ABSTRACT Purpose: To assess Meibomian gland dysfunction using meibography in patients with xeroderma pigmentosum and correlate with ocular surface changes. Methods: This cross-sectional study evaluated patients with xeroderma pigmentosum. All patients underwent a comprehensive and standardized interview. The best-corrected visual acuity of each eye was determined. Detailed ophthalmic examination was conducted, including biomicroscopy examination of the ocular surface, Schirmer test type I, and meibography, and fundus examination was also performed when possible. Meibomian gland dysfunction was assessed by non-contact meibography using Oculus Keratograph® 5M (OCULUS Inc., Arlington, WA, USA). Saliva samples were collected using the Oragene DNA Self-collection kit (DNA Genotek Inc., Ottawa, Canada), and DNA was extracted as recommended by the manufacturer. Factors associated with abnormal meiboscores were assessed using generalized estimating equation models. Results: A total of 42 participants were enrolled, and 27 patients underwent meibography. The meiboscore was abnormal in the upper eyelid in 8 (29.6%) patients and in the lower eyelid in 17 (62.9%). The likelihood of having abnormal meiboscores in the lower eyelid was 16.3 times greater than that in the upper eyelid. In the final multivariate model, age (p=0.001), mutation profile (p=0.006), and presence of ocular surface malignant tumor (OSMT) (p=0.014) remained significant for abnormal meiboscores. For a 1-year increase in age, the likelihood of abnormal meiboscores increased by 12%. Eyes with OSMT were 58.8 times more likely to have abnormal meiboscores than eyes without ocular surface malignant tumor. Conclusion: In the final model, age, xeroderma pigmentosum profile, previous cancer, and clinical alterations on the eyelid correlated with a meiboscore of ≥2. Meibomian gland dysfunction was common in patients with xeroderma pigmentosum, mainly in the lower eyelid. The severity of Meibomian gland dysfunction increases with age and is associated with severe eyelid changes. Purpose changes Methods crosssectional cross sectional interview bestcorrected best corrected determined conducted I possible noncontact non contact Keratograph M OCULUS Inc Inc. Arlington WA USA. USA . USA) Selfcollection Self collection Ottawa Canada, Canada , Canada) manufacturer models Results 4 enrolled 2 29.6% 296 29 6 (29.6% 1 62.9%. 629 62.9% 62 9 (62.9%) 163 16 3 16. model p=0.001, p0001 p p=0.001 0 001 (p=0.001) p=0.006, p0006 p=0.006 006 (p=0.006) (OSMT p=0.014 p0014 014 (p=0.014 1year year 12 12% 588 58 58. Conclusion cancer ≥2 29.6 (29.6 62.9 (62.9% p000 p=0.00 00 (p=0.001 (p=0.006 p=0.01 p001 01 (p=0.01 5 ≥ 29. (29. 62. (62.9 p00 p=0.0 (p=0.00 (p=0.0 (29 (62. p0 p=0. (p=0. (2 (62 p=0 (p=0 ( (6 p= (p= (p

