Three novel germ-line VHL mutations in Hungarian von Hippel-Lindau patients, including a nonsense mutation in a fifteen-year-old boy with renal cell carcinoma.

BACKGROUND: Von Hippel-Lindau disease is an autosomal dominantly inherited highly penetrant tumor syndrome predisposing to retinal and central nervous system hemangioblastomas, renal cell carcinoma and phaeochromocytoma among other less frequent complications. METHODS: Molecular genetic testing of the VHL gene was performed in five unrelated families affetced with type I VHL disease, including seven patients and their available family members. RESULTS: Molecular genetic investigations detected three novel (c.163 G > T, c.232A > T and c.555C > A causing p.Glu55X, p.Asn78Tyr and p.Tyr185X protein changes, respectively) and two previously described (c.340 + 1 G > A and c.583C > T, resulting in p.Gly114AspfsX6 and p.195GlnX protein changes, respectively) germline point mutations in the VHL gene. Molecular modeling of the VHL-ElonginC-HIF-1alpha complex predicted that the p.Asn78Tyr amino acid exchange remarkably alters the 77-83 loop structure of VHL protein and destabilizes the VHL-HIF-1alpha complex suggesting that the mutation causes type I phenotype and has high risk to associate to renal cell carcinoma. The novel p.55X nonsense mutation associated to bilateral RCC and retinal angioma in a 15-year-old male patient. CONCLUSION: We describe the earliest onset renal cell carcinoma in VHL disease reported so far in a 15-year-old boy with a nonsense VHL mutation. Individual tailoring of screening schedule based on molecular genetic status should be considered in order to diagnose serious complications as early as possible. Our observations add to the understanding of genotype-phenotype correlation in VHL disease and can be useful for genetic counseling and follow-up of VHL patients.

Graves' orbitopathy results in profound changes in tear composition: a study of plasminogen activator inhibitor-1 and seven cytokines.

BACKGROUND: Secretion of cytokines and expression of cytokine receptors have been reported in the orbital connective tissue in Graves' orbitopathy (GO). Lacrimal glands are putative autoimmune targets, and changes in tear film and ocular surface have also been described. Our aim was to characterize the cytokine profile of tears in patients with Graves' disease (GD) with and without orbitopathy. METHODS: Tear samples were collected from 54 eyes of GO patients (age 43.4±15.2 years), 18 eyes of GD patients (age 46.8±11.7 years), and 24 control eyes (age 38.6±13.8 years). Patients underwent ophthalmological examination including Clinical Activity Score (CAS). The level of interleukin (IL)-1ß, IL-6, IL-13, IL-17A, IL-18, tumor necrosis factor (TNF)-α, and RANTES (regulated upon activation, normal T-cell expressed, and secreted) as well as plasminogen activator inhibitor-1 (PAI-1) were measured by multiplex bead array and release values were calculated. RESULTS: The release of IL-1ß, IL-6, IL-13, IL-17A, IL-18, TNF-α, and RANTES were significantly higher in GO patients compared to controls (p<0.05). There was a 2.5-fold increase of IL-6 release. No significant differences were found in cytokine release between the GO and GD groups. In the GO group, significant positive correlation was found between CAS and the release of IL-6 and PAI-1 into tears (r=0.27, p<0.05 and r=0.24, p<0.05, respectively). PAI-1 release was significantly higher in GO than in GD patients and was increased in both the GD and GO groups compared to controls. CONCLUSIONS: Impaired cytokine balance has been observed in tears of GO patients. Secretion of IL-6 into tears might be a useful indicator of disease activity in GO.

Retrobulbar 99mTc-diethylenetriamine-pentaacetic-acid uptake may predict the effectiveness of immunosuppressive therapy in Graves' ophthalmopathy.

BACKGROUND: In Graves' ophthalmopathy (GO), only patients with immunologically active disease respond to immunosuppressive therapy. Previous studies and theoretical considerations suggest that elevated orbital (99m)Tc-diethylenetriamine-pentaacetic-acid (DTPA) single photon emission computed tomography (SPECT) reflects inflammatory disease activity. We studied whether corticosteroid treatment causes a substantial decrease in DTPA uptake in GO, a result consistent with successful immunosuppressive treatment of GO and referred to as a favorable treatment outcome. METHODS: One hundred fourteen orbits in 57 patients with active GO (CAS >or= 4) were entered into the study. All patients received corticosteroid treatment. Orbital DTPA uptakes were numerically quantified for the entire orbit as well as the anterior and posterior segments separately. DTPA SPECT was performed before, and 2 to 9 months after the initiation of immunosuppressive treatment. The normal range for DTPA uptake was established in 34 orbits of 17 patients who were being worked up for Raynaud's phenomenon and had no thyroid disease. RESULTS: The mean DTPA uptake of the 114 orbits of GO patients was higher prior to corticosteroid therapy than after this treatment (11.03 +/- 4.26 MBq/cm(3) and 9.84 +/- 3.51 MBq/cm(3), respectively, p < 0.001) but a substantial decline in DTPA uptake was seen in only 39.5% of GO patients. The positive predictive value of an initial DTPA >12.28 MBq/cm(3) for a substantial decline in DTPA uptake (favorable treatment outcome) was 76%, while a negative predictive value of a pretreatment DTPA

Imaging of disease activity in Graves' orbitopathy with different methods: comparison of (99m)Tc-DTPA and (99m)Tc-depreotide single photon emission tomography, magnetic resonance imaging and clinical activity scores.

BACKGROUND: The immunosuppressive treatment of Graves' orbitopathy (GO) influences the course of the disease in the early, active, retrobulbar inflammatory phase (active GO), which cannot be detected by direct clinical examination. AIM: To evaluate the clinical effectiveness of a newly developed method for the detection of intraorbital inflammatory activity in patients suffering from Graves' orbitopathy, utilizing a four-headed single photon emission tomograph (SPET) camera and (99m)Tc-diethylenetriamine pentaacetic acid (Tc-DTPA). METHODS: The magnetic resonance imaging (MRI) T2 relaxation time score, as a measure of ongoing orbital inflammation (reference method), was compared with the uptake activities (UA) of (99m)Tc-DTPA orbital SPET and the more specific (99m)Tc-Neospect ((99m)Tc-depreotide) SPET, as well as the clinical activity scores (CAS), in 21 patients (42 orbits). RESULTS: By visual inspection, the 'eye SPET' frames of patients suffering from active GO could be easily distinguished from those with inactive GO. Although the distributions of the two radiopharmaceuticals were mildly different, the information obtained was essