Submitted by Renata Lopes (renatasil82@gmail.com) on 2016-05-20T17:53:34Z No. of bitstreams: 1 flaviabatistabarbosadesa.pdf: 1013858 bytes, checksum: 1e5c85e290deeb77a83c4367e5c61db1 (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2016-07-02T11:27:01Z (GMT) No. of bitstreams: 1 flaviabatistabarbosadesa.pdf: 1013858 bytes, checksum: 1e5c85e290deeb77a83c4367e5c61db1 (MD5) / Made available in DSpace on 2016-07-02T11:27:01Z (GMT). No. of bitstreams: 1 flaviabatistabarbosadesa.pdf: 1013858 bytes, checksum: 1e5c85e290deeb77a83c4367e5c61db1 (MD5) Previous issue date: 2012-03-26 / Objetivo: Analisar os fatores que influenciam o processo de doação de córneas entre os potenciais doadores de um Hospital Público e de um Hospital Privado de Juiz de Fora. Métodos: estudo seccional do tipo descritivo. Foram rastreados os óbitos ocorridos em um Hospital Público e um Hospital Privado de Juiz de Fora no período de 01 janeiro de 2010 a 31 de dezembro de 2010, identificados os potenciais doadores de córneas, verificado se os óbitos foram notificados à Central de Notificação, Captação e Distribuição de Órgãos (CNCDO) da Zona da Mata de Juiz de Fora e se esta notificação resultou ou não em doação. Caso não tenha ocorrido doação, foi justificada a causa da não efetivação da mesma, e caso tenha ocorrido, foi analisado o destino da córnea captada (transplante ou perda). Os dados foram processados e discutidos utilizando-se a análise estatística feita através do Programa SPSS (Statistical Package for the Social Sciences), versão 13.0. Resultados: A população em estudo se caracteriza por serem em sua maioria pacientes do sexo masculino (53,1%), com idade entre 2 a 80 anos (69,1%), com causas mortis que não contraindicaram o transplante de córneas (59,1%). Dos 863 óbitos, 138 (16%) não foram notificados à CNCDO-Zona da Mata e destes, 54 eram potenciais doadores. Do total de óbitos, 210 (24,3%) foram classificados como potenciais doadores. O percentual de captação efetiva dos casos entrevistados foi de 34,9%. Foram realizadas 30 doações de córneas (19,2%). Observou-se que das 60 córneas captadas (30 doadores), 75,0% (45) foram descartadas. A não efetivação da doação foi justificada pela recusa familiar em 58 casos (7,0%), pelos problemas logísticos ou estruturais em 125 casos (15,0%) e pelas contraindicações médicas registradas na CNCDO-Zona da Mata em 650 casos (78,0%). Conclusão: Da população estudada, 24,3% eram potenciais doadores de córneas. A contraindicação médica foi a maior justificativa para a não efetivação da doação de córneas, correspondendo a 78,0% das causas. Considerando que apenas 25% das córneas captadas foram transplantadas, conclui-se que há uma grande desigualdade entre o número de doadores potenciais e o número de doadores reais para o transplante de córneas, pois muito se perde durante todo o processo de doação. / Objective: To analyze factors that influence the process of cornea donation among potential donors for a Public Hospital and a Private Hospital in Juiz de Fora. Methods: A descriptive cross sectional study. We traced the deaths occurred in a Public Hospital and a Private Hospital in Juiz de Fora in the period from January 1, 2010 to December 31, 2010, identified the potential donors of corneas, checked whether the deaths were reported to the Central Notification, Procurement and Distribution organs (CNCDO) from Zona of Mata in Juiz de Fora and whether or not this notification resulted in donation. If there were no donation, it was justified the cause of not being done, and if it has occurred, we analyzed the fate of the captured cornea (transplantation or loss). The data were analyzed and discussed using the statistical analysis performed using the SPSS (Statistical Package for Social Sciences) version 13.0. Results: The mortality profile of the institutions under study is characterized by being mostly male (53.1%), aged from 2 to 80 years (69.1%), with cause of death that did not prevent corneal transplantation to be done (59.1%). Of the 863 deaths, 138 (16%) were not notified to CNCDO-Zona of Mata and of these, 54 were potential donors. Of the total deaths, 210 (24.3%) were classified as potential donors. The percentage of effective capture of the cases surveyed was 34.9%. 30 donations of corneas were performed (19.2%). It was observed that from 60 corneal grafts (30 donors), 75.0% (45) were discarded. The effectiveness of the donation was not justified by family refusal in 58 cases (7.0%), by logistical or structural problems in 125 cases (15.0%) and recorded in the medical contraindications CNCDO-Zona da Mata in 650 cases (78.0 %). Conclusion: The study population, 24.3% were potential donors of corneas. The medical contraindication was the biggest reason for not effecting the donation of corneas, corresponding to 78.0% of the causes. Whereas only 25% of corneal grafts were transplanted, it is concluded that a considerable difference between the number of potential donors and the number of actual donors for transplantation of corneas, since much is lost during the donation process.

O conhecimento da morfologia endotelial nas diferentes regiões da córnea é de suma importância para avaliação de endotélios corneanos saudáveis e doentes e de suas respostas ao uso de medicações. O objetivo do presente estudo foi descrever a morfologia do endotélio nas regiões central e periférica superior da córnea saudável de equinos, medir a área média das células pentagonais, hexagonais e heptagonais presentes nas regiões avaliadas, calcular o polimegatismo de cada tipo celular e correlacionar estes parâmetros entre os diferentes formatos celulares. Foram estudados dez equinos, machos ou fêmeas, de diferentes idades, provenientes de um abatedouro licenciado. Imagens da superfície posterior do endotélio da córnea foram obtidas com uso de microscopia eletrônica de varredura. A morfologia do endotélio de diferentes regiões da córnea foi avaliada. Além disso, foi correlacionada a variabilidade do tamanho celular médio com a morfologia endotelial. A análise estatística foi conduzida usando o teste de análise de variância (ANOVA) seguido do teste de Tukey (Post-Hoc), com nível de significância de 5%. As amostras avaliadas foram compostas em sua maioria por células hexagonais (60,5%), pentagonais (21,4%) e heptagonais (16,9%), células com quatro, oito ou nove lados compuseram 1,3%. A área celular média das células pentagonais foi 203,58 μm², das hexagonais foi 223,84 μm² e nas heptagonais foi 270,54 μm². O polimegatismo encontrado foi de 16% nas células pentagonais e hexagonais e de 20% nas heptagonais. A morfologia das células endoteliais de equinos saudáveis não diferiu entre as regiões central e periférica superior da córnea, sugerindo que a região central é representativa da região periférica. As células com sete lados apresentaram polimegatismo maior em relação às células de seis e de cinco lados. / The knowledge of the endothelial morphology in the different regions of the cornea is important for the evaluation of healthy and sick corneal endothelium and its response to drugs. In order to describe the endothelial morphology in the central and superior peripheral regions of the equines’ cornea, evaluate area from pentagonal, hexagonal and heptagonal cells present in the evaluated regions, calculate the polimegathism and correlate these parameters, two healthy corneas were collected of ten equine, male or female, of different ages. Images of the posterior surface of the corneal endothelium were taken with scanning electron microscope. The endothelial morphology was studied in the different regions of the equines’ cornea. In addition, the polimegathism and morphology was also correlated. A statistical analysis was conducted using the Analysis of Variance (ANOVA) followed by Tukey's Test (Post-Hoc), with a 5% level of significance. In the central region, the endothelium consisted of 58.8% hexagonal cells, 22.6% pentagon, 17.1% heptagonal and 1.4% cells with either four, eight or nine sides. In the superior peripheral region, 62.1% of the cells were hexagonal, 20.2% pentagon, 16.6% heptagonal and 1.1% cells with four or eight sides. The average cell area of the pentagonal cells was 210.77 ± 31.53 μm² in the central region and 196.39 ± 34.35 μm² in the superior peripheral region; in hexagonal cells, the average cell area found in the central region was 216.12 ± 37.09 μm² and 231.56 ± 34.95 μm² in the superior peripheral region; and in cells with seven sides, the average cell area was 261.76 ± 55.29 μm² in the central region and in the superior peripheral region was found 279.32 ± 52.37μm². When not considering the corneal regions, the polymegathism found was 16% in the pentagonal and hexagonal cells and 20% in the heptagonal cells. The morphology results obtained did not differ between the central and peripheral regions of the cornea, suggesting that the central region is representative of the peripheral region. The highest coefficient of variation was seen in cells with seven sides.

Purpose: The aim of this work was to characterize the healthy human cornea and the cornea of patients suffering from posterior polymorphous corneal dystrophy (PPCD) using different antibodies. Despite the fact that PPCD is a very rare disorder, one of the largest groups of PPCD patients in the world comes from the Czech Republic. This offers us the opportunity to investigate the changes on the clinical, cellular and molecular levels. Material and Methods: A collection of 25 control corneas as well as 16 pathological corneas from PPCD patients were used. Epithelial (cytokeratins) and mesothelial markers (mesothelin, calbindin 2, HBME-1 protein) were detected in all layers of the healthy corneas using immunocyto- and immunohistochemistry. The expression of all markers was confirmed using molecular methods as well (RT-PCR and Western blot). Changes in the expression of cytokeratins and changes in the extracellular matrix structure (collagen IV and VIII) were studied in the PPCD corneas. Combined fluorescent immunohistochemistry with fluorescence in situ hybridization were used in order to characterize the origin of abnormal cells on the posterior graft surface, which cause the recurrence of the PPCD after penetrating keratoplasty surgery. Results: Changes in the cytokeratin expression (strong...

