Vasco Bravo-Filho

Sebaceous adenomas of the eyelid and Muir-Torre Syndrome

Background/aims Sebaceous adenomas (SAs) are rare, benign sebaceous gland tumours of the eyelid. SAs may be associated with primary internal malignancies. This association is known as Muir-Torre Syndrome (MTS). The purpose of this study was to approximate the prevalence of SAs, to determine the reliability of the clinical diagnosis of SAs and to demonstrate immunohistochemical staining of DNA mismatch repair proteins mutL homologue 1 (MLH1) and mutS homologue 2 (MSH2) for a case of MTS. Methods We reviewed the histopathology reports from all eyelid specimens collected between 1993 and 2013 at the Henry C Witelson Ocular Pathology Laboratory to determine the proportion of SAs. For the SAs identified on histopathology, we looked at patient charts to see what diagnosis was originally suspected on clinical examination. Immunohistochemical staining for MLH1 and MSH2 was performed on all SAs to screen for MTS. Results Of the 5884 eyelid specimens collected, 9 were SAs (6 women, 3 men; 42–72 years old). The diagnosis of SA was suspected clinically in only one of the nine cases based on the gross appearance of the eyelid lesion. Immunohistochemistry revealed one SA case with positive MLH1 expression and negative MSH2 expression. These findings prompted systemic work-up and this patient was diagnosed with MTS after discovery of a colon adenocarcinoma T2M0N0. Conclusions The diagnosis of eyelid SA is rare. The importance of this benign eyelid tumour stems from its association with internal malignancies in MTS. Immunohistochemical staining of mismatch repair proteins MLH1 and MSH2 is a valid and accessible strategy for investigating MTS in patients with SAs.

Superficial epithelioma with sebaceous differentiation involving the eyelid: a case report

IntroductionSuperficial epithelioma with sebaceous differentiation is a rare benign epithelial neoplasm. It usually involves the head, neck or the back of a middle-age person. To the best of our knowledge, two ocular cases have been reported in the literature.Case presentationA 46-year-old man of Italian descent, with a known history of testicular seminoma treated by orchiectomy with chemotherapy and radiotherapy, presented with a tan-colored lesion measuring 4mm in diameter in his right upper lid that had been growing over 10 months. It was clinically diagnosed as papilloma. An excisional biopsy was done. On histological examination, the lesion was a well-circumscribed and sharply demarcated epithelial tumor attached to the overlying epidermis and characterized by plate-like proliferation of basaloid to squamous cells with clusters of mature sebaceous cells and foci of ductal differentiation. After a follow-up period of 5 months, no recurrence of the lesion has been documented.ConclusionsSuperficial epithelioma with sebaceous differentiation is part of the differential diagnoses of eyelid lesions. Arguments in the literature about the correct nomenclature of superficial epithelioma with sebaceous differentiation have resulted in under-diagnosed cases. The benign histological features and the lack of recurrence support its benign nature. Although no clear association has linked superficial epithelioma with sebaceous differentiation with Muir–Torre syndrome, further clinical correlation and close follow up for patients are recommended.

High frequency of squamous intraepithelial neoplasia in pterygium related to low ultraviolet light exposure

Purpose The aim of this study was to determine the frequency of clinically unsuspected ocular surface squamous neoplasia (OSSN) in cases of biopsied pterygium (PT). Methods We reviewed 15,016 cases presented at the Henry C. Witelson Ocular Pathology Laboratory during the period 1993–2013. All cases with a clinical diagnosis of PT were included. Histopathological diagnoses were reviewed and demographic data were retrieved from histopathological request forms. All cases associated with OSSN were re-evaluated independently by two ocular pathologists. The classification of OSSN in PT cases was made based on the Armed Forces Institute of Pathology (AFIP) recommendations. Results Two hundred and fifteen cases were diagnosed clinically as PT (1.43%) and 54% were from male patients. The average age at diagnosis was 53.4 ± 15.5 years. OSSN was identified in five cases (2.33%), and four of these cases were from female patients (80%). The average age of patients with PT and OSSN was similar to PT patients without OSSN (P > 0.05). Cases with OSSN were diagnosed as conjunctival intraepithelial neoplasia (CIN) I (60%), CIN II (20%), and CIN III (20%). There was complete agreement between the two pathologists (100%). Conclusions The relatively high rate of dysplasia in a low ultraviolet light index area challenges the main cause of this disease in our population, a hypothesis that should be evaluated in future studies. We suggest that all PT samples should be sent for histopathological evaluation even in areas with low ultraviolet light index.

SIRT2 Expression Is Higher in Uveal Melanoma than In Ocular Melanocytes

Purpose: Sirtuins (SIRTs) are the family of proteins associated with the cell cycle and that correlate with cancer development and progression. SIRTs have never been studied in uveal melanocytes. The aim of this study is to characterize the expression of SIRT2 in uveal melanoma (UM) cases and compare it with the expression of SIRT2 in melanocytes of the uveal tract of normal human eyes (NHE). Methods: Twenty-one formalin-fixed, paraffin-embedded human UM cases were immunostained for SIRT2, along with 15 NHE obtained from the Eye Bank of Canada. Results: SIRT2 expression was higher in melanomas than in normal melanocytes of both tumor and donor eyes (p < 0.0001). No significant difference in SIRT2 expression was found when comparing normal melanocytes in UM and NHE cases. Conclusions: SIRT2 expression is significantly stronger in UM cells than in normal ocular melanocytes. This finding may indicate an important role of SIRT2 as a prognostic marker in UM progression. SIRT2 should also be investigated as a possible therapeutic target.