ABSTRACT Objective: To evaluate the epidemiological, clinical, and imaging profile of lesions that mimic choroidal melanoma. Methods: Retrospective study of medical records of suspected choroidal melanoma lesions referred to the Ocular Oncology Service from the Universidade Federal de São Paulo, from 2014 to 2020. Demographic data, clinical history, and exams were evaluated. Results: A total of 104 patients (mean age: 65.57 ± 13.18; 49.04% female) were referred to our service with suspected choroidal melanoma. Of these, 32 (30.77%) were classified as pseudomelanoma, while 72 (69.23%) had a confirmed diagnosis of choroidal melanoma. Pseudomelanoma cases manifested in older individuals (p < 0.001), with smaller lesions in height (p < 0.001), anteroposterior diameter (p = 0.008), and lateral diameter (p = 0.003) on ultrasound. Pseudomelanoma cases were associated with higher frequencies of vitreous hemorrhage (p = 0.014) and lower rates of the presence of a mass (p = 0.001) and retinal detachment (p < 0.001). The main diagnoses of pseudomelanoma cases were choroidal nevus (40.63%), subretinal hemorrhage (18.75%) and choroidal neovascular membrane (18.75%). Conclusion: Almost one third of the cases referred with suspected choroidal melanoma were pseudomelanomas, which demonstrates that there is still a considerable path to improve the ability of general ophthalmologists to clinically discriminate melanoma from other conditions that can mimic it. Objective epidemiological Methods Paulo 201 2020 data history evaluated Results 10 mean age 6557 65 57 65.5 13.18 1318 13 18 4904 49 04 49.04 female these 3 30.77% 3077 30 77 (30.77% 7 69.23% 6923 69 23 (69.23% p 0.001, 0001 0.001 , 0 001 0.008, 0008 0.008 008 0.008) 0.003 0003 003 ultrasound 0.014 0014 014 0.001. . 40.63%, 4063 40.63% 40 63 (40.63%) 18.75% 1875 75 (18.75% 18.75%. Conclusion pseudomelanomas it 20 202 1 655 6 5 65. 13.1 131 490 4 49.0 30.77 307 (30.77 69.23 692 2 (69.23 000 0.00 00 0.01 01 406 40.63 (40.63% 18.75 187 (18.75 13. 49. 30.7 (30.7 69.2 (69.2 0.0 40.6 (40.63 18.7 (18.7 30. (30. 69. (69. 0. 40. (40.6 18. (18. (30 (69 (40. (18 (3 (6 (40 (1 ( (4

RESUMO Objetivo: Avaliar a frequência e o perfil epidemiológico, clínico e de imagem das lesões que simulam o melanoma de coroide. Métodos: Trata-se de estudo de revisão retrospectiva de prontuários de suspeita de lesões de melanoma de coroide de 2014 a 2020 no Setor de Oncologia Ocular da Universidade Federal de São Paulo. Foram avaliados dados demográficos, dados clínicos e exames complementares. Resultados: Um total de 104 pacientes (média de idade: 65,57 ± 13,18; 49,04% do sexo feminino) foram encaminhados ao nosso serviço com suspeita de melanoma de coroide. Destes, 32 (30,77%) foram classificados como pseudomelanoma, enquanto 72 (69,23%) tiveram diagnóstico confirmado de melanoma de coroide. Os casos de pseudomelanoma manifestaram-se em indivíduos mais velhos (p < 0,001) e apresentaram lesões menores em altura (p < 0,001), diâmetro anteroposterior (p = 0,008) e diâmetro lateral (p = 0,003) na ultrassonografia. Os casos de pseudomelanoma estão associados a maiores frequências de hemorragia vítrea (p = 0,014) e menores taxas de presença de massa (p = 0,001) e descolamento de retina (p < 0,001). Os principais diagnósticos dos casos de pseudomelanoma foram nevo (40,63%), hemorragia sub-retiniana (18,75%) e membrana neovascular coroidal (18,75%). Conclusão: Quase um terço dos casos encaminhados com suspeita de melanoma de coroide foram pseudomelanomas, o que demonstra que ainda há um caminho considerável para melhorar a habilidade do oftalmologista geral em discriminar clinicamente o melanoma de outras condições que o simulam. Objetivo epidemiológico Métodos Tratase Trata se 201 202 Paulo demográficos complementares Resultados 10 média idade 6557 65 57 65,5 13,18 1318 13 18 4904 49 04 49,04 feminino Destes 3 30,77% 3077 30 77 (30,77% 7 69,23% 6923 69 23 (69,23% manifestaramse manifestaram p 0,001 0001 0 001 0,001, , 0,008 0008 008 0,003 0003 003 ultrassonografia 0,014 0014 014 0,001. . 40,63%, 4063 40,63% 40 63 (40,63%) subretiniana sub retiniana 18,75% 1875 75 (18,75% 18,75%. Conclusão pseudomelanomas 20 1 655 6 5 65, 13,1 131 490 4 49,0 30,77 307 (30,77 69,23 692 2 (69,23 0,00 000 00 0,01 01 406 40,63 (40,63% 18,75 187 (18,75 13, 49, 30,7 (30,7 69,2 (69,2 0,0 40,6 (40,63 18,7 (18,7 30, (30, 69, (69, 0, 40, (40,6 18, (18, (30 (69 (40, (18 (3 (6 (40 (1 ( (4

