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Influence of sex hormones and genetic predisposition in dry eye in Sjèogren's syndrome: a new clue to the immunopathogenesis of dry eye diseaseUnknown Date (has links)
Sjèogren's syndrome (S) is a chronic autoimmune disease characterized by ocular and oral dryness and primarily affects post menopausal women. In the present study we investigated the time course of lymphocytic infiltration, apoptosis, caspase-3 activity and different cytokines levels in the lacrimal glands of both genetically predisposed and control mice to elucidate immunopathological mechanism leading to dry eye. The results of our experiments showed that ovariectomy accelerated pathological findings of SS by increasing lympocytic infiltration, cytokine production, lacrimal gland cell death and cleaved caspase-3 activity, and these effects were more pronounced and persistent in the genetically predisposed mouse model of SS. In addition, we observed that lymphocytic infiltration occurred earlier compared to apoptosis which may perpetuate immune mediated destruction of lacrimal epithelial cells. Furthermore, treatment with physioloigical doses of 17-B Estradiol (E2) or DIhydrotestosterone (DHT) prevented all these pathological events observed after ovariectomy. / by Safinaz Mostafa. / Thesis (M.S.)--Florida Atlantic University, 2011. / Includes bibliography. / Electronic reproduction. Boca Raton, Fla., 2011. Mode of access: World Wide Web.
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Resultados funcionais da ressecção supramáxima do levantador da pálpebra superior em pacientes com blefaroptose congênita / Functional results in supramaximal levator resection for congenital ptosisMendonça, Ana Karina Téles Silveira de 28 May 2015 (has links)
O tratamento cirúrgico da blefaroptose congênita é controverso e, embora a ressecção supramáxima do levantador palpebral tenha sido advogada por alguns autores como eficaz em diminuir a assimetria palpebral, existe o risco de que a cirurgia possa estar associada à restrição dos movimentos palpebrais e à ocorrência de olho seco. Com o objetivo de quantificar os movimentos palpebrais nos pacientes submetidos à cirurgia e avaliar a eventual relação entre supostas alterações funcionais dos movimentos palpebrais e a ocorrência de complicações relacionadas à superfície ocular, foram caracterizadas, mediante a utilização da técnica magnetic search coil, as amplitudes e as velocidades dos movimentos do piscar e dos sacádicos palpebrais dos olhos operados de dezoito pacientes e, em seguida, foram comparadas com as amplitudes e velocidades obtidas para os olhos contralaterais não operados dos mesmos pacientes e também com as obtidas para os olhos dos pacientes do grupo controle, não operados. Também foram avaliados os resultados cosméticos nos pacientes operados e a ocorrência de complicações relacionadas à superfície ocular entre os olhos operados e os controles, não operados. A cirurgia acarretou aumento da distância margem-reflexo da pálpebra superior e diminuição da assimetria palpebral, logrando bom resultado estético. Porém, tanto a amplitude quanto a velocidade, seja das piscadelas, ou dos movimentos sacádicos palpebrais dos olhos operados, foram inferiores às dos controles e a cirurgia esteve relacionada com maior risco de ocorrência de olho seco. Por esse motivo, os autores advogam que a ressecção supramáxima do levantador da pálpebra superior permanece uma opção eficaz para o tratamento da ptose palpebral congênita, mas contraindicam a sua utilização em pacientes com ausência do reflexo de Bell ou outras alterações prévias documentadas ou predisponentes à ceratoconjuntivite seca. / Surgical treatment of congenital blepharoptosis remains controversial and, although supramaximal resection of the levator palpebrae superioris has been advocated as effective in reducing eyelid asymmetry, there is a risk that the surgery may be associated with restriction of eyelid movements and the occurrence of dry eye. In order to quantify eyelid movements in patients undergoing surgery and to assess the possible relationship between the alleged functional changes of eyelid movements and the occurrence of complications related to the ocular surface, we measured by magnetic search coil technique the amplitude and the maximal descent speed of blinks and lid saccades in eighteen patients who undergone surgery and in ten patients of the control group, with no eyelid diseases. We compared the amplitude and speed of the operated eyes with that of the non-operated eyes. We also evaluate the cosmetic results in operated patients and the occurrence of dry eye between operated and non-operated eyes. The surgery resulted in increased upper lid margin-reflex distance and consequent decreased eyelid asymmetry, achieving good aesthetic result. However, both amplitude and speed of eyelid movements in operated eyes were lower than those for the control eyes and the surgery was associated with increased risk of dry eye. For this reason, the authors advocate that supramaximal levator resection of the levator palpebrae superioris remains an effective option for the treatment of congenital blepharoptosis, but contraindicate its use in patients with absence of Bell\'s reflex or with previous ocular surface diseases.
