Orientadora: Márcia Rita Fernandes Machado / Banca: José Antonio Thomazini / Banca: Luiza da Silva Lopes / Banca: Áureo Evangelista Santana / Banca: Marcos Lania de Araujo / Resumo: Diversos estudos mostraram que a ingestão de dieta com níveis inadequados de ferro pode causar, no sistema nervoso central (SNC) de ratos, alterações morfológicas, bioquímicas e comportamentais no animal. Esses estudos têm ainda indicado que animais deficientes em ferro apresentam redução no número de lamelas de mielina e prejuízos na aprendizagem. A deficiência de ferro é uma das mais comuns desordens nutricionais em pacientes pediátricos e adultos e atinge cerca de 2,5 a 5 bilhões de pessoas em todo mundo. A consequência mais explícita da deficiência de ferro é a anemia. O ferro está relacionado ao desenvolvimento de fibras nervosas mielínicas, as quais constituem mais de 80% do nervo óptico. Objetivou-se, na presente investigação, avaliar com o auxílio de microscopia eletrônica de transmissão, os possíveis efeitos da anemia ferropriva na estrutura do nervo óptico de ratos Wistar durante os períodos de lactação e pós-lactação. Os animais foram divididos em 2 grupos: Controle e Anêmico. Os anêmicos receberam uma dieta com 4 mg de ferro/Kg, e os controle, uma dieta com 35 mg de ferro/Kg. Avaliações do peso corpóreo, hemoglobina e hematócrito foram feitas para checar os efeitos da deficiência de ferro. Os animais foram anestesiados com cloridrato de quetamina IM (22 mg/Kg) e então sacrificados por perfusão transcardíaca com PBS 0,05M, pH 7,4, seguido da mistura fixadora paraformaldeído 2% e glutaraldeído 1% diluída em tampão fosfato. Um segmento do nervo óptico foi retirado e pós-fixado em solução de tetróxido de ósmio a 1% por duas horas a 4ºC, desidratado em acetona e incluído em araldite. Cortes ultra-finos com 60 nanômetros de espessura foram montados em grades de cobre, contrastados com acetato de uranila e citrato de chumbo, observados e fotografados ao microscópio eletrônico de transmissão para detalhada análise ultraestrutural... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Several studies showed that ingestion of diets with inadequate iron levels can cause morphological, biochemical and behavioral changes in the central nervous system (CNS) of rats. These studies have also shown that iron-deficient animals have reduced number of myelin lamellae and prejudice on learning. Iron deficiency is one of the most common nutritional disorders in pediatric patients and adults and affects about 2.5-5 billion people around the world. The most explicit result of iron deficiency is anemia. The iron is related to the development of myelinated nerve fibers, which constitute more than 80% of the optic nerve. The aim of this research is to evaluate, with transmission electronic microscopy, the possible effects of iron deficiency anemia in Wistar rats optic nerve structure during lactation and pos-lactation period. The animals were divided into 2 groups: Control and Anemic. The anemic group received 4 mg iron/Kg, the control group received 35 mg iron/Kg. Evaluation of body weight, hemoglobin and hematocrit were made to check the iron deficiency effects. The animals were anesthetized with ketamine 22 mg/Kg and then sacrificed by transcardiac perfusion with PBS 0.05 M, pH 7.4, followed by paraformaldehyde fixative mixture 2% and 1% glutaraldehyde. An optic nerve segment was removed and post-fixed in a solution of osmium tetroxide for two hours at 4°C, dehydrated in acetone and embedded in Araldite. Ultrathin sections with 60 nanometers thick were mounted on copper grids, contrasted with uranyl acetate and lead citrate, observed and photographed by transmission electronic microscope for detailed ultrastructural analysis of nerve fibers, blood vessels and glial cells. Both hematological and body weight were smaller in the anemic group. The ultrastructural analysis showed damaged myelinated and unmyelinated fibers, and glial cells of the anemic animals when compared with... (Complete abstract click electronic access below) / Doutor

Introdução: As doenças que envolvem a órbita representam um complexo problema cirúrgico, principalmente as localizadas no ápice orbitário, por onde passam estruturas críticas e um espaço pequeno. O uso do endoscópio por via endonasal para abordagem cirúrgica das lesões do ápice orbitário é uma técnica recente, com poucas citações na literatura. É necessário o estudo de referências anatômicas objetivas que tornem a cirurgia mais segura. Objetivo: Descrever os parâmetros anatômicos utilizados na abordagem cirúrgica endonasal endoscópica, assim como avaliar a concordância entre os hemicrânios do mesmo cadáver e as diferenças conforme o gênero. Casuística e métodos: Estudo anatômico em 30 cadáveres adultos, ambas as fossas nasais foram dissecadas (n=60 hemicrânios). Sob visibilização endoscópica endonasal, realizou-se a dissecção do ápice orbitário. Mensuramos a distância entre a crista etmoidal e o arco coanal para o forame óptico e para a fissura orbitária superior. Os resultados foram registrados na ficha de protocolo do estudo. Resultados: Foram dissecados 30 cadáveres, 60 hemicrânios ou lados. O sexo masculino foi mais prevalente, representando 63,3% dos cadáveres (19/30), enquanto o sexo feminino representou 36,7% (11/30). 