The eye contains closely related but widely different tissues, offering a unique opportunity to investigate the phenotype and function of monocyte-derived cell populations within functionally unique microenvironments in a single complex organ. The uveal tract and retina contain rich networks of immune cells that reside and traffic through the eye, these cells having been implicated in various ocular inflammatory processes and immune-mediated diseases. One such inflammatory condition is human posterior uveitis, an autoimmune disease mainly affecting the retina. As current treatments for posterior uveitis only serve to slow down disease progression, studies using animal models, namely, experimental autoimmune uveoretinitis (EAU), have focused on determining the key cellular and molecular mediators involved in disease initiation in order to expand the potential for novel therapeutic applications. The overall purpose of experiments in this thesis was to explore monocyte-derived cell populations of the uveal tract and retina, this being achieved by utilising a novel transgenic mouse model. Cx3cr1gfp/gfp transgenic mice on both BALB/c and C57Bl/6 backgrounds contain an enhanced green fluorescent protein (eGFP) encoding cassette knocked into the Cx3cr1 gene, disrupting its expression but facilitating GFP expression under the control of the Cx3cr1 promoter. Heterozygous (Cx3cr1+/gfp) mice were generated by crossing Cx3cr1gfp/gfp mice to wild-type (WT) mice. This transgenic model allowed for the exquisite visualisation of Cx3cr1-bearing monocyte-derived dendritic cells (DC) and macrophages in ocular tissues, whilst also enabling the investigation of a potential role for Cx3cr1 in recruiting monocyte-derived cells to the eye in steady-state and inflammatory conditions.

L'orientation des lentilles sclérales devient extrêmement importante pour maximiser la stabilité et la vision chez des patients qui portent des lentilles ayant une face antérieure torique. Les techniques cliniques actuelles pour déterminer la rotation d'une lentille sclérale impliquent l'utilisation de lentilles diagnostiques; cependant, ces méthodes nécessitent beaucoup de temps de chaise pour obtenir un ajustement optimal. Des appareils novateurs existent maintenant pour évaluer la forme de la surface oculaire et peuvent s’avérer utiles pour prédire la rotation des lentilles sclérales et augmenter l'efficacité globale des ajustements. Le but de cette étude comparative et randomisée était d'évaluer la rotation des lentilles sclérales en se basant sur des valeurs fournies par deux topographes. Quinze participants ont été recrutés et leurs deux yeux ont été imagés à l'aide de deux topographes: le Eye Surface Profiler (ESP) et le Cornea Sclera Profile (CSP). Les participants ont ensuite été ajustés avec la lentille OneFit MED dans un oeil et la Zenlens dans l'autre. La rotation de chaque lentille a été évaluée à l’aide de la lampe à fente et comparée à l’axe le plus cambré de l’astigmatisme conjonctival identifié par chaque topographe. Bien que les rotations des deux instruments ne soient pas comparables, l'ESP a prédit une rotation à moins de 15° de l'observation à la lampe à fente, ce qui la rend cliniquement acceptable pour les lentilles avec de faibles valeurs de puissance cylindrique. L'acuité visuelle des patients portant des lentilles avec des quantités élevées de toricité pouvant être plus affectée par la quantité de rotation, les ajustements empiriques semblent, à ce stade, être déconseillés pour ces cas. / Scleral lens orientation becomes extremely important to maximize vision and lens stability in patients who are fitted in front-toric lenses. Current clinical techniques to determine the rotation of a scleral lens involve the use of diagnostic lenses; however, these methods require a substantial amount of chair time to achieve an optimal fit. Contemporary equipment exists to evaluate ocular surface shape, which could be useful in predicting scleral lens rotation and increasing the overall efficiency of scleral lens adjustments. The goal of this comparative and randomized study was to evaluate scleral lens rotation based on the values provided by two scleral topographers. Fifteen participants were recruited and both eyes were imaged using two scleral topographers: the Eye Surface Profiler (ESP) and the Cornea Sclera Profile (CSP). Participants were fitted with the OneFit MED on one eye and the Zenlens on the other. Each lens’s rotation was evaluated at the slit lamp and compared to the steep axis of conjunctival astigmatism identified by each topographer. While the rotations from both instruments are not comparable, the ESP predicted rotation within 15˚ from slit lamp observation, which makes it clinically acceptable for lenses with low values of cylindrical power. The visual acuity of patients wearing lenses with a high amount of toricity may be affected by the amount of rotation and are not suitable for empirical fittings at this point in time.

