Histopathological analysis of residual lens cells in capsular opacities after cataract surgery using objective software.

Purpose: Remnant lens epithelial cells (LECs) within the capsular bag (CB) undergo epithelial-to-mesenchymal transition (EMT) and acquire a myofibroblast phenotype, depositing extracellular matrix (ECM) components, leading to posterior capsular opacification (PCO). This study histopathologically analyzes the LEC-to-myofibroblast transition and de novo ECM component deposition (i.e., smooth muscle actin (SMA) and fibronectin (FN) expression) and determines the intraocular lens (IOL) and patient factors associated with these changes. Methods: In total, 190 CBs with IOLs were removed from donor eyes. Digital images were obtained, and PCO was graded using published software (ADOS, Medical Parachute). Automated immunohistochemistry was performed using anti-SMA to detect EMT and anti-FN to document ECM remodeling. Slides were digitized and analyzed using the Positive Pixel Count v9 algorithm. Linear regression and Poisson regression were performed (P < 0.05). Results: SMA positive expression decreased as the time of IOL implantation increased (P < 0.0001). Positivity of SMA and FN demonstrated a positive correlation (P = 0.0002). Controlling for confounding factors in Poisson regression, hydrophobic and hydrophilic materials showed higher FN and SMA expression when compared to silicone material lenses (FN; P = 0.018; P < 0.0001, SMA; P = 0.001; P = 0.003, respectively). The square optic design had 29% higher SMA positivity compared to the opti-edge design (P = 0.042). One-piece haptic lenses had higher SMA expression compared to three-piece haptic (P = 0.042). A higher risk of expression of SMA and FN was seen in patients with a history of smoking, hypertension, and glaucoma (P < 0.05). Conclusion: This study demonstrated that SMA and FN expression is different according to IOL design and patient factors, thus indicating that LEC changes depend on lens biocompatibility. Therefore, by analyzing the histopathological composition of PCO by using LECs, further insight into the characteristics of IOLs that are important for biocompatibility can be ascertained.

Acetylsalicylic Acid Exerts Potent Antitumor and Antiangiogenic Effects in Cutaneous and Uveal Melanoma Cell Lines.

INTRODUCTION: Acetylsalicylic acid (ASA) has been investigated for a potential anticancer role in several cancers, such as colorectal, ovarian, and endometrial cancer. Moreover, ASA has been shown to abrogate various processes that contribute to tumor growth and progression. OBJECTIVE: The aim of this study was to evaluate the effects of ASA on cutaneous melanoma (CM) and uveal melanoma (UM). METHODS: Human CM and UM cells were treated with 5 mM ASA and assessed for changes in cellular functions. Antiangiogenic effects of ASA were determined using an ELISA-based assay for 10 proangiogenic cytokines, and then validated by Western blot. Finally, proteomic analysis of ASA-treated cells was performed to elucidate the changes that may be responsible for ASA-mediated effects in melanoma cells. RESULTS: Treatment with ASA significantly inhibited the proliferation, invasion, and migration capabilities, and caused a significant decrease in angiogenin and PIGF secretion in both CM and UM. Mass spectrometry revealed 179 protein changes associated with ASA in the CM and UM cell lines. CONCLUSIONS: These results suggest that ASA may be effective as an adjuvant therapy in metastatic CM and UM. Future studies are needed to determine the regulating targets that are responsible for the antitumor effects of ASA.

Freeze-drying as an alternative method of human sclera preservation.

PURPOSE: To compare the effect of preserving sclera samples in either 95% ethanol or freeze-dried. METHODS: Ninety-six samples of human sclera were studied. Half of them were freeze-dried and half preserved in 95% ethanol. Preservation periods of 18, 45, 90 or 174 days were studied. Automated immunostaining was carried out in the Ventana BenchMarkR LT platform using collagen 1 and fibronectin antibodies. Histological staining was also performed with hematoxilin-eosin and Masson trichrome. Samples were classified according to the degree of collagen fiber parallelism (0-2), intensity of Masson staining (0-2), and the expression of both antibodies (0-3). Friedman and Wilcoxon tests were applied to compare preservation methods and p-values below 0.05 were considered to ensure statistical significance. RESULTS: Relevant results were found in three situations: (i) Friedman's test showed better collagen fiber integrity in the freeze-dried group rehydrated after 174-days as compared to the 90-day group; (ii) Wilcoxon's test showed better collagen fiber integrity in the freeze-dried group after 18 and 174 days as compared to the ethanol group; (iii) the freeze-dried group disclosed higher immunohistochemical expression for COL-1 antibody in the sclera samples rehydrated after 45, 90 and 174 days as compared to the ones rehydrated after 18 days. CONCLUSION: Histological and immunohistochemical analysis showed freeze-drying to be a superior method for sclera preservation as compared to 95% ethanol. This technique provides an easy method to manipulate tissue, with longer shelf life, and storage at room temperature.

