   MeSH
• Check Tag
• dítě MeSH
• kojenec MeSH
• lidé MeSH
• mladiství MeSH
• předškolní dítě MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH

Osteoblasts orchestrate bone formation through the secretion of type I collagen and other constituents of the matrix on which hydroxyapatite crystals mineralize. Here, we show that TENT5A, whose mutations were found in congenital bone disease osteogenesis imperfecta patients, is a cytoplasmic poly(A) polymerase playing a crucial role in regulating bone mineralization. Direct RNA sequencing revealed that TENT5A is induced during osteoblast differentiation and polyadenylates mRNAs encoding Col1α1, Col1α2, and other secreted proteins involved in osteogenesis, increasing their expression. We postulate that TENT5A, possibly together with its paralog TENT5C, is responsible for the wave of cytoplasmic polyadenylation of mRNAs encoding secreted proteins occurring during bone mineralization. Importantly, the Tent5a knockout (KO) mouse line displays bone fragility and skeletal hypomineralization phenotype resulting from quantitative and qualitative collagen defects. Thus, we report a biologically relevant posttranscriptional regulator of collagen production and, more generally, bone formation.

• MeSH
• buněčná diferenciace MeSH
• fyziologická kalcifikace genetika   MeSH
• kolagen typu I, řetězec alfa 1 genetika   metabolismus   MeSH
• kolagen typu I genetika   metabolismus   MeSH
• lidé MeSH
• messenger RNA genetika   metabolismus   MeSH
• modely nemocí na zvířatech MeSH
• myši knockoutované MeSH
• myši MeSH
• neurotrofní faktory genetika   metabolismus   MeSH
• nukleotidyltransferasy genetika   metabolismus   MeSH
• oční proteiny genetika   metabolismus   MeSH
• osteoblasty metabolismus   patologie   MeSH
• osteogenesis imperfecta genetika   metabolismus   patologie   MeSH
• osteogeneze genetika   MeSH
• osteonektin genetika   metabolismus   MeSH
• poly(A)-polymerasa genetika   metabolismus   MeSH
• polyadenylace MeSH
• protein - isoformy nedostatek   genetika   MeSH
• sekvenční analýza RNA MeSH
• serpiny genetika   metabolismus   MeSH
• signální transdukce MeSH
• vývojová regulace genové exprese MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

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.

• MeSH
• aniridie * genetika   MeSH
• dítě MeSH
• dospělí MeSH
• fenotyp MeSH
• lidé středního věku MeSH
• lidé MeSH
• mutace MeSH
• oční proteiny genetika   MeSH
• rodokmen MeSH
• sestřih RNA MeSH
• transkripční faktor PAX6 genetika   MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH

Gentiana lutea is a bitter herb that is traditionally used to improve gastric disorders. Recently, we have shown that Gentiana lutea extract (GE) also modulates the lipid metabolism of human keratinocytes in vitro and in vivo. In the present study, we investigated the role of GE on ceramide synthesis in human primary keratinocytes (HPKs) and psoriasis-like keratinocytes. We could demonstrate that GE increased the concentrations of glucosylceramides and the ceramide AS/AdS subclass without affecting the overall ceramide content in HPKs. The expression of ceramide synthase 3 (CERS3) and elongases (ELOVL1 and 4) was reduced in psoriasis lesions compared to healthy skin. Psoriasis-like HPKs, generated by stimulating HPKs with cytokines that are involved in the pathogenesis of psoriasis (IL-17, TNF-α, IL-22 and IFN-γ) showed increased levels of IL-6, IL-8 and increased expression of DEFB4A, as well as decreased expression of ELOVL4. The treatment with GE partly rescued the reduced expression of ELOVL4 in psoriasis-like HPKs and augmented CERS3 expression. This study has shown that GE modulates ceramide synthesis in keratinocytes. Therefore, GE might be a novel topical treatment for skin diseases with an altered lipid composition such as psoriasis.

• MeSH
• ceramidy metabolismus   MeSH
• elongasy mastných kyselin genetika   metabolismus   MeSH
• Gentiana chemie   MeSH
• keratinocyty cytologie   účinky léků   metabolismus   MeSH
• kultivované buňky MeSH
• lidé MeSH
• membránové proteiny genetika   metabolismus   MeSH
• metabolismus lipidů účinky léků   MeSH
• oční proteiny genetika   metabolismus   MeSH
• primární buněčná kultura MeSH
• psoriáza genetika   metabolismus   MeSH
• regulace genové exprese účinky léků   MeSH
• rostlinné extrakty chemie   farmakologie   MeSH
• sfingosin-N-acyltransferasa genetika   metabolismus   MeSH
• studie případů a kontrol MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Background: Congenital cataract is the most common cause of blindness in the world. Congenital cataracts are clinically and genetically heterogeneous and are mostly inherited in an autosomal dominant fashion. We identified the genetic cause of isolated autosomal dominant cataract in a four-generation British family and a Czech family.Methods: Whole exome sequencing (WES) was performed on one affected member in the British family and two affected members in the Czech family.Results: A novel missense variant c.388C > T; p.(R130C) was identified in the Lens integral membrane protein (LIM2) and found to co-segregate with disease in both families.Conclusions: Here we report the first autosomal dominant congenital cataract variant p.(R130C) in LIM2, causing a non-syndromic pulverulent and nuclear phenotype in European families.

