A diabetes mellitus (DM) causa alterações em todas as camadas da córnea. Córneas de pacientes diabéticos têm controle de sua hidratação prejudicado e são mais propensas à descompensação após injúria. A uveíte faco-induzida (UFI) ocorre em grandes proporções associada à catarata hipermatura em cães. A UFI também acarreta danos ao endotélio da córnea e pode levar ao edema estromal, usualmente transitório. Cães diabéticos são particularmente predispostos ao desenvolvimento de catarata e UFI associada. Os efeitos da DM e UFI podem potencialmente vir a se somar em córneas de cães com catarata diabética. A pressão intra-ocular (PIO) e paquimetria podem ser ferramentas úteis na determinação dessas alterações. O objetivo deste estudo foi determinar a relação entre pressão intra-ocular e paquimetria nos diferentes estágios de desenvolvimento das cataratas diabéticas e não diabéticas em cães da raça Poodle. Cento e vinte e dois cães adultos da raça Poodle, 134 fêmeas e 99 machos, com idades de 2 a 16 anos, foram admitidos no Serviço de Oftalmologia do Hospital Veterinário da Universidade de São Paulo e incluídos no trabalho. Afecções oculares concomitantes, ou doenças sistêmicas com manifestação ocular, foram consideradas fatores de exclusão. Após exame clínico oftalmológico, coletou-se dados referentes ao tempo de leucocoria e tempo de desenvolvimento da DM. Os cães tiveram a PIO mensurada por meio do TonoPen-XL® e a paquimetria mensurada com o uso do PachPen®. Para isso realizou-se anestesia tópica (colírio de Proximetacaína a 0,5% - Anestalcon®, Alcon). As comparações estatísticas entre as variáveis foram realizadas utilizando os testes de Spearman, Mc-Nemar e Mann-Whitney quando apropriados. As distribuições das variáveis entre os grupos foram avaliadas pelo test de Kruskal-Wallis, e quando significantes, o teste de Dunn foi utilizado para descriminar as diferenças encontradas. O nível de significância foi estabelecido em p<0,05. Dos 122 cães, 233 olhos foram incluídos e classificados em grupo controle (n=39), catarata incipiente (n=20), imatura (n=29), matura (n=20), hipermatura (n=80), catarata diabética imatura (n=9), diabética matura (n=11) e diabética hipermatura (n=25). As distribuições das variáveis (sexo, idade, peso, presença de uveíte, tempo de leucocoria, PIO e paquimetria) foram estatisticamente diferentes entre os diversos grupos. Fêmeas apresentaram-se mais acometidas por catarata diabética. A idade média dos animais variou de 7,03 a 10,73 anos entre os grupos, e os animais diabéticos foram estatisticamente mais velhos. A PIO foi estatisticamente menor e os sinais clínicos de UFI mais prevalentes nos grupos de cataratas hipermaturas, tanto diabéticas, quanto não diabéticas. A PIO correlacionou-se negativamente com a idade e a paquimetria. Houve correlação positiva da paquimetria com a idade e o peso; e correlação negativa com o tempo de leucocoria e a PIO. Olhos acometidos por UFI tiveram PIO estatisticamente menor e paquimetria maior, com diferença de 2 mmHg e 60 µm, respectivamente. Nos cães acometidos por catarata diabética, a paquimetria encontrou-se particularmente aumentada, e a UFI particularmente presente. Esse aumento possivelmente reflete a somatória dos efeitos da DM e UFI sobre a córnea. A hipermaturidade foi correlacionada à presença da UFI. A PIO e paquimetria mostraram-se instrumentos valiosos na determinação da UFI, principalmente se forem negativamente correlacionadas entre si. / Diabetes mellitus (DM) causes different alterations in all corneal layers, such as deficient control of hydration, what makes them prone to decompensate after injury. Lens induced uveitis (LIU) occurs mainly in dogs presenting hypermature cataracts, what causes irreversible damages to the corneal endothelium and may lead to stromal edema, usually transient. DM dogs are particularly predisposed to cataract with LIU associated. Intraocular pressure (IOP) and pachymetry measurements may be useful procedures capable to detect such alterations. The aim of this study was to determinate the relationship between intraocular pressure and pachymetry on diabetic and nondiabetic Poodles, presenting different stages of cataracts. One hundred twenty-two Poodles, 134 females and 99 males, with ages varying from 2 to 16 years, were admitted at the Ophthalmology Service, of the Veterinary Hospital, of the University of São Paulo. After complete identification of the animals, anamnese (time of DM and cataract formation) and ophthalmological exam, dogs were included in the study. Dogs affected by other systemic diseases inducing ocular manifestation or other primary ocular alteration were excluded. LIU was diagnosed by observation of congestion of the episcleral vases and resistence to midriasis after induction with Mydriacyl®. IOP was measured by TonoPen-XL® and pachymetry by PachPen® after instillation of anesthetic eyedrop (Proximetacaine 0,5% - Anestalcon, Alcon). Statistical comparison between factors were made using Spearman, Mc-Nemar and Mann-Whitney tests when appropriated. The distributions of the factors among the groups