Altered serum levels of autophagy proteins Beclin-1 and mTOR in patients with exudative age-related macular degeneration.

Autophagy is a key process in the maintenance of cellular survival and homeostasis. Inhibition of autophagy results in degenerative changes resembling ageing. We wondered if autophagy can contribute to the pathogenesis of age-related macular degeneration (AMD). We aimed to investigate the serum concentrations of two key autophagy regulators, Beclin-1 and mechanistic target of rapamycin (mTOR), in patients with exudative AMD. This retrospective case-control study included 38 patients with exudative AMD and 36 sex- and age-matched controls selected among senile cataract patients. Circulating Beclin-1 and mTOR were assessed using an enzyme-linked immunosorbent assay. The proteins levels were correlated with age, sex, duration of ocular symptoms, as well as angiographic and optical coherence tomography findings. Serum Beclin-1 levels were much lower in patients with AMD than in controls (median, 0.100 ng/ml versus 1.123 ng/ml; p = 0.0033), while mTOR levels did not differ (median, 4.377 ng/ml versus 3.608 ng/ml; p = 0.4522). Participants of the study older than 70 years had lower Beclin-1 levels than younger ones (p = 0.0444). However, this difference was the most evident in patients with AMD (p = 0.0024). Serum mTOR levels increased with age. In patients with AMD, lower mTOR levels were associated with drusen, while higher levels were observed in those with a fibrous scar in the contralateral eye (p = 0.0212). Our findings suggest that circulating Beclin-1 decreases with age and that is downregulated in patients with AMD.

Effect of photodynamic therapy with verteporfin on tumor thickness and best corrected visual acuity in patients with circumscribed choroidal hemangioma: a single-center experience.

The management of circumscribed choroidal hemangioma (CCH) with photodynamic therapy (PDT) using verteporfin has resulted in significant functional and clinical improvement compared with the pre-PDT era. Literature data on factors influencing clinical outcomes and predictors of response to PDT in symptomatic CCH are inconsistent. The aim of this study was to investigate the efficacy of PDT with verteporfin in patients with CCH depending on symptom duration and tumor thickness at baseline. We analyzed the medical records of 37 patients with symptomatic CCH divided into 3 groups according to symptom duration (≤ 50 weeks, 51 - 100 weeks, and > 100 weeks) and into 2 groups according to tumor thickness (≤ 2.3 mm and > 2.3 mm). Patients were subjected to PDT with verteporfin at a concentration of 6 mg/m2 body surface area and a light dose of 50 J/cm2 at a wavelength of 689 nm. The mean number of treatment sessions was 1.57 (range, 1 - 3). Tumor thickness, the transverse and longitudinal diameters of the tumor base, and best corrected visual acuity (BCVA) were evaluated at baseline and at 12 - 15 months after treatment. After PDT, the mean tumor thickness in the whole study group decreased by 1.19 ± 0.66 mm (from 3.14 mm to 1.95 mm). Subgroup analyses revealed no significant differences between the 2 groups divided according to tumor thickness (p = 0.49). However, tumor thickness differed significantly between the 3 groups divided according to symptom duration (p < 0.05). BCVA increased in 22 patients (59.5%), remained unchanged in 12 patients (16.2%), and decreased in 3 patients (10.1%). Our study provides evidence for the efficacy of PDT with verteporfin in terms of improving or stabilizing visual function as well as reducing tumor thickness in patients with CCH, including those with long-lasting disease.

Central serous chorioretinopathy induced by drugs metabolized by cytochrome P450 3A4.

