Pathogenicity reclassification of the RPE65 c.1580A>G (p.His527Arg) - a case report.

BACKGROUND: It is of utmost importance to define the molecular diagnosis of patients with retinitis pigmentosa (RP) due to existing targeted therapeutic option: voretigene neparvovec.We provide clinical evidence for pathogenicity reclassification of variants of uncertain significance (VUSs) RPE65 c.1580A>G (p.His527Arg). MATERIALS AND METHODS: A case report of a 10-year-old boy with progressive vision loss. The patient manifested disease highly suggestive of RPE65 retinal dystrophy: nyctalopia, fairly good central vision, severely depressed full-field electroretinography responses and complete loss of peripheral fundus aut ofluorescence. RESULTS: Invitae Inherited Retinal Disorders Panel identified likely pathogenic mutation RPE65 c.499G>T (p.Asp167Tyr) and RPE65 c.1580A>G (p.His527Arg), variant of uncertain significance. Segregation analysis confirmed that these variants are in trans. CONCLUSIONS: We conclude that the variant RPE65 c.1580A>G (p.His527Arg) has contributed to the pathologic phenotype, demonstrating its significance clearly in the case presented, and should be reclassified according to the criteria of evidence as pathogenic. Therefore, patients with this specific variant in homozygous or compound heterozygous form would likely benefit from genetic treatment based on recombinant adeno-associated virus vector, providing a working RPE65 gene to act in place of a mutated RPE65 gene.

RPE65 c.353G>A, p.(Arg118Lys): A Novel Point Mutation Associated with Retinitis Pigmentosa and Macular Atrophy

Precise genetic diagnosis in RPE65-mediated retinitis pigmentosa (RP) is necessary to establish eligibility for genetic treatment with voretigene neparvovec: a recombinant adeno-associated viral vector providing a functional RPE65 gene. This case report aims to report a novel RP-related point mutation RPE65 c.353G>A, p.(Arg118Lys), a variant of uncertain significance associated with a severe clinical presentation and the striking phenotypic feature of complete macular atrophy. We report the case of a 40-year-old male with inherited retinal dystrophy, all features typical for the RPE65-associated RP, and marked macular atrophy. Genetic testing identified that the patient was a compound heterozygote in trans form with two heterozygous variants: RPE65 c.499G>T, p.(Asp167Tyr) and RPE65 c.353G>A, p.(Arg118Lys). Furthermore, short-wavelength and near-infrared autofluorescence patterns exhibited deficiencies specific to mutations in the visual cycle genes. To the best of our knowledge, RPE65 c.353G>A, p.(Arg118Lys) is the first described point mutation on this locus, among all other reported insertional mutations, currently classified as likely benign and of uncertain significance. We concluded that this variant contributed to the pathological phenotype, demonstrating its significance clearly to be reclassified as likely pathogenic. This being the case, patients with this specific variant in homozygous or compound heterozygous form would be likely candidates for genetic treatment with voretigene neparvovec.

RPE65 c.393T>A, p.(Asn131Lys): Novel Sequence Variant Detected.

Background: Leber congenital amaurosis (LCA) is a monogenic, but genetically heterogenous disease, and at least 27 genes are implicated. This case report is aimed at providing evidence to link the novel variant RPE65 c.393T>A, p.(Asn131Lys), variant of uncertain significance (VUS), to clinical phenotype and to set the ground for objective assignment of pathogenicity confidence. Case Presentation. A case report of a female patient with LCA who manifested with nystagmus, night blindness, profound visual deficiency, and peripheral involvement of the retina consistent with RPE65 dystrophy. A thorough clinical examination, diagnostic evaluation, and genetic testing were performed. The patient was a compound heterozygote in trans form: RPE65 c.304G>T, p.(Glu102∗) pathogenic, and RPE65 c.393T>A, p.(Asn131Lys), VUS. The latter variant is absent in healthy controls and is considered harmful on in silico prediction. Conclusions: We conclude that RPE65 c.393T>A, p.(Asn131Lys) contributed to the pathologic phenotype, demonstrating its significance clearly in the case presented, and should be reclassified according to the criteria of evidence as likely pathogenic. This being the case, patients with this specific variant are likely candidates for genetic treatment.

Do Gender, Age, Body Mass and Height Influence Eye Biometrical Properties in Young Adults? A Pilot Study.