ABSTRACT Purpose: To describe cellular alterations detected by impression cytology of the ocular surface in patients with xeroderma pigmentosum. The secondary objective was to assess the reliability of impression cytology in diagnosing ocular surface squamous neoplasia. Methods: Patients with xeroderma pigmentosum underwent a single-day complete ophthalmological examination and impression cytology for ocular surface evaluation using 13 mm diameter mixed cellulose esters membrane filters and combined staining with Periodic Acid Schiff, Hematoxylin and Eosin, and Papanicolaou stains followed by microscopic analysis. The cytological findings were correlated with the clinical diagnosis. The impression cytology findings at baseline and one-year follow-up were correlated with the clinical course (no tumor, treated tumor, residual tumor recurrent tumor, new tumor). Results: Of the 42 patients examined, impression cytology was performed in 62 eyes of 34 participants (65% females). The mean age of patients was 29.6 ± 17 years (range 7-62). Fifteen eyes had a clinical diagnosis of ocular surface squamous neoplasia. Impression cytology showed goblet cells (47, 75%), inflammatory cells (12, 19%), keratinization (5, 8%), and squamous metaplasia (30, 48%). Impression cytology was positive for atypical cells in 18 patients (12 with and 6 without ocular surface squamous neoplasia). The sensitivity, specificity, positive predictive value, and negative predictive value of impression cytology (at baseline) for diagnosis of ocular surface squamous neoplasia were 80%, 87%, 67%, and 93%, respectively, using clinical diagnosis of ocular surface squamous neoplasia as the reference standard. Conclusion: Impression cytology has a moderate positive predictive value for the diagnosis of ocular surface squamous neoplasia in patients with xeroderma pigmentosum. However, the lack of detection of atypical cells on impression cytology has a high negative predictive value for ocular surface squamous neoplasia. Integration of impression cytology in the long-term management of high-risk patients, such as patients with xeroderma pigmentosum, can avoid unnecessary diagnostic biopsies. Purpose Methods singleday single day 1 Schiff Eosin analysis oneyear one year followup follow up no tumor. . tumor) Results 4 examined 3 65% 65 (65 females. females females) 296 29 29. range 762. 762 7 7-62) 47, 47 (47 75%, 75 75% , 75%) 12, 12 19%, 19 19% 19%) 5, 5 (5 8%, 8 8% 8%) 30, 30 (30 48%. 48 48% 48%) (1 neoplasia) sensitivity specificity 80 80% 87 87% 67 67% 93 93% respectively standard Conclusion However longterm long term highrisk risk biopsies (6 2 76 7-62 (4 ( (3 9 7-6 7-