43,3% dos cadáveres eram da raça branca (13/30), 20%, pardos (6/30), e 36,7%, negros (11/30). A correlação entre os valores conforme o lado nas seguintes aferições foi observada: Crista etmoidal - Forame óptico, (r=0,748, p=0.0001); Crista etmoidal - Fissura Orbitária Superior (r=0.785, p=0.0001), Arco coanal - Forame óptico (r=0,835, p=0.0001); Arco coanal - Fissura orbitária superior (r=0.820, p=0.0001). Foi obtido um Kappa de 0,444 na avaliação da concordância entre os lados em relação ao posicionamento da artéria etmoidal anterior no forame óptico. Conclusões: A sistematização da abordagem do ápice orbitário facilita seu acesso cirúrgico e a compreensão da anatomia. A crista etmoidal e o arco coanal se mostraram estruturas relevantes e com medidas constantes nos cadáveres estudados. Os valores do coeficiente de correlação de Spearman (r) foram maiores que 0,7, o que revela uma boa correlação entre as medidas dos hemicrânios do mesmo indivíduo. Ao analisarmos a concordância do posicionamento da artéria oftálmica entre os hemicrânios de um mesmo cadáver, podemos observar que a concordância foi moderada, o que representa assimetria e variação de localização da artéria. Ao compararmos as medidas aferidas entre os lados, observou-se que os valores são semelhantes e não houve diferença estatística das distâncias em nenhuma das referências anatômicas propostas para o estudo / Introduction: Diseases that affect the orbit pose a complex surgical challenge, particularly those involving the orbital apex, a small space through which critically important structures course. Endoscopic endonasal approaches to the surgical treatment of orbital apex lesions are a recent technique, with few citations in the literature. Research is still needed into objective anatomic landmarks that can improve surgical safety. Objective: To describe the anatomic landmarks used in endoscopic endonasal surgical approaches and assess agreement between placement of these landmarks in midsagittal sections of cadaver skulls and potential gender differences. Materials and methods: In this anatomic study, the nasal fossae of 30 adult cadavers were dissected (n=60 half-skulls). The orbital apex was dissected under endoscopic endonasal visualization. The distances between the ethmoidal crest and choanal arch to the optic foramen and to the superior orbital fissure were measured and recorded. Results: Overall, 30 cadavers were dissected for a total of 60 half-skulls or sides. The sample was predominantly male (63.3%, 19/30 cadavers); females accounted for the remaining 36.7% (11/30). Regarding skin color, 43.3% of cadavers were white (13/30), 20% were brown (6/30), and 36.7% were black (11/30). The following correlations between measurements according to side were observed: ethmoidal crest to optic foramen, r=0.748 (p=0.0001); ethmoidal crest to superior orbital fissure, r=0.785 (p=0.0001); choanal arch to optic foramen, r=0.835 (p=0.0001); choanal arch to superior orbital fissure, r=0.820 (p=0.0001). Analysis of the agreement of ophthalmic artery location within the optic foramen between skull halves revealed a kappa of 0.444. Conclusions: The approach systematization to the orbital apex will facilitate surgical access and improve understanding of the anatomy. In the cadavers studied in this sample, the ethmoidal crest and choanal arch were relevant structures and exhibited consistent measurements. Spearman correlation coefficients (r) were greater than 0.7, which is indicative of good correlation between measurements obtained in the skull halves of each cadaver. Analysis of the position of the ophthalmic artery in each skull half of the same cadaver revealed moderate agreement, which indicates asymmetry and variation in the location of this artery. Comparison of the measurements obtained in different sides showed similar values, with no statistically significant differences in the distances between any of the proposed anatomic landmarks

Orientador: Edi Lúcia Sartorato / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-25T22:58:29Z (GMT). No. of bitstreams: 1 Miranda_PauloMauriciodoAmorDivino_D.pdf: 3949549 bytes, checksum: 8cd1e0647087ac34cd2bc0ed5d096d0b (MD5) Previous issue date: 2014 / Resumo: A Neuropatia Óptica Hereditária de Leber (LHON) e a Atrofia Óptica Autossômica Dominante (ADOA ou OPA1) são doenças caracterizadas pela perda da visão bilateral, devido a uma degeneração do nervo óptico. Ambas as doenças apresentam também acuidade visual reduzida, discromatopsia, palidez do nervo óptico e escotoma central ou centrocecal. A LHON é causada por mutações no DNA mitocondrial (mtDNA), onde três dessas mutações representam 95% dos casos (mutações primárias principais G11778A, T14484C e G3460A) e as mutações subsequentes representam apenas 5% do total (mutações raras). A ADOA é causada por mutações (mais de 300) no gene nuclear OPA1. Embora os mecanismos moleculares precisos envolvidos no desenvolvimento das duas doenças ainda não são bem compreendidos, foi demonstrado que LHON e ADOA possuem um defeito comum de acoplamento da fosforilação oxidativa. Nesses dois casos, a hipótese é de que as mutações no mtDNA e no gene OPA1 afetariam a integridade mitocondrial, resultando em uma diminuição do fornecimento de energia para os neurônios do nervo óptico. É possível que no Brasil a presença e frequência das alterações da relacionadas à LHON e ADOA sejam diferentes das encontradas em outras partes do mundo. Por isso, o presente estudo teve como principais objetivos rastrear mutações e haplogrupos associadas à LHON e detectar mutações no gene OPA1 em pacientes brasileiros com hipótese diagnóstica de LHON e com Neuropatia Óptica de etiologia a esclarecer. Também foi objetivo otimizar o método de PCR Multiplex Alelo-Específico e padronizar as plataformas de alto rendimento TaqMan® OpenArray® e Iplex Gold/Maldi TOF MS para o rastreamento da LHON. Foram avaliados 101 pacientes, sendo 67 com hipótese diagnóstica de LHON e 34 com neuropatia óptica de etiologia a esclarecer. As mutações da LHON foram detectadas por meio de PCR-RFLP e PCR Multiplex Alelo-Específico. As mutações raras da LHON e do gene nuclear OPA1 (10 principais éxons) foram rastreadas por sequenciamento direto. Foram encontradas mutações da LHON em 36 casos (83.3% com a mutação G11778A e 16.7% com a mutação T14484C). Não foi encontrada a mutação G3460A. Também não foram encontradas mutações raras da LHON e nem mutações relacionadas à ADOA. Haplogrupos de origem africana (L1/L2 e L3) foram mais frequentes no estudo. Foi otimizado o método de PCR Multiplex Alelo-Específico e padronizadas as plataformas TaqMan® OpenArray® e Iplex Gold/Maldi TOF, os quais se mostraram reprodutivos, eficientes e eficazes. A análise molecular das mutações da LHON e do OPA1 foi importante para a confirmação do diagnóstico de 35% dos casos clínicos típicos de LHON e para a elucidação 35% casos de neuropatia óptica de etiologia a esclarecer / Abstract: The Leber Hereditary Optic Neuropathy (LHON) and Autosomal Dominant Optic Atrophy (ADOA) are diseases characterized by loss of vision in both eyes due a degeneration of the optic nerve. Both diseases also exhibit reduced visual acuity, dyschromatopsia, optic nerve and central scotoma or centrocecal. The LHON is caused by mutations in mitochondrial DNA (mtDNA), where three of these mutations account for 95% of cases (major primary mutations G11778A, T14484C and G3460A) and subsequent mutations account for only 5% of the total (rare mutations). The ADOA is caused by mutations (more than 300) in the nuclear gene OPA1. Although the precise molecular mechanisms involved in the development of the two diseases are not well understood, it was shown that LHON and ADOA have a common defect coupling of oxidative phosphorylation. In both cases, the hypothesis is that mutations in mtDNA and OPA1 gene affect mitochondrial integrity, resulting in a decrease in the supply of energy to the neurons of the optic nerve. It is possible that in Brazil the presence and frequency of changes related to LHON and ADOA be different from those found in other parts of the world. Therefore, the present study had two main objectives track haplogroups and mutations associated with LHON and detect mutations in the OPA1 gene in Brazilian patients with a diagnosis of LHON and optic neuropathy of unknown etiology. Another objective was to optimize the method of PCR Multiplex allele-specific and standardize platforms high throughput TaqMan® OpenArray® and Iplex Gold/Maldi TOF MS for screening of LHON. 101 patients were evaluated, 67 with a diagnosis of LHON and 34 with optic neuropathy of unknown etiology. LHON mutations were detected by PCR-RFLP and allele-specific multiplex PCR. Rare mutations of LHON and nuclear gene OPA1 (top 10 éxons) were screened by direct sequencing. LHON mutations were found in 36 cases (83.3% with the G11778A mutation and 16.7% with the T14484C mutation). Not the G3460A mutation. Nor rare mutations of LHON and ADOA or related mutations were found. Haplogroups of African origin (L1/L2 and L3) were more frequent in the study. The method of allele-specific multiplex PCR was optimized and standardized the TaqMan® OpenArray® and iPLEX Gold/Maldi TOF platforms which are shown reproductive, efficient and effective. Molecular analysis of mutations of LHON and ADOA was important to confirm the diagnosis of 35% of the typical clinical cases of LHON and to elucidate 35% cases of optic neuropathy of unknown etiology. Besides being useful also in the prognosis of each patient, for the phenotypic expression of the LHON and ADOA may vary with different genetic background in our population of individuals / Doutorado / Genetica Animal e Evolução / Doutor em Genetica e Biologia Molecular

Abstract Glaucoma is a group of eye diseases characterized by a chronic, progressive optic neuropathy. During the disease process, the axon damage of the retinal ganglion cells leads to changes in the retinal nerve fiber layer, causing optic nerve head, and visual field defects typical of glaucoma. The Heidelberg Retina Tomograph (HRT) is a confocal scanning laser imaging device acquiring and analysing three-dimensional data of the ocular fundus wit good accuracy and reproducibility. Conventional planimetric measurements were compared to those taken with the HRT in a pilot study of 12 eyes with early glaucomatous optic disc, retinal nerve fiber layer and/or visual field abnormalities. The neuroretinal rim area measurements and cup-to-disc area ratio did not differ statistically from each other when using these two different methods. The effect of four different reference levels on the HRT parameter measurement values was tested in two separate studies. In the first study there were 67 eyes, 40 of the eyes were healthy and 27 eyes had glaucoma of different stages. Then, 279 eyes, 180 of which were non-glaucomatous and 99 glaucoma eyes, were included in another study. The flexible reference level gave the most reliable HRT parameter measurement values in both non-glaucomatous and glaucomatous eyes. The ability of the HRT parameters to separate between non-glaucomatous and glaucomatous eyes was tested in 77 eyes, 40 of the eyes were non-glaucomatous, 10 ocular hypertensives and 27 eyes had different stages of glaucoma. The reference level dependent HRT parameters cup-to-disc area ratio, vertical linear cup-to-disc ratio, mean retinal nerve fiber layer thickness (RNFLt) and rim volume as well as the reference level non-dependent HRT parameter, cup shape measure (CSM), separated best between the clinical groups. The best combination of the HRT and other structural and functional parameters in separating between non-glaucomatous and glaucomatous eyes was studied in 55 eyes. There were 32 non-glaucomatous eyes and 23 eyes with ocular hypertension or glaucoma. CSM, RNFLt, together with age- and lens coloration-corrected mean deviation of the B/Y perimetry showed good discrimination (ROC area 0.91) between non-glaucomatous and glaucomatous eyes. / Tiivistelmä Glaukooma koostuu joukosta hitaasti eteneviä näköhermon rappeumasairauksia. Sairausprosessin aikana verkkokalvon gangliosolujen aksonivaurio johtaa muutoksiin verkkokalvon hermosäiekerroksessa ja näköhermon päässä aiheuttaen glaukoomalle tyypillisiä näkökenttämuutoksia. The Heidelberg Retina Tomograph (HRT) on konfokaali laserskanneritekniikkaan perustuva kuvantamislaite, joka tuottaa ja analysoi silmänpohjasta saatua kolmiulotteista mittaustietoa tarkasti ja toistettavasti. Tavanomaisen planimetrian antamia mittaustuloksia verrattiin HRT:n antamiin tuloksiin 12:ssa silmässä, joissa oli todettu varhaisia glaukoomamuutoksia. Näköhermon pään hermoreunan (rim) pinta-ala ja keskuskuopan suhde papillan läpimittaan eivät poikenneet tilastollisesti toisistaan näitä kahta menetelmää käytettäessä. Neljän eri referenssitason vaikutusta HRT-parametrien mittausarvoihin testattiin kahdessa eri tutkimuksessa. Ensimmäisen tutkimusaineisto koostui yhteensä 67:stä silmästä, joista 40 oli terveitä ja 27:ssä eriasteisia glaukoomamuutoksia. Toisessa tutkimuksessa oli yhteensä 279 silmää, joista 180 oli terveitä ja 99:llä oli glaukooma. Papillomakulaarisäikeisiin tukeutuva, fleksiibeli referenssitaso antoi luotettavimmat HRT-parametrien mittaustulokset sekä terveissä että glaukoomasilmissä. HRT-parametrien kykyä erottaa terveet silmät glaukomatoottisista testattiin yhteensä 77:ssä silmässä, joista 40 oli terveitä, 10 oli korkeapaineisia ilman glaukoomamuutoksia, ja 27:ssä oli glaukoomamuutoksia. Referenssitasosta riippuvaiset HRT-parametrit, keskuskuopan suhde papillan läpimittaan, vertikaali-lineaarinen keskuskuopan suhde papillan läpimittaan, keskimääräinen verkkokalvon hermosäiekerroksen paksuus (RNFLt) ja `rim´:in tilavuus samoin kuin referenssitasosta riippumaton keskuskuopan ´vinous´-mitta (CSM) erottelivat parhaiten nämä kliiniset ryhmät toisistaan. Terveitä ja glaukoomasilmiä erottelevaa HRT:n ja muiden rakenteellisten ja toiminnallisten parametrien kombinaatiota etsittiin 55:n silmän aineistosta. Silmistä 32 oli terveitä ja 23 korkeapaineisia ja/tai glaukoomavaurioisia. CSM ja RNFLt, yhdessä iän ja mykiövärjäytymisen suhteen korjatun sinikeltaperimetrian keskipoikkeaman kanssa osoittivat hyvää erottelukykyä (ROC area 0.91) terveiden ja glaukoomasilmien välillä.