La greffe de Membrane Amniotique Humaine (MAH) permet la cicatrisation des ulcères pré-perforants de la cornée et de sauver un nombre significatif d'yeux victimes de brûlures chimiques. La MAH est un matériel biologique, son utilisation pour le traitement des maladies de la surface oculaire donne de bons résultats en raison de sa capacité à réduire l'inflammation et promouvoir une épithélialisation rapide. Pour son utilisation en clinique, la MAH doit bien évidemment être stérile, mais aussi facile à transporter du centre préleveur au centre greffeur, et stockable longtemps et facilement. Actuellement en routine à la banque de tissus de Rouen, la membrane amniotique est séparée de l’amnios puis dénudée de leur couche spongieuse. Par la suite cette membrane est conservée par cryopréservation (congélation à –80°C) ce qui complique potentiellement l’acheminement des membranes. Par conséquent, dans le cadre de cette étude avec la Banque Normande de Cornées du CHU de Rouen nous avons développé la lyophilisation des MAH pour faciliter son utilisation et sa distribution. L’étude du mapping de la MAH permettra également de déterminer si le taux de facteurs de croissance est homogène dans la MAH ou s’il dépend sa distance par rapport au cordon ombilical. L’étude de la biocompatibilité in vivo d’un deuxième matériau composé de collagène nous permet également d’envisager une alternative pour une implantation du stroma. Nos analyses protéiques (ELISA et Label-free) des MAH la lyophilisées ne montrent pas de différence significative en terme de quantité et de qualité protéique. L’approche protéomique est complétée par l’analyse de la capacité des cellules épithéliales de cornée humaines (CEC) à se multiplier sur la membrane amniotique lyophilisée in vitro. Nous n’avons pas observé de différence entre la croissance des cellules épithéliales sur la MAH lyophilisée ou congelée. L’analyse du total de protéines extraites montre également que la lyophilisation ne dégrade pas les MAH au niveau protéique.Au niveau structurel les résultats de la microscopie électronique ont montré que la structure du stroma de la MAH est impactée par la lyophilisation. La greffe des MAH a été réalisée sur les ulcères cornéens chez le lapin. Au cours de l’expérimentation les lapins n’ont pas montré de signe d’inflammation, les analyses histologiques ont mis en évidence l’épithélialisation de la surface oculaire. Ce projet est en collaboration avec l‘association ophtalmo sans frontière pour qu’à terme le développement de l’utilisation clinique des MAHL répondant notamment à des besoins humanitaires (Cameroun). Notre étude de la cartographie de la MAH a également montré qu’une variabilité en termes de quantité de protéine existe entre les différents donneurs. Dans cette étude nous avons également montré que la couche spongieuse est une source de facteurs de croissance importante dans le processus de cicatrisation des ulcères cornéens. / The Human Amniotic Membrane (HAM) graft allows the healing of corneal ulcers and rescues a significant number of eyes with chemical burn. HAM is a biological material, its use for the treatment of ocular surface diseases gives good results because of its ability to reduce inflammation and promote rapid epithelialization. For its clinical use, the HAM must of course be sterile, but also easy to transport from the sampling center to the transplant center, and easily storable and for a long time. Currently on routine in the tissue bank of Rouen, the amniotic membrane is separated from the amnion and denuded of its spongy layer. Subsequently this membrane is stored by cryopreservation (freezing at -80 ° C) which potentially complicates the delivery of membranes. Consequently, as part of this study with the Banque Normande de Cornées of Rouen University Hospital, we have developed freeze-drying of HAM to facilitate its use and distribution. The HAM mapping study will also determine whether the level of growth factors is homogeneous in the HAM or whether it depends on its distance from the umbilical cord. The study of the in vivo biocompatibility of a second material composed of collagen also allows us to consider an alternative for implantation at the level of the stroma. Our protein analyzes (ELISA and Label-free) of freeze-dried HAM do not show any significant difference in terms of quantity and protein quality. The proteomic approach is complemented by the analysis of the ability of human corneal epithelial cells (CECs) to multiply on the freeze-dried amniotic membrane in vitro. We did not observe any difference between the epithelial cells growths on freeze-dried or frozen HAM. The analysis of the extracted protein total also shows that freeze-drying does not degrade the HAM at the protein level. At the structural level the electron microscopy results showed that the structure of the MAH stroma is impacted by freeze-drying. The MAH transplant performed on corneal ulcers in rabbits was performed. During the experiment the rabbits did not show any sign of inflammation, the histological analyzes highlighted the epithelialization of the ocular surface.This project is in collaboration with OSF association for the development of the clinical use of MAHL responding in particular to humanitarian needs (Cameroon). Our study of HAM mapping also showed that variability in terms of amount of protein exists between different donors. We have also shown that the spongy layer is an important source of important growth factor in the healing process of corneal ulcers.