RESUMO Objetivo: Avaliar o impacto do isolamento social devido à pandemia COVID-19 sobre o número de casos novos e abordagens terapêuticas. Métodos: Estudo retrospectivo por revisões de prontuários de novos pacientes tratados antes da pandemia, de março de 2019 a setembro de 2019 (grupo pré-pandemia), e durante a pandemia, de março de 2020 a setembro de 2020 (grupo pandemia). Dados analisados incluíram idade, sexo, etnia, local de origem, diagnóstico clínico, tempo desde o encaminhamento e terapia proposta. Resultados: Um total de 186 novos casos foram analisados, sendo 122 do grupo pré-Pandemia e 64 do grupo Pandemia, representando uma redução de 47,54%. Não houve alteração estatisticamente signi ficativa quanto a sexo, raça, estado de origem, história do câncer, idade ou tempo de encaminhamento (p>0,05). Observou-se maior frequência de malignidades no grupo pandemia (68%) quando comparado com o grupo pré-pandemia (48,48%). Os tumores benignos foram os casos mais diagnosticados no grupo pré-pandemia (41,80%), enquanto no grupo pandemia o diagnóstico mais frequente foi o carcinoma de células escamosas de conjuntiva (31,25%). Houve tendência (p=0,097) de diminuição no número de cirurgias (-7,63%) e de aumento no tratamento tópico (+10,68%). Houve também uma tendência a diminuição de indicação cirúrgica em tumores benignos e diminuição dos retornos imediatos. Conclusão: Nossos achados mostram uma diminuição significativa no número de novos casos encaminhados ao setor de Oncologia Ocular. Além disso, a pandemia levou a uma mudança na abordagem terapêutica com preferência a tratamentos não invasivos, a fim de evitar o uso de salas de cirurgia. Um aumento drástico de casos, talvez em estágios avançados, pode ser esperado como resultado da diminuição observada nos primeiros 6 meses de quarentena. Objetivo COVID19 COVID 19 COVID-1 terapêuticas Métodos 201 prépandemia, prépandemia pré , pré-pandemia) 202 pandemia. . pandemia) sexo etnia origem clínico proposta Resultados 18 12 préPandemia Pandemia 4754 47 54 47,54% raça câncer p>0,05. p005 p p>0,05 0 05 (p>0,05) Observouse Observou se 68% 68 (68% 48,48%. 4848 48,48% 48 (48,48%) 41,80%, 4180 41,80% 41 80 (41,80%) 31,25%. 3125 31,25% 31 25 (31,25%) p=0,097 p0097 097 (p=0,097 7,63% 763 7 63 (-7,63% +10,68%. 1068 +10,68% 10 (+10,68%) imediatos Conclusão Ocular disso invasivos cirurgia avançados quarentena COVID1 1 COVID- 20 475 4 5 47,54 p00 p>0,0 (p>0,05 (68 484 48,48 (48,48% 418 41,80 8 (41,80% 312 31,25 3 2 (31,25% p=0,09 p009 09 (p=0,09 7,63 76 (-7,63 106 +10,68 (+10,68% 47,5 p0 p>0, (p>0,0 (6 48,4 (48,48 41,8 (41,80 31,2 (31,25 p=0,0 (p=0,0 7,6 (-7,6 +10,6 (+10,68 47, p>0 (p>0, ( 48, (48,4 41, (41,8 31, (31,2 p=0, (p=0, 7, (-7, +10, (+10,6 p> (p>0 (48, (41, (31, p=0 (p=0 (-7 +10 (+10, (p> (48 (41 (31 p= (p= (- +1 (+10 (p (4 (3 + (+1 (+