Optic nerve head and macula are important structures in fundus images. Detection and measurement plays crucial role in several diagnosis methods of optic disease. This work is focused on the detection of the central point of macula and optic nerve head, where the inner border is detected too. There are many methods for extracting this structure in retinal images. Due to the unique properties of each acquisition technique, a single generally acknowledged detection algorithm does not exist. The whole detection process is described from preprocessing through segmentation towards postprocessing. Presented methods are based on the combination of correlation techniques, Hough transform, active contours and morphological operations. The detected contours of the optic nerve head are evaluated and quantitatively compared with the contour drawn by experienced ophthalmologist. The master thesis contains quantity of images that help to describe detection methods.

The main target of this work is a design of the program for border localization of the optic disc in OCT data. Part of this program is also localization of the „optic cup“ and calculation of several values that characterize optic disc. The work is also dealing with a description of the optical coherence tomography principle which is used in ophthalmology. One chapter describes some published methods of optic cup localization. Success of the program and its comparison with OCT Zeiss Stratus 3000 is evaluated at the end of the work.

<p>Cilj ovog istraživanja bio je da se utvrdi razlika u debljini sloja retinalnih nervnih vlakana (RNFL) i parametara glave očnog živca kod pacijenata sa preperimetrijskim glaukomom i pacijenata sa glaukomom otvorenog ugla (POAG) u odnosu na zdravu populaciju, kao i da se utvrdi razlika u debljini RNFL i parametara glave očnog živca kod pacijenata sa POAG u odnosu na stepen progresije bolesti. Materijal i metode: U ovu kliničku, analitičku i opservacionu, po tipu &bdquo;slučaj-kontrola&ldquo; studiju, bilo je uključeno 120 pacijenata. Na osnovu kliničkog nalaza formirane su četiri grupe. Prva grupa (grupa zdravih): 30 pacijenata bez glaucoma i drugih očnih bolesti. Druga grupa (grupa sa početnim POAG): 30 pacijenata sa POAG, sa karakterističnim o&scaron;tećenjem glave očnog živca i RNFL, kod kojih je srednja vrednost devijacije standardizovane automatske perimetrije MD&lt;-6dB (prema Hodap klasifkaciji) sa karakterističnim glaukomskim ispadima u vidnom polju. Treća grupa (grupa sa srednje uznapredovalim POAG): 30 pacijenata sa POAG, kod kojih je srednja vrednost devijacije standardizovane automatske perimetrije MD od -6dB do -12dB (prema Hodap klasifkaciji). Četvrta grupa (grupa sa preperimetrijskim glaukom): 30 pacijenata sa promenama na glavi očnog živca karakterističnim za glaukomsku neuropatiju, kod kojih ne postoje funkcionalni ispadi tj. standardna automatizovana perimetrija pokazuje normalne vrednosti MD parametara (od -2 dB do +2dB). Kod svih pacijenata bio je urađen kompletan oftalmolo&scaron;ki pregled, kompjuterizovano vidno polje i optička koherentna tomografija peripapilarne regije RNFL i glave očnog živca (na aparatu Stratus OCT 3000, Carl Zeiss Meditec). Rezultati su pokazali da je debljina RNFL-a kod pacijenata sa početnim POAG manja u odnosu na zdravu populaciju. Najveće sniženje debljine RNFL je u sektorima 1,6,7 i 8h. Jedino u sektoru 4h i 9h ne dolazi do smanjenja debljine RNFL-a. Najveće smanjenje debljine RNFL je u gornjem i donjem kvadrantu, te oni imaju visoku specifičnost za diskriminaciju između zdravih i pacijenata sa početnim POAG. Parametri glave očnog živca: volumen ekskavacije, vertikalni C/D, horizontalni C/D i ukupni C/D odnos kod pacijenata sa početnim POAG povećani su u odnosu na zdravu populaciju. Parametri glave očnog živca: povr&scaron;ina neuroretinalnog oboda i volumen neuroretinalnog oboda, kod pacijenata sa početnim POAG smanjeni su u odnosu na zdravu populaciju. Debljina RNFL kod pacijenata sa srednje uznapredovalim POAG smanjena je i u odnosu na pacijente sa početnim POAG i u odnosu na zdravu populaciju (59,69&plusmn;10,63 &mu;m vs 73,44&plusmn;12,16&mu;m vs 105,57&plusmn;11,34 &mu;m). Parametri glave očnog živca prate ove promene. Ukupna povr&scaron;ina glave očnog živca se statistički značajno ne menja između zdravih osoba, pacijenata sa početnim i srednje uznapredovalim glaukomom otvorenog ugla i kod pacijenata sa preperimetrijskim glaukomom, te ovaj parametar ne determini&scaron;e glaukomsku bolest. Postojanje i napredovanje glaukoma kod pacijenata dovodi do istanjenja&nbsp; peripapilarnog RNFL &scaron;to je praćeno povećanjem ekskavacije glave očnog živca. Sa smanjenjem MD vrednosti dolazi do sledstvenih promena većine parametara. Postoji pozitivna korelacija između uznapredovalosti galukoma i srednje vrednosti debljine RNFL. Promena ove vrednosti najbolje pokazuje da dolazi do progresije POAG. Parametri glave očnog živca koji najbolje oslikavaju progresiju glaukoma su: ukupni C/D, vertikalni