Search

Purpose: To investigate the presence of KIT gene mutations and immunoreactivity in 85 conjunctival melanocytic tumours and to clarify the role of KIT as a potential therapeutic target in this group of patients. Methods: Eighty-five conjunctival pigmented tumours [27 melanomas, 12 primary acquired melanosis (PAMs) and 46 nevi] were immunostained for KIT. Intensity and pattern of expression were evaluated. Molecular analysis to identify KIT mutations was performed in 15 selected cases (tumour-rich areas >50%). KIT immunostaining score and pattern were statistically related to patients’ age, sex, diagnostic category, presence of relapse, disease-free survival, presence of metastases, metastasis-free survival, limbal versus nonlimbal tumour location and thickness of melanomas. Results: KIT stains were documented in 48% of melanomas, 50% of PAMs and 24% of nevi. The mean score of KIT staining in the melanomas/PAMs group was significantly different from nevi (p = 0.0076). No statistically significant differences were detected between either c-kit immunostaining score or pattern and each of the other clinico-pathologic parameters considered. No KIT gene mutations were detected in melanomas and nevi. A silent mutation/polymorphism in KIT exon 13 was found in one PAM. Conclusions: Despite the high level of KIT immunostains in PAMs and melanomas, this parameter seems not to be a good predictor of the presence of molecular mutations. KIT-activating mutations should be considered an uncommon event in this tumour.

Purpose: To evaluate prospectively, on a long-term range, the clinical outcomes of transpupillary thermotherapy (TTT) as primary treatment of selected choroidal melanoma. Methods: Seventy-seven eyes of 77 patients affected by small posterior choroidal melanoma were treated with TTT as a sole treatment, using an infrared diode laser at 810 nm according to a standard procedure. Follow-up was longer than 36 months. Results: Seventeen tumours (22%) were parapapillary (PP) and 60 tumours (78%) were non-parapapillary (NPP) in location. Mean follow-up was 55.2 ± 17.9 months in PP tumours and 44.3 ± 23.7 months in NPP tumours. Thirteen (76%) PP tumours and 55 (92%) NPP tumours regressed (P > 0.05). Nine tumours recurred: seven were retreated using Iodine-125 brachytherapy and two were enucleated (both parapapillary). Four patients (5%) developed liver metastasis and died during follow-up. Tumour thickness was found to be predictive of recurrence (odds ratio: 4.3). Complications were found in 20 eyes (26%): macular pucker in 11 (14%), macular oedema in three (4%), retinal vein occlusion in six (8%), vitreous and subretinal haemorrhage in two (3%) and neovascular glaucoma in three (4%). PP tumours had more local complications (but this was not statistically significant; P > 0.05). Complications appeared more frequently in tumours treated with more than one TTT session (P = 0.01), and time-risk to develop intraocular complications seems longer in the PP group, without reaching statistical significance (P = 0.07). Conclusion: TTT may be a clinically effective method for conservative treatment of selected, non-parapapillary, small posterior choroidal melanoma.

Summary The pathophysiology of macular complications in diabetic retinopathy is multifactorial. Diabetic macular edema and ischemia (both macular and peripheral) are the major drivers of macular involvement in diabetes. Macular edema may be both intra and extra-cellular and it is clinically characterized by different aspects, mainly detected by an accurate qualitative analysis of spectral domain OCT data. This approach allows to identify different phenotypes of diabetic macular edema, providing a track to a personalized treatment. Macular ischemia has always been considered a negative prognostic factor in the treatment of macular edema, and this aspect has been recently re-confirmed. Peripheral retinal ischemia, easily documented by wide field retinal angiography, is emerging as a “new” driver of macular edema. All these new aspects, better defined by a multimodal imaging approach, are the essential to plan an adequate treatment.