The purpose of this study was investigate whether replacing or discontinuing drugs that are inhibitors or substrates of cytochrome P450 3A4 (CYP3A4) may improve the clinical course of central serous chorioretinopathy (CSC). A retrospective observational study included 43 patients with active CSC. Twenty seven patients (32 eyes, group 1) were using drugs that act as substrates or inhibitors of CYP3A4. In 25 of these 27 patients, treatments including steroids, calcium channel blockers, anticoagulants, statins, beta-adrenolytics, angiotensin receptor antagonists, antidepressants, muscarinic receptor antagonists, phosphodiesterase type 5 inhibitors, and others were discontinued or replaced with medications not affecting CYP3A4. Sixteen patients (19 eyes, group 2) not using any medication that affects CYP3A4, were given eplerenone, rifampicin, or laser treatment. Main outcomes measures were assessed by functional and anatomical images obtained using multimodal imaging techniques. The average follow-up time was 12 months. In group I after discontinuing or replacing substrates or inhibitors of CYP3A4, improvements were observed in 18 patients (22 eyes). None of the patients that were using drugs affecting CYP3A4 improved with eplerenone therapy, however, all 18 patients improved after discontinuing the drugs. All these drugs had a blocking effect on eplerenone therapy. Best corrected visual acuity (BCVA) improved in 14 eyes, remained unchanged in 5 eyes, and worsened in 3 eyes. In 21 of the 22 eyes, subretinal fluid absorption was observed with optical coherence tomography (OCT). Mean central retinal thickness decreased from 361 µm to 219 µm. One patient (2 eyes) was unable to change treatment (due to neoplasm), one patient (1 eye) did not agree to change or stop treatment, and seven patients (7 eyes) were lost to follow-up. Of the 16 patients (19 eyes) who were treated with eplerenone, rifampicin, or laser, improvements were observed in 14 patients (16 eyes), two patients (2 eyes) were lost to follow-up, and CSC worsened in 1 eye. We concluded that patients with CSC should not take substrates or inhibitors of CYP3A4. These drugs should be replaced with alternatives that act through other metabolic pathways.

Differences in anti-endothelial and anti-retinal antibody titers: implications for the pathohysiology of acute and chronic central serous chorioretinopathy.

The aim of the study was to evaluate the prevalence of serum anti-retinal (ARAs) and anti-endothelial cell antibodies (ACEAs) in patients with acute and chronic central serous chorioretinopathy (CSC). We enrolled 28 patients with acute CSC, 42 patients with chronic CSC, and 40 healthy controls. The presence of ARAs was determined by indirect immunofluorescence using monkey retina as an antigen substrate, while the presence of AECAs was determined using cultivated human umbilical vein endothelial cells (HUVECs) and primate skeletal muscle according to the manufacturer's instructions (Euroimmun AG). There were no differences in the prevalence of antibodies against rods, cones, cytoplasmic components of retinal nuclear layer cells, and retinal vessels between the acute and chronic CSC groups and the control group (P = 0.27, P = 0.16, P = 0.71, and P = 0.06, respectively). However, AECAs reactive with HUVECs were observed in 46% of patients with acute CSC, 45% of those with chronic CSC, and 22% of controls, whereas AECAs reactive with the skeletal muscle were present in 46%, 45%, and 15%, respectively (difference between groups: P = 0.045 for HUVECs and P = 0.005 for the skeletal muscle). Furthermore, AECA titers were higher in CSC patients than in controls (P = 0.004). This study provides evidence for the possible involvement of an autoimmune process directed against vessel antigens in the pathogenesis of CSC. AECAs may be more important than ARAs in this disease and may be involved in endothelial damage in the choroidal vessels and choriocapillaris, leading to hyperpermeability, which is central to the pathophysiology of CSC.

Mydrane intracameral injection site can be an alternative to mydriatic drops instillation in cataract surgery.

The aim of this study is to compare Mydrane (combination of tropicamide 0.02%, phenylephrine 0.31% and lidocaine 1%) and mydriatic drops (tropicamide 1% and phenylephrine 10%) used for cataract surgery in terms of efficacy in pupil dilation and impact on corneal endothelial cell density (CD) and central corneal thickness (CCT). Prospective study including 64 eyes of 64 patients that underwent phacoemulsification with intraocular lens implantation. Patients were randomized into two groups: Mydrane group received: tropicamide and phenylephrine one day preoperatively and Mydrane during the surgery. Reference group received: tropicamide and phenylephrine preoperatively. Pupil size was measured only in Mydrane group, in the same eye of the same patient one day preoperatively after mydriatic drops were given and during the surgery, after intracameral Mydrane injection. CD and CCT were evaluated one day preoperatively and one month postoperatively in all patients and compared between Mydrane and reference groups. The results show CCT and CD significantly decreased after surgery in both groups. There is no difference in this decrease between groups. In Mydrane group there was no difference in dilated pupil diameter between Mydrane and mydriatic drops. Gender, diabetes mellitus, POAG, alpha-1 blocker treatment failed to affect pupil dilation obtained with Mydrane. Cataract surgery affects CCT and CD regardless of which mydriatic protocol had been used. Pupil diameter was similar after drops instillation and after Mydrane injection in all patients from the Mydrane group.

Involvement of serum anti-retinal antibodies in the pathophysiology of diabetic retinopathy: a pilot study.

The aim of this single-center cross-sectional study was to identify tissue targets for circulating anti-retinal antibodies (ARAs) in the serum of patients with diabetic retinopathy (DR). The study included 61 participants with DR (30 with nonproliferative diabetic retinopathy and 31 with proliferative diabetic retinopathy) and 30 healthy controls. An indirect immunofluorescence (IIF) test was used to detect serum ARAs and identify their targets in the tissue. Four types of ARAs were found in serum samples from DR patients: antibodies against the outer segments of the rods, antibodies against the outer segments of the cones, antibodies against the cytoplasmic components of retinal nuclear layer cells, and antibodies against retinal nerve fibers. A significant difference was noted between groups in the prevalence of antibodies against the outer segments of the rods (NPDR, 40%; PDR, 74.2%; and controls, 40%; P = 0.008) as well as antibodies against the outer segments of the cones (NPDR, 23.3%; PDR, 61.3%; and controls, 30%; P = 0.005). Interestingly, the distribution of immunofluorescence intensity for the outer segments of the rods and cones differed significantly between study groups (P ≢ 0.001 and P = 0.019, respectively). In conclusion, the results of our pilot study showed that in most patients with DR, the outer segments of photoreceptors tend to be the tissue target for serum ARAs. This may indicate their possible involvement in the pathogenesis of DR. However, further research is needed to fully elucidate whether these antibodies participate in photoreceptor damage in the course of DR.

Non-responsiveness and tachyphylaxis to anti-vascular endothelial growth factor treatment in naive patients with exudative age-related macular degeneration.

Age-related macular degeneration (AMD) is a leading cause of central visual loss in people aged over 50 years in well developed countries. Although the anti-vascular endothelial growth factor (VEGF) therapy has become a standard treatment for exudative AMD, its effectiveness may be limited in some cases. We aimed to assess the prevalence of non-responsiveness and tachyphylaxis to anti-VEGF drugs in patients with exudative AMD. The study included 63 initially treatment-naive AMD patients who were analyzed for non-responsiveness and tachyphylaxis to intravitreal injections (IVI) of ranibizumab and aflibercept. The participants were enrolled in a National Healthcare Fund (NHF) Therapeutic Program for the Treatment of Exudative AMD. Best-corrected visual acuity (BCVA) and morphological features of a disease activity assessed in optical coherence tomography (OCT) were evaluated during a 12-month follow-up. The percentage of non-responders achieved 22.2% (14 eyes). No significant correlation was found between the type of VEGF inhibitor and a negative response to therapy. Eight patients (12.7%) developed early tachyphylaxis, which was more common in eyes treated with aflibercept (P = 0.04). The presence of serous pigment epithelium detachment (sPED) at baseline was associated with non-responsiveness as determined by both BCVA (OR 18.2, 95% CI 2.86 - 248; P = 0.021) and OCT features (OR 23.0, 95% CI 1.80 - 321; P = 0.030). Eyes treated with aflibercept showed statistically significant greater BCVA improvement (P = 0.0034) and central retinal thickness (CRT) reduction (P = 0.0129) as compared to ranibizumab group during a loading phase of therapy. In a maintain phase of treatment the differences in BCVA and CRT between these two groups were not statistically significant, however eyes treated with aflibercept still showed better functional and anatomical results. Anti-VEGF therapy is an effective method of treatment for exudative AMD, however some patients may show week or no positive reaction or may develop tachyphylaxis. Awareness of these possible negative effects is an important clinical problem in the long-term management of AMD patients with VEGF inhibitors.