yes / Confocal microscopy is employed as a fast and non-invasive way to capture a sequence of images from different layers and membranes of the cornea. The captured images are used to extract useful and helpful clinical information for early diagnosis of corneal diseases such as, Diabetic Peripheral Neuropathy (DPN). In this paper, an automatic corneal subbasal nerve registration system is proposed. The main aim of the proposed system is to produce a new informative corneal image that contains structural and functional information. In addition a colour coded corneal image map is produced by overlaying a sequence of Cornea Confocal Microscopy (CCM) images that differ in their displacement, illumination, scaling, and rotation to each other. An automatic image registration method is proposed based on combining the advantages of Fast Fourier Transform (FFT) and phase correlation techniques. The proposed registration algorithm searches for the best common features between a number of sequenced CCM images in the frequency domain to produce the formative image map. In this generated image map, each colour represents the severity level of a specific clinical feature that can be used to give ophthalmologists a clear and precise representation of the extracted clinical features from each nerve in the image map. Moreover, successful implementation of the proposed system and the availability of the required datasets opens the door for other interesting ideas; for instance, it can be used to give ophthalmologists a summarized and objective description about a diabetic patient’s health status using a sequence of CCM images that have been captured from different imaging devices and/or at different times

The main aim of this thesis is to evaluate the effect of meridional blur, using refractive induced astigmatism, on visual performance at far and close distances. Visual performance was evaluated using letter discrimination tasks at distance and near (visual acuity, VA) and a reading task at near on subjects with pharmacologically blocked (young) or absent accommodation (presbyopic and pseudophakic). The effect of astigmatism was tested using positive cylindrical lenses oriented at 180 and 90 degrees, these simulating with- (WTR) and against-the-rule (ATR) astigmatism. Other refractive status were also evaluated, namely, in-focus and spherical defocus. The visual performance data were correlated with biometric measurements (pupil size, anterior chamber depth (ACD), corneal and ocular aberrations, corneal multifocality, patient age, axial length). Further, the functionality of meridional blur was evaluated for alphabets in addition to the standard Roman alphabet using a VA task. The results confirm that myopic astigmatism contributes to a better visual performance at closer distances, with ATR astigmatism providing higher performance for reading tasks compared to other forms of astigmatism. Anatomical factors such as pupil size, corneal multifocality and ACD were significantly correlated visual performance, while other ocular characteristics were not. Ray tracing modelling using wavefront data was a moderate predictor of VA and reading acuity. The results of the effect of meridional blur orientation on alphabets other than the Roman alphabet, suggest that visual performance is dependent on the interaction between blur orientation and letter¿s spatial characteristics. In conclusion, pseudoaccommodation is a multifactorial phenomenon with pupil size being the major contributor for the improvement in visual performance. Against-the-rule shows advantages over WTR astigmatism, by providing higher reading performance, however extending the present and previous findings for clinical application will require further investigation on the effect of meridional blur in common and socio-culturally adapted tasks. / Bradford School of Optometry and Vision Sciences

Corneal diseases are one of the major causes of visual impairment and blindness worldwide. Used for diagnoses, a laser confocal microscope provides a sequence of images, at incremental depths, of the various corneal layers and structures. From these, ophthalmologists can extract clinical information on the state of health of a patient’s cornea. However, many factors impede ophthalmologists in forming diagnoses starting with the large number and variable quality of the individual images (blurring, non-uniform illumination within images, variable illumination between images and noise), and there are also difficulties posed for automatic processing caused by eye movements in both lateral and axial directions during the scanning process. Aiding ophthalmologists working with long sequences of corneal image requires the development of new algorithms which enhance, correctly order and register the corneal images within a sequence. The novel algorithms devised for this purpose and presented in this thesis are divided into four main categories. The first is enhancement to reduce the problems within individual images. The second is automatic image classification to identify which part of the cornea each image belongs to, when they may not be in the correct sequence. The third is automatic reordering of the images to place the images in the right sequence. The fourth is automatic registration of the images with each other. A flexible application called CORNEASYS has been developed and implemented using MATLAB and the C language to provide and run all the algorithms and methods presented in this thesis. CORNEASYS offers users a collection of all the proposed approaches and algorithms in this thesis in one platform package. CORNEASYS also provides a facility to help the research team and Ophthalmologists, who are in discussions to determine future system requirements which meet clinicians’ needs. / The data and image files accompanying this thesis are not available online.