were compared by Krusskal-Walliss test, and when significant, Dunns test was used to describe the differences found. The level of significance was set at p<0,05. Two hundred thirty-three eyes of 122 Poodles were evaluated. Eyes were classified in different groups: healthy dogs without cataracts (n=39); dogs presenting incipient cataract (n=20); with immature cataracts (n=29); mature cataracts (n=20); or hypermature cataracts (n=80); DM dogs presenting immature cataract (n=9); DM with mature cataracts (n=11); and DM with hypermature cataracts (n=25). Distribution of factors, such as age, sex, weight, time of DM and cataract, presence of LIU, IOP and pachymetry were statistically different between groups. Female Poodles were more affected by diabetic cataract and diabetic dogs were older than the other ones. IOP was statistically lower and LIU alterations more prevalent in the hypermature cataract groups (diabetic and no diabetics). IOP was negatively correlated to age and pachymetry measures. Pachymetry was positively correlated to age and weight. Negative correlation was observed between pachymetry and time of cataracts formation. LIU eyes presented lower IOP and higher pachymetry. The difference found at pachymetry and IOP measurements in LIU and non LIU eyes were 60 µm and 2 mmHg respectively. In diabetic dogs, pachymetry was particularly higher and LIU particularly present. The higher corneal thickness may possible be related to the sum of alterations caused by DM and LIU to those corneas. Hypermaturity was directly related to LIU. We concluded that pachymetry and IOP measurements are important procedures to the LIU diagnosis, mainly when negatively correlated.

O tratamento da catarata em cães têm passado por algumas mudanças. A facoemulsificação reduziu o tempo cirúrgico e a necessidade de incisão extensa; o implante de lentes intra-oculares (LIOs) permitiu restabelecer a emetropia após a remoção do cristalino. O processo inflamatório induzido pelas cirurgias intra-oculares leva a um estado de estresse oxidativo, com efeitos deletérios às estruturas oculares. O presente estudo teve como objetivo avaliar comparativamente o estresse oxidativo após facoemulsificação experimental em cães, com e sem implante de LIO. Doze cães (três machos e nove fêmeas), sem raça definida, com peso médio de 15,8kg, foram submetidos à facoemulsificação bilateral, sendo doze olhos sem implante de LIO (Grupo 1) e doze olhos com implante de LIO de polimetilmetacrilato no saco capsular (Grupo 2). Os animais foram medicados nos períodos pré-operatório e pós-operatório com antiinflamatórios, antibióticos e midriáticos. As amostras de humor aquoso (HA) foram obtidas por paracentese de câmara anterior, sob anestesia geral, nos períodos pré-operatório (T0) e pós-operatório com 1 dia (T1), 2 dias (T2), 3 dias (T3), 7 dias (T7) e 14 dias (T14). O estresse oxidativo foi avaliado por meio da determinação quantitativa de proteínas no HA (método de Lowry), avaliação da atividade antioxidante total do HA utilizando o AAPH, cloreto de 2,2'azobis (2-metilamidinopropano) e avaliação da concentração de ácido ascórbico no HA e no plasma (cromatografia líquida de alta eficiência). Os resultados foram submetidos à análise estatística, utilizando-se o teste t de Student, quando as variáveis apresentaram distribuição normal, e pelo teste de Wilcoxon, quando não houve distribuição normal. Os níveis de proteína total no HA aumentaram nos dois grupos, sendo de forma mais acentuada no Grupo 2 em todos os tempos pós-operatórios: T1 (p= 0,004), T2 (p= 0,002), T3 (p= 0,004), T7 (p= 0,003) e T14 (p= 0,009). A atividade antioxidante total do HA apresentou uma diminuição nos dois grupos, mais acentuada, entretanto, no Grupo 2, com diferença estatisticamente significante nos tempos T1 (p= 0,013), T2 (p= 0,016), T3 (p= 0,002) e T14 (p= 0,033). No tempo T7 (p= 0,155) essa diferença não foi observada. A concentração de ácido ascórbico no HA apresentou uma diminuição marcante no primeiro dia do pós-operatório, seguida por uma recuperação progressiva nos dois grupos. O Grupo 2 mostrou resultados mais baixos, com diferença estatisticamente significante nos tempos T1 (p= 0,003), T3 (p= 0,015), T7 (p= 0,004) e T14 (p= 0,007), exceto em T2 (p= 0,051), quando não houve diferença. A concentração de ácido ascórbico no plasma não variou em função dos procedimentos. A comparação entre os procedimentos sugere que o implante de LIO contribui para a condição de estresse oxidativo. / The cataract treatment in dogs has undergone some changes. Phacoemulsification has reduced the surgical time of cataract extraction and the need of large incision. Intraocular lens (IOL) implantation has allowed emetropic vision recovery. The inflammatory process induced by intraocular surgery leads to an oxidative stress, with ocular structures damage. The purpose of this study was to comparatively evaluate the oxidative stress following phacoemulsification in dogs with and without IOL implantation. Twelve dogs, three male and nine female, mixed breed, with an average weight of 15.8kg were submitted to bilateral phacoemulsification, being twelve eyes without IOL implantation (Group 1) and twelve eyes with polymethylmethacrylate IOL in the capsular bag (Group 2). In the pre-operative and post-operative periods, the medication consisted of anti-inflammatories, antibiotics and mydriatics. Aqueous humor (AH) samples were taken by anterior chamber paracentesis, under general anesthesia in the pre-operative (T0) and post-operative periods after 1 (T1), 2 (T2), 3 (T3), 7 (T7) and 14 (T14) days of the surgery. The oxidative stress was evaluated by quantitative protein levels in the AH (Lowry’s method), total antioxidant activity in the AH using 2,2’ azobis (2-methylpropionamidine) dihydrochloride (AAPH) and ascorbic acid concentration in the AH and plasma (high performance liquid chromatography). Results were submitted to statistical analysis using the t-student test when the variables had a normal distribution and Wilcoxon test when they did not. Aqueous humor total protein levels increased in both groups, more intensively in Group 2 in all post-operative periods: T1 (p= 0.004), T2 (p= 0.002), T3 (p= 0.004), T7 (p= 0.003) and T14 (p= 0.009). Total antioxidant activity in the AH decreased in both groups; however, it was more impressive in Group 2, with statistical significant difference in T1 (p= 0.013), T2 (p= 0.016), T3 (p= 0.002) e T14 (p= 0.033). In T7 (p= 0.155), this difference was not observed. Ascorbic acid concentration in the AH presented a remarkably decrease in the first post-operative day, then started to recover progressively in the two groups. Group 2 showed lower results with statistically significant differences in T1 (p= 0.003); T3 (p= 0.015); T7 (p= 0.004) and T14 (p= 0.007), except in T2 (p= 0.051) when there was no difference. Ascorbic acid concentration in plasma did not vary as a result of the procedures. The comparison between procedures suggests that IOL implantation contributes to an oxidative stress condition.

INTRODUÇÃO: O blefaroespasmo essencial é uma distonia focal caracterizada por contrações involuntárias, espasmódicas e bilaterais dos músculos protratores das pálpebras. O glaucoma é a principal causa de cegueira irreversível em adultos de países desenvolvidos e a elevação da pressão intra-ocular é o maior fator de risco para o desenvolvimento da doença. A contração voluntária forçada das pálpebras pode causar aumentos da pressão intra-ocular de até 90 mmHg. Uma vez que pacientes com blefaroespasmo essencial apresentam contrações palpebrais freqüentes e de forte intensidade, eles poderiam compor um grupo de risco para o desenvolvimento do glaucoma. MÉTODOS: Vinte e oito pacientes com blefaroespasmo essencial e 28 pacientes de grupo controle, formado por indivíduos com doenças palpebrais ou no seu pós-operatório, foram submetidos a um exame oftalmológico completo. No grupo de pacientes com blefaroespasmo, a avaliação foi realizada em 8 a 11 dias após o tratamento com toxina botulínica. Efetuou-se exame de refração e medida de melhor acuidade visual corrigida com tabela de Snellen, biomicroscopia em lâmpada de fenda, tonometria de aplanação, campo visual computadorizado, teste de sobrecarga hídrica e biomicroscopia de fundo para avaliação da papila óptica. RESULTADOS: 1) A prevalência de glaucoma nos pacientes com blefaroespasmo foi significativamente maior do que nos indivíduos do grupo controle, sendo diagnosticada em 14,3% e 3,6% dos casos, respectivamente (p=0,008). 2) A pressão intra-ocular foi significativamente maior nos pacientes com blefaroespasmo (15,80 ± 3,80 mmHg) do que nos indivíduos do grupo controle (13,90 ± 2,75 mmHg) (p= 0,004). 3) O pico da pressão intra-ocular na prova de sobrecarga hídrica foi significativamente maior nos pacientes com blefaroespasmo essencial (18,82 ± 4,47 mmHg) do que nos indivíduos do grupo controle (16,27 ± 2,69 mmHg) (p=0,0421). DISCUSSÃO: Diversos estudos descreveram a influência da compressão palpebral sobre a pressão intra-ocular. A prova de sobrecarga hídrica tem sido considerada uma ferramenta indireta para se avaliar a capacidade do fluxo de drenagem do trabeculado, além de exibir correlação com os picos de pressão intra-ocular que muitas vezes não são detectados em exames de rotina. Os pacientes com blefaroespasmo essencial apresentaram pressões intra -oculares e picos de pressão intra-ocular na prova de sobrecarga hídrica mais elevados do que os pacientes do grupo controle, o que poderia indicar um baixo fluxo de drenagem. Estas altíssimas variações de pressão intra -ocular a que estes indivíduos são submetidos constantemente, em pacientes com menor fluxo de drenagem poderiam causar um aumento crônico da pressão intra -ocular e o desenvolvimento do glaucoma. CONCLUSÕES: Os resultados observados sugerem que estes pacientes compõem um grupo de risco para o desenvolvimento de glaucoma e esta doença deve ser pesquisada de maneira sistemática na avaliação inicial e durante o seguimento desses casos / INTRODUCTION: Essential blepharospasm is a focal distonia characterised by involuntary, spasmodic, bilateral contractions of eyelid protractors. Glaucoma is the most important cause of irreversible blindness in adults in developed countries and high intraocular pressure is the major risk factor for development of the disease. Voluntary forced eyelid closure can produce an intraocular pressure increase of 90 mmHg. As patients with essential blepharospasm present frequent and strong eyelid contractions, they could be at risk for glaucoma development. METHODS: Twenty eight patients with essential blepharospasm and 28 patients of a control group, with eyelid diseases or in the post-operative period, were submitted to a complete ophthalmic examination. In the group of patiens with blepharospasm, the evaluation was done between 8 to 11 days after botulinum toxin treatment. We performed refractometry and best corrected visual acuity with Snellen chart, slitlamp biomicroscopy, applanation tonometry, automated perimetry, water drinking test and dilated funduscopy to evaluate optic discs. RESULTS: 1) Prevalence of glaucoma in patients with blepharospasm (14,3%) was higher than in the individuals from control group (3,6%) (p=0,008). 2) Intraocular pressure was higher in patients with blepharospasm (15,80 ± 3,80 mmHg) than in the individuals from control group (13,90 ± 2,75 mmHg) (p=0,004). 3) Intraocular pressure peaks in the water drinking test were higher in patients with essential blepharospasm (18,82 ± 4,47 mmHg) than in the individuals from control group (16,27 ± 2,69 mmHg) (p=0,0421). DISCUSSION: Several studies reported the influence of eyelid compression over intraocular pressure. The water drinking test has been considered as an indirect tool to measure outflow facility and it has good correlation with intraocular pressure peaks which are frequently missed in routine examinations. Patients with essential blepharospasm had higher mean intraocular pressures and intraocular pressure peaks in the water drinking test than patients from control group, which could indicate low outflow facility. These high intraocular pressure variations in patients with low outflow facility could increase the intraocular pressure chronically and lead to development of glaucoma. CONCLUSION: Our results suggest that patients with essential blepharospasm be at higher risks for development of glaucoma and this disease should be always investigated at presentation and during follow-up of these cases

Objetivos: Comparar a performance visual dos pacientes submetidos ao implante das lentes intraoculares multifocais difrativas Tecnis® MF ZM900, Acrysof® Restor® SN60D3, Acrysof® SN60WF e Acrysof® SN60AT. Métodos: Estudo prospectivo comparativo, não randomizado, que incluiu 142 olhos de 71 pacientes com catarata, provenientes do ambulatório de oftalmologia do HC- FMUSP. A avaliação oftalmológica contou com medida da acuidade visual para longa, intermediária e curta distância, sem correção e com a melhor correção óptica, teste de sensibilidade ao contraste, pupilometria e análise de frente de onda por meio do aberrômetro. Um questionário de satisfação foi realizado para avaliar a acuidade visual, glare e halos. Todos os exames foram realizados com seis meses de pós-operatório. Resultados: A média de idade dos pacientes foi de 60,7 ± 6,6 anos no grupo Tecnis, 63,1 ± 4,4 anos no grupo Restor e 63,7 ± 4,2 anos no grupo monofocal (SN60AT no olho contralateral SN60WF). A acuidade visual para perto não corrigida e corrigida para longe foi estatisticamente superior nos grupos multifocal Restor e multifocal Tecnis em comparação ao grupo monofocal SN60AT/SN60WF (p<0,001). Não houve diferença estatística entre os grupos na comparação da acuidade visual para longe (p=0,56). A sensibilidade ao contraste fotópica monocular foi estatisticamente inferior nos grupos Restor e Tecnis (p<0,001). A SN60AT apresentou maior aberração esférica comparada a todas as outras lentes (p<0,001). A Tecnis se mostrou com menores valores médios de aberrações esféricas na comparação com a Restor (p<0.001). O questionário de satisfação apresentou maior independência de óculos a favor da Tecnis e Restor, mas com mais queixas de halos e glare do que no grupo monofocal SN60AT/SN60WF. Conclusão: A Restor e Tecnis apresentaram melhor acuidade visual para perto do que o grupo monofocal SN60AT/SN60WF. Todas as lentes intraoculares promoveram boa visão para longe. O grupo da Tecnis apresentou melhor sensibilidade de contraste, menos aberrações óptica e melhor visão intermediária que o grupo Restor. As lentes Restor e Tecnis apresentaram maior satisfação na visão de perto e independência do uso de óculos e glare e halos do que as lentes monofocais / Purpose: Comparison of the visual performance between patients with Tecnis® MF ZM900, Acrysof® Restor® SN60D3, Acrysof® SN60AT and Acrysof® SN60WF intraocular lenses. Patients and methods: This prospective comparative study included 142 eyes of 71 patients in Sao Paulo University. The ophthalmologic evaluation performed included near, intermediate and distance corrected and uncorrected visual acuity, contrast sensitivity measurement and wavefront analysis. All patient underwent a quality of life questionnaire related to visual acuity, glare and halos. The minimum follow-up was 6 months. Results: The mean age of patients was 60.7 ± 6.6 years in the Tecnis® MF, 63.1 ± 4.4 years in-group Restor® 63.7 ± 4.2 years in monofocal group (SN60AT in the fellow eye SN60WF). The Restor and Tecnis groups has uncorrected near and corrected distance visual acuity statistically superior compared to SN60AT/SN60WF group (p <0.001). There was no statistical difference between groups when compared uncorrected and best-corrected distance visual acuity (p = 0.56). Contrast sensitivity in photopic conditions was significantly lower in the Restor and Tecnis group (p <0.001). The SN60AT showed higher spherical aberration compared to all other lenses (p <0.001). The Tecnis showed a lower mean values of spherical aberration compared to Restor lenses (p <0.001). Quality of life questionnaire showed greater independence from glasses for the Tecnis and Restor groups, but with more complaints of halos and glare than the monofocal SN60AT/SN60WF group. Conclusion: Restor and the Tecnis had better near visual acuity than the SN60AT/SN60WF group. All intraocular lenses promoted good distance vision. The Tecnis group showed better contrast sensitivity, less aberrations and better intermediate vision than the Restor group. Restor and Tecnis lenses showed higher satisfaction with near vision and independence from glasses, glare and halos than the monofocal lens

A mensuração da pressão intraocular (PIO) é essencial para o exame oftálmico, pois permite o diagnóstico de doenças oftálmicas como uveíte e glaucoma, além de auxiliar no monitoramento das cirurgias intraoculares. Objetivou-se comparar a tonometria de rebote com a tonometria de aplanação em equinos. Além disso, determinar a reprodutibilidade dos valores obtidos em ambos os aparelhos. A PIO foi aferida em 30 equinos da raça Crioula, machos ou fêmeas, com idade entre um e 16 anos utilizando o tonômetro de rebote (TonoVet®) e o tonômetro de aplanação (Tonopen Avia®). Previamente a aferição da PIO, todos os animais foram submetidos ao exame oftálmico incluindo, avaliação dos reflexos pupilares, teste da lágrima de Schirmer, prova da fluoresceína e biomicroscopia com lâmpada de fenda portátil. Foram utilizados apenas bulbos oculares hígidos. A análise estatística foi feita com ANOVA e teste t de Student. A reprodutibilidade foi avaliada pelo cálculo do coeficiente de correlação intraclasse (ICC). A média da PIO foi de 26,16 ± 5,13 mmHg utilizando o tonômetro de rebote e de 22,55 ±7,32 mmHg com o tonômetro de aplanação. A média da PIO de acordo com o avaliador utilizando o tonômetro de rebote foi de 25,27 ± 4,9 mmHg para o examinador A, 25,80 ± 5,2 mmHg para o examinador B e de 27,40 ± 5,1 mmHg para o examinador C. Com o tonômetro de aplanação as médias obtidas foram de 20,03 ± 4,5 mmHg com o examinador A, 19,67 ± 6,0 mmHg com o examinador B e de 27,97 ± 10,3 mmHg com o examinador C (p<2,3). O ICC foi de 0,18 para o tonômetro de aplanação e de 0,41 para o tonômeto de rebote. Os valores da pressão intraocular média obtidos com o tonômetro de rebote são superestimados quando comparados aos valores obtidos com o tonômetro de Tonopen Avia® em equinos saudáveis. Comparativamente ao Tonopen Avia® os valores obtidos com o Tonovet® demonstraram maior reprodutibilidade. / The measurement of intraocular pressure (IOP) is essential for the ophthalmic examination, as it allows the diagnosis of eye diseases such as uveitis and glaucoma, as well as aid in the monitoring of intraocular surgery. This study aimed to compare the mean intraocular pressure (IOP) values in horses obtained by three different examiners using the rebound and applanation tonometer’s. Each examiner measured the IOP of 30 “Crioulo” horses, males or females, aged one to 16 years old, using rebound tonometer and applanation tonometer. Previous to the experiment an ophthalmic examination was performed in horses. Only animals without alterations were selected. Three veterinarians measured IOP with rebound (Tonovet®) and applanation tonometer’s (TonoPen Avia®). Comparisons were made using ANOVA and student t test. Intraclass correlation coefficient (ICC) was calculated for reproducibility. IOP measurements resulted in mean values of 26.16 ± 5.13 mmHg for rebound tonometer and 22.55 ±7.32 mmHg for applanation tonometer. These values were different on statistical analysis. Rebound tonometer mean values, according with the examiner, were: A 25.27 ± 4.9 mmHg, B 25.80 ± 5.2, C 27.40 ± 5.1 mmHg. For applanation tonometer the mean IOP for examiner A (20.03 ± 4.5 mmHg), B (19.67 ± 6.0 mmHg), C (27.97 ± 10.3 mmHg). With the Tonopen Avia® tonometer examiner C measured higher IOP’s than examiners A and B. Intraclass correlation coefficient was 0.41 for rebound tonometer was 0.18 for applanation tonometer. It was observed that the values obtained with the Tonovet® were overestimated compared to Tonopen Avia® in healthy horses. When compared with Tonopen Avia® values obtained with the Tonovet® showed a higher reproducibility.