ABSTRACT Purpose: To evaluate the impact of social isolation due to the COVID-19 pandemic on the number of new cases and therapeutic approaches at the Ocular Oncology division from the Universidade Federal de São Paulo (UNIFESP). Methods: A retrospective study was conducted by medical records review of new patients treated before the pandemic from March 2019 to September 2019 (pre-pandemic group) and during the pandemic from March 2020 to September 2020 (pandemic group). Data regarding age, sex, ethnicity, place of origin, clinical diagnosis, time since referral, and proposed therapy were analyzed. Results: We analyzed 186 new cases, 122 from the pre-pandemic group and 64 from the pandemic group, representing a decrease of 47.54% in new cases. There was no statistically significant change in sex, race, state of origin, history of cancer, age, or time with suspected cancer (p>0.05). A higher frequency of malignancies was observed in the pandemic group (68%) when compared to the pre-pandemic group (48.48%). Benign tumors were the most common diagnosis in the pre-pandemic group (41.80%), while conjunctival squamous cell carcinoma was the modal diagnosis in the pandemic group (31.25%). There was a decreasing trend (p=0.097) in the number of surgeries (-7.63%) and an increase in topical treatment (+10.68%). There was also a tendency to perform fewer surgeries in benign tumors and decreased follow-up visits. Conclusion: Our findings showed a significant decrease in the number of new cases referred to the Ocular Oncology service. Moreover, the pandemic led to a switch in the therapeutic approach with preference to non-invasive treatments that would demand operating rooms. A drastic increase of cases perhaps in advanced stages might be expected because of the decrease observed in the first six months of quarantine. Purpose COVID19 COVID 19 COVID-1 UNIFESP. UNIFESP . (UNIFESP) Methods 201 prepandemic pre 202 group. age sex ethnicity origin referral Results 18 12 6 4754 47 54 47.54 race p>0.05. p005 p p>0.05 0 05 (p>0.05) 68% 68 (68% 48.48%. 4848 48.48% 48 (48.48%) 41.80%, 4180 41.80% , 41 80 (41.80%) 31.25%. 3125 31.25% 31 25 (31.25%) p=0.097 p0097 097 (p=0.097 7.63% 763 7 63 (-7.63% +10.68%. 1068 +10.68% 10 (+10.68%) followup follow up visits Conclusion service Moreover noninvasive non invasive rooms quarantine COVID1 1 COVID- (UNIFESP 20 475 4 5 47.5 p00 p>0.0 (p>0.05 (68 484 48.48 (48.48% 418 41.80 8 (41.80% 312 31.25 3 2 (31.25% p=0.09 p009 09 (p=0.09 7.63 76 (-7.63 106 +10.68 (+10.68% 47. p0 p>0. (p>0.0 (6 48.4 (48.48 41.8 (41.80 31.2 (31.25 p=0.0 (p=0.0 7.6 (-7.6 +10.6 (+10.68 p>0 (p>0. ( 48. (48.4 41. (41.8 31. (31.2 p=0. (p=0. 7. (-7. +10. (+10.6 p> (p>0 (48. (41. (31. p=0 (p=0 (-7 +10 (+10. (p> (48 (41 (31 p= (p= (- +1 (+10 (p (4 (3 + (+1 (+

RESUMO Tumores sebáceos da conjuntiva e carúncula são condições raras, sendo mais encontrados nas pálpebras. O ade noma sebáceo é responsável por 1% das lesões carunculares e ainda menos frequente nas lesões da conjuntiva. Esse diagnóstico é de suma importância pois quase 50% desses pacientes podem ser diagnosticados com a Síndrome de Muir-Torre, uma condição autossômica dominante rara que é caracterizada pela presença de ao menos um tumor de pele sebáceo e uma neoplasia visceral (gastrointestinal, câncer geniturinário e de mama). O estudo tem como objetivo relatar 3 casos de adenoma sebáceo com diferentes apresentações, que foram submetidos a biópsia excisional e estudo imuno-histoquímico. O diagnóstico desses tumores deve levantar suspeitas e aconselhar a investigação clínica para descartar outras neoplasias, principalmente porque em até 41% dos casos esse pode ser o primeiro sinal da Síndrome de Muir-Torre.

ABSTRACT Sebaceous tumors of the conjunctiva and caruncle are rare conditions, accounting for 1% of caruncle lesions and even lower among conjunctival lesions. Almost 50% of cases are associated with Muir-Torre syndrome, a rare autosomal-dominant condition characterized by at least one sebaceous skin tumor and one visceral malignancy. We report 3 cases of sebaceous adenoma with different presentations that were submitted to excisional biopsy and immunohistochemical study. Diagnosis of these tumors should increase the level of suspicion and lead to clinical investigation to rule out neoplasms, particularly because in up to 41% of cases, these can be the first sign of the disease.