Univerzita Karlova 1. lékařská fakulta Studijní program: Zobrazovací metody v lékařství (P5150) Studijní obor: Zobrazovací metody v lékařství (5103V000) MUDr. Adam Kopecký Změny na předním segmentu po 25G+ PPV Changes in the Anterior Segment after 25G+ PPV Abstract Vedoucí závěrečné práce/Školitel: doc. MUDr. Jiří Pašta, CSc., FEBO Praha, 2019 Abstract: Pars plana vitrectomy is a dominant surgical method in posterior segment surgery of the eye. The principle of this method is the surgical approach via the pars plana anatomical region. With this approach, surgeon performs the posterior segment surgery without directly affecting the anterior segment of the eye. Short-term effect of pars plana vitrectomy on the anterior segment of the eye has been repeatedly proven. That might be an important factor both in assessing the complications of this method and planning the surgery in complicated patients. Generally, it is presumed that we do not observe long-term effects on anterior segment after pars plana vitrectomy. The aim of our work is to evaluate the long-term changes in the anterior segment after the uncomplicated 25G + PPV with gas tamponde. The group of 20 patients was selected according to strict indication criteria. Using a series of examinations, we examined specific parameters such as: intraocular...

Proper eye development depends on expression and mutual regulation of many genes. Anterior segment dysgenesis (ASD) are a highly heterogeneous group of diseases exhibiting all types of Mendelian inheritance, which manifest as combination of congenital abnormalities of the cornea, iris, anterior chamber angle or lens. Screening of genes associated with ASD does not often lead to the identification of the underlying genetic cause implying that there are still novel variants or genes to be discovered. Molecular genetic analysis in 12 probands with ASD using Sanger and whole-exome sequencing were performed. Functional analysis by Exon trapping assay was provided in variants predicted to effect pre-mRNA splicing. Four PAX6 mutations evaluated as pathogenic or likely pathogenic in a heterozygous state were found in four probands c.183C˃G; p.(Tyr61*), c.1032+1G>A, c.1183+1G>T and c.622C>T; p.(Arg208Trp). One proband was found to be a compound heterozygote for c.244A>G; p.(Met82Val) and c.541delG; p.(Glu181Lysfs*26) mutations in FOXE3. In 7 probands, no potentially pathogenic variants were identified. Exon trapping assay confirmed that mutations c.1032+1G>A and c.1183+1G>T have an effect on pre-mRNA splicing of the PAX6 gene. Detailed molecular-genetic analysis in patients with ASD may contribute to...

Proper eye development depends on expression and mutual regulation of many genes. Anterior segment dysgenesis (ASD) are a highly heterogeneous group of diseases exhibiting all types of Mendelian inheritance, which manifest as combination of congenital abnormalities of the cornea, iris, anterior chamber angle or lens. Screening of genes associated with ASD does not often lead to the identification of the underlying genetic cause implying that there are still novel variants or genes to be discovered. Molecular genetic analysis in 12 probands with ASD using Sanger and whole-exome sequencing were performed. Functional analysis by Exon trapping assay was provided in variants predicted to effect pre-mRNA splicing. Four PAX6 mutations evaluated as pathogenic or likely pathogenic in a heterozygous state were found in four probands c.183C˃G; p.(Tyr61*), c.1032+1G>A, c.1183+1G>T and c.622C>T; p.(Arg208Trp). One proband was found to be a compound heterozygote for c.244A>G; p.(Met82Val) and c.541delG; p.(Glu181Lysfs*26) mutations in FOXE3. In 7 probands, no potentially pathogenic variants were identified. Exon trapping assay confirmed that mutations c.1032+1G>A and c.1183+1G>T have an effect on pre-mRNA splicing of the PAX6 gene. Detailed molecular-genetic analysis in patients with ASD may contribute to...