Glaucoma affects tens of millions of people and is the leading cause of irreversible blindness worldwide. Virtually all current glaucoma therapies target elevated intraocular pressure (IOP); however, the contribution of intracranial pressure (ICP) to glaucoma has recently garnered interest. Strain at the optic nerve head is now known to depend on the translaminar pressure difference (TLPD), which is the difference between IOP and ICP, rather than IOP alone. A better understanding of how IOP and ICP relate to glaucoma development and progression is essential for developing improved therapies and diagnostic tests. Glaucoma is commonly modeled in rats, yet aqueous humor dynamics are not well-documented in healthy nor diseased rat eyes. Moreover, because rats do not develop glaucoma spontaneously, it is essential to develop low-cost, reliable, and relevant models of glaucomatous pathology in the animal. The purpose of this dissertation work is to achieve the following goals: i) quantitatively assess aqueous humor dynamics in healthy, living rat eyes, ii) develop an ideal model of experimental glaucoma in rats, iii) quantitatively characterize aqueous humor dynamics throughout experimental glaucoma in living rats, and iv) investigate the effects of ICP manipulations on aqueous humor dynamics in living rats. Chapter 2 reports physiological parameters of aqueous humor dynamics for the first time in the eyes of living, healthy Brown-Norway rats, and presents a novel perfusion technique for efficiently and accurately estimating these parameters. Chapter 3 introduces the constant-pressure perfusion model of experimental glaucoma: a powerful new model which overcomes several limitations of existing techniques. The constant-pressure perfusion model induces IOP elevations which are prescribable and easily manipulated, does not directly target the trabecular meshwork or its vasculature, and offers continuous records of IOP rather than requiring regular animal handling and tonometry. Chapter 3 characterizes IOP-induced optic neuropathies in rats and demonstrates their resemblance to human glaucoma. Chapter 4 evaluates whether the constant-pressure perfusion model affects ocular physiology, specifically showing that resting IOP and conventional outflow facility are not permanently nor significantly altered in the model. Chapter 5 examines the effect of ICP manipulations on aqueous outflow physiology in living rats, and reports for the first time a graded effect of intracranial hypertension on conventional outflow facility. Evidence for a neural feedback mechanism that may serve to regulate the TLPD is also presented. Chapter 6 summarizes the results of this dissertation, provides recommendations for future work, and gives closing remarks. These collective projects provide insight into IOP regulation in both healthy and diseased rat eyes, advancing our understanding of glaucomatous development and damage in rats. A novel model of experimental glaucoma and several perfusion systems have been developed which are distinctly tailored for use in future glaucoma studies and will allow future investigators to study the disease with enhanced efficiency and exactitude. The results of this dissertation work suggest that detecting and correcting impairments of either IOP or ICP homeostatic capabilities may be of utmost importance for improving clinical outcomes in human glaucoma.

The murine eye naturally undergoes post-natal changes in eye size. This dissertation quantifies longitudinal structural and functional changes in control mice (C57BL/6J (B6), D2-Gpnmb+/SjJ) and in DBA/2J (D2) mice, which spontaneously develop elevated intraocular pressure (IOP). IOP elevation results in abnormal eye elongation, retinal nerve fiber layer (RNFL) thickness thinning and retinal ganglion cell (RGC) dysfunction and demise resembling human glaucoma. I measured structural changes with Optical Coherence Tomography (OCT), and RGC function with Pattern Electroretinogram (PERG). I also developed and refined provocation approaches (IOP elevation with changes in body posture; metabolic load with flickering light) to probe susceptibility of RGC function in D2 mice prone to glaucoma. Finally, I developed a novel system for recording, simultaneously but independently, the PERG from both eyes using asynchronous visual stimuli and deconvolution analysis. Simultaneous PERG recording from each eye was hitherto impossible due to the interocular cross-talk of the PERG signal. Altogether, the combination of these measures (OCT, PERG) and provocative conditions may represent powerful tools for glaucoma research using mouse models.