RESUMO Objetivo: Avaliar a segurança e o efeito de 12 meses de tratamento com fotocoagulação pelo pattern scanning laser para neoplasia escamosa da superfície ocular em um ambiente com poucos recursos e acesso extremamente limitado a um tratamento cirúrgico. Métodos: Pacientes adultos com diagnóstico de neoplasia escamosa de superfície ocular foram submetidos a exame oftalmológico completo. Após anestesia tópica e instilação de azul de toluidina 1%, a lesão foi tratada com laser por um tempo de duração que variou de 20 a 100 ms e potência de 600 a 1800 mW. Os pacientes foram examinados semanalmente durante o primeiro mês e foram retratados semanalmente das lesões restantes, conforme necessário. Os pacientes tiveram um seguimento mínimo de 12 meses. Resultados: Trinta e oito pacientes (38 olhos) foram incluídos no estudo. Todos os pacientes apresentaram neoplasia escamosa da superfície ocular clínica, confirmada por citologia de impressão. A idade dos pacientes variou entre 40 e 83 anos (média de 65.5 anos) e 28 deles eram do sexo masculino (74%). Os pacientes foram divididos em dois grupos: Grupo I (imunocompetente) e grupo II (imunossuprimido). No grupo I, 23 pacientes (74%) apresentaram resposta completa com o controle da lesão após o tratamento com laser. No grupo II, dois dos sete pacientes (28%) apresentaram resposta ao tratamento durante o acompanhamento. A média de aplicações de laser foi de 2,5 (1 a 6 aplicações). Os procedimentos foram bem tolerados. Conclusões: Os resultados a curto prazo da abordagem de fotocoagulação a laser para o tratamento das lesões conjuntivais de neoplasia escamosa de superfície ocular foram favoráveis, com uma taxa de sucesso de 74% observada em pacientes imunocompetentes. Essa nova estratégia é uma alternativa menos intensiva em recursos que pode demonstrar sua utilidade em ambientes com escassez de salas cirúrgicas e em casos recorrentes. Estudos com acompanhamentos mais longos e amostras maiores são necessários para confirmar nossos achados e avaliar a eficácia do tratamento a laser associado à quimioterapia tópica.

ABSTRACT Purpose: To evaluate the safety and 12-month effect of treatment with pattern scanning laser photocoagulation for ocular surface squamous neoplasia in a low-resource setting with extremely limited access to an operating room. Methods: Adult patients with a clinical diagnosis of ocular surface squamous neoplasia underwent a complete ophthalmologic examination. After topical anesthesia and instillation of toluidine blue 1%, the lesion was treated using pattern scanning photocoagulation for a duration time that varied from 20 to 100 ms and power from 600 to 1,800 mW. Patients were examined on a weekly basis for the first month and underwent weekly retreatment of the remaining lesions, as necessary. Patients had a minimum follow-up of 12 months. Results: Thirty-eight patients (38 eyes) were included. All patients had clinical ocular surface squamous neoplasia that was confirmed by impression cytology. The age of patients ranged from 40 to 83 years (average: 65.5 years) and 28 of them were males (74%). The patients were divided into two groups: group I (immunocompetent) and group II (immunosuppressed). In group I, 23 patients (74%) presented complete response with lesion control after laser treatment alone. In group II, two of seven patients (28%) showed treatment response during the follow-up. The average number of treatments was 2.5 (one to six laser treatments). Procedures were well tolerated. Conclusion: Short-term results of the laser photocoagulation approach for the treatment of ocular surface squamous neoplasia conjunctival lesions were favorable, with a 74% success rate observed in immunocompetent patients. This novel strategy is a less resource-intensive alternative that could demonstrate its usefulness in settings with shortages in operating rooms and in recurrent cases. Studies with longer follow-ups and larger sample sizes are warranted to confirm our findings and evaluate the effectiveness of laser treatment in association with topical chemotherapy.

RESUMO Objetivo: Avaliar os efeitos do ranibizumabe em associação com o amfenac nas células de melanoma uveal humano e explorar a capacidade desses compostos em sensibilizar as células de melanoma uveal à radioterapia. Métodos: Células de melanoma uveal humano do tipo 92.1 foram cultivadas e submetidas ao tratamento proposto (ranibizumabe, amfenac e a combinação de ambos). Ensaios de proliferação, migração e invasão com as células de melanoma uveal do tipo 92.1 foram avaliados após tratamento com ranibizumabe (125 mg/ml), amfenac (150 nM) e a combinação de ambos. Além disso, as taxas de proliferação foram avaliadas após tratamento com ranibizumabe e amfenac com subsequente exposição das células a diferentes doses de radiação (0 Gy, 4 Gy e 8 Gy). Resultados: Ensaio de proliferação: células tratadas com ranibizumabe e amfenac combinados apresentaram taxas de proliferação inferiores em comparação ao grupo controle (p=0,016), do que as tratadas apenas com ranibizumabe (p=0,033). Ensaio de migração: foi observada uma taxa de migração significativamente mais baixa nas células tratadas com amfenac do que no grupo controle (p=0,014) e do que nas tratadas com ranibizumabe (p=0,044). Ensaio de invasão: não houve diferenças significativas entre os grupos estudados. Exposição à irradiação: no grupo da dose de 4 Gy, não houve diferença significante entre os grupos. No grupo da dose de 8 Gy, o tratamento com ranibizumabe, afenac e sua combinação antes da aplicação da radiação de 8 Gy levou a uma redução acentuada nas taxas de proliferação (p=0,009, p=0,01 e p=0,034, respectivamente) em comparação aos grupos controle. Conclusão: A combinação de ranibizumabe e amfenac reduziu a taxa de proliferação das células de melanoma uveal; no entanto, apenas o amfenac diminuiu significativamente a migração celular. A radiossensibilidade das células de melanoma uveal do tipo 92.1 aumentou após a administração de ranibizumabe, amfenac e sua combinação. Mais investigações são necessárias para determinar se esta é uma estratégia de pré-tratamento viável para tornar grandes tumores passíveis de radioterapia.

ABSTRACT Purpose: To evaluate the effects of ranibizumab and amfenac in human uveal melanoma cell lines and to explore the ability of these compounds to sensitize uveal melanoma cells to radiation therapy. Methods: The 92.1 human uveal melanoma cell line was cultured and subjected to the proposed treatment (ranibizumab, amfenac, and a combination of both). Proliferation, migration, and invasion assays of the 92.1 uveal melanoma cell line were assessed after pretreatment with ranibizumab (125 mg/mL), amfenac (150 nM), or a combination of both. In addition, proliferation rates were assessed after treatment with ranibizumab and amfenac, and the cells were subsequently exposed to various radiation doses (0, 4, and 8 Gy). Results: Proliferation assay: cells treated with a combination of ranibizumab and amfenac had lower proliferation rates than controls (p=0.016) and than those treated with only ranibizumab (p=0.033). Migration assay: a significantly lower migration rate was observed in cells treated with amfenac than the control (p=0.014) and than those treated with ranibizumab (p=0.044). Invasion assay: there were no significant differences among the studied groups. Irradiation exposure: in the 4 Gy dose group, there were no significant differences among any groups. In the 8 Gy dose group, treatment with ranibizumab, amfenac, and their combination prior to application of the 8 Gy radiation led to a marked reduction in proliferation rates (p=0.009, p=0.01, and p=0.034, respectively) compared with controls. Conclusion: Combination of ranibizumab and amfenac reduced the proliferation rate of uveal melanoma cells; however, only amfenac monotherapy significantly decreased cell migration. The radiosensitivity of the 92.1 uveal melanoma cell line increased following the administration of ranibizumab, amfenac, and their combination. Further investigation is warranted to determine if this is a viable pretreatment strategy to render large tumors amenable to radiotherapy.

RESUMO Objetivos: Este estudo visa determinar a origem do retinoblastoma em um número de casos e correlacionar essos achados com fatores prognósticos e histopatológicos conhecidos. Métodos: Trinta e nove casos de retinoblastoma foram diagnosticados e analisados com imuno-histoquímica usando marcadores de anticorpos monoclonais contra as células de retina imaturas (SOX-2: SRY-box containing gene 2), contra as células da retina maturas (MAP2: microtubule -associated protein 2) e contra as células gliais maturas (GFAP: glial fibrillar acidic protein). Foram avaliadas características microscópicas dos casos (grau de diferenciação, presença de semeadura vítrea, invasão de coroide/esclera, nervo óptico e câmara anterior). Duas linhas celulares de retinoblastoma (WERI-1 e Y79) também foram testadas, utilizando os três marcadores. Resultados: A expressão de SOX-2 foi positiva em 97,4% dos casos de retinoblastoma, enquanto MAP2 foi positivo em 59% dos casos. GFAP foi apenas positivo no estroma (astrócitos reativos). Não houve correlação entre preditores histopatológicos e marcadores imunohistoquímicos avaliados. As linhagens celulares mostraram positividade para SOX-2 (90% em WERI-1 e 70% das células Y79). Ambas as linhagens celulares se mostraram fortemente positivas con MAP2 (90%), enquanto não houve expressão de GFAP em nenhuma das linhas celulares estudadas. Conclusões: A maioria das células de retinoblastoma desta série de casos expressa marcadores de células retinianas imaturas, além de marcadores de células maduras. As linhas celulares Y79 e WERI-1 apresentaram imunomarcação para ambos os marcadores neurais em percentagens semelhantes a dos casos avaliados. Portanto, estes resultados confirmam a origem neural do tumor em particular. Alem disso, a ausência de células positivas para GFAP no tumor descarta diferenciação de astrócitos em retinoblastoma.