Expression of the Metastasis Suppressor KAI1 in Uveal Melanoma

Introduction. Uveal melanoma (UM) is an intraocular tumor that leads to metastatic disease in approximately 50% of afflicted patients. There is no efficacious treatment for metastatic disease in this cancer. Identification of markers that can offer prognostic and therapeutic value is a major focus in this field at present. KAI1 is a metastasis suppressor gene that has been reported to play a role in various human malignancies, although it has not previously been evaluated in UM. Purpose. To investigate the expression of KAI1 in UM and its potential value as a prognostic marker. Materials and Methods. 18 cases of human primary UM were collected and immunostained for KAI1 expression. A pathologist evaluated staining intensity and distribution semiquantitatively. Each case was categorized as group 1 (low staining) or group 2 (high staining). Results. In group 2, two of the 12 cases presented with metastasis. Conversely, in group 1, five out of 6 cases had metastasis. The mean follow-up of patients who did not develop metastasis was 81.81 months (median: 75 months) versus 42.14 months (median: 44 months) for patients with metastasis. Conclusions. KAI1 is a promising candidate marker that may offer prognostic value in UM; it may also represent a therapeutic target in metastatic disease.

Pretreatment of RPE Cells with Lutein Can Mitigate Bevacizumab-Induced Increases in Angiogenin and bFGF

Purpose: To determine whether pretreatment of retinal pigmented epithelial (RPE) cells with lutein can affect the response of cells to bevacizumab therapy. Methods: One human RPE cell line (ARPE-19) was used for all experiments. The cells were treated with lutein in different concentrations (0.01, 0.1, 1, 10, or 100 μg/ml). After 24 h, all plates were treated with bevacizumab (0.25 mg/ml). Media were harvested 24 h later for sandwich ELISA-based angiogenesis arrays. A Quantibody Human Angiogenesis Array was used in order to quantify the secretion of the following 10 proangiogenic cytokines: angiogenin, ANG2, EGF, bFGF, HB-EGF, PDGF-BB, leptin, PIGF, HGF and VEGF. Results: Treatment with bevacizumab alone led to a significant decrease in VEGF, as well as a significant increase in angiogenin and bFGF. Pretreatment with 0.1 and 1.0 μg/ml of lutein led to significant decreases in both bFGF and angiogenin following treatment with bevacizumab compared to bevacizumab treatment alone. Lutein alone did not modify the secretion of proangiogenic cytokines. Conclusions: Pretreatment of human RPE cells in culture with specific doses of lutein prior to bevacizumab treatment mitigated the increase in bFGF and angiogenin caused by bevacizumab monotherapy.

Pericyte Status in Routinely Discarded Vitrectomy Samples May Be an Early Marker of Diabetic Retinopathy

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; Spearmans 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.

Usefulness of Discarded Vitreous Samples from Routine Vitrectomy

Purpose. To describe the histopathological features of vitreous samples obtained after vitrectomy surgery from diabetic and nondiabetic patients. Methods. Vitreous specimens from 137 patients who underwent vitrectomy for different clinical conditions were analysed. All samples were centrifuged and each resulting pellet was fixed and processed as part of routine paraffin section histopathology. The histopathological features were categorized in a semiquantitative fashion. The samples from diabetic and nondiabetic patients were compared. Results. The 125 included patients (58 diabetic, 60% males) were aged 64.2 ± 13.9 years. The presence of hemorrhage, inflammatory cells, and histiocytes was significantly higher in the diabetic group (P < 0.001, P = 0.028, and P = 0.016, resp.), showing more vessels (P < 0.001) and ghost vessels (P = 0.049). The presence of inflammatory cells was the feature with the highest sensitivity for detecting diabetes mellitus (98%) and also the highest negative predictive value (89%). In the multivariate analysis, three variables emerged as independent significant predictors of diabetes in vitrectomy samples: hemorrhage, endothelial-lined vessels, and age (P < 0.001, P < 0.001, and P = 0.019, resp.). Conclusions. Different histopathological features can be found in vitreous samples from diabetic patients. Analysis of vitrectomy samples may serve as a tool for diabetes management.

Analysis of HSP90 Expression Is Valuable in the Differential Diagnosis of Ocular Surface Squamous Lesions

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.

Diagnosing Pathological Prognostic Factors in Retinoblastoma: Correlation between Traditional Microscopy and Digital Slides

Objective: It was the aim of this study to determine the diagnostic accuracy of high-risk prognostic factors and morphological characteristics of retinoblastomas using digital whole slide images (WSI) generated by a scanner. Methods: Forty-seven H&E sections of glass slides with high-risk morphological features of retinoblastoma were analyzed. Slides were scanned as WSI and reviewed. The results were compared with those obtained after reviewing the slides using a regular microscope as the gold standard. McNemars test (MT), the percentage of agreement (POA), and sensitivity (S) and specificity (Sp) were evaluated between WSI and conventional microscopy. Results: There were no differences with respect to multicentricity, growth type, rosette formation, choroidal invasion, anterior chamber invasion, extraocular extension, scleral extension, optic nerve invasion, necrosis, or Azzopardi effect between WSI analysis and light microscopy (MT, p = 1.0; POA = 100%; S = 100%, and Sp = 100%). Discordance was found in 1 case where calcification could not be found using WSI (MT, p = 1.00; POA = 97.9%; S = 100%, and Sp = 97.8%). Conclusion: To the best of our knowledge, this is the first report using digital pathology (WSI) to evaluate prognostic factors in eyes containing retinoblastomas. Using WSI, the pathologist was able to detect high-risk morphological features in retinoblastoma. To date, WSI is an important tool, in particular for ophthalmic pathologists examining enucleation and exenteration specimens.