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OBJECTIVE: To evaluate the capsulorrhexis structure in postmortem eyes and determine factors associated with posterior capsular opacification (PCO). DESIGN: Experimental study. PARTICIPANTS: Postmortem pseudophakic human eyes (nâ¯=â¯420). METHODS: Postmortem eyes were obtained and examined. Photographs were taken of the eyes in Miyake-Apple view and of the extracted lens-capsule complexes. PCO and Soemmering's ring (SR) were quantified using automated detector opacification software as factors of intensity and area. Miyake-Apple views and ImageJ were used to assess capsulorrhexis diameter, area of anterior capsule-optic overlap, length of the shortest anterior capsular leaflet, and area and angle of capsulorrhexis-optic nonoverlap. Linear regression analysis and Welch's t test were used to determine the relationship of these factors with PCO and SR. All analyses were repeated in sample groups specific to the 5 most common intraocular lens models. RESULTS: Capsule-optic overlap was positively correlated with PCO (P < 0.0001) and SR (Pâ¯=â¯0.0016). Capsulorrhexis diameter was negatively correlated with PCO (P < 0.0001) and SR (Pâ¯=â¯0.014). Leaflet length was positively correlated with PCO (Pâ¯=â¯0.009). Area and angle of capsulorrhexis-optic nonoverlap were not correlated with PCO or SR. Slopes and coefficients of determination were relatively low for all significant results. CONCLUSIONS: The pathogenesis of PCO development after cataract surgery is multifactorial. This study shows that with modern operating technology, capsulorrhexis factors have at best a modest influence on PCO formation. Factors such as time from surgery to death and intraoperative techniques such as laser capsule polishing, posterior capsule vacuuming, and cortical cleanup are likely to play a more significant role.
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Uveal melanoma is the most common intraocular tumor in adults. Our group has previously developed a human uveal melanoma animal model; however, adverse effects caused by the immunosuppressive agent, cyclosporine A, prevented animals from surviving more than 12 weeks. In this study, we tested multiple cyclosporine A doses over an extended disease course up to 20 weeks, providing complete clinical imaging of intraocular tumors, histopathological analysis and liquid biopsy biomarker analysis. Twenty albino rabbits were divided into four groups with different daily cyclosporine A schedules (0-10â mg/kg) and inoculated with human uveal melanoma cell lines, 92.1 or MP41, into the suprachoroidal space. Rabbits were monitored with fundoscopy, ultrasound and optical coherence tomography. Intraocular tumors (macroscopic or microscopic) were detected in all study animals. Tumor size and growth were correlated to cyclosporine A dose, with tumors regressing when cyclosporine A was arrested. All tumors expressed HMB-45 and MelanA; however, tumor size, pigmentation and cell morphology differed in 92.1 vs. MP41 tumors. Finally, across all groups, circulating tumor DNA from plasma and aqueous humor was detected earlier than tumor detection by imaging and correlated to tumor growth. In conclusion, using three clinically relevant imaging modalities (fundoscopy, ultrasonography and optical coherence tomography) and liquid biopsy, we were successfully able to monitor tumor progression in our rabbit xenograft model of human uveal melanoma.
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Melanoma , Neoplasias Uveais , Animais , Neoplasias Uveais/patologia , Coelhos , Melanoma/patologia , Humanos , Biópsia Líquida/métodos , Modelos Animais de Doenças , Ensaios Antitumorais Modelo de Xenoenxerto , Linhagem Celular TumoralRESUMO
OBJECTIVE: This study aims to objectively measure the degree of zonular dehiscence in postmortem eyes and to assess for clinical and anatomic correlates. DESIGN: Cross-sectional study. MATERIALS: Four hundred and twenty-seven postmortem pseudophakic human eyes. METHODS: Eyes were obtained from the Lions Gift of Sight Eye Bank. Microscope photographs were taken of the eyes in Miyake-Apple view, and region-of-interest analysis was performed using ImageJ to measure the area, circumference, and diameter of the capsular bag, the ciliary ring, and the capsulorhexis. Clinical and anatomic parameters were assessed using simple linear regression analysis and one-way analysis of variance with post hoc Bonferroni testing. Zonular dehiscence was measured via 2 surrogates: capsule area over ciliary ring area ratio (CCR) and capsule-ciliary ring decentration (CCD). Low CCR and high CCD indicate more zonular dehiscence. RESULTS: CCR was significantly inversely correlated with smaller capsulorhexi (pâ¯=â¯0.012), lower intraocular lens power (p < 0.00001), younger age at death (pâ¯=â¯0.00002), and a longer cataract-to-death time (pâ¯=â¯0.00786). CCR also was significantly lower in glaucomatous cases (pâ¯=â¯0.0291). CCD was significantly correlated with longer cataract-to-death time (pâ¯=â¯0.000864), larger ciliary ring area (pâ¯=â¯0.001), more posterior capsule opacification (pâ¯=â¯0.0234), and higher Soemmering's ring opacity (pâ¯=â¯0.0003). There was also significantly more decentration in male eyes than in female eyes (pâ¯=â¯0.00852). CONCLUSIONS: CCR and CCD are novel measures of zonular dehiscence in postmortem eyes, with many interesting correlates. An enlarged ciliary ring area is possibly associated with zonular dehiscence in pseudophakic eyes and may be a quantifiable surrogate in vivo.
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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.
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Opacificação da Cápsula , Cristalino , Lentes Intraoculares , Opacificação da Cápsula/diagnóstico , Opacificação da Cápsula/etiologia , Células Epiteliais/metabolismo , Humanos , Cristalino/patologia , Lentes Intraoculares/efeitos adversos , SoftwareRESUMO
Cytokinesis, the process that partitions the mother cell into two daughter cells, requires the assembly and constriction of an equatorial actomyosin network. Different types of non-motor F-actin crosslinkers localize to the network, but their functional contribution remains poorly understood. Here, we describe a synergy between the small rigid crosslinker plastin and the large flexible crosslinker spectrin in the C. elegans one-cell embryo. In contrast to single inhibitions, co-inhibition of plastin and the ßH-spectrin (SMA-1) results in cytokinesis failure due to progressive disorganization and eventual collapse of the equatorial actomyosin network. Cortical localization dynamics of non-muscle myosin II in co-inhibited embryos mimic those observed after drug-induced F-actin depolymerization, suggesting that the combined action of plastin and spectrin stabilizes F-actin in the contractile ring. An in silico model predicts that spectrin is more efficient than plastin at stabilizing the ring and that ring formation is relatively insensitive to ßH-spectrin length, which is confirmed in vivo with a sma-1 mutant that lacks 11 of its 29 spectrin repeats. Our findings provide the first evidence that spectrin contributes to cytokinesis and highlight the importance of crosslinker interplay for actomyosin network integrity.
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Actomiosina , Citocinese , Actinas/metabolismo , Actomiosina/metabolismo , Animais , Caenorhabditis elegans/metabolismo , Glicoproteínas de Membrana , Proteínas dos Microfilamentos , Espectrina/genéticaRESUMO
The sandy beaches of the Brazilian Amazon coast have enormous touristic potential and attract thousands of visitors each year, principally in July, at the beginning of the summer vacation period. In 2020, beach going was affected worldwide by the Covid-19 pandemic, but how did the pandemic affect recreational beach use on the Amazon coast in the Brazilian state of Pará? Given this, the objective of this work is to identify public perception in compliance with health safety established by authorities due to COVID-19 pandemic in two popular beaches on the Brazilian Amazon. Recreational patterns on the beaches of this region were analyzed from the user's perspective in July, through semi-structured interviews and direct observations. Recreational activities were prohibited on the beaches located within protected areas. At other locations, however, including Ajuruteua and Atalaia, the beaches were reopened on July 1 st, and both sites received thousands of visitors in July, despite the fact that interviewees believed the pandemic to be dangerous, and considered the situation in Pará state to be at least as bad or worse than in previous months. Agglomerations were verified on both study beaches and social distancing and other protective measures were limited. The increase in the new cases recorded in August was due to the relaxation of restrictions on social, recreational, and economic activities by the local authorities in July 2020, including the reopening of public access to beaches. As Pará state has been hard-hit by the pandemic, prohibitions on recreational beach use should clearly not have been lifted during this period. A number of management measures were presented in this study. These measures should contribute to the prevention of the spread of the virus during the upcoming public holidays, as long as the pandemic continues.
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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.
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BACKGROUND: Melanoma is a life-threatening group of cancers mainly affecting the skin (cutaneous melanoma, CM) and the eyes (uveal melanoma, UM). Nearly half of patients with UM develop liver metastases regardless of the primary treatment. For this reason, adjuvant therapy to prevent disease progression is essential to improve survival of patients with melanoma. Beta-adrenoceptors (ß-AR) have emerged as novel targets to inhibit tumor growth and dissemination in CM, but have not been investigated in UM. METHODS: The aim of this study was to comprehensively evaluate the effects of a non-selective ß-blocker in UM and CM. Propranolol was tested on four UM and two CM cell lines to determine the effects of this beta-blocker. The expression of ß-AR in UM was assessed in enucleated eyes of 36 patients. RESULTS: The results showed that propranolol exerts potent anti-proliferative effects, attenuates migration, reduces VEGF and induces cell cycle arrest and apoptosis in both UM and CM in a dose-dependent manner. Furthermore, levels of cell-free DNA released from the cells correlated to propranolol treatment and may be an indicator of treatment response. Finally, immunohistochemical analysis revealed the expression of ß1 and ß2 adrenoceptors in all UM patients, with higher expression seen in the more aggressive epithelioid versus less aggressive spindle cells. CONCLUSIONS: Collectively our data suggest that a nonselective beta-blocker may be effective against melanoma. For the first time, we show potent anti-tumor effects in UM cells following propranolol administration and expression of ß1 and ß2 adrenoceptors in patient tissue.
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Antagonistas Adrenérgicos beta/farmacologia , Melanoma/tratamento farmacológico , Propranolol/farmacologia , Neoplasias Cutâneas/tratamento farmacológico , Neoplasias Uveais/tratamento farmacológico , Antagonistas Adrenérgicos beta/uso terapêutico , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Melanoma/patologia , Melanoma/cirurgia , Cultura Primária de Células , Propranolol/uso terapêutico , Receptores Adrenérgicos beta 1/análise , Receptores Adrenérgicos beta 1/metabolismo , Receptores Adrenérgicos beta 2/análise , Receptores Adrenérgicos beta 2/metabolismo , Neoplasias Cutâneas/patologia , Úvea/patologia , Úvea/cirurgia , Neoplasias Uveais/patologia , Neoplasias Uveais/cirurgiaRESUMO
Programmed cell death-1/ligand (PD-1/PD-L1) interaction negatively regulates T cell activity. PD-L1 expression in tumor cells, antigen-presenting cells, and lymphocytes of the tumor microenvironment is associated with response to treatment with PD-1/PD-L1 inhibitors, but there is still debate on the cutoff value that correlates with responders. In uveal melanoma (UM), 40% of patients will develop liver metastases and, amongst them, 90% will succumb to their disease. The aim of this study was to analyze PD-L1 expression as a prognostic marker and as a possible therapeutic target for UM. Sixty-seven enucleated eyes from UM patients with relevant clinical information were analyzed. Univariate and multivariate analysis were used to evaluate association of PD-L1 with survival. PD-L1 expression was positive relatively to tumor cells, immune cells, and the tumor and tumor-infiltrating immune cell group scoring in 46, 34 and 55% of the cases, respectively. On univariate analysis, tumor cells and the tumor and tumor-infiltrating immune cell group PD-L1 expression was associated with a longer metastasis-free survival (P = 0.04 and P = 0.007). However, on multivariate analysis, only the tumor and tumor-infiltrating immune cell group positivity was associated with longer metastasis-free survival (P = 0.01). Furthermore, tumor cells and the tumor and tumor-infiltrating immune cell group PD-L1 expression was associated with decreased tumor-infiltrating lymphocytes (P = 0.02). PD-L1, when expressed in uveal melanoma, is associated with better patient outcome and decreased tumor-infiltrating lymphocytes. These results support the consideration of anti-PD-1/PD-L1 therapy in uveal melanoma. To determine the best cutoff value, further studies from patients enrolled in clinical trials treated with PD-1/PD-L1 inhibitors are necessary.
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Antígeno B7-H1/biossíntese , Biomarcadores Tumorais/análise , Linfócitos do Interstício Tumoral/imunologia , Melanoma/imunologia , Melanoma/patologia , Neoplasias Uveais/imunologia , Neoplasias Uveais/patologia , Adulto , Idoso , Intervalo Livre de Doença , Feminino , Humanos , Estimativa de Kaplan-Meier , Linfócitos do Interstício Tumoral/patologia , Masculino , Melanoma/mortalidade , Pessoa de Meia-Idade , Prognóstico , Neoplasias Uveais/mortalidadeRESUMO
PURPOSE: Superior limbic keratoconjunctivitis (SLK) is a chronic and recurrent condition of unknown etiology. It is often managed conservatively, but there is a high rate of success with surgical management for severe or recalcitrant cases. The purpose of this article is to describe and analyze clinicopathological features of patients with SLK who underwent surgical treatment and their association with the clinical outcome. METHODS: A total of 22 eyes from 18 patients who underwent surgical SLK management were retrospectively analyzed. Clinicopathological data were collected including details of follow-up and patient satisfaction (n = 15). Moreover, 12 cases had specimens available for review of histopathologic findings and COX-2 expression analysis by immunohistochemistry. RESULTS: From a clinical perspective, 66.7% of the SLK eyes had nonmechanical factors contributing to SLK, and 66.7% of eyes demonstrated significant symptomatic improvement after surgery. Histopathological analysis of all the lesions showed acanthosis and goblet cell loss. Unexpectedly, in 93% of the eyes, dilated lymphatic vessels were found. Furthermore, a high inflammatory infiltrate correlated with minimal symptomatic improvements (P = 0.013). Moreover, COX-2 expression was higher in patients with SLK than in a normal conjunctiva (P = 0.001). CONCLUSIONS: In this study, the most common systemic association with SLK was the patient's autoimmune status. Histopathological evaluation revealed that high inflammatory infiltration in the biopsy might be predictive of minimal symptomatic improvement with surgical management. Finally, the higher COX-2 expression in patients with SLK compared with that in individuals with a normal conjunctiva supports the use of anti-COX-2 drugs as a possible therapeutic target.
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Inflamação/patologia , Ceratoconjuntivite , Adulto , Idoso , Idoso de 80 Anos ou mais , Ciclo-Oxigenase 2/metabolismo , Feminino , Células Caliciformes/patologia , Humanos , Imuno-Histoquímica , Ceratoconjuntivite/metabolismo , Ceratoconjuntivite/patologia , Ceratoconjuntivite/cirurgia , Vasos Linfáticos/patologia , Masculino , Pessoa de Meia-Idade , Satisfação do Paciente , Estudos RetrospectivosRESUMO
BACKGROUND: Retinoblastoma (RB) is the most common primary intraocular malignancy. Current therapies are associated with high morbidity in the short- and long-term. Human epidermal growth factor receptor 2 (HER2) is a transmembrane protein detected in 15-30% of breast cancers, but it has also been described in other malignancies. Recently, it has been claimed that a truncated version of this protein is expressed in RB, responsive to directed therapies in vitro. We scored HER2 overexpression in RB tissue samples and discussed its potential clinical utility. METHODS: HER2 overexpression was investigated using immunohistochemistry; the overexpression was evaluated with a score ranging from 0 to 3+ according to the membranous staining pattern in archival formalin-fixed, paraffin-embedded RBs. RESULTS: A total of 60 RB cases and a RB cell line (Y79) were considered. The mean age at enucleation was 31.6 ± 31.5 months. The mean time from diagnosis to enucleation was 11.8 ± 11.2 months (range 1-44). Five (8%) cases were multifocal. HER2 overexpression was negative in all RB cases (49 cases scored 0 and 11 scored 1+) and in the Y79 cell line. CONCLUSIONS: Overall, we were not able to demonstrate the overexpression of HER2. Further studies should clarify and better elucidate the potential role of HER2-targeted therapies in RB.
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BACKGROUND: Diabetic retinopathy (DR) is the leading cause of blindness among the working-age population. The earliest morphological manifestation of the disease is pericyte loss, as shown by animal models. AIMS: The purpose of this study was to evaluate the presence of pericytes in vitreous samples (VS) from diabetic and nondiabetic patients. METHODS: VS from 125 patients with and without diabetes were analyzed. Thirty-three of the VS contained blood vessels and were therefore included in further analysis. Pericyte status was evaluated using α-smooth muscle actin and quantified using the following scoring system: total loss (3), >50% loss (2), <50% loss (1), and no loss (0). RESULTS: Of the 33 VS, 29 samples were from patients with diabetes and 4 from nondiabetic patients. Six diabetic cases had a score of 1, 8 diabetic cases had a score of 2, and 15 cases had a score of 3. A positive correlation between glycemia levels and pericyte loss was observed (p = 0.0016; Spearman's r = 0.61). Moreover, all nondiabetic cases had a score of 0 (sensitivity and specificity = 100%). CONCLUSION: Pericyte loss in VS might be a sensitive and specific marker of DR that correlates with glycemia levels. Furthermore, VS, which are currently discarded, may contain valuable information for diabetic management.
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Retinopatia Diabética/cirurgia , Diagnóstico Precoce , Pericitos/patologia , Vitrectomia/métodos , Corpo Vítreo/patologia , Adulto , Idoso , Retinopatia Diabética/patologia , Retinopatia Diabética/fisiopatologia , Progressão da Doença , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Prognóstico , Estudos Retrospectivos , Fatores de Tempo , Adulto JovemRESUMO
OBJECTIVES: The aim of this study was to evaluate heat shock protein 90 (HSP90) expression in squamous lesions (SLs) and to assess its diagnostic value for different lesions within the SL spectrum. METHODS: A total of 70 conjunctival SLs, including 19 papillomas, 22 cases of conjunctival intraepithelial neoplasia (ConINs) I, 11 cases of ConIN II, six cases of ConIN III, and 12 squamous carcinomas (sqCAs), were evaluated using the German immunoreactive score against HSP90. RESULTS: Cytoplasmic HSP90 expression differed between low- and high-grade lesions (P < .001). Among high-grade lesions, the nuclear HSP90 score was higher in the ConIN III-sqCA group than in the ConIN II group (P = .0162). A percentage of total thickness staining of less than 73% differentiated between ConIN III and sqCA. CONCLUSIONS: The expression of HSP90 is particularly useful to differentiate low-grade from high-grade lesions of the conjunctiva. HSP90 may play an important role in the malignant transformation of SLs and could be a new target for therapy.
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Biomarcadores Tumorais/metabolismo , Carcinoma in Situ/diagnóstico , Carcinoma de Células Escamosas/diagnóstico , Neoplasias da Túnica Conjuntiva/diagnóstico , Proteínas de Choque Térmico HSP90/metabolismo , Papiloma/diagnóstico , Adulto , Idoso , Idoso de 80 Anos ou mais , Carcinoma in Situ/classificação , Carcinoma de Células Escamosas/classificação , Estudos de Casos e Controles , Túnica Conjuntiva/patologia , Neoplasias da Túnica Conjuntiva/classificação , Citoplasma/metabolismo , Diagnóstico Diferencial , Feminino , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Gradação de Tumores , Papiloma/classificação , QuebequeRESUMO
OBJECTIVES: SOX-10 has been shown to be a sensitive marker of cutaneous melanoma. This study aimed to evaluate Sox-10 expression in uveal melanoma. METHODS: A total of 40 tissue blocks of enucleated eyes with uveal melanoma were cut and stained using an anti-SOX-10 mouse monoclonal antibody and HMB-45 antibody. RESULTS: SOX-10 showed exclusive nuclear positivity in 100% of the uveal melanoma cases (38/38). HMB-45 showed cytoplasmic positivity in 97.3 (37/38). Positivity for SOX-10 was also noted in the inner and outer nuclear layers of the retina in 78% of the enucleated eyes. CONCLUSIONS: SOX-10 expression proved to be the most sensitive marker for uveal melanoma, and therefore, we propose a modified panel for the diagnosis of uveal melanoma that includes both SOX-10 and HMB-45. The observation of distinct, diffuse nuclear SOX-10 expression in retinal inner and outer nuclear layers is a finding that warrants further investigation as a marker for retinoblastoma.
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PURPOSE: To evaluate the ability of novel dyes to stain lens capsule (LC), internal limiting membrane (ILM), epiretinal membrane (ERM), and vitreous. DESIGN: Experimental study in animal and human donor eyes. METHODS: Thirteen dyes, methyl violet, crystal violet, eosin Y, sudan black B, methylene blue, toluidine blue, light green, indigo carmine, fast green, congo red, evans blue, brilliant blue, and bromophenol blue, were injected onto the LC and ILM of enucleated porcine eyes. The vitreous was stained with 2 mL of dyes for 1 minute. Six dyes (indigo carmine, evans blue, fast green, light green, bromophenol blue, and brilliant blue) were selected for experiments in human donor eyes and freshly removed ERM. RESULTS: In the porcine eyes, ILM staining with methylene blue, toluidine blue, indigo carmine, evans blue, bromophenol blue, and fast green was moderate, and methyl violet, crystal violet, brilliant blue, or sudan black resulted in strong staining. Methyl violet, crystal violet, sudan black, toluidine blue, and methylene blue caused histologic damage in porcine retinas. Vitreous examination revealed moderate staining with congo red, crystal violet, fast green, eosin Y, methylene blue, toluidine blue, brilliant blue, bromophenol blue, and methyl violet and strong staining with light green and evans blue. ERMs showed strong staining with 0.5% evans blue and moderate staining with 0.5% light green, fast green, brilliant blue, and bromophenol blue. Evaluation of donor eyes disclosed moderate staining with evans blue, light green, and bromophenol blue and strong staining with 0.5% brilliant blue. Moderate or strong staining of the vitreous occurred with most dyes. LC evaluation showed moderate staining with 0.5% evans blue, fast green, and brilliant blue, whereas 0.5% light green produced strong LC staining. CONCLUSIONS: Brilliant blue shows the best ILM staining, whereas bromophenol blue, evans blue, and light green also stain ILM. Most dyes bind well to LC, vitreous, and ERM.
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Membrana Basal/anatomia & histologia , Corantes , Membrana Epirretiniana/diagnóstico , Cápsula do Cristalino/anatomia & histologia , Corpo Vítreo/anatomia & histologia , Animais , Membrana Basal/efeitos dos fármacos , Corantes/toxicidade , Membrana Epirretiniana/tratamento farmacológico , Humanos , Cápsula do Cristalino/efeitos dos fármacos , Coloração e Rotulagem/métodos , Suínos , Doadores de Tecidos , Corpo Vítreo/efeitos dos fármacosRESUMO
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.
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Anti-Infecciosos Locais/química , Etanol/química , Liofilização , Esclera , Anticorpos/imunologia , Colágeno Tipo I/imunologia , Colágeno Tipo I/metabolismo , Fibronectinas/imunologia , Liofilização/métodos , Liofilização/normas , Humanos , Imuno-Histoquímica , Estudos Prospectivos , Esclera/imunologia , Esclera/metabolismo , Coloração e Rotulagem , Estatísticas não Paramétricas , Fatores de TempoRESUMO
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.
Assuntos
Humanos , Anti-Infecciosos Locais/química , Etanol/química , Liofilização , Esclera , Anticorpos/imunologia , Colágeno Tipo I/imunologia , Colágeno Tipo I/metabolismo , Fibronectinas/imunologia , Liofilização/métodos , Liofilização/normas , Imuno-Histoquímica , Estudos Prospectivos , Coloração e Rotulagem , Estatísticas não Paramétricas , Esclera/imunologia , Esclera/metabolismo , Fatores de TempoRESUMO
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.
Assuntos
Corioide/patologia , Nervo Óptico/patologia , Neoplasias da Retina/patologia , Retinoblastoma/patologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , Terapia Combinada , Enucleação Ocular , Feminino , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Invasividade Neoplásica/patologia , Neovascularização Patológica/patologia , Prognóstico , Neoplasias da Retina/terapia , Retinoblastoma/terapiaRESUMO
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.
Assuntos
Humanos , Masculino , Feminino , Lactente , Pré-Escolar , Criança , Adolescente , Adulto , Pessoa de Meia-Idade , Idoso de 80 Anos ou mais , Corioide/patologia , Neoplasias da Retina/patologia , Nervo Óptico/patologia , Retinoblastoma/patologia , Terapia Combinada , Enucleação Ocular , Invasividade Neoplásica/patologia , Neoplasias da Retina/terapia , Neovascularização Patológica/patologia , Prognóstico , Retinoblastoma/terapiaRESUMO
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.