• MeSH
• fenotyp MeSH
• katarakta vrozené   etiologie   patologie   MeSH
• lidé MeSH
• membránové proteiny genetika   MeSH
• mutace * MeSH
• mutační analýza DNA MeSH
• oční proteiny genetika   MeSH
• prognóza MeSH
• rodokmen MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

PURPOSE: The aim of this study was to determine the molecular genetic basis of an early-onset severe retinal dystrophy in three unrelated consecutive patients of Czech origin and to describe their ocular phenotype. METHODS: DNA samples from two probands were analyzed using a genotyping microarray (Asper) followed by either target analysis of 43 genes implicated in retinal disorders by next generation sequencing or whole-exome sequencing, respectively. The third proband underwent conventional Sanger sequencing of CRB1 based on her ocular findings. RESULTS: All three probands harboured a known disease-causing mutation c.2843G>A; p.(Cys948Tyr) in the CRB1 gene. One individual was homozygous for this mutation, while in the other two probands c.2308G>A; p.(Gly770Ser) and c.3121A>G; p.(Met1041Val) were also identified in the heterozygous state, respectively. Both variants were novel and evaluated by in silico analysis as pathogenic. A false-negative result was observed in one of the two samples examined by the genotyping microarray. Disease onset in all patients was before the age of 7 years. Hypermetropic refractive error, bilateral nummular retinal pigmentation, retinal thickening and cystoid spaces in the macula were observed in two probands, aged 6 and 7 years. The third proband, aged 28 years, had bone spicule-like pigmentary changes associated with increased retinal nerve fiber layer. CONCLUSIONS: The first study reporting on the molecular genetic cause of non-syndromic early-onset severe retinal dystrophy in Czech patients identified one homozygous and two compound heterozygote probands with CRB1 mutations. Retina nerve fibre layer measurements should be considered an integral part of the clinical evaluation of retinal dystrophies. Detailed clinical examination and imaging can both direct molecular screening and help to confirm or refute disease causation of identified variants.

• MeSH
• dědičné nemoci očí diagnóza   genetika   metabolismus   MeSH
• dítě MeSH
• dospělí MeSH
• elektroretinografie MeSH
• fenotyp MeSH
• genotyp MeSH
• homozygot MeSH
• lidé MeSH
• membránové proteiny genetika   metabolismus   MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mutace * MeSH
• mutační analýza DNA MeSH
• oční proteiny genetika   metabolismus   MeSH
• proteiny nervové tkáně genetika   metabolismus   MeSH
• retina diagnostické zobrazování   metabolismus   patofyziologie   MeSH
• retinální dystrofie diagnóza   genetika   metabolismus   MeSH
• rodokmen MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

Wnt/β-catenin signaling plays an essential role in the retinal pigment epithelium (RPE) determination. Since activity of Pax6 (together with Pax2) is also required for the RPE determination, we investigated a possible genetic interaction between Pax6 and Wnt/β-catenin signaling pathway by analyzing Pax6, β-catenin, and Pax6/β-catenin conditional knockout mice. Although Pax6 inactivation alone had no impact on initial specification determined by the expression of Mitf and Otx2, melanin pigmentation was reduced in the RPE. This suggests that along with Mitf and Otx2, Pax6 is required for the full differentiation of RPE. Reporter gene assays in vitro suggest that hypopigmentation is at least in part due to the direct regulation of genes encoding enzymes involved in melanin synthesis by Pax6, Mitf, and β-catenin. The RPE of a β-catenin/Pax6 double mutant was differentiated into the neural retina; however, the tissue was thinner than that of the conditional β-catenin mutant due to reduced proliferation. Together, our data demonstrate that Pax6 is required for the RPE differentiation by regulating pigmentation and accountable for hyperproliferation in the transdifferentiated RPE. In this context, Pax6 appears to function as a pleiotropic regulator, directing development of ocular tissues in concert with the signaling pathway and, at the same time, regulating expression of structural component of the eye, such as shielding pigment.