Freeze-drying as an alternative method of human sclera preservation / Liofilização como alternativa à preservação de esclera humana

PURPOSE: To compare the effect of preserving sclera samples in either 95 percent ethanol or freeze-dried. METHODS: Ninety-six samples of human sclera were studied. Half of them were freeze-dried and half preserved in 95 percent ethanol. Preservation periods of 18, 45, 90 or 174 days were studied. Automated immunostaining was carried out in the Ventana BenchMarkR LT platform using collagen 1 and fibronectin antibodies. Histological staining was also performed with hematoxilin-eosin and Masson trichrome. Samples were classified according to the degree of collagen fiber parallelism (0-2), intensity of Masson staining (0-2), and the expression of both antibodies (0-3). Friedman and Wilcoxon tests were applied to compare preservation methods and p-values below 0.05 were considered to ensure statistical significance. RESULTS: Relevant results were found in three situations: (i) Friedman's test showed better collagen fiber integrity in the freeze-dried group rehydrated after 174-days as compared to the 90-day group; (ii) Wilcoxon's test showed better collagen fiber integrity in the freeze-dried group after 18 and 174 days as compared to the ethanol group; (iii) the freeze-dried group disclosed higher immunohistochemical expression for COL-1 antibody in the sclera samples rehydrated after 45, 90 and 174 days as compared to the ones rehydrated after 18 days. CONCLUSION: Histological and immunohistochemical analysis showed freeze-drying to be a superior method for sclera preservation as compared to 95 percent ethanol. This technique provides an easy method to manipulate tissue, with longer shelf life, and storage at room temperature.

OBJETIVO: Comparar dois métodos de preservação de esclera humana, liofilização e álcool 95 por cento, em diferentes períodos de tempo. MÉTODOS: Foram avaliados 96 fragmentos de seis escleras humanas. Metade das amostras foi submetida ao processo de liofilização e metade conservada em álcool 95 por cento. Dois fragmentos de cada grupo foram avaliados pelas colorações de hematoxilina-eosina e tricrômio de Masson e submetidos a técnica de imuno-histoquímica para os anticorpos fibronectina e colágeno 1, após 18, 45, 90 e 174 dias de preservação. Os espécimens foram classificados de acordo com o paralelismo (PF:0-2) e integridade (IF:0-1) das fibras de colágeno e expressão imuno-histoquímica para os anticorpos fibronectina (FIB:0-3) e colágeno 1 (COL-1:0-3). A análise estatística foi realizada por meio dos testes de Friedman e Wilcoxon e o valor de p menor que 0,05 foi considerado estatisticamente significante. RESULTADOS: Verificaram-se diferenças significantes em três situações: (i) maior integridade das fibras de colágeno das escleras liofilizadas após 174 dias quando comparado aos 90 dias; (ii) maior expressão imuno-histoquímica para o anticorpo COL-1 nas amostras de escleras liofilizadas após os 18 dias iniciais de preservação; (iii) maior integridade das fibras de colágeno das escleras liofilizadas após 18 e 174 dias quando comparado às escleras preservadas em álcool. CONCLUSÕES: A preservação de tecido escleral por liofilização mostrou-se técnica tão eficaz quanto a preservação em álcool, apresentado vantagem quando considerada a integridade das fibras de colágeno. A liofilização mostra-se benéfica por permitir a estocagem do tecido em temperatura ambiente e com prazo de validade estendido.

[Histopathologic findings in retinoblastoma]. / Achados histopatológicos em retinoblastoma.

OBJECTIVE: To study histopathological findings of enucleated eyes with retinoblastoma. METHODS: Twenty-eight cases of retinoblastoma treated by enucleation at the Federal University of São Paulo from December 2000 to October 2002 were histopathologically reviewed. Clinical data included age, gender, race, unilateral or bilateral involvement and previous treatment. The histopathological review evaluated the presence of iris and/or angle neovascularization, tumor differentiation and optic nerve and choroidal invasion according to Khelfaoui's classification. RESULTS: Of 27 patients, 13 (48.5%) were boys and 14 (59.3%) were girls, 16 were white, 6 were black and 5 were asiatic, age ranging from 2 to 96 months (mean, 22.7 months). 13 cases were bilateral and 14 cases were unilateral. All tumors were histologically characterized by a proliferation of small cells with high nuclear-to-cytoplasmic ratios and 20 (71.4%) of them were well differentiated. Choroidal involvement was observed in 18 (64.2%) cases (degree II, III) and optic nerve invasion in 8 (28.5%) cases (degree III, IV, V). CONCLUSION: Neovascularization, necrosis and calcification were the most commonly observed feature. The invasion into the optic nerve and choroid, which are the two most important predictors of patient outcome were found in 28.5% and 64.2% of the cases, respectively.