El glaucoma es una de las principales causas de ceguera parcial o total e irreversible en el mundo. Los avances en su estudio revelan la compleja naturaleza multifactorial de la enfermedad que consiste finalmente en un daño del nervio óptico (neuropatía).El incremento de la presión intraocular (PIO) prolongado en el tiempo es uno de los factores de riesgo más significativo y reconocido para el desarrollo de esta enfermedad. Este incremento puede causar lesiones en el nervio óptico por deformación mecánica y/o cambios fisiopatológicos en una zona de la retina dónde se sitúan las células ganglionares del nervio. La PIO es resultado del balance entre la producción y drenaje del humor acuoso. El humor acuoso es un fluído derivado de plasma sanguíneo, producido en los procesos ciliares, que baña la cámara anterior del ojo para alimentar sus estructuras avasculares y mantener la tensión mecánica adecuada del globo ocular, y es drenado, principalmente, a través de la malla trabecular o trabéculo. Se sabe que el incremento de la contracción de este tejido implica una disminución en la facilidad de drenaje del humor acuoso y un aumento de la PIO. La presencia incrementada de endotelina-1 (ET-1), potente vasoconstrictor, en humor acuoso de pacientes glaucomatosos plantea algunas cuestiones que abordamos en este trabajo de tesis doctoral. Estas sustancias (endotelina-1, bradiquinina) son agonistas de receptores acoplados a proteína Gq que activan la vía de la fosfolipasa C (vía PLC- inositol trifosfato y diacilglicerol) y podrían tener un efecto contráctil, también, sobre la malla trabecular dado que activan la depleción de Ca2+ intracelular. El objetivo principal de este trabajo de tesis doctoral ha sido identificar y caracterizar un tipo de corriente de entrada de calcio extracelular (vía principal de entrada de calcio en células no excitables), llamada corriente 'capacitativa' de Ca2+ o dependiente del vaciado de Ca2+ de los depósitos intracelulares (ICCE o ISOC) y su posible implicación en la función de contracción/relajación de las células trabeculares y en consecuencia del tejido. Identificamos distintos tipos de corrientes capacitativas de Ca2+ en cultivos primarios de células trabeculares en modelo bovino y humano, tras el estímulo con sustancias vasoconstrictoras y tapsigargina. Para la caracterización farmacológica usamos bloqueadores típicos de la ICCE (2-APB, lantánidos, SKF96365) y registramos la evolución de la activación de ICCE mediante registros electrofisiológicos, testeando nuevamente bloqueadores conocidos de este tipo de corrientes. Observamos cómo en algunos casos las corrientes capacitativas son selectivas para el Ca2+ y en otros (vía PLC), en cambio, aparecían formas de corrientes catiónicas no selectivas.Detectamos mediante western blot la presencia de proteínas de canal tipo TRPC (TRPC1 y TRPC4) que forman canales catiónicos en membrana y pueden ser activados vía PLC, posibles mediadores de la ICCE en células trabeculares.La técnica de microscopía de tracción permitió estudiar la evolución de la contracción de la célula por estímulo con sustancias vasoconstrictoras. Obtuvimos medidas cuantitativas y la distribución de fuerzas en la célula durante la contracción. Estos resultados revelaron que la contracción inducida por estas sustancias depende primeramente de la depleción de Ca2+ intracelular y curiosamente, el Ca2+ extracelular regula, disminuyendo, la intensidad de esa contracción. Mostramos en este trabajo, la existencia de la ICCE en células trabeculares y sugerimos el papel de esta vía de entrada de Ca2+ como señal autoreguladora del proceso de contracción celular y su posible efecto en la función evacuadora del tejido. / High concentrations of endothelin-1 (ET-1), one of the most potent vasoconstrictor peptide, has been found in the aqueous humor of glaucomatous eyes. It is said that vasoconstrictor substances as ET-1 or bradykinin could induce contraction of trabecular meshwork tissue (TM) and decreases the aqueous humour outflow facility increasing the intraocular pessure (IOP) and the risk of glaucoma. The aim of this study is the calcium signalling, mainly, the characterization of the calcium influx/entry in these cells, the identification of capacitative calcium entry (ICCE) or, also called, store-operated calcium entry (ISOC) in trabecular cells and their role on cell contraction.We found these currents in cell culture of bovine and human trabecular cells and in a human cell line, too, using fluorescence techniques and electrophysiology recordings. They are stimulated by the intracellular Ca2+ depletion from stores via IP3 (bradykinin, endothelin) and blocking the Ca2+-pump of endoplasmic reticulum (thapsigargin). These currents are blocked or inhibited by certain lanthanids ions, 2-APB or SKF96365. Western blots showed the presence of certain members of the Transient Receptor Potential (TRP) channels subfamily C (TRPC1 and TRPC4). The pharmacology of inhibition of the Ca2+ influx suggests the TRPC channels and store-operated calcium channels as mediators. The measurements of cell traction evolution by traction microscopy maps stimulating with vasoconstrictor substances were useful to determine store-operated Ca2+ entry may not be necessary for cell contraction in TM cells but may contribute to modulate cell contractility and so, the aqueous humour outflow facility.