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Avaliação do desempenho de colírio usando soro alogênico e fatores de crescimento derivados de plaquetas em cultura de células do anel córneo-escleral /Martins, Juliana Ravelli Baldassarre. January 2014 (has links)
Orientador: Elenice Deffune / Banca: Magda Massae Hata Viveiros / Banca: Jaqueline de Camargo Rinaldi / Resumo: Introdução: Muitas patologias podem acometer a córnea, entre elas a síndrome do olho seco e as doenças relacionadas. Tratamentos inovadores para a síndrome do olho seco têm surgido, entre eles, o uso de colírio produzido com soro autólogo de uso ocular, soro de sangue de cordão umbilical, meio condicionado obtido pela expansão de células tronco do epitélio limbal (modelo ex vivo), entre outros. Pacientes com provável indicação do colírio de soro autólogo podem ter a contra-indicação na produção oriunda do status imune para doenças virais como HIV, hepatites B e C, sífilis, doença de Chagas e HTLV I e II, que do ponto de vista de produção do autocolírio, encontram barreiras. Para estes casos, o colírio de soro alogênico, oriundo de doadores saudáveis, é uma alternativa viável. Objetivos: Analisar o efeito in vitro da ação de colírio alogênico em cultura de células da região córneo-escleral, estabelecer a cultura e expansão de células aderentes obtidas da região córneo-escleral, comparar a cultura das células aderentes em meio de cultura contendo colírio e fatores de crescimento derivados de plaquetas, avaliar as células por meio da técnica de imunohistoquímica e analisar os dados médicos dos pacientes que utilizam autocolírio por meio de prontuários. Casuística e Métodos: Os anéis córneo-esclerais foram obtidos no centro cirúrgico após cirurgia de doação de córnea, sendo que estes anéis são materiais de descarte após a cirurgia. Em seguida, o tecido foi encaminhado ao Laboratório de Engenharia Celular do Hemocentro de Botucatu, onde foi fragmentado em pequenas partes e digerido com colagenase tipo I. As células foram plaqueadas em frascos de 25 cm² com meio DMEM Knockout, em uma primeira parte do experimento, e em meio Keratinocyte na segunda parte do trabalho. Após confluência de 80%, as células passaram pelo processo de tripsinização para desprendimento ... / Abstract: Introduction: Many diseases can affect cornea, including dry eye syndrome and related diseases. Innovative treatments for dry eye syndrome have emerged , among them the use of autologous serum eye drops produced with the use of eye serum, umbilical cord blood, conditioned medium obtained by expansion of limbal epithelial stem cells (ex vivo model) among others. Patients with probable indication of autologous serum eyedrops may be contraindicated in production originating immune status for viral diseases such as HIV, hepatitis B and C, syphilis , Chagas disease and HTLV I and II, from the point of view of eye drops's production, have barriers . For these cases, eye drops from healthy donors allogeneic serum can be an alternative. Objectives: Evaluate in vitro effect of the action of allogeneic eye drops in corneal-scleral cell culture, established the culture and expansion of adherent cells obtained from the corneal- scleral region, compare the culture of adherent cells in culture medium containing eye drops and platelets derived growth factors, evaluating the cells by immunohistochemistry and analyze medical data of eye drops's patients. Materials and Methods: The corneal- scleral rings were obtained in the operating room after surgery of tissue donor, and these rings were discarded after surgery. Then, tissue was sent to the Laboratory of Cell Engineering of Botucatu Blood Center, which was fragmented into small pieces and digested with collagenase type I. Cells were plated in 25 cm² flasks with DMEM Knockout in the first part of the experiment, and Keratinocyte in the second part of the experiment. After 80% confluence, cells passed through the trypsinization procedure for detachment of the cells from the culture flask. Mounting plaque experiment, cells were placed to control, or untreated cells, treated cells with eye drops and cells treated with platelets derived growth factors was performed. After 5 days, immunohistochemistry ... / Mestre
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Ocular Discomfort Upon Tear DryingVarikooty, Jalaiah January 2003 (has links)
<b>Purpose:</b> Assess the relationship between tear film drying and sensation between blinks. <b>Methods:</b> MATLAB sampled a slitlamp video camera, a potentiometer and a microphone while subjects kept one eye open for as long as possible. 23 subjects rated the intensity of the ocular sensation while video and voice data were collected simultaneously. The tear drying on the cornea was measured. <b>Results:</b> The sensation was triphasic. Two linear functions described the latter 2 parts of the data (r ≥ 0. 95). The correlation between TBUT and the elbow in the time-discomfort function was 0. 72. Extent of tear film drying was linearly correlated to time (median correlation = 0. 88). The correlation between the discomfort elbow and image elbow was 0. 93 with single data pair for each subject. Analysis of sensation characteristics showed significant differences between itching and burning for both intensity and time (p = 0. 03 and p = 0. 02 respectively). <b>Conclusions:</b> Simultaneous recording of ocular surface appearance, discomfort intensity and attributes of sensation provide novel information about the development of discomfort during ocular surface drying. The rapid increase in discomfort proceeding blinking has been quantified and the relationship between the time course of drying and discomfort is elucidated.
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Effects of Computer Usage on Ocular HealthMoy, Alexa J 01 January 2014 (has links)
The 2010 National Education Technology Plan steadily replaces paper textbooks with study materials on computers. One risk of increased computer usage is increased Computer Vision Syndrome (CVS) symptoms. I researched multiple studies to analyze the effects of computer use on dry eye and eye strain and how these symptoms can progress to musculoskeletal pain, headaches, decreased quality of life, loss of confidence and even anxiety and depression. Currently, there is not much data on tablet use so I propose two future experiments to determine if tablets can also cause CVS ocular symptoms.
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Avaliação do desempenho de colírio usando soro alogênico e fatores de crescimento derivados de plaquetas em cultura de células do anel córneo-escleralMartins, Juliana Ravelli Baldassarre [UNESP] 28 February 2014 (has links) (PDF)
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000786133.pdf: 2473732 bytes, checksum: 2718404dbc454aa4d3963273c33b3ecc (MD5) / Introdução: Muitas patologias podem acometer a córnea, entre elas a síndrome do olho seco e as doenças relacionadas. Tratamentos inovadores para a síndrome do olho seco têm surgido, entre eles, o uso de colírio produzido com soro autólogo de uso ocular, soro de sangue de cordão umbilical, meio condicionado obtido pela expansão de células tronco do epitélio limbal (modelo ex vivo), entre outros. Pacientes com provável indicação do colírio de soro autólogo podem ter a contra-indicação na produção oriunda do status imune para doenças virais como HIV, hepatites B e C, sífilis, doença de Chagas e HTLV I e II, que do ponto de vista de produção do autocolírio, encontram barreiras. Para estes casos, o colírio de soro alogênico, oriundo de doadores saudáveis, é uma alternativa viável. Objetivos: Analisar o efeito in vitro da ação de colírio alogênico em cultura de células da região córneo-escleral, estabelecer a cultura e expansão de células aderentes obtidas da região córneo-escleral, comparar a cultura das células aderentes em meio de cultura contendo colírio e fatores de crescimento derivados de plaquetas, avaliar as células por meio da técnica de imunohistoquímica e analisar os dados médicos dos pacientes que utilizam autocolírio por meio de prontuários. Casuística e Métodos: Os anéis córneo-esclerais foram obtidos no centro cirúrgico após cirurgia de doação de córnea, sendo que estes anéis são materiais de descarte após a cirurgia. Em seguida, o tecido foi encaminhado ao Laboratório de Engenharia Celular do Hemocentro de Botucatu, onde foi fragmentado em pequenas partes e digerido com colagenase tipo I. As células foram plaqueadas em frascos de 25 cm² com meio DMEM Knockout, em uma