C/D i horizontalni C/D odnos. Debljina RNFL-a kod pacijenata sa preperimetrijskim glaukomom je značajno manja u od nosu na zdravu populaciju (83,65&plusmn;9,24&mu;m vs 105,57&plusmn;11,34&mu;m). To se posebno izražava u gornjem kvadrantu, dok u temporalnom kvadrantu ne dolazi do promena. Parametar S zajedno sa srednjom vrednosti debljine RNFL predstavljaju najbolje pokazatelje nastajanja preperimetrijskog glaukoma. Sektor 1h je sektor sa visokom specifično&scaron;ću za diskriminaciju izmeĎu zdravih i pacijenata sa preperimetrijskim glaukomom. Parametri glave očnog živca: volumen ekskavacije, vertikalni C/D, horizontalni C/D i srednji C/D odnos, kod pacijenata sa preperimetrijskim glaukomom statistički su značajno povećavani i u odnosu na zdravu populaciju. Parametri glave očnog živca: povr&scaron;ina neuroretinalnog oboda i volumen neuroretinalnog oboda, kod pacijenata sa preperimetrijskim glaukomom statistički su značajno manji u odnosu na zdravu populaciju. Najbolji prediktori nastanka i napredovanja glaukomske bolesti su sledeći parametri: AvgThic, debljina RNFL po kvadrantima-S,I,N; parametric debljine RNFL: Smax, Savg, Iavg; kao i parametri PNO: RimArea, RimVol, DiscArea, CupAear, C/DHorRat, C/DVertRat, C/DAreaRat. ROC kriva je pokazala da su sledeću parametri lo&scaron;i marker za progresiju bolesti: debljine RNFL kavdranta T, Imax i upVol. Zaključak: Određivanje parametara glave očnog živca i debljine peripapilarnih RNFL kod pacijenata sa glaukomom, optičkom koherentnom tomografijom, predstavlja metodu koja izdvaja pacijente sa preperimetrijskim glaukomom od zdrave populacije. Ono posebno ukazuje na sektore, kvadrante i parametre koji su najosetljiviji na glaukomsku noksu i koji prvi postaju patolo&scaron;ki pri nastanku glaukoma. Takođe, ukazuje i na razliku između pojedinih stepena glaukomske bolesti. Na ovaj način se omogućuje sigurna i rana dijagnoza glaukoma, njegovo pravovremeno lečenje i bolja prognoza kod pacijenata sa POAG.</p> / <p>Aim: The aim of this study was to determine the difference in thickness of retinal nerve fibre layer (RNFL) and parameters of optic nerve head in patients with preperimetric glaucoma and in patients with open angle glaucoma (POAG) in comparison to healthy population, as well as to determine the difference in thickness of RNFL and parameters of optic nerve head in patients with POAG according to progression of the disease. Material and methods: This clinical study was analytical and opservational, &bdquo;case-control&ldquo; type of study. 120 patients were included. On the basis of clinical finding 4 groups were formed. First group (healthy): 30 patients without glaucoma and with no other ocular disease. Second group (group of patients with mild POAG): 30 patients with POAG, with characteristical optic nerve head and RNFL damage, in whom the value of standard deviation of standardised automatic perimetry is MD&lt;-6dB (according to Hodap classification) with typical glaucomatous visual field defects. Third group (group of patients with moderate POAG): 30 patients with POAG in whom the mean value of standard deviation of standardised automatic perimetry, MD is from -6dB to -12dB (according to Hodap classification). Fourth group (group of patients with preperimetric glaucoma): 30 patients with changes of optic nerve head that are typical of glaucomatous neuropathy in whom there are no functional changes and with normal values of MD parameters of standardised automatic perimetry. In all patients complete ophthalmological examination, complete visual field and optic coherent tomography of peripapillar region of RNFL and optic nerve head (using Stratus OCT 3000, Carl Zeiss Meditec) were performed.The results showed that thickness of RNFL in patients with mild POAG is lesser than in healthy subjects. The greatest decrease in RNFL thickness is in sectors 1,6,7 and 8h. Only in sectors 4h and 9h there is no decrease in RNFL thickness. The greatest decrease in RNFL thickness is in upper and lower quadrant, so they are highly specific in determination between healthy subjects and patients with mild POAG. Parameters of optic nerve head such as: excavation volume, vertical C/D, horisontal C/D and total C/D ratio in patients with mild POAG are higher comparing to healthy population. Parameters of optic nerve head such as: neuroretinal rim area and neuroretinal rim volume in patients with mild POAG are lower than in healthy population. RNFL thickness in patients with moderate POAG is lesser than in patients with mild POAG, as well as in healthy subjects. Optic nerve head parameters follow these changes. Total optic nerve head area does not change in healthy subjects, in patients with mild and moderate open angle glaucoma and in patients with preperimetric glaucoma, so this parameter does not determine glaucomatous disease. The existence and progression of glaucoma in patients leads to thinning of peripapillar RNFL which is followed by increase of excavation of optic nerve