BACKGROUND: Do gender, age, body mass and height influence eye biometrical properties in young adults? METHODS: A total of 155 eyes (92 female, 63 male) of healthy subjects between the ages of 18 and 39 years were included in the study. The subjects' gender and age were recorded, and their body mass, height and biometrical properties of the eyes were measured. RESULTS: The male subjects had significantly thicker and flatter corneas and lower minimal rim-to-disk ratios than the female subjects did. In both genders, age showed strong, negative correlations with anterior chamber depth and pupil diameter and a positive correlation with lens thickness. We also found significant, negative correlations between body height and mass with keratometry measurements, negative correlations between body height and optic disk rim area and rim volume, and positive correlations between body mass and axial length in both genders. CONCLUSIONS: Biometric eye parameters differ among people. In addition to age and gender, which are usually taken into consideration when interpreting ocular biometry findings, we strongly suggest that body height and mass should be also routinely considered when interpreting eye biometry data, as these factors have an impact on ocular biometry.

Single-Stage Orbital Socket Reconstruction Using the Oversized Dermis Fat Graft and the 22 mm Silicone Orbital Implant after an Extended Enucleation.

We would like to present a surgical technique of orbital socket reconstruction using oversized dermis fat graft and 22 mm silicone orbital implant in a single-stage after extended enucleation in two patients with massive local recurrence of anteriorly located choroidal melanoma previously treated with endoresection. Orbital tissues en bloc were removed leaving conjunctival lining only at the fornices. Simultaneously, the 22 mm silicone sphere was implanted deeply into the orbit and covered with the oversized dermis fat graft of 30 mm in height and 35 mm in length with 20 mm of the fat thickness. The graft was sutured to the residual forniceal conjunctiva with interrupted 6/0 absorbable sutures overlapping conjunctiva with the graft edge for 2 mm to facilitate the epithelization. Epithelization was completed in two months, leaving well-formed fornices with good fitting of the prosthesis. The key point of orbital socket reconstruction after extended enucleation is to restore conjunctival lining prior to volume. Thus, whenever facing a massive volume and conjunctival lining loss, simultaneous insertion of the 22 mm silicone sphere deep into the orbit combined with oversized dermis fat graft is, in our opinion, the method of choice. It proved to be safe and effective with favourable long-term results.

Expression and prognostic value of putative cancer stem cell markers CD117 and CD15 in choroidal and ciliary body melanoma.

AIMS: The aim of the present study was to immunohistochemically investigate the expression and prognostic significance of putative cancer stem cell markers CD117 (c-kit), CD34, CD20 and CD15 in a cohort of patients with primary choroidal and ciliary body melanoma. METHODS: The immunohistochemical expression of these markers was evaluated using 3,3'-diaminobenzidine tetrahydrochloride (DAB) and 3-amino-9-ethylcarbazole (AEC) chromogens on paraffin-embedded tissue samples from 40 patients who underwent enucleation in the period from 1985 through 2000. Thirty-one patients had adequate tissue specimens for the analysis. RESULTS: CD117 overexpression was observed in 12 of the 31 samples (39%) when AEC chromogen was used and in 14 of 26 (54%) samples when DAB was used. CD15 positivity was seen in three out of 30 (10%) samples with AEC and in six out of 26 (23%) samples with DAB. CD20 and CD34 exhibited no positivity in the tested samples. During the average follow-up time of 8.7 years (range 0.5-22 years), 17 patients (55%) died due to metastatic disease. The Kaplan-Meier plots showed a significantly shorter overall and disease-free survival in CD117-positive patients when the AEC chromogen was used. CD15 expression was not associated with patients' survival. In multivariate analysis, patients expressing the CD117 AEC had 4.13 times higher risk of lethal outcome in comparison with CD117 AEC negative patients. CONCLUSIONS: Our retrospective cohort study has for the first time demonstrated a small proportion of CD15-positive uveal melanomas. CD117 AEC overexpression was associated with a worse outcome in patients with choroidal and ciliary body melanoma. Further studies should confirm the validity of these observations and their potential for targeted treatment modalities.

Oxidative stress impact on growth hormone secretion in the eye.