Achados histopatológicos em retinoblastoma / Histopathologic findings in retinoblastoma

OBJETIVO: Descrever e analisar os principais achados histopatológicos no retinoblastoma com ênfase na invasão de nervo óptico e túnicas oculares, correlacionando-os ao tratamento realizado. MÉTODOS: Vinte e oito olhos com diagnóstico de retinoblastoma tratados por enucleação na Universidade Federal de São Paulo entre dezembro de 2000 e outubro de 2002 foram submetidos à revisão histopatológica. Os dados clínicos incluiram idade, sexo, raça, lateralidade e tratamento realizado. Na revisão histopatológica foram avaliados neovascularização da íris e seio camerular, grau de diferenciação do tumor e invasão do nervo óptico e coróide de acordo com a classificação de Khelfaoui. RESULTADOS: De 27 pacientes, 13 (48,1 por cento) eram do sexo masculino e 14 (51,9 por cento) eram do sexo feminino, 16 (59,3 por cento) eram da raça branca, 5 (18,5 por cento) eram da raça amarela, 4 (14,8 por cento) eram pardos e 2 (7,4 por cento) eram negros. A média da idade dos pacientes foi de 22,7 meses, sendo 13 casos bilaterais e 14 casos unilaterais. Com relação ao tratamento utilizado, 13 (46,4 por cento) olhos foram submetidos a enucleação primária e 15 (53,6 por cento) tiveram tratamento prévio. Na avaliação histopatológica, 15 (53,6 por cento) tumores eram bem diferenciados. Neovascularização da íris e/ou seio camerular foram observados em 60,6 por cento e necrose e calcificação em 89,2 por cento dos casos. O envolvimento da coróide foi observado em 18 (64,2 por cento) dos casos (grau II e III) e a invasão de nervo óptico em 8 (28,5 por cento) (graus II, III e IV). Dos casos submetidos ao tratamento prévio 40 por cento tinham algum grau de invasão das túnicas oculares e apenas 6,67 por cento apresentavam simultaneamente invasão do nervo óptico e das túnicas oculares. Entretanto nos olhos que foram submetidos à enucleação primária 30,7 por cento apresentavam invasão das túnicas oculares e 53,8 por cento apresentavam simultaneamente invasão do nervo óptico e das túnicas oculares. CONCLUSAO: Neovascularização, necrose e calcificação foram achados mais comumente observados. A invasão do nervo óptico e coróide foram os dois fatores prognósticos mais importantes, encontrados em 28,5 por cento e 64,2 por cento dos casos respectivamente. Os casos tratados com enucleação primária apresentavam maior comprometimento ocular.

A evolução do mercado farmacêutico brasileiro no tratamento do glaucoma nos últimos 30 anos / Evolution of the Brazilian pharmaceutical market regarding glaucoma treatment during the last 30 years

OBJETIVO: Analisar as variações de custo das drogas para tratamento do glaucoma no mercado farmacêutico, nos últimos 30 anos, em intervalos de 10 anos. MÉTODOS: A partir dos Dicionários de Especialidades Farmacêuticas, dos anos de 1972, 1982, 1992 e 2002 nos quais foram anotadas as drogas utilizadas no tratamento do glaucoma, suas apresentações e laboratórios. Foram analisados os valores dos medicamentos no mercado brasileiro, nos diferentes anos, por meio do Guia Farmacêutico Brasíndice. Para análise dos dados, os preços foram convertidos para o dólar americano na época e estudada a relação percentual entre o preço dos medicamentos e o salário mínimo vigente na época, considerando-se como unidade de referência do produto os valores por mililitro, comprimido ou grama, dependendo da apresentação do fármaco. RESULTADOS: O número de laboratórios e medicações antiglaucomatosas variaram nestes últimos trinta anos. Drogas como a pilocarpina e acetazolamida continuam sendo utilizadas. Há predominância do uso de mióticos nos primeiros anos do estudo, e de beta-bloqueadores nos últimos anos. O custo médio das medicações disponíveis no mercado aumentou principalmente na última década. CONCLUSAO: O preço dos medicamentos antiglaucomatosos variou de forma considerável no período estudado, bem como o tipo de medicação usada no tratamento.