primeira parte do experimento, e em meio Keratinocyte na segunda parte do trabalho. Após confluência de 80%, as células passaram pelo processo de tripsinização para desprendimento ... / Introduction: Many diseases can affect cornea, including dry eye syndrome and related diseases. Innovative treatments for dry eye syndrome have emerged , among them the use of autologous serum eye drops produced with the use of eye serum, umbilical cord blood, conditioned medium obtained by expansion of limbal epithelial stem cells (ex vivo model) among others. Patients with probable indication of autologous serum eyedrops may be contraindicated in production originating immune status for viral diseases such as HIV, hepatitis B and C, syphilis , Chagas disease and HTLV I and II, from the point of view of eye drops´s production, have barriers . For these cases, eye drops from healthy donors allogeneic serum can be an alternative. Objectives: Evaluate in vitro effect of the action of allogeneic eye drops in corneal-scleral cell culture, established the culture and expansion of adherent cells obtained from the corneal- scleral region, compare the culture of adherent cells in culture medium containing eye drops and platelets derived growth factors, evaluating the cells by immunohistochemistry and analyze medical data of eye drops´s patients. Materials and Methods: The corneal- scleral rings were obtained in the operating room after surgery of tissue donor, and these rings were discarded after surgery. Then, tissue was sent to the Laboratory of Cell Engineering of Botucatu Blood Center, which was fragmented into small pieces and digested with collagenase type I. Cells were plated in 25 cm² flasks with DMEM Knockout in the first part of the experiment, and Keratinocyte in the second part of the experiment. After 80% confluence, cells passed through the trypsinization procedure for detachment of the cells from the culture flask. Mounting plaque experiment, cells were placed to control, or untreated cells, treated cells with eye drops and cells treated with platelets derived growth factors was performed. After 5 days, immunohistochemistry ...
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Organização estrutural da cromatina em células epiteliais da conjuntiva palpebral de cães com ceratoconjuntivite seca, antes e após tratamento local com ciclosporina A / Chromatin structure organization in palpebral conjunctival epithelial cells from dogs with sicca keratoconjunctivitis, before and after local treatment with cyclosporine ASantos, Daniela Moura 28 February 2018 (has links)
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Previous issue date: 2018-02-28 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Alterações na organização estrutural da cromatina estão sendo associadas ao desenvolvimento e à fisiopatologia de diversas afecções oftálmicas. Com o advento da epigenética, emergiu o conceito de que parte dessas alterações pode ser revertida por fármacos. Visando-se avaliar a organização estrutural da cromatina em células da conjuntiva palpebral de cães com ceratoconjuntivite seca (CCS), antes e após tratamento com pomada de ciclosporina A (CsA) 0,2%, a presente pesquisa foi dividida em duas fases. Na fase I estudaram-se núcleos de células epiteliais e de linfócitos colhidos da conjuntiva palpebral de cães com e sem CCS. Foram incluídos na pesquisa 64 olhos de 32 cães domésticos, distribuídos em dois grupos: grupo controle, composto por 16 cães saudáveis (32 olhos, valores de Schirmer ≥ 15 mm/min); grupo CCS, abrangendo 16 cães (32 olhos, valores de Schirmer ≤ 10 mm/min) com CCS bilateral nunca antes tratados com imunomoduladores, sem oftalmopatias concorrentes e livres de alterações sistêmicas. As células foram colhidas por citologia esfoliativa e coradas pela reação de Feulgen. Os seguintes parâmetros foram estudados por vídeo-análise de imagens: área nuclear, perímetro nuclear, fator de circularidade relativa do núcleo (RNRF), densidade óptica integrada (IOD = conteúdo de DNA), densidade óptica (OD = compactação de cromatina); e desvio padrão de valores densitométricos (SDtd = textura de cromatina). Os resultados mostraram que a CCS enseja alterações nos parâmetros nucleares das células epiteliais e dos linfócitos da conjuntiva palpebral. Comparativamente aos controles, as células epiteliais foram mais afetadas pela CCS (alterações em área, perímetro, RNRF, IOD, e OD) do que os linfócitos (alterações em OD, apenas). Tanto as células epiteliais quanto os linfócitos do grupo CCS apresentaram cromatina mais descompactada do que as células do grupo controle. Padrões aberrantes de cromatina, como a "snake-like-chromatin", comumente vistos em pacientes humanos, não foram detectados. Na fase II estudou-se se as alterações nucleares detectadas na fase I regrediriam após tratamento local com pomada de CsA 0,2% a intervalos regulares de 12 horas e lacrimomimético a base de ácido poliacrílico 0,2%, instilado localmente a cada 4 horas. O mesmo grupo controle composto por cães hígidos foi adotado e as células também foram colhidas por citologia esfoliativa aos 30 e 60 dias de tratamento. As preparações citológicas foram coradas pela reação de Feulgen ou submetidas ao bandamento AgNOR. Estudaram-se parâmetros de video-análise de imagens relacionados com a funcionalidade da cromatina, notadamente a fração de área nuclear coberta pela cromatina mais condensada (Sc%), a taxa média de absorbância (AAR), e a entropia. Após bandamento AgNOR, estudaram-se os tamanhos das regiões organizadores de nucléolo e as frações de áreas nucleares ocupadas por elas. Os resultados mostraram que o tratamento com CsA e lacrimomimético (30/60 dias) enseja remodelação cromatínica e desfaz parcialmente as alterações associadas à CCS. / Changes in the structural organization of chromatin are being associated with the development and pathophysiology of various ophthalmic conditions. With the advent of epigenetics, the concept emerged that part of these alterations can be reversed by drugs. To evaluate the structural organization of chromatin in palpebral conjunctival cells of dogs with keratoconjunctivitis sicca (KCS), before and after treatment with 0.2% cyclosporine A (CsA) ophthalmic ointment, the present research was divided into two phases. In stage I, epithelial cells and lymphocyte from the palpebral conjunctiva of dogs with and without KCS were studied. The study included 64 eyes of 32 domestic dogs, distributed into two groups: control group, composed of 16 healthy dogs (32 eyes, Schirmer values ≥ 15 mm / min); KCS group, formed of 16 dogs (32 eyes, Schirmer values ≤ 10 mm / min) with bilateral CCS never previously treated with immunomodulatory drugs, and without concurrent ophthalmic or systemic disorders. Cells were collected by brush cytology and stained by the Feulgen reaction. The following parameters were studied by video image analysis: nuclear area, nuclear perimeter, relative nuclear roundness factor (RNRF), integrated optical density (IOD = DNA content), optical density (OD = chromatin compaction). The results showed that KCS causes alterations in the nuclear parameters of the epithelial cells and the lymphocytes from the palpebral conjunctiva. Compared with the control, the epithelial cells were more affected by the disease (alterations in area, perimeter, RNRF, IOD, and OD) than lymphocytes (changes in OD only.) Both epithelial cells and lymphocytes from the KCS group showed more decompressed chromatin than cells from the control group. "Snakelike-chromatin" commonly seen in human patients was not detected. In phase II it was studied whether the nuclear detected in stage I, regress after treatment with 0.2% CsA at regular intervals of 12 h and 0.2% polyacrylic acid-based artificial tears at regular intervals of 4h. The same control group composed of healthy dogs was adopted and cells were collected by brush cytology after 30 and 60 days of treatment. Cytological preparations were stained by the Feulgen reaction or submitted to the AgNOR banding. We studied video image analysis parameters related to the functionality of chromatin, notably the fraction of nuclear area covered by more condensed chromatin (Sc %), the average absorption ratio (AAR), and entropy. After AgNOR banding, the samples were studied for the sizes of the nucleolar organizer regions and the fractions of nuclear areas occupied by them. Results show that treatment with CsA and lacrimomimetic (30/60 days) leads to chromatin remodeling and partially reverses the chromatin changes elicited by KCS.
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The Development of a Validated Clinically Meaningful Endpoint for the Evaluation of Tear Film Stability as a Measure of Ocular Surface Protection for Use in the Diagnosis and Evaluation of Dry Eye DiseaseJanuary 2012 (has links)
abstract: This dissertation presents methods for the evaluation of ocular surface protection during natural blink function. The evaluation of ocular surface protection is especially important in the diagnosis of dry eye and the evaluation of dry eye severity in clinical trials. Dry eye is a highly prevalent disease affecting vast numbers (between 11% and 22%) of an aging population. There is only one approved therapy with limited efficacy, which results in a huge unmet need. The reason so few drugs have reached approval is a lack of a recognized therapeutic pathway with reproducible endpoints. While the interplay between blink function and ocular surface protection has long been recognized, all currently used evaluation techniques have addressed blink function in isolation from tear film stability, the gold standard of which is Tear Film Break-Up Time (TFBUT). In the first part of this research a manual technique of calculating ocular surface protection during natural blink function through the use of video analysis is developed and evaluated for it's ability to differentiate between dry eye and normal subjects, the results are compared with that of TFBUT. In the second part of this research the technique is improved in precision and automated through the use of video analysis algorithms. This software, called the OPI 2.0 System, is evaluated for accuracy and precision, and comparisons are made between the OPI 2.0 System and other currently recognized dry eye diagnostic techniques (e.g. TFBUT). In the third part of this research the OPI 2.0 System is deployed for use in the evaluation of subjects before, immediately after and 30 minutes after exposure to a controlled adverse environment (CAE), once again the results are compared and contrasted against commonly used dry eye endpoints. The results demonstrate that the evaluation of ocular surface protection using the OPI 2.0 System offers superior accuracy to the current standard, TFBUT. / Dissertation/Thesis / Ph.D. Industrial Engineering 2012
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Resultados funcionais da ressecção supramáxima do levantador da pálpebra superior em pacientes com blefaroptose congênita / Functional results in supramaximal levator resection for congenital ptosisAna Karina Téles Silveira de Mendonça 28 May 2015 (has links)
O tratamento cirúrgico da blefaroptose congênita é controverso e, embora a ressecção supramáxima do levantador palpebral tenha sido advogada por alguns autores como eficaz em diminuir a assimetria palpebral, existe o risco de que a cirurgia possa estar associada à restrição dos movimentos palpebrais e à ocorrência de olho seco. Com o objetivo de quantificar os movimentos palpebrais nos pacientes submetidos à cirurgia e avaliar a eventual relação entre supostas alterações funcionais dos movimentos palpebrais e a ocorrência de complicações relacionadas à superfície ocular, foram caracterizadas, mediante a utilização da técnica magnetic search coil, as amplitudes e as velocidades dos movimentos do piscar e dos sacádicos palpebrais dos olhos operados de dezoito pacientes e, em seguida, foram comparadas com as amplitudes e velocidades obtidas para os olhos contralaterais não operados dos mesmos pacientes e também com as obtidas para os olhos dos pacientes do grupo controle, não operados. Também foram avaliados os resultados cosméticos nos pacientes operados e a ocorrência de complicações relacionadas à superfície ocular entre os olhos operados e os controles, não operados. A cirurgia acarretou aumento da distância margem-reflexo da pálpebra superior e diminuição da assimetria palpebral, logrando bom resultado estético. Porém, tanto a amplitude quanto a velocidade, seja das piscadelas, ou dos movimentos sacádicos palpebrais dos olhos operados, foram inferiores às dos controles e a cirurgia esteve relacionada com maior risco de ocorrência de olho seco. Por esse motivo, os autores advogam que a ressecção supramáxima do levantador da pálpebra superior permanece uma opção eficaz para o tratamento da ptose palpebral congênita, mas contraindicam a sua utilização em pacientes com ausência do reflexo de Bell ou outras alterações prévias documentadas ou predisponentes à ceratoconjuntivite seca. / Surgical treatment of congenital blepharoptosis remains controversial and, although supramaximal resection of the levator palpebrae superioris has been advocated as effective in reducing eyelid asymmetry, there is a risk that the surgery may be associated with restriction of eyelid movements and the occurrence of dry eye. In order to quantify eyelid movements in patients undergoing surgery and to assess the possible relationship between the alleged functional changes of eyelid movements and the occurrence of complications related to the ocular surface, we measured by magnetic search coil technique the amplitude and the maximal descent speed of blinks and lid saccades in eighteen patients who undergone surgery and in ten patients of the control group, with no eyelid diseases. We compared the amplitude and speed of the operated eyes with that of the non-operated eyes. We also evaluate the cosmetic results in operated patients and the occurrence of dry eye between operated and non-operated eyes. The surgery resulted in increased upper lid margin-reflex distance and consequent decreased eyelid asymmetry, achieving good aesthetic result. However, both amplitude and speed of eyelid movements in operated eyes were lower than those for the control eyes and the surgery was associated with increased risk of dry eye. For this reason, the authors advocate that supramaximal levator resection of the levator palpebrae superioris remains an effective option for the treatment of congenital blepharoptosis, but contraindicate its use in patients with absence of Bell\'s reflex or with previous ocular surface diseases.