AIM: To evaluate the influence of oxidative stress on extrapituitary growth hormone (GH) secretion in the eye and to analyze the interdependence between eye and serum GH levels under normal and hypoxic conditions. METHODS: Pars plana vitrectomy (PPV) was performed in 32 patients with developed proliferative diabetic retinopathy (PDR) and 49 non-diabetic controls, both of whom required this procedure as part of their regular treatment in the period from April 2013 to December 2014. During PPV, vitreous samples were taken and blood was simultaneously collected from the cubital vein. GH levels in serum and vitreous samples were measured by electrochemical luminescence assay. Oxidative stress was measured by enzyme-linked immunosorbent assay of advanced oxidation protein products (AOPP) and lipid hydroperoxide (LPO) in serum and vitreous. RESULTS: Serum AOPP levels were significantly higher than vitreous levels in both groups (P<0.001 for each group) and LPO levels were significantly higher only in PDR group (P<0.001). There was a significant positive correlation between serum and vitreous LPO levels in PDR group (r=0.909; P<0.001). Serum GH levels were significantly higher than vitreous levels in both groups (P<0.001 for each group). Serum GH levels were significantly higher in PDR group than in controls (P=0.012). Vitreous GH values were slightly higher in PDR group, but the difference was not significant. CONCLUSION: Our study confirms that GH production in the eye is autonomous and independent of oxidative stress or pituitary GH influence.

Levels of selected oxidative stress markers in the vitreous and serum of diabetic retinopathy patients.

PURPOSE: In diabetes, an impaired antioxidant defense system contributes to the development of diabetic retinopathy. The main objective of this paper was to find correlations of oxidative stress parameters within and between the vitreous and serum in patients with type 2 diabetes who had developed proliferative diabetic retinopathy. METHODS: The study included and compared two groups of patients who underwent vitrectomy: 37 patients with type 2 diabetes and proliferative retinopathy (PDR), and 50 patients with non-diabetic eye disorders (NDED). Vascular endothelial growth factor (VEGF), advanced oxidized protein product (AOPP), and oxidative stress markers (direct lipid hydroperoxidation (LPO), malondialdehyde (MDA), total superoxide dismutase (SOD), and glutathione (GSH)) were measured in the vitreous and serum of both groups and correlated with one another, between humoral compartments and with gender, age, and serum glucose levels. RESULTS: In the vitreous of PDR patients, VEGF, LPO, and MDA (p<0.05) were increased and SOD values were slightly lowered (p<0.05) than in NDED patients. Vitreous AOPP and GSH showed no differences between the groups. In the serum, AOPP, MDA, and SOD were increased (p<0.05) and VEGF was slightly increased (p<0.05) in the PDR group compared to NDED. With regard to gender, similar changes were recorded for both groups, except for the lower serum MDA in males than females in the NDED group. Advanced age showed no significant effect on changes of measured parameters in the vitreous. In the serum, VEGF was positively correlated (p<0.05) and MDA and SOD negatively correlated (p<0.05) with increasing age. Among measured parameters within and between the vitreous and serum, several correlative links occurred in the PDR group that were not present in the NDED group. The most prominent correlation changes were between serum LPO and vitreal LPO, serum SOD and vitreal LPO, serum LPO and serum SOD, and vitreal VEGF and serum SOD. CONCLUSIONS: Among the selected oxidative stress markers, SOD and LPO were highly correlative in both the vitreous and serum in PDR compared to patients without metabolic disorders. Their correlations suggested that monitoring their mutual alterations might be informative during PDR development and should be considered in further research.

Is the effect of intravitreal triamcinolone acetonide on diabetic macular edema dose-dependent?

PURPOSE: To determine whether the effect of intravitreal triamcinolone acetonide in diffuse diabetic macular edema is dose-dependent regarding the velocity and duration of macular edema reduction, visual acuity improvement, and induction of predictable complications. METHODS: This clinical prospective study included 32 consecutive patients (32 eyes) with diffuse diabetic macular edema. All patients had visual acuity, fluorescein angiogram, and optical coherence tomography performed at the initial visit (baseline). Single triamcinolone acetonide injection was applied intravitreally in different high doses using sterile technique followed by timolol-maleate 0.5%/acetazolamide fixed combination eyedrops twice a day for 6 consecutive months. Macular edema, intraocular pressure, and best-corrected distance visual acuity were reviewed after 5 days and 1, 3, and 6 months. RESULTS: Intraocular pressure did not change significantly during 6 months and was maintained within normal levels in all patients using prescribed antiglaucoma therapy. Best-corrected distance visual acuity and macular edema were better than preoperatively even 6 months after treatment while the highest improvement was recorded 1 month after treatment. There was a slight correlation of macular edema reduction and triamcinolone dose, especially in central 1-mm diameter zone 6 months post triamcinolone acetonide intravitreal injection, but it was not statistically significant. CONCLUSIONS: Triamcinolone intravitreal injection is a relatively safe, inexpensive, and effective method used for diabetic macular edema reduction but its effect is not proven to be dose-dependent, at least in 10-32 mg dose range. Macular edema reduction rate correlated significantly with preoperative edema.

Early satellite metastatic uveal melanoma to the lower eyelid--the first report.