Translation and validation of the Polish version of the EORTC QLQ-OPT30 module for the assessment of health-related quality of life in patients with uveal melanoma.

The purpose of this study was to translate into Polish and validate the European Organization for Research and Treatment of Cancer (EORTC) Quality of Life Questionnaire Ophthalmic Module (QLQ-OPT30). The QLQ-OPT30 was translated and pilot-tested according to EORTC guidelines. Patients were qualified based on the histological or ophthalmoscopic and ultrasonographic diagnosis of uveal melanoma. Each patient filled out the Polish version of the EORTC Quality of Life Questionnaire (EORTC QLQ-C30), the QLQ-OPT30 module and a personal questionnaire. Patients were divided into three groups according to treatment type, melanoma localisation and possible negative outcomes of the disease or treatment. Reliability and validity test were performed and patients' comments were analysed. Our study included 94 patients--50 women (53.2%) and 44 men (46.8%) in the mean age of 58.4 (±13) years. Treatment type and melanoma location did not influence the health-related quality of life. The QLQ-OPT30, apart from two scales, showed positive internal consistency (Cronbach's alpha coefficients range: 0.70-0.88). We found the Polish version of the EORTC QLQ-OPT30 module a useful tool for measuring health-related quality of life in uveal melanoma patients. Although its discriminative validity is somewhat limited it still can be recommended for clinical use in the Polish population.

Angiomorphology of the pigmented Bomirski melanoma growing in hamster eye.

An melanotic line of Bomirski Hamster Melanoma (BHM Ma) was implanted into the anterior chamber of the hamster eye and the ensuring vascular system was studied using scanning electron microscopy (SEM) of vascular corrosion casts. The tumor vasculature, induced mainly from the host iris vessels, showed generally disorganized and irregular patterns. Tortuous tumor capillaries of uneven contour with local dilatations and constrictions were drained by markedly dilated, thin-walled, venous vessels connected with the eye veins. Vascular sprouting and, less frequently, intussusceptive formation of new vessels were observed at the periphery and also within the tumor mass. The presence of numerous nodular outgrowths, varying in size, on the surface of dilated venules and venous vessels represent morphological evidence for continuous remodeling of the tumor vasculature. The observed features of the vascular system seem to provide a pathway for further tumor expansion. Our study showed that BHM Ma line, originating from an aggressive skin melanotic melanoma, implanted to the eye anterior chamber gave rise to rapidly growing tumors with the capability of inducing abundant vasculature which allows metastasis to the lungs, kidneys and regional lymph-nodes.

[Failures of brachytherapy in posterior uveal melanoma]. / Niepowodzenia brachyterapii w czerniaku naczyniówki.

PURPOSE: To estimate the risk factors responsible for unsuccessful treatment of melanoma with episcleral plaque brachytherapy. MATERIAL AND METHODS: Based on 23 cases of choroidal melanoma, in which the eyes had to be enucleated after earlier brachytherapy, clinical characteristics of the tumors, type of plaque therapy and reasons of enucleation were analysed. Histopathologic examination of enucleated eyes was performed by standard techniques and immunohistochemical stains on formalin-fixed paraffinembedded tumor tissues. RESULTS: Tumor size is the most valuable risk factor for prognosis in plaque brachytherapy of melanoma. The direct indications for enucleation were: increase of tumor mass, optic disc invasion, consecutive glaucoma and extrascleral extension. Histopathological examination revealed 17 mixed-cell melanoma, 4 spindle-cell, 1 epithelioid and 1 necrotic. Post-irradiation necrosis was found in only 6 tumors. CONCLUSIONS: It is very difficult to predict the results of conservative treatment of melanomas basing on their clinical characteristics. The aggressive nature of the tumors and their resistance to irradiation seem to be the likely explanation of failures.

Experimental ruthenium plaque therapy of amelanotic and melanotic melanomas in the hamster eye.