head. With decrease of MD value there are consecutive changes in most parameters. There is positive correlation between progression of glaucoma and average thickness of RNFL. The change of this value shows the best if there is progression of POAG. Paremeters of optic nerve head that are the best determinants of progression of glaucoma are: total C/D, vertical C/D and horisontal C/D ratio. Thickness of RNFL in patients with preperimetric glaucoma is significantly lesser than in healthy subjects. It is particularly seen in upper quadrant, while in temporal quadrant there are no changes. Parameter S together with mean value of RNFL thickness is the best parameter of appearance of preperimetric glaucoma. Sector 1h is the sector that is highly specific in discrimination between healthy subjects and patients with preperimetric glaucoma. Optic nerve head parameters such as: volume of excavation, vertical C/D, horizontal C/D and C/D mean ratio in patients with preperimetric glaucoma are statistically significantly higher than in healthy population. Optic nerve head parameters such as: neuroretinal rim area and neuroretinal rim volume in patients with preperimetric glaucoma are statistically significantly lower than in healthy population. The best predictors of appearance and progression of glaucomatous disease are: AvgThic, RNFL thickness in quadrants: S,I,N; RNFL:Smax, Savg, Iavg; as well as PNO: RimArea, RimVol, DiscArea, CupAear, C/DHorRat, C/DVertRat, C/DAreaRat. ROC curve has shown that the following parameters are bad markers for progression of the disease: RNFL thickness in quadrant T, Imax and CupVol. Conclusion: Determination of parameters of optic nerve head and peripapillar RNFL in patients with glaucoma using optical coherent tomography represents the method that distinguishes the patients with preperimetric glaucoma from healthy subjects. It particularly points the sectors, quadrants and parameters that are the most sensitive to glaucomatous disease and that first become pathological when disease appears. It also indicates the difference between certain levels of glaucomatous disease. In this way safe and early diagnosis of glaucoma is provided, as well as adequate therapy and better prognosis in patients with POAG.</p>

Durchtrennte Axone adulter retinaler Ganglienzellen (RGZ) können in periphere Nerventransplantate (PNT) einwachsen, die als “bypass” des distalen Sehnervenstumpfes verwendet werden. Das Transplantationsmodell, bei dem der durchtrennte Sehnerv durch ein Ischiasnervsegment ersetzt wird, ist in der Regenerationsforschung ein seit Jahren fest etabliertes Verfahren. In dieser Arbeit soll der Frage nachgegangen werden, ob a) der Ersatz des Sehnervs durch ein peripheres Nervensegment RGZ über einen langen Zeitraum hinweg morphologisch und funktionell stabilisiert, ob b) Unterschiede der Stabilisierung in Abhängigkeit von der Hirnregion, mit der das PNT in Kontakt tritt, zu beobachten sind und c) inwieweit regenerierende RGZ dadurch selbst peripher-nervöse Eigenschaften annehmen. Der Sehnerv adulter Ratten wurde zunächst komplett intraorbital durchtrennt. Der okuläre Stumpf wurde über ein autologes Ischiasnervsegment mit verschiedenen visuellen Zentren (Kortex, Mittelhirn) oder mit Fremdzielgebieten (z. B. Muskel) verbunden. Weitere Kontrollgruppen bestanden in der Quetschung des Sehnervs, der Durchtrennung ohne Transplantation und der Transplantation mit blind endendem Transplantat. Die Netzhautintegrität wurde pupillometrisch und elektroretinographisch regelmäßig überprüft, um eine eventuelle, funktionelle Wiederherstellung der visuellen Bahn zu erfassen. Nach einem, sechs und neun Monaten wurden die regenerierenden bzw. axotomierten oder gequetschten RGZ mit 4-(4-(didecylamino)styryl)-N-methylpyridinium (4-Di-10-ASP) retrograd markiert und morphometrisch quantifiziert (Fluoreszenz-, Konfokal- und Elektronenmikroskopie sowie Differentialinterferenzkontrast). Zusätzlich wurden immunhistochemische und anterograde Markierungsuntersuchungen durchgeführt. Regenerierende Ganglienzellen bleiben bis neun Monate nach der Transplantation am Nervus opticus stabil. Es gibt quantitative sowie morphometrisch erfassbare Unterschiede zwischen den experimentellen Gruppen und den Kontrollen, wobei die wieder verbundenen Ganglienzellen morphologisch am besten zu klassifizieren sind. Quantitativ zeigen die Retinae mit gequetschtem Sehnerv nach sechs Monaten die höchste Überlebensrate der RGZ. Die Effektivität dieses Verfahrens als Modell der zentralen Nervenläsion darf in Folge dieser Ergebnisse in Frage gestellt werden. Nach neun Monaten sind in den Retinae mit Rekonnektion zum Mittelhirn die meisten Ganglienzellen vorhanden. Elektrophysiologisch zeigen die Augen mit Verbindung zum Muskelgewebe die besten funktionellen Ergebnisse. Schlussfolgernd zeigt sich, dass adulte RGZ der Ratte über ein peripher-nervöses Transplantat, welches mit visuellen Zentren in Verbindung steht, über lange Zeit stabilisiert werden können.