Purpose: To locate protein sorption on the surface and inside the matrix of soft contact lens materials and intraocular lenses (IOL). Methods: The proteins albumin and lysozyme were investigated as they are highly abundant in blood serum and tears, respectively. Proteins were conjugated with organic fluorescent probes and using confocal laser scanning microscopy (CLSM) the sorption profile to contact lenses and IOL could be determined. Radiolabeled protein was used for quantification purposes. • Albumin sorption to etafilcon A and lotrafilcon B was determined (Chapter 3) • Different fluorescent probes were used for conjugation and the impact on albumin sorption behaviour was investigated (Chapter 4) • Lysozyme sorption to nine different pHEMA-based and silicone hydrogel contact lenses was determined using two fluorescent probes (Chapter 5) • The efficiency of protein removal from contact lenses using contact lens care regimens was investigated (Chapter 6) • Albumin sorption to IOL materials was quantified and imaged using a modified CLSM technique (Chapter 7) Results: Albumin and lysozyme sorption profiles differed between materials, and were influenced by the fluorescent probes used for conjugation. After one day of incubation, both proteins could be located within all contact lens materials, except for lotrafilcon A and lotrafilcon B, which primarily allowed deposition on the lens surface. An increase in protein accumulation was found for most materials over the maximum investigated period of 14 days, using CLSM and radiolabel techniques. The efficiency of contact lens care regimens to remove lysozyme and albumin depended on the lens material, care regimen and protein type investigated. PMMA and silicone IOLs showed protein exclusively on the surface, while a hydrophilic acrylic IOL allowed penetration into the lens matrix over time. Despite the albumin penetration depth into hydrophilic acrylic, the highest albumin levels were determined for the silicone IOL. Conclusions: CLSM provides detailed information that can describe the protein distribution in transparent biomaterials, with scanning depths up to a few hundred microns. However, the CLSM data are primarily of qualitative value, which necessitates a quantitative technique (e.g. radiolabeling) to determine the total protein content.

Purpose: To locate protein sorption on the surface and inside the matrix of soft contact lens materials and intraocular lenses (IOL). Methods: The proteins albumin and lysozyme were investigated as they are highly abundant in blood serum and tears, respectively. Proteins were conjugated with organic fluorescent probes and using confocal laser scanning microscopy (CLSM) the sorption profile to contact lenses and IOL could be determined. Radiolabeled protein was used for quantification purposes. • Albumin sorption to etafilcon A and lotrafilcon B was determined (Chapter 3) • Different fluorescent probes were used for conjugation and the impact on albumin sorption behaviour was investigated (Chapter 4) • Lysozyme sorption to nine different pHEMA-based and silicone hydrogel contact lenses was determined using two fluorescent probes (Chapter 5) • The efficiency of protein removal from contact lenses using contact lens care regimens was investigated (Chapter 6) • Albumin sorption to IOL materials was quantified and imaged using a modified CLSM technique (Chapter 7) Results: Albumin and lysozyme sorption profiles differed between materials, and were influenced by the fluorescent probes used for conjugation. After one day of incubation, both proteins could be located within all contact lens materials, except for lotrafilcon A and lotrafilcon B, which primarily allowed deposition on the lens surface. An increase in protein accumulation was found for most materials over the maximum investigated period of 14 days, using CLSM and radiolabel techniques. The efficiency of contact lens care regimens to remove lysozyme and albumin depended on the lens material, care regimen and protein type investigated. PMMA and silicone IOLs showed protein exclusively on the surface, while a hydrophilic acrylic IOL allowed penetration into the lens matrix over time. Despite the albumin penetration depth into hydrophilic acrylic, the highest albumin levels were determined for the silicone IOL. Conclusions: CLSM provides detailed information that can describe the protein distribution in transparent biomaterials, with scanning depths up to a few hundred microns. However, the CLSM data are primarily of qualitative value, which necessitates a quantitative technique (e.g. radiolabeling) to determine the total protein content.