Purpose To investigate NRAS and BRAF mutations and BRAF immunohistochemical expression in a group of melanocytic conjunctival tumors previously tested for KIT status, and to verify the correlation between KIT/BRAF mutational statuses with miRNAs involved in the RAS-RAF-MAPK pathway. Methods 85 non-consecutive cases of conjunctival melanocytic proliferation whit no remarkable KIT gene mutations were included. BRAF was analyzed for the V600E mutation, whereas NRAS molecular analysis was conduced for exons 1 and 2. Expression profiling of mature miRNAs was performed with the NCode miRNA qRT-PCR method. Results The nucleic acids amount was suitable for the analyses in 25 cases (29%). Of these, two melanomas (15%) presented a NRAS point mutation (Q61K) and other two melanomas (15%) and one nevus (10%) resulted V600E BRAF-mutated. The sensitivity and specificity of VE1 antibody in detecting BRAF V600E mutation were, respectively, 100% and 82%. Expression levels of miR-15b and miR-16 detected in BRAF mutated cases were higher than in wild type cases (p=0.05). MiR-222 was associated with tumor thickness (p=0.01) and tumor stage (p=0.02) whereas miR-15b was related to tumor stage (p=0.04). Let-7a, miR-15a/b and miR-16 were significantly down-regulated in melanomas and PAMs compared with nevi. Conclusion VE1 immunohistochemical staining is an accurate, rapid, and cost-effective method in selecting cases that should undergo molecular testing. Specific miRNAs have been identified as a potential diagnostic tool to differentiate between benign and malignant melanocytic lesions of the conjunctiva, and as prognostic markers among malignant ones.

Purpose To evaluate in vivo the presence of intraretinal hyperreflective spots in eyes with radiation induced maculopathy. Methods 25 subjects affected by maculopathy secondary to eye irradiation for a primary uveal melanoma were enrolled. All eyes previously underwent I-125 brachytherapy with a standard apex dose (85 Gy). Full ophthalmic examination, including spectral domain-OCT (SD-OCT) and fluorescein angiography, was performed in all eyes. After segmentation of retinal layers, retinal images were analyzed for reflectivity changes (hyperreflectivity spots) at the level of: internal limiting membrane plus retinal nerve fiber layer (ILM+RNFL), inner nuclear layer plus outer plexiform layer (INL+OPL) and outer nuclear layer (ONL). Hyperreflective spots were analyzed in a full retinal section both in the foveal area, and between 500 and 1500 µm from the foveal center. All examinations were performed twice, by two independent masked graders. Results The inter-grader agreement was at least substantial for all measurements. In all studied eyes hyperreflective spots were detected by SD-OCT. Hyperreflective spots were mainly located in the inner retina (ILM+RNFL and INL+OPL) compared to the outer retina (ONL) (p

Aim: The aim of this study is to evaluate long-term safety and efficacy of in-vivo transcleral Fine Needle Aspiration Biopsy (FNAB) for cytogenetic testing of posterior uveal melanoma (UM). Methods: One hundred thirty-nine consecutive patients affected by UM underwent in-vivo 25-G transcleral FNAB just before brachitherapy. Sampled material underwent both FISH (chromosome 3 and 6) and MLPA analysis using standard procedures. Follow-up was longer than 36 months. Results: The use of 25-G transcleral FNAB appears a long-term safe and effective procedure for in-vivo cytogenetic testing of posterior uveal melanoma. Combined analysis of both arms of uveal melanoma bifurcated pathway (-3 and +6p) increase predictive value of FISH technique. MLPA allows obtaining more information than standard FISH in UM prognostication (partial chromosomes deletion). Conclusions: The biological and prognostic value of partial chromosome 3 deletion, as well as others subtle chromosomes alterations or complex MLPA results, remains unclear.

Purpose To define the specific clinical characteristics of uveal metastases from carcinoids and the minimal invasive therapeutic approach. Methods Four consecutive cases of symptomatic uveal metastases from carcinoid tumors were included in this prospective analysis. Each patient underwent full ophthalmological examination, including A-B scan ultrasonography (US), fluorescein (FA) and indocyanine green angiography (IGA), and optical coherence tomography (OCT) at baseline and during follow-up. Follow-up examination was performed at 1 month and every 3 months thereafter. Results Mean follow-up was 17±6 months. Tree patients were affected by choroidal metastases (75%) whereas one patients by iris metastasis (25%). Choroidal metastases were ipo-pigmented, mainly located at the posterior pole and characterized by medium internal reflectivity at US, prominent vascular net at ICG and perilesional serous retinal detachment at OCT. Each patient was treated by photodynamic therapy (PDT) with verteporfine as sole focal treatment. Mean PDT sessions were 1.6 (1-3 sessions). Two patients (50%) showed clinical regression of the treated lesions, whereas in the others two patients (50%) tumor size remained stable during follow-up. Subretinal fluid resolved in all choroidal lesions. BCVA increased in three patients (75%) and remained unchanged in one patient (25%). Conclusion Uveal metastases from carcinoids are characteristically ipo-pigmented and highly vascularized. Photodynamic therapy appears to be an effective, safe and well-tolerated treatment option for the management these metastases.