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Deutschsprachige Zusammenfassung Dissertation zur Erlangung des akademischen Grades Dr. med. Titel: Mögliche Korrelation zwischen dem Stadium der altersabhängigen Makuladegeneration und der zentralen Hornhautdicke eingereicht von: Christian Koch angefertigt an der: Klinik und Poliklinik für Augenheilkunde Universität Leipzig betreut von: Professor Dr. med. Peter Wiedemann Klinik und Poliklinik für Augenheilkunde Universität Leipzig Monat und Jahr: Juni 2015 Annahme: Es soll ein statistischer Zusammenhang zwischen der zentralen Hornhautdicke in der Pupillenmitte mit dem nicht exsudativen frühen Stadium der AMD, dem exsudativen späten Stadium der AMD und einer Kontrollgruppe überprüft werden. Methodik: Die erste Studiengruppe stellten 48 Augen von 48 Patienten mit einer Form der frühen AMD dar (Durchschnittsalter 75,4 Jahre, 70,8% der Probanden waren Frauen). Die zweite Studiengruppe bildeten 49 Augen von 49 Patienten mit einer Form der späten AMD (Durchschnittsalter 79,8 Jahre, 59,2% der Probanden waren Frauen). Als Kontrollgruppe wurden 49 Augen von 49 Individuen ohne retinale oder korneale Erkrankungen genutzt (Durchschnittsalter 68,9 Jahre, 59,2% der Probanden waren Frauen). Die Vermessung der Hornhautdicke in Pupillenmitte im Sinne der Hornhautmittendicke erfolgte als Pachymetrie mit der Oculus Pentacam. Ergebnisse: Die durchschnittliche zentrale Hornhautdicke betrug bei der Kontrollgruppe 552,76 μm (SD 36,27 μm), bei der nicht exsudativen Gruppe 552,19 μm (SD 31,27 μm) und bei der exsudativen Gruppe 553,73 μm (SD 33,11 μm). Die Extrempunkte der Kontrollgruppe lagen bei 483 und 640 μm, der Gruppe der nicht exsudativen AMD bei 480 und 617 μm und Minimum und Maximum der exsudativen Gruppe bei 473 und 617 μm. Es gab keine statistisch signifikanten Unterschiede im arithmetischen Mittel der zentralen Hornhautdicke in Pupillenmitte bei der Studiengruppe mit früher AMD im Vergleich mit der Studiengruppe mit später AMD und jeweils im Vergleich zur Kontrollgruppe (P > 0,05). Schlussfolgerung: Die Messwerte der zentralen Hornhautdicke bei Patienten mit früher AMD, später AMD und Kontrollindividuen unterscheiden sich nicht. Die Erhebung der Hornhautmittendicke bietet somit keine Screeningmöglichkeit zur Erkennung einer AMD. / English Abstract Central corneal thickness in patients with AMD Koch, C.; Jochmann, C.; Wiedemann, P. University of Leipzig, Department of Ophthalmology and Eye Hospital, Leipzig, Germany Purpose: To evaluate central corneal thickness in patients with AMD in the early and late phase and a control group. Method: The first study group was made up of 48 eyes of 48 patients with early AMD (mean age 75.4 years, 70.8 % women), the second study group was made up of 49 eyes of 49 patients with late AMD (mean age 79.8 years, 59.2 % women). 49 eyes of 49 individuals without any retinal or corneal damage (mean age 68.9 years, 59.2 % women) were used as control group. The central corneal thickness was measured with the Oculus Pentacam pachymetry. Results: The mean central corneal thickness in early non-exudative AMD group was found to be 552.19 μm (SD 31.27 μm), while the mean central corneal thickness in the late exudative AMD group was measured as 553.73 μm (SD 33.11 μm). The control group had a mean central corneal thickness of 552.76 μm (SD 36.27 μm). There were no statistically significant differences in the mean central corneal thickness in the early non-exudative AMD group in comparison with the late exudative AMD group and each of them compared to the control group (P > 0,05). Conclusion: The central corneal thickness measurements do not differ in patients with early non-exudative AMD, late exudative AMD and control subjects.