• MeSH
• beta-katenin genetika   metabolismus   MeSH
• homeodoménové proteiny genetika   metabolismus   MeSH
• myši transgenní MeSH
• myši MeSH
• oční proteiny genetika   metabolismus   MeSH
• represorové proteiny genetika   metabolismus   MeSH
• retina cytologie   metabolismus   MeSH
• retinální pigmentový epitel metabolismus   MeSH
• signální dráha Wnt * MeSH
• transdiferenciace buněk MeSH
• transkripční faktory paired box genetika   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Diabetes has become one of the major diseases affecting human health. Diabetic cataracts (DCs) are considered a common complication in diabetic patients. The present study investigated differences in lens proteomic profiles between DCs and age-related cataracts (ACs) to determine the mechanism underlying the formation of DCs. Intrasurgical samples were collected from eight DC patients and 12 AC patients, and lens proteins were extracted by lysis and separated using two-dimensional gel electrophoresis (2-DE). The electrophoretic bands were analysed using PD-Quest software 8.0.1. Differentially expressed proteins were identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and peptide mass fingerprinting combined with protein database searching. In the 2-DE maps, the DC and AC lens proteins migrated in the region of pH 5-9 with a relative molecular weight (RMW) of 14-97 kDa, whereas the RMW of more abundant crystallin was 20-31 kDa. Approximately three protein spots with differential intensity were detected. Two crystallin proteins (αB and βB1) were identified using MALDI-TOF-MS. Proteomic analysis of the crystalline humour is feasible, and the proteins can be well separated; moreover, differentially expressed lens proteins can be analysed using 2-DE and mass spectrometry to compare DC and AC. The present results indicate that the αB and βB1 crystallins may accelerate the development of DCs. These techniques offer new avenues for mechanistic evaluation and future prevention or therapy of DCs.

• MeSH
• 2D gelová elektroforéza MeSH
• alfa-krystaliny - řetězec B biosyntéza   genetika   MeSH
• beta-krystaliny - řetězec B biosyntéza   genetika   MeSH
• diabetes mellitus 1. typu komplikace   MeSH
• dospělí MeSH
• hmotnostní spektrometrie MeSH
• katarakta etiologie   genetika   metabolismus   MeSH
• komplikace diabetu etiologie   genetika   metabolismus   MeSH
• lidé středního věku MeSH
• lidé MeSH
• oční čočka chemie   MeSH
• oční proteiny biosyntéza   genetika   MeSH
• proteom MeSH
• regulace genové exprese * MeSH
• stanovení celkové genové exprese MeSH
• stárnutí genetika   metabolismus   patologie   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH

Phosducin (Pdc), a highly conserved phosphoprotein involved in the regulation of retinal phototransduction cascade, transcriptional control, and modulation of blood pressure, is controlled in a phosphorylation-dependent manner, including the binding to the 14-3-3 protein. However, the molecular mechanism of this regulation is largely unknown. Here, the solution structure of Pdc and its interaction with the 14-3-3 protein were investigated using small angle x-ray scattering, time-resolved fluorescence spectroscopy, and hydrogen-deuterium exchange coupled to mass spectrometry. The 14-3-3 protein dimer interacts with Pdc using surfaces both inside and outside its central channel. The N-terminal domain of Pdc, where both phosphorylation sites and the 14-3-3-binding motifs are located, is an intrinsically disordered protein that reduces its flexibility in several regions without undergoing dramatic disorder-to-order transition upon binding to 14-3-3. Our data also indicate that the C-terminal domain of Pdc interacts with the outside surface of the 14-3-3 dimer through the region involved in Gtβγ binding. In conclusion, we show that the 14-3-3 protein interacts with and sterically occludes both the N- and C-terminal Gtβγ binding interfaces of phosphorylated Pdc, thus providing a mechanistic explanation for the 14-3-3-dependent inhibition of Pdc function.