Aim Macular edema is evaluated using pure morphologic imaging tests and visual acuity as sole visual function test. Unfortutanetly, visual acuity just partly reflects visual function. A new integrated morpho-functional approach seems useful. Methods Patients affected by macular edema were investigated using a multimodal/integrated imaging approach including currently available pure morphological retinal imaging techniques and microperimetry. Results The correlation between morphologic and functional imaging modalities strictly depends on the different involvement of retinal layers by the pathologic process. Data shows that visual function impairment is better quantified when microperimetry is added to the currently available diagnostic armamentarium. Conclusions Both in clinical trials and in daily clinical practice visual function needs to be better investigated to cope with patient’s requests. Adding reliable, topographically related visual functional data, currently obtained by microperimetry, to other pure retinal imaging investigations allows not only to better identify different “clinical phenotypes “ of macular disorders, but also to provide invaluable information about treatment and retreatment criteria.

Purpose To evaluate safety and efficacy of combined neo-adjuvant topical chemotherapy with 1%5-Fluoruracil (1%5-FU), followed by extensive surgery and adjuvant (post-surgical) 1%5-FU topical chemotherapy in the management of corneo-congiuntival squamous cell carcinoma (SCC). Methods Twenty-nine consecutive patients, with histological evidence of SCC, were included in this prospective study. Each patient was treated by combined neo-adjuvant topical chemotherapy (1%5FU, four time daily for four week, 1 cycle), followed by extensive surgery (5 mm of free conjunctival surgical margins) with contemporary amniotic membrane reconstruction. Adjuvant topical chemotherapy was started 3 months after surgery (1%5-FU, four time daily for four week, 1 cycle). Follow-up was performed every 3 months for the first year and every 6 months thereafter, including clinical confocal microscopy. Follow-up was longer than 36 months. Results Mean follow up was 56±11(range, 36-78 years). Good functional, anatomic and aesthetic results were obtained by surgery. Two patients (5%) needed short-term surgical amniotic membrane repositioning due to early sutures dehiscence. 1%5FU, used both as an adjuvant or neo-adjuvant treatment caused low-to-mild temporary local irritation. No-one patient discontinued therapy because of 5-FU side effects. No other long- or short-term side effects were documented. No local recurrences were documented during follow up. Conclusion Combined neo-adjuvant topical chemotherapy with 1%5FU, extensive surgery and adjuvant 1%5FU topical chemotherapy appear a safe and effective combined treatment in the management of conjunctival SCC.

Purpose to investigate the presence of circulating melanoma cells (CMCs) in patients affected by posterior uveal melanoma in different stages of the disease and to determine their prognostic relevance. Methods Blood samples from 14 healthy donors and 23 patients affected by posterior uveal melanoma were collected. Fourteen patients were included at the time of initial treatment without any evidence of metastatic disease using liver ultrasonography and Pet-CT (non-metastatic group). Nine patients were included at time of initial treatment of liver metastasis (metastatic group). mRNA expression of tyrosinase, MelanA/MART1 and GP100 as a surrogate marker for the presence of CMCs was analyzed by real-time RT-PCR and compared with patient characteristics. Results There was no significant difference on tyrosinase, MelanA/MART1 and GP100 levels between healthy donors and uveal melanoma patients (p>0.05). There was also no significant difference between non-metastatic vs metastatic group (p>0.05). High levels of tyrosinase, MelanA/MART1 and GP100 in non-metastatic patients were not related to the development of metastasis in a median follow-up time of 24 months (p>0.05). Conclusion PCR-based detection of tyrosinase, MelanA/MART1 and GP100 in uveal melanoma patients appear unrelated with presence/absence of the disease and with the disease stage.