PURPOSE: To present a unique case of an early satellite metastatic uveal melanoma to the ipsilateral lower eyelid. METHODS: Retrospective review of the medical records of a single patient. RESULTS: A 71-year-old white male developed a fast growing, painless, solitary, subcutaneous, nodular mass in the medial half of his right lower eyelid 13 months after enucleation of the right eye for inferonasally located uveal melanoma of the spindle cell type. Microscopically excised eyelid tumor proved to be malignant melanoma of the epitheloid type. The patient underwent complete systemic examination including positron emission tomography that ruled out the primary cutaneous or visceral melanoma. CONCLUSION: The paper is, according to our knowledge, the first documented case of an early satellite metastatic uveal melanoma to the lower eyelid in the patient with no evidence of further metastases.

Intravitreal bevacizumab (Avastin) in treatment of neovascular age-related macular degeneration.

To evaluate the efficacy and safety of intravitreal bevacizumab in treatment of minimally classic and occult choroidal neovascularization secondary to age-related macular disease 48 eyes of 48 patients (mean age of 74.8) included in this prospective, noncomparative, interventional case series. Median follow-up was 18 weeks (6-24). Intravitreal bevacizumab injection of 0.05 mL (1.25 mg) was administered at baseline and in 6 week intervals until leakage resolved, and repeated in case of leakage recurrence. Visual acuity (VA) improved in the majority of patients (mean baseline VA = 1.078 log MAR) by mean increase of 1.32 lines (last follow-up) (p = 0.001). Central foveal thickness and total macular volume decreased by 51 microm (p = 0.01) and 0.84 mm3 (p < 0.0001) respectively. No serious complications were observed. As initial therapy, intravitreal bevacizumab appears to be safe and effective. A significant functional and anatomical improvement was noted in majority of patients and maintained during follow-up.

[Acute retinal necrosis]. / Akutna retinalna nekroza.

AIM: To draw attention to this relatively common disease, which may cause major visual function impairment, and to present our own experience in the diagnosis, treatment and follow-up of acute retinal necrosis patients. METHODS: The manifestation, detection, treatment options and complications of unilateral acute retinal necrosis are illustrated by six case reports. RESULTS: Five patients were immunocompetent, whereas the sixth one suffered from chronic leukemia. In two patients the disease developed in association with herpes zoster, whereas the remaining four showed no signs of herpes disease. Systemic therapy with acyclovir, corticosteroids, salicylates and photocoagulation produced favorable response in five patients. Therapy had to be discontinued in leukemia patient for complications, which resulted in the disease recurrence with retinal detachment and proliferative vitreoretinopathy. Other patients remained stable with preserved visual function and only minor complications. DISCUSSION: In all six patients, the accurate diagnosis was reached relatively late. Obviously, anterior uveitis alone attracted ophthalmologists' attention, and their diagnostic and therapeutic efforts had mostly been focused on anterior uveitis impairment in the visual acuity developed. Appropriate therapy with virostatics, which is necessary in the treatment of this disease, was introduced relatively late, however, all patients responded favorably to this therapy, which also held for the immunocompromised patients administered a half usual dose of acyclovir. Our experience confirmed that virostatic therapy for acute retinal necrosis should be administered for at least 6 weeks. Retinal photocoagulation for prevention of rhegmatogenous retinal detachment appers to be beneficial, as this type of retinal detachment did not develop in these patients. The patient who did not undergo laser photocoagulation of retina did not develop rhegmatogenous retinal detachment but did develop proliferative vitreoretinopathy. CONCLUSION: Acute retinal necrosis is a relatively common disease affecting visual function, which occurs in both immunocompetent and immunocompromised subjects. The disease with its typical clinical picture and many complications should be taken in consideration on the differential diagnosis of uveitis. Therapy for acute retinal necrosis is complex, long-term and associated with frequent systemic complications that may threaten the patient's general health status.

Axial eye length after retinal detachment surgery.

Changes in the eye axial diameter were studied to assess the eye globe impact of conventional operation for retinal detachment. The study included 69 eyes in 69 patients operated on for rhegmatogenous retinal detachment. There were 46.4% of men and 53.6% of women, mean age 52.7 (+/- 15.21) years. Results of preoperative and postoperative ultrasonographic measurement of axial diameter are presented. The mean preoperative and postoperative eye axial diameter was 23.69 (+/- 1.84) mm and 24.43 (+/- 1.91) mm, respectively. Postoperative results showed the axial eye length to increase by a mean of 0.74 (+/- 0.44) mm, yielding a statistically significant difference from the preoperative measurement (p < 0.001). The mean myopia induced by this eyeball elongation was 1.77 D. The encircling band with and without segmental buckling used in surgical repair of retinal detachment creates circular and segmental indentation of the eyeball, thus increasing its axial length. The myopia induced by elongation of the eyeball results in considerable myopia, which requires appropriate correction in the early postoperative period to achieve favorable vision rehabilitation.