• MeSH
• fosfoproteiny chemie   genetika   metabolismus   MeSH
• fosforylace MeSH
• krysa rodu rattus MeSH
• lidé MeSH
• molekulární modely MeSH
• oční proteiny chemie   genetika   metabolismus   MeSH
• proteiny 14-3-3 chemie   genetika   metabolismus   MeSH
• proteiny vázající GTP - regulátory chemie   genetika   metabolismus   MeSH
• sekvence aminokyselin MeSH
• terciární struktura proteinů MeSH
• vazba proteinů MeSH
• vazebná místa MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Úvod: vrozená izolovaná aniridie je vzácné postižení struktur obou očí projevující se především chyběním duhovky, fotofobií a sníženým viděním. V klinickém obrazu může být přítomný nystagmus, strabismus, ptóza víčka, amblyopie, vyšší refrakční vada, anisometropie, rohovkové změny, patologie čočky, dysgeneze struktur komorového úhlu, hypoplazie makuly a zrakového nervu, kongenitální nebo sekundární glaukom. Tento typ dysgeneze oka je nejčastěji podmíněn mutací PAX6. Cílem naší práce bylo popsat oftalmologické nálezy členů rodiny s výskytem dědičné aniridie (MIM #106210), diskutovat jejich závažnost, prognózu a možnosti terapie. Materiál a metodika: zhodnocení předchozích klinických záznamů a oftalomogické vyšetření proběhlo u 4 osob z tří generací rodiny s výskytem izolované vrozené aniridie. Dle možností a compliance byla u pacientů provedena tato vyšetření: vízus, nitrooční tenze, refrakce, vyšetření štěrbinovou lampou, biomikroskopické vyšetření, pachymetrie, OCT. Dále bylo provedeno klinicko genetické vyšetření a následně sekvenace PAX6. Výsledky: u všech vyšetřených osob byla zjištěna plně vyjádřená závažná symptomatologie, byla pozorována přítomnost aniridie a fotofobie. Pozitivní korelaci s věkem vykazoval progresivní pokles vizu k praktické slepotě v důsledku keratopatie asociované s aniridií, sekundárním glaukomem a kataraktou. DNA analýza u všech postižených osob odhalila přítomnost mutace p.Gln180X v PAX6. Tato mutace vede k tvorbě zkráceného atak nefunkčního PAX6 proteinu. Závěr: mutace v genu PAX6, které vedou k předčasné terminaci tvorby bílkoviny, jsou spojené často s těžkým průběhem onemocnění as malou variabilitou závažnosti postižení intrafamiliárně. Tento fakt dokumentuje irodina popsaná v této práci, kdy projevy upacientů jsou závažné aprogredují s věkem. Terapie je svízelná, často jen parciálně úspěšná, jako je tomu iv případě sekundárního glaukomu mladé pacientky. Každý operační zákrok je nutno individuálně vždy velmi pečlivě uvážit. Navíc utéto nemoci dochází kneúprosné progresi očního nálezu spostupným zhoršováním itak již většinou chabé zrakové ostrosti až do praktické slepoty. Klíčová slova: aniridia, PAX6, hypoplazie makuly, glaukom

Background: inborn isolated aniridia is rare bilateral impairment of several eye structures manifesting mainly by absence of iris, photophobia and decreased visual acuity. There are also others ocular symptoms associated with aniridia such as nystagmus, strabismus, eyelid ptosis, amblyopia, serious refractive errors, anisometropia, corneal changes, impairment of the lens, chamber angle dysgenesis, optic nerve and macular hypoplasia and congenital or secondary glaucoma. The most frequent aetiology of this eye dysgenesis is mutation in PAX6. Aim of this report is to describe ocular findings in the family with familial aniridia (MIM #106210), to debate their severity, prognosis and therapy options. Material and methods: assessment of previous medical history and actual ophthalmological findings in 4 persons of 3 generation family with aniridia. According to the compliance, the patients underwent these tests: assessment of the visual acuity, intraocular pressure, refraction test, slit-lamp examination and biomicroscopy, pachymetry test and OCT examination. The genetic counselling was performed with subsequent PAX6 mutation analysis. Results: all of the examined aniridia family members showed severe symptoms of the disease, the aniridia and photophobia were present. Positive age related correlation showed progressive visual acuity decrease to the practical blindness due to aniridia-associated keratopathy, secondary glaucoma and cataract. DNA analysis revealed presence of p.Gln180X PAX6 mutation in all of the affected persons. The mutation leads to shortened and therefore non-functional protein. Conclusions: PAX6 mutations leading to premature termination of protein translation are frequently associated with severe symptoms of aniridia and small intrafamilial variability of ocular impairment. This fact is also well demonstrated in members of family described by this report, the symptoms are severe and progressing with age. Therapy is difficult and often with partial success, such in case of secondary glaucoma in young girl from this family. Any eye surgery must be individually judged due to risk of several post-operative complications. And more, the poor vision in aniridia patients is progressively worsening in time to practical blindness. Key words: aniridia, PAX6, macular hypoplasia, glaucoma

• Klíčová slova
• hypoplazie makuly,
• MeSH
• aniridie * diagnóza   genetika   patofyziologie   terapie   MeSH
• diagnostické techniky oftalmologické MeSH
• dítě MeSH
• dospělí MeSH
• fotofobie etiologie   MeSH
• glaukom etnologie   farmakoterapie   MeSH
• homeodoménové proteiny genetika   MeSH
• katarakta etiologie   MeSH
• lidé MeSH
• mutace MeSH
• nemoci rohovky etiologie   MeSH
• oční proteiny * genetika   MeSH
• patologický nystagmus etiologie   MeSH
• progrese nemoci MeSH
• represorové proteiny genetika   MeSH
• retina patologie   MeSH
• rodokmen MeSH
• rohovka patologie   MeSH
• sekvenční analýza DNA MeSH
• senioři MeSH
• strabismus etiologie   MeSH
• transkripční faktory paired box genetika   MeSH
• zraková ostrost MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• kazuistiky MeSH