Purpose To discuss the use of propranolol (and other β-blockers) in the treatment of pediatric orbital and periocular capillary hemangiomas, and perform literature review. Methods Two babies (3 and 5 months) affected by orbital capillary haemangioma with cutaneous periocular involvement were treated by oral propranolol (2 mg/kg/day) as a sole treatment. Complete baseline local and systemic examination revealed no contraindication to treatment. A strict follow-up protocol was performed to monitor tumor changes and any side effect. Follow-up was longer than 12 months. Results Both patients had significant improvement with resolution of the orbital component and slower resolution of the cutaneous side. No local or general (mainly hypotension, hypoglycaemia and asthma) side effects of propranolol were observed. Conclusion Propranolol appears to be a safe and effective option in the management of infantile capillary haemangioma. The role of other β-blockers, even topically applied, should be considered.

Purpose Purpose:To evaluate safety and efficacy of different treatments of choroidal neovascularization (CNV) associated with choroidal nevus at the posterior pole. Methods Methods: Six patients affected by choriodal nevus complicated by CNV were treated with photodynamic therapy with verteporfin (PDT; 50 J/cm2, 83 seconds) (five cases), or intravitreal anti-VEGF (bevacizumab, 1.25 mg) (one case). All patients underwent an ophthalmologic evaluation, including fluorescein and indocyanine green angiography, A-and B scan ultrasonography and OCT at presentation and at each follow-up examination. CNV was extrafoveal in all PDT treated cases, and a foveal serous detachment was detectable. CNV was subfoveal in the eye treated with anti-VEGF. Results Results: Mean follow-up was 15.5 months (range: 6 – 24). Visual acuity improved in four of the PDT treated cases and in the anti-VEGF treated eye, while it remained unchanged in the remaining PDT treated lesion. In all eyes resolution of the foveal serous detachment was detectable. In all PDT treated eyes a single treatment was performed with no recurrence during follow-up. Meanwhile anti-VEGF was repeated after three months for CNV recurrence. Conclusion Conclusions: PDT and anti-VEGF seem both effective in the treatment of CNV secondary to choroidal nevus but a larger study is required to evaluate long-term efficacy and safety expecially of anti-VEGF therapy.

Purpose To investigate the effects of different 5-Fluorouracil (5-FU) concentrations, exposure times and application techniques on in vitro cultured human corneal cells. Methods Human corneal epithelial cells (HCEC) and human corneal keratocytes (HCK) cultures were exposed to different 5-FU concentration (0.025% to 1%) and incubation times (single 5’ to 2 hrs). The cytostatic effect was evaluated as ratio of inhibition of migration towards controls. The cytotoxic effect evaluation included both contrast phase microscopic observations, and viability measures performed using a MTT[3-(4,5-dimethylthiazo-2-yl)-2,5-diphenyltetrazolium bromide; thiazolyl blue] colorimetric assay. The results were expressed as ratio of optical density (OD) reduction 24 hrs after exposures. Results Cytostatic effect is time and dose dependent. Minimal inhibiting dose (ID50) was 0,55% after 1 hour incubation for HCEC; ID50 was 0,5% after 2 hrs incubation for HCK . A 100% inhibitory effect was never observed at any concentration or incubation interval. No cytotoxic changes were observed at 5-FU < 1%; 5-FU 1% showed time-dependent cytotoxic changes just in HCEC cultures. MTT analysis showed no OD reduction at 5-FU concentration < 1%, whereas 1% 5-FU showed OD reduction < 50% at any tested exposure time. HCEC showed higher reduction in OD than HCK. Conclusion 5-FU shows no definite signs of irreversible toxicity to normal cultured corneal epithelial and keratocyte cells at examined concentrations and exposure time.