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Rôle pro-inflammatoire des cellules épithéliales de la conjonctive dans un modèle in vitro de sécheresse oculaire / Proinflammatory role of epithelial cells from the conjunctiva in an in vitro model of dry eye diseaseWarcoin, Elise 25 January 2016 (has links)
La sécheresse oculaire est une pathologie fréquente qui peut impacter fortement la qualité de vie des patients. D’origine multifactorielle complexe, elle présente de nombreuses étiologies et son diagnostic est difficile en raison de présentations cliniques variées, en termes de symptômes et de tests cliniques, et en l'absence d'un marqueur spécifique. En outre, elle ne bénéficie actuellement que d’une seule molécule thérapeutique, cependant non curative, la ciclosporine. Devenant un réel problème de santé publique, la recherche visant à une meilleure compréhension des mécanismes sous-jacents à son apparition reste indispensable. L'état actuel des connaissances sur la sécheresse oculaire reconnaît à l'inflammation et à l'hyperosmolarité un rôle central. Ce travail de thèse a eu pour objectif d'étudier le comportement pro-inflammatoire des cellules conjonctivales dans un modèle in vitro classique de sécheresse oculaire induite par une hyperosmolarité saline. Nous avons ainsi montré que les cellules conjonctivales sécrètent une chimiokine pro-inflammatoire majeure, CCL2/MCP-1, et que cette induction est totalement sous la dépendance du facteur de transcription osmoprotecteur NFAT5/TonEBP ainsi que partiellement liée à la voie des MAPKs et de NFκB. Nous avons également observé que cette induction était en partie inhibée par la ciclosporine et la dexaméthasone. Nous avons ensuite complété ce travail par l'étude de l'inflammasome à la fois sur ces cellules et sur des monocytes, type cellulaire majeur ciblé par CCL2, en utilisant différents activateurs classiques dans le modèle d’hyperosmolarité. Nos résultats ont montré l’absence d’activation de l’inflammasome par l'hyperosmolarité seule. En revanche, l'hyperosmolarité aurait un rôle inhibiteur de l’inflammasome dans les monocytes. Ces résultats ouvrent des pistes nouvelles d’exploration afin de mieux comprendre les relations entre monocytes et cellules épithéliales de la conjonctive. Ils nous ont permis d'identifier le rôle incontournable de cellules épithéliales conjonctivales dans l'apparition ou le maintien de l'inflammation dans la pathologie et de caractériser certaines voies intracellulaires impliquées dans ce processus, ouvrant de nouvelles perspectives en termes de thérapeutiques. / Dry eye disease is a common condition that significantly impacts the quality of life of patients. It is a complex multifactorial disease with many etiologies and a diagnosis recognized as difficult due to the multiple clinical presentations in terms of symptoms and clinical tests, and lacking of a reliable and specific marker. There is currently only one therapeutic molecule to treat this disease, cyclosporin. However it is not curative. Dry eye disease is becoming an important public health problem and it is necessary to focus research on its underlying mechanisms, as they remain largely unknown. The current state of knowledge in dry eye disease recognizes inflammation and hyperosmolarity as central elements in the pathology. This work aimed to study the pro-inflammatory behavior of conjunctival cells in an in vitro model of dry eye disease by NaCl-induced hyperosmolarity. We showed that conjunctival cells secrete a major pro-inflammatory chemokine known as CCL2/MCP-1. This induction depends entirely on the osmoprotectant transcription factor NFAT5/TonEBP and is partially related to the MAPKs and the NFκB pathways. We showed that the process could be partially inhibited by cyclosporin and dexamethasone. We also studied the inflammasome in conjunctival cells and monocytes, the major cell type targeted by CCL2, using various classical inflammasome activators in this hyperosmolarity model. We did not observe any inflammasome activation induced by hyperosmolarity alone. On the other hand, hyperosmolarity shows signs of inhibiting inflammasome activation in monocytes. These results open new ways of understanding the relations between monocytes and conjunctival epithelial cells. They highlight the crucial role of the conjunctival epithelial cell type in the inflammation process in dry eye disease and make it possible to characterize intracellular pathways involved in this process, opening new therapeutic prospects.
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