Purpose Compare the basic protein composition of the aqueous humor from retinoblastoma (RTB) patients with aqueous humor from patients with cataract who served as controls. Methods The study was carried out on 18 hospitalized RTB patients, who had undergone ocular enucleation (Reese-Ellsworth stage V or ABC classification group E RB), and 10 cataract patients. Five out of 18 RTB patients presented with associated secondary glaucoma and 5 out of 13 with no secondary glaucoma received chemotherapeutical treatment with Melphalan. The total protein concentration and electrophoresis pattern (SDS PAGE) of the aqueous humor was analysed. Results The aqueous humor from RTB patients had significantly higher total protein concentrations than controls (p<0.01); patients with secondary glaucoma presented with the highest total protein concentrations, and they were significantly different from controls (p<0.05), while those treated with Melphalan presented a 70% decreasing in protein concentration and, in respect to all non-treated patients, were significantly different (p<0.01). Besides, the protein content of controls was not significantly different from treated patients. The SDS PAGE pattern of RTB patients was very different from controls; yet, after chemotherapeutical treatment, patterns were similar. Conclusion This study represents a preliminary step towards a more accurate 2DE pattern, which will be combined with mass spectrometry analysis to clarify the potential role of specific proteins in tumor development and progression; although this results suggest that aqueous humor protein pattern in RTB is characteristic, several aspects of the study are still under investigation

Purpose Proteomics in ocular fluids is gaining rapidly relevance.Since 2009 we work on the analysis of proteic composition of the aqueous humor (AH) from retinoblastoma (RTB) patients and,in this work,we use the 2D‐DIGE coupled to mass spectrometer to find new RTB markers. Methods We enrolled 22 RTB patients(Reese‐Ellsworth stage V or ABC classification group E)undergoing enucleation and 20 normal subjects undergoing cataract surgery(CTR).Five of 22 patients presented with associated secondary glaucoma whereas 17 had no secondary glaucoma;8 of 17 patients with no secondary glaucoma received chemotherapeutical treatment with melphalan.Pools were analyzed using 2D‐DIGE coupled to mass spectrometer.DIA and BVA were fully automated with 2D DeCyder® and statistical analysis was performed with EDA® module.Significant spots were In‐Gel digested and identified with Maldi‐TOF/TOF. Results PCA analysis of the data underlines high distance between CTR pool and RTB pools.In particular the analysis of single protein shows that all the RTB pools are characterized by a high level of Crystallin and pro‐inflammatory proteins and CTR pool are characterized by a high level of Epithelium Growth Factor. Conclusion This study is the last step for the clarification of AH protein pattern in patients with RTB.Although we learn from literature that the presence of inflammatory processes is widely documented in late stages of pathology,however the presence of crystalline proteins could be either a new marker or an active part of the late stage tumor‐induced panorama in RTB AH samples.Due to this,we think that several molecular biology studies are necessary to better understand involvement of this protein family in RTB pathological pathway.

Purpose Iridociliary tumors are usually uncommon in childhood. Our aim is to describe cases of juvenile xanthogranuloma, iris tuberculoma and medulloepithelioma in young patients and to differentiate them from other similar lesions Methods Retrospective cases study. Four patients ( two girls and two boys) were referred at the Referral Center for Retinoblastoma of the University of Siena. They underwent all the ophthalmological examination including standard procedures, MRI of the orbits, CT, UBM, immunological and molecular analysis. In three cases organ‐preserving operations were performed. Results Two tumors were histologically identified as medulloepitheliomae. Two tumor‐like lesions were determined as juvenile xanthogranuloma and iris tuberculous granuloma. In the last case, antituberculous therapy was performed Conclusion Iridociliary tumors and pseudotumors are rather rare in childhood. Nevertheless, they should be taken in consideration in differential diagnosis with other pediatric intraocular tumors, particularly retinoblastoma, ciliary body adenoma and adenocarcinoma. An executive checkup including pathology, immunohistochemistry and immunology is to be performed.

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Purpose Proteomics can be a useful tool to understand how therapies act and affect on the molecular composition of ocular fluids. In this retrospective study we focus on how the aqueous humor (AH) proteic pattern from retinoblastoma (RTB) patients is modified by intra‐arterial(IA)treatment. Methods We enrolled 22 RTB patients(Reese stage VB or ABC group E)undergoing enucleation and 20 normal subjects undergoing cataract surgery(CTR).Five of 22 patients presented with associated secondary glaucoma (SG) whereas 17 had no secondary glaucoma (NSG);8 of 17 NSG received IA Melphalan (CHM). Pools were firstly investigated with Bradford method and SDS‐PAGE and then,analyzed using 2D‐DIGE coupled to mass spectrometer.DIA and BVA were fully automated with 2D DeCyder® and statistical analysis was performed with EDA® module.Significant spots were In‐Gel digested and identified with Maldi‐TOF/TOF. Results According to Bradford method,CTR protein content was not significantly different from CHM.After melphalan treatment, total protein contents decreased about 70% while in respect to all non‐treated patients were significantly different(Tamhane p<0.01).PCA analysis underlines high distance either between CTR pool and SG/NSG pool or CHM and RTB.In particular the analysis of single protein shows that CHM pool is characterized by a high level of Crystallin and pro‐inflammatory proteins and CTR pool is characterized by a high level of Epithelium Growth Factor. Conclusion IA chemotherapy with Melphalan modifies the AH proteic pattern revealing the presence of Crystallin family and pro‐inflammatory proteins. This opens new horizons to further explore local pharmacological effects of new drugs.