Purpose To evaluate safety and efficacy of photodynamic therapy (PDT) for the treatment of choroidal metastases. Methods Five cases of selected, symptomatic choroidal metastases were included in this prospective study. Each patient underwent a single-spot standard-fluence PDT with verteporfin (50 J/cm2, 83 sec.). BCVA, fundus photography and optical coherence tomography were performed at baseline and at each follow-up examination (at 1 month and every 3 months thereafter). Fluorescein angiography and indocyanine green angiography were performed at baseline. Results In two patients choroidal metastases were secondary to carcinoid tumors, in two cases to lung and in one case to breast adenocarcinoma. Mean follow-up was 17±6 months. Mean PDT sessions were 1.6 (1-3 sessions). Three patients (60%) showed clinical regression of the treated lesions and in two patients (40%) tumor size remained stable during follow-up. Subretinal fluid resolved in al cases. BCVA increased in four patients (80%) and remained unchanged in one patient (20%). Conclusion Photodynamic therapy appears as an effective and well-tolerated treatment option for the management of selected choroidal metastasis

Purpose To analyze cytogenetic profile of locally invasive posterior uveal melanoma (UM). Methods Twenty consecutive cases of large posterior UM with histopathologically confirmed extrascleral extension were included in this non-comparative cases series. Fine needle aspiration biopsy (FNAB) of the intraocular tumor portion was performed using 25-gauge trans-scleral approach, immediately after enucleation of the globe. FNAB of the extrascleral tumor portion was also performed when it was > 1mm in thickness. Sampled material underwent fluorescence in situ hybridization (FISH). Follow-up was longer than 12 months. Results Six tumors (30%) had both intraocular and extraocular tumor samplings, whereas 14 tumors (70%) showed extrascleral extension less than 1 mm in thickness. Monosomy 3 was found in 5 tumors (25%), whereas disomy 3 in 15 tumors (75%). Cytogenetic profile of the intraocular tumor portion appears to be maintained in the extrascleral extension in all cases (100%). Five patients (25%) developed metastatic disease during follow-up (all had monosomy 3 tumors). Conclusion Extrascleral extension appears more frequent in disomy 3 tumors. Cytogenetic profile of locally invasive posterior UM is maintained in the extrascleral tumor portion and must be considered the most important prognostic factor in locally invasive tumors.

Purpose To evaluate the role of micropulse diode laser trabeculoplasty (MDLT) in the treatment of secondary glaucoma due to intravitreal injection of corticosteroid. Methods Five consecutives patients previously treated with intravitreal triamcinolone (4mg) for refractory diabetic macular edema developed secondary glaucoma in the treated eye. Each eye underwent 360° micropulse diode laser trabeculoplasty (2000 mw power, 200 msec duration, 15% duty cicle, 300 µm diameter per spot). Patients were followed at 1, 7, 15, 28 days and monthly for 12 months. Results Mean age of the patients was 56 ± 2 years. Mean baseline intraocular pressure (IOP) was 30 ± 3 mmHg. IOP significantly decreased at 7 days post operative (19 ± 4 mmHg; p < 0.001) and maintained at normal level in 3 of 5 eyes. After 28 days two eyes needed retreatment and IOP maintained normal in one eye during follow up. No pain or side effect was documented. Conclusion Secondary glaucoma due to intravitreal injection of corticosteroid may be safely treated with micropulse diode laser trabeculoplasty.

Purpose Aim of this study was to analyze in-vivo structural and cellular features of ocular surface squamous (OSS) neoplasia using clinical confocal microscopy (CM). Methods Ten consecutive cases of OSS neoplasia were investigated using in-vivo corneo-conjunctival CM (ConfoScan4, Nidek, Gamagori, Japan) with a 40X surface- noncontact objective. Data obtained using CM were also compared to standard histopathology or cytologic samples obtained by scraping technique. Results CM analysis of OSS neoplasia showed good agreement with correspondent tumor cytology or histopathology, revealing some structural and cellular features of the lesions, including: loss of the normal epithelium structure in 10 cases (100%), high-reflective areas in 10 cases (100%), inhomogeneous cell density in 8 cases (80%), cells debris in 8 cases (80%), abrupt demarcation at the lateral edge of the lesion in 7 cases (70%), nodular zone of cell proliferation in 6 cases (70%), fibrovascular structures in 3 cases (30%), keratin pearls in 1 case (10%) and pre-Bowman space involvement in 1 case (10%). No anterior stroma involvement was documented. Cells anisocytosis, pleocytosis and anisonucleosis were also documented in 9 of 10 tumors (90%). Conclusion In-vivo CM seems a reliable, non-invasive method useful for in-vivo diagnosis of OSS neoplasia.

Purpose Clinically significant diabetic macular edema (CSME) shows variable retinal structural and functional impairment. The aim of this study was to assess the correlation between different macular parameters, mainly fundus autofluorescence (FAF) and macular sensitivity, in diabetics with CSME. Methods Sixty diabetic patients with untreated CSME (104 eyes) underwent: best corrected visual acuity determination (BCVA, logMAR), slit lamp biomicroscopy, fluorescein angiography, OCT, FAF and microperimetry. Results Thirty eyes had normal FAF , whereas 74 increased (IFAF) pattern. Retinal sensitivity over areas with IFAF was significantly lower than over areas with normal FAF (p

Diabetic retinopathy (DR) is the most frequent ocular complication in patients with diabetes and the main cause of blindness in the working age population in industrialized and developing countries. The most important risk factors that contribute to the pathogenesis of DR are chronic hyperglycaemia, hyperlipidemia and hypertension. Epidemiologic data show that each year 12 percent of new cases of legal blindness are attributed to diabetic retinopathy (DR) and that its prevalence increases with both increased duration of diabetes and the age of patients. Of insulin-dependent patients with diabetes for 30 or more years, 12% are blind. Diabetic macular edema (DME) is a frequent manifestation of DR and the leading cause of legal blindness in patients with type 2 diabetes. Over a 10-year period, non clinically significant DME and clinically significant DME will respectively develop in 14% and 10% of known diabetic patients. Approximately half of patients with DME will loose two or more lines of visual acuity within 2 years. Overall social costs (expenditures and lost productivity) caused by DR are about $174 billion in the USA. Approximately 20% of health care expenditures for persons with DM (50% with DM as a co-morbidity), 10% of expenditures for complications of DM and 15% of expenditures for health care for DM are ophthalmic. Therefore, DR accounts for significant economic costs that can be reduced with early diagnosis, achieved with screening methods. Screening of DR allows for timely treatment, vision preservation and consequent saving costs. Implementation of screening systems in different environments worldwide is the first necessary step in order to prevent blindness caused by diabetes mellitus.

Purpose To investigate the different pattern of fundus autofluorescence imaging of choroidal tumors generated with short-wavelength and near-infrared Methods Thirty-one eyes of 31 consecutive patients affected by choroidal tumor performed standard fundus autofluorescence with short-wavelength (SW FAF) and fundus autofluorescence with near-infrared (NIR FAF). Fundus photography, A and B scan ultrasound and OCT were performed. Autofluorescence features of choroidal tumor and overlying retinal pigment epithelium (RPE) were correlated with clinical features. Results Twelve of 31 choroidal tumors were choroidal melanoma, 8 choroidal nevus, 5 circumscribed choroidal hemangioma and one was choroidal granuloma. Different pattern of SW FAF and NIR FAF were detected related to the presence of pigment, drusen, RPE atrophy and hyperplasia, RPE detachment and subretinal fluid over or around the lesion. Conclusion Standard autofluorescence (SW FAF) and NIR FAF provide different information on intrinsic autofluorescence of choroidal tumor and on the related RPE and retinal changes

Purpose To describe the technique and results of modified (enlarged) enucleation via lateral orbitotomy for choroidal melanomas with macroscopic periopticposterior extrascleral extension. Methods 5 cases of choroidal melanoma with macroscopic periopticposterior entrascleral extension underwent modified enucleation via lateral orbitotomy. After lateral orbitotomy and orbital mass exposure with direct tumor visualization, a long optic nerve stump was cut and the orbital component of the tumor was completely (macroscopically) removed along with the globe in all cases. After haemostasis and orbital reconstruction an orbital implant was than placed. Follow up was longer than 12 months Results After pathologic examination, the tumor result completely removed in 5 patients (100%). Excellent cosmetic outcome was achieved in all cases (100%) without operative or postoperative complications. Conclusion Modified (enlarged) enucleation via lateral orbitotomy for choroidal melanomas with macroscopic perioptic/posterior extrascleral extension allows placement of an orbital implant avoiding the long healing process of orbital exenteration with excellent clinical and cosmetic outcome.

Purpose To compare standard versus bolus photodynamic therapy (PDT) in the treatment of symptomatic circumscribed choroidal hemangioma (CCH). Methods Twenty consecutive cases of CCH were included in this prospective randomized study. Each patient was randomly assigned to receive either standard PDT (10-minutes 6mg/mq2 verteporfin infusion; treatment at 15 min; 50 J/cm2; 83 sec) or bolus-PDT (6mg/mq2 verteporfin intravenously bolus in 2 min; treatment at 5 min; 100 J/cm2; 166 sec). All patients were treated with a single PDT application. Best correct visual acuity, fundus photography, optical coherence tomography, fluorescein and indocyanine green angiography were performed at baseline and at each follow-up examination. Retinal sensitivity was tested with microperimetry after treatment. Follow-up was longer than 24 months. Results Mean follow-up was 28 ± 4 months. All cases (100%) showed clinical regression of the treated lesion, complete resolution of subretinal fluid and reduction of choroidal leakage on angiograms. Cases treated with bolus-PDT showed retinal pigment epithelium (RPE) hyperplastic changes over treated area. No similar RPE changes were found in patients treated with standard PDT. Two patients treated with bolus-PDT developed macular pucker, respectively 4 and 5 months after treatment. Microperimetry examination revealed areas of reduced sensitivity after bolus-PDT compare to standard PDT (p

Purpose: Some evidence suggests that uveal melanomas develop following one of two ways in a pathogenetic model, one starting with a loss of chromosome 3 and the other with extra copies of 6p (+6p), heaving different prognostic outcome. The aim of this study was to analyze the role of Fluorescence In Situ Hybridization (FISH) applied on Fine Needle Aspiration Biopsy (FNAB) specimen to in-vivo verify the presence/ absence of this bifurcated pathogenetic and prognostic model. Methods: Thirty-five consecutive patients, affected by posterior uveal melanoma (>3,5 mm in thickness), scheduled for Iodine-125 brachitherapy, underwent intraoperative trans-scleral FNAB just before plaque implantation. Specimens underwent Fluorescent in Situ Hybridisation (FISH) to detect chromosome 3 and 6 (as +6p and 6q-) status. Results: Minimum follow-up was 12 months (range: 12-40 months). All cases gave enough material for cytogenetic analysis. +6p and monosomy 3 were mutually exclusive in 29 cases (83%). Three cases (8.5 %) showed no chromosome 3 and 6 abnormalities and three cases (8.5%) showed both monosomy 3 and +6p. Monosomy 3 was not significantly related (p>.05) to tumour dimensions and tumour location (choroid/ciliary body). No major complications or extrascleral extension were documented. Three patients (8,5%) with monosomy 3 developed metastatic disease during the follow up. Conclusions: In-vivo detection of cytogenetic prognostic-related characteristics in posterior uveal melanoma is possible analyzing cytologic specimens obtained by transcleral FNAB before episcleral plaque implantation. FISH analysis for chromosome 3 and 6 status could offer more prognostic information than monosomy 3 alone.

Purpose: To validate the Helsinki University Central Hospital (HUCH) Working Formulation, the first substaging system designed to predict prognosis of metastatic uveal melanoma and to facilitate design, reporting and interpretation of controlled clinical trials. Methods: Data of 226 patients who died of disseminated uveal melanoma were collected from members of the European Ophthalmic Oncology Group (OOG). Karnofsky or WHO performance status, largest diameter of largest metastasis and serum alkaline phosphatase (AP) level at diagnosis of metastases, and time to death were requested. The working formulation is based on a multivariate model that identified these variables as independent predictors of survival, modeling time on chemotherapy as a confounding variable (Eskelin et al. Cancer 2003; 97: 65–75). Patients whose predicted survival was less than 6 months were assigned to stage IVc, between 6 and 12 months to stage IVb and over 12 months to stage IVa. Observed survival time was calculated for each stage. Results: The median AP level was 0.69 (range, 0.24-11) times upper normal limit, median largest metastasis was 3.5 cm (range, 0.8-19), and percentage of patients representing WHO performance index 0, 1-2 and 3 was 61%, 38% and 1%, respectively. Of the 226 patients, 100 (44%) were assigned to stage IVa, 97 (43%) to IVb and 29 (13%) to IVc. The median observed survival times were 18.3, 10.0 and 4.6 months, respectively. In Kaplan-Meier analysis, the stages reflected best to worst survival (P

Purpose: To evaluate the ability to determine single lesions and to grade clinical levels of diabetic retinopathy (DR)using nonmydriatic(NM)digital color retinal images compared to Early Treatment Diabetic Retinopathy Study(ETDRS)7 standard 35-mm stereoscopic color fundus photos. Methods: 80 eyes of 42 diabetic patients with wide spectrum of DR lesions were enrolled.Nonsimultaneous 45°-field non stereoscopic digital color images were taken from 1 central field(1F) and from 3 fields (3F) with a NM automatic fundus camera(Nidek Technologies, Italy).After dilatation standard 30° ETDRS photos were obtained with Topcon TRC50IA(Japan).The images obtained were analysed by 2 independent, masked readers evaluating lesion-by-lesion to assess the ETDRS grading and the clinical severity of the disease according the International proposed 5-stage disease severity classification. Results: Comparison of single lesions among 1F or 3F images and ETDRS fields revealed good agreement for hard exudates(k= 0.74 and k=0.75 respectively);moderate agreement for hemorrhages and/or microaneurysms(k=0.49, k= 0.60),cotton wool spots(k=0.41, K=0.49)and retinal new vessels(k=0.40, k=0.43).The agreement was fair for clinically significant macular edema(k=0.40, k=0.39).There was substantial agreement between 3F and ETDRS images for moderate non-proliferative and proliferative DR(k=0.70 and k= 0.89 respectively),while it was moderate between 1F and ETDRS images. Conclusions: The automatic non-mydriatic 3 field system may be an effective tool to determine the level of DR and to identify the need for prompt referral to the ophthalmologist, even when applied as a telemedicine tool. It is still controversial if one 45° image is adequate for DR screening.

Purpose: Diabetic macular edema (DME) determines central retinal sensitivity reduction with a functional correlate to increased macular thickness. Data about fixation are controversial. The aim of this study was to determine fixation characteristics in diabetic patients with or without clinically significant macular edema (CSME). Methods: 120 consecutive diabetic patients (165 eyes) underwent best corrected visual acuity (BCVA) determination (ETDRS charts), and a complete ophthalmologic examination. Diagnosis and classification (focal, diffuse and cystoid) of CSME were made on 30 degrees stereo retinal photographies (field 2 of ETDRS) and fluorescein angiography and OCT. The presence and site of hard exudates was evaluated. Retinal fixation and macular sensitivity were investigated with an advanced, automatic microperimeter (MP1, Nidek Technologies, Italy). Macular thickness and foveal neuroretinal detachment were determined by Optical Coherence Tomography (OCT Stratus, Zeiss Humphrey Instruments, Germany). Results: Of 141 examined eyes 24 (17.14%) had focal CSME, 37 (26.43%) had diffuse CSME, 51 (36.43%) cystoid CSME, 29 (17.6%). Mean BCVA was 0.6+0.36 logMAR. There was no significant correlation between the type of edema and the site and the stability of fixation (P>0.05). Fixation was eccentric or unstable in case of subfoveal hard exudates (Site P=0.032; Stability P= 0.0113). The presence subfoveal neuroretinal detachment did not significantly influence fixation pattern (P>0.05). Conclusions: Retinal fixation patterns in DME vary diferently based on clinical characteristics of foveal involvement. Fixation may be often located in areas undergoing laser treatment. The functional impact of focal photocoagulation should be investigated also from the fixation point of view