The effects of beta-radiation on melanoma implanted into the hamster's eye were investigated. Two Bomirski hamster melanomas (BHMs), differing in their melanin content, were compared with regard to their radiosensitivity to ruthenium-106 (106Ru) radiation. Tumours growing in the iris were irradiated with 3, 6 or 10 Gy of 106Ru given as a single dose or in four fractions at 24 h Intervals. Tumour growth kinetics and distant metastases were studied, and the eyeballs were examined histologically. Dose-dependent delay of tumour growth was observed in both melanomas. After treatment with a dose of 6 Gy, the Ab amelanotic tumours grew 2.6 times slower, and the Ma melanotic tumours 1.4 times slower than untreated ones. The location of metastases differed in the two tested lines--pigmented metastases were found mainly in the lungs, while unpigmented metastases were found mainly in the kidneys. Histopathological analysis showed signs of blood vessel damage such as endothelial cells swelling, erythrocyte extravasation and tumour necrosis. This last finding increased with the rising dose of beta-radiation. Pigmented tumours were found to be two times more resistant to beta-radiation than amelanotic ones. The pattern of metastases of BHMs is determined by the type of melanoma (Ab or Ma). Exposure to beta-radiation from 106Ru did not significantly affect either the number or size of metastases except at a dose of 10 Gy. This dose caused a statistically significant decrease in the number of metastases in the Ma melanotic subline.

[Trans-scleral resection of the ciliary body and choroidal melanoma]. / Przeztwardówkowa resekcja czerniaka ciala rzeskowego i naczyniówki.

PURPOSE: To present our preliminary results of trans-scleral resection of ciliary body and choroidal melanoma. MATERIAL AND METHODS: 16 patients aged 20-73 years (mean 56 years) were treated from February 1999 to March 2000. There were 10 patients with ciliary body tumors, 2 with choroidal tumors and in 4 patients the tumor involved ciliary body and choroid. The mean basal diameter of tumors was 9.4 mm and the mean thickness was 7.2 mm. Trans-scleral resection was performed in all cases. Adjunctive radiotherapy using 106Ru plaques was applied in 7 cases. RESULTS: Histopathologically the diagnosis of melanoma was confirmed in all cases. In 10 cases there were spindle cell melanomas and in 6 mixed type. The intra-operative complications were vitreous hemorrhage in 3 patients, vitreous loss in three and both complications in one patient. Post-operative complications occurred in 3 cases of single instances each of retinal detachment, retinal edema and haemophthalmus. After surgery visual acuity did not change in 9 eyes, improved in 2 and deteriorated in 5 eyes. CONCLUSIONS: Our preliminary results confirm the opinion that trans-scleral resection could be a valuable method in the treatment of intraocular melanoma.

[Transpupillary thermotherapy combined with 106Ru as a method of managing choroidal melanoma]. / Przezzreniczna termoterapia w skojarzeniu z brachyterapia 106Ru jako metoda postepowania w czerniaku naczyniówki.

BACKGROUND: A study was carried out to evaluate the effectiveness of transpupillary thermotherapy (TTT) with diode laser applied as adjuvant treatment to 106Ru brachytherapy in choroidal melanoma. These two methods coined as "sandwich therapy" are complementary and allow to decrease the dose of radiation. MATERIAL AND METHODS: Between November 1996 and December 1997, 45 patients with choroidal melanoma were treated simultaneously with 106Ru and TTT. There were 18 males and 27 females, aged 20 to 78 (mean 57). There were 7 tumors with height of 5 mm or less, 28 ranging 5.43-8.63 mm and 10 with height 9 mm or more. The anterior tumor margin was located in the equator area or posterior to it in 71.1% of cases. The laser treatment was delivered using spots 2-3 mm in size, at a mean power 600 mW, for about 1 minute. The mean radiation dose was 60 Gy to the tumor apex. The follow-up ranged from 12 to 24 months. RESULTS: Reduction in tumor height by at least 30% was exhibited in 35 (77.8%) tumors, in one evaluated as positive result it was 27%. There was no improvement after treatment in 4 cases, 2 eyes were enucleated. A significant relationship was found between tumor thickness and the result of treatment. CONCLUSION: Our preliminary observations suggest that TTT combined 106Ru brachytherapy is an effective method of treatment. A longer follow-up is needed to assess final local results and the impact of treatment on survival.