Obesity--a risk factor for diabetic retinopathy in type 2 diabetes?

The aim of the study was to investigate whether obesity, independently or associated with other risk factors, increases the risk for the diabetic retinopathy in type 2 diabetic persons. Data of 156 diabetic persons that have consecutively attended the Outpatient Department in the Vuk Vrhovac Institute in Zagreb during two months period were studied. According to their body mass index (BMI) they were divided into three groups: group 1 (BMI < or = 25; n = 49), group 2 (BMI 26-29.9; n = 52) and group 3 (BMI > or = 30; n = 55). The three groups did not differ in age, duration of diabetes, treatment, cholesterol, HDL-cholesterol and triglycerides. With increase in BMI, we observed a significant deterioration of HbA1c and a significant increase in LDL-cholesterol, systolic and diastolic blood pressure. Statistical analyses shown that the prevalence of retinopathy increased significantly with higher body weight (gr. 1: 40.8%, gr. 2: 63.4%, gr. 3: 63.6%;p < 0.05), but also with correlation to quality of metabolic control (HbA1c) and systolic blood pressure. Therefore, obesity may be, because of its significant correlation to quality of metabolic control (HbA1c) and systolic blood pressure, considered as risk factor for diabetic retinopathy in type 2 diabetic persons.

Ocular blood flow parameters in patients with rhegmatogenous retinal detachment.

The aim of the present study is to evaluate the potential statistically significant differences in the ocular blood flow parameters in eyes with rhegmatogenous retinal detachment (RD). Eleven patients, 5 females and 6 males, mean age 46 years (range 22-70), with the unilateral rhegmatogenous RD were enrolled in the study. Colour Doppler Ultrasound was used to measure ocular blood flow velocities in the ophthalmic artery (OA), posterior ciliary's arteries (PCA) and ophthalmic vein (OV). The contralateral eye served as a control. All Doppler examinations were performed 1 day before and exactly 3 days after the retinal detachment surgery. The measurements of the peak systolic velocity (Vmax), diastolic velocity (Vd), minimum velocity (Vmin), time-averaged velocity (TAV), resistive index (RI) and pulsatility index (PI) showed no statistically significant difference (by paired Student's t-test, p > 0.05) between the OA, PCA and OV in healthy eyes and eyes with RD before operation, as well as between the eyes with RD before and after the operation. Only was increased RI in OV of eyes with RD after the surgery (p < 0.05). All these parameters were not related with 2 or more quadrants of RD, but the difference in duration of retinal detachment in days is statistically significant (by Wilcoxon t-test p > 0.05). Pearson correlation method gave statistically significant correlation between RI and PI of the OA in healthy eyes (r = 0.826, p < 0.01), eyes with RD before operation (r = 0.847, p < 0.01) and eyes with RD after the operation (r = 0.856, p < 0.01). Formula for the calculation of PI by RI was derived using linear regression analysis in all three cases. Scleral buckling surgery leaves the ocular blood parameters in OA unchanged. The correlation between RD and logarithm of duration of RD in days is statistically significant.

Choanal stenosis, hypothelia, deafness, recurrent dacryocystitis, neck fistulas, short stature, and microcephaly: report of a case.

We report on a 11-year-old girl with bilateral choanal stenosis, hypothelia, hearing loss, recurrent dacryocystitis, neck fistulas, short stature, and microcephaly. Only three individuals with choanal atresia from a consanguineous family have been reported. One of the patients also had hypoplastic nipples, hypotonia, and delay in speech development. Similar clinical features were seen in two children reported by Greenberg [1987: Am J Med Genet 28:931-934] and Wilson et al. [1998: Am J Med Genet 75:220-222]. They were prenatally exposed to methimazole because of maternal Graves disease. Neck fistulas and microcephaly noted in our patient were not previously reported as features of the syndrome or in the patients prenataly exposed to methimazole. Our patient and those reported by Qazi et al. [1982: Am J Med Genet 13:413-416] most probably have a rare syndrome characterized by this distinctive combination of symptoms. Prenatal exposure to methimazole can cause a phenocopy of the syndrome, which was probably the case in the patients reported by Greenberg and Wilson et al.