Traumatic brain injury (TBI) affects millions of people worldwide, leading to significant mortality and disability rates. Elevated intracranial pressure (ICP) resulting from TBI can cause severe complications and requires early detection to improve patient outcomes. While invasive methods are commonly used to measure ICP accurately, non-invasive techniques such as optic nerve sheath diameter (ONSD) measurement show promise. This study aims at the creation of a tool that can automatically perform a segmentation of the ONS from a head computed tomography (CT) scan, and extracts meaningful measures from the segmentation mask, that can be used by radiologists and medics when treating people affected by TBI. This has been achieved using a deep learning model called ”nnU-Net”, commonly adopted for semantic segmentation in medical contexts. The project makes use of manually labeled head CT scans from a public dataset named CQ500, to train the aforementioned segmentation model, using an iterative approach. The initial training using 33 manually segmented samples demonstrated highly satisfactory segmentations, with good performance indicated by Dice scores. A subsequent training, combined with manual corrections of 44 unseen samples, further improved the segmentation quality. The segmentation masks enabled the development of an automatic tool to extract and straighten optic nerve volumes, facilitating the extraction of relevant measures. Correlation analysis with a binary label indicating potential raised ICP showed a stronger correlation when measurements were taken closer to the eyeball. Additionally, a comparison between manual and automated measures of optic nerve sheath diameter (ONSD), taken at a 3mm distance from the eyeball, revealed similarity between the two methods. Overall, this thesis lays the foundation for the creation of an automatic tool whose purpose is to make faster and more accurate diagnosis, by automatically segmenting the optic nerve and extracting useful prognostic predictors. / Traumatisk hjärnskada (TBI) drabbar miljontals människor över hela världen, vilket leder till betydande dödlighet och funktionshinder. Förhöjt intrakraniellt tryck (ICP) till följd av TBI kan orsaka allvarliga komplikationer och kräver tidig upptäckt för att förbättra patientens resultat. Medan invasiva metoder vanligtvis används för att mäta ICP exakt, icke-invasiva tekniker som synnervens höljediameter (ONSD) mätning ser lovande ut. Denna studie syftar till att skapa ett verktyg som automatiskt kan utföra en segmentering av ONS från en datortomografi skanning av huvudet, och extraherar meningsfulla åtgärder från segmenteringsmasken, som kan användas av radiologer och läkare vid behandling av personer som drabbats av TBI. Detta har uppnåtts med hjälp av en deep learning modell som kallas ”nnU-Net”, som vanligtvis används för semantisk segmentering i medicinska sammanhang. Projektet använder sig av manuellt märkta datortomografi skanningar från en offentlig datauppsättning som heter CQ500, för att träna den tidigare nämnda segmenteringsmodellen, med hjälp av en iterativ metod. Den inledande träningen med 33 manuellt segmenterade prov visade tillfredsställande segmentering, med god prestation indikerad av Dice-poäng. En efterföljande utbildning, i kombination med manuella korrigeringar av 44 osedda prover, förbättrade segmenteringskvaliteten ytterligare. Segmenteringsmaskerna möjliggjorde utvecklingen av ett automatiskt verktyg för att extrahera och räta ut optiska nervvolymer, vilket underlättade utvinningen av relevanta mått. Korrelationsanalys med en binär märkning som indikerar potentiellt förhöjd ICP visade en starkare korrelation när mätningar gjordes närmare ögongloben. Dessutom avslöjade en jämförelse mellan manuella och automatiserade mätningar av optisk nervmanteldiameter (ONSD), tagna på ett avstånd på 3 mm från ögongloben, likheten mellan de två metoderna. Sammantaget lägger denna avhandling grunden för skapandet av ett automatiskt verktyg vars syfte är att göra snabbare och mer exakta diagnoser, genom att automatiskt segmentera synnerven och extrahera användbara prognostiska prediktorer.

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