ABSTRACT Purpose: The cellular origin of retinoblastoma is uncertain as constituent tumor cells heterogeneously express markers of both immature and mature retinal cells. An immunohistochemical analysis of cellular origin may yield valuable insights into disease progression and treatment options. This study aimed to determine the cellular origin of retinoblastoma in a large case series and correlate these findings with histopathological prognostic factors. Methods: Thirty-nine retinoblastoma cases were histopathologically diagnosed and analyzed by immunohistochemistry using monoclonal antibodies against the immature neural cell marker SRY-box containing gene 2 (SOX-2), the mature neuronal cell marker microtubule-associated protein 2 (MAP2), and the mature glial cell marker glial fibrillary acidic protein (GFAP). Histopathological features were also evaluated, including patterns of growth, differentiation, vitreous seeding, and choroidal/scleral, optic nerve, and anterior chamber invasion. Two retinoblastoma cell lines, WERI-1 and Y79, were studied by immunocytochemistry using the same antibodies. Results: Expression of SOX-2 was strong in 97.4% of retinoblastoma cases, while MAP-2 was expressed in 59% of cases. Immunostaining for GFAP was positive only in reactive stromal astrocytes interspersed amongst tumor cells and in peritumoral tissue. There was no correlation between histopathological prognostic factors and immunohistochemical markers. Retinoblastoma cell lines showed strong positivity for SOX2 (90% of WERI-1 cells and 70% of Y79 cells) and MAP2 (90% of cells in both lines). GFAP was completely negative in both cell lines. Conclusion: The majority of retinoblastomas and both RB cell lines expressed an immature neural and/or a mature neuronal cell marker, but not a glial marker. These results indicate a typical neuroblast or neuronal origin and eliminate astrocyte differentiation from neural stem cells as the source of retinoblastoma.

Mulher de 53 anos apresentou proptose lentamente progressiva no olho esquerdo. Tomografia computadorizada mostrou uma lesão na região temporal inferior da órbita esquerda, bem delimitada, arredondada, homogênea. O tumor era composto de células com citoplasma granular eosinofilico. Algumas das células possuíam grandes grânulos eosinofílicos circundados por um halo claro, conhecidos como corpos ovoides-pustulares de Milian or corpos de Bangle. O diagnóstico de tumor de células granulares foi estabelecido, confirmado pela imuno-histoquímica. Tumor de células granulares são neoplasias incomuns com predileção da região da cabeça e pescoço. O conhecimento das características histopatológicas típicas são cruciais para o correto diagnóstico.

A 53 year-old woman presented with a slowly progressive, painless proptosis OS. Computed tomography disclosed a round, homogeneous, well-delimited lesion in the inferior-temporal orbit. The tumor was composed of round cells with eosinophilic granular cytoplasm. Some of the cells had larger eosinophilic granules surrounded by a clear halo; known as pustulo-ovoid bodies of Milian or Bangle bodies. The diagnosis of a granular cell tumor was then established and confirmed by immunohistochemistry. Granular cell tumors are uncommon benign soft tissue neoplasms that have a predilection for the head and neck region. Awareness of the typical histopathological features is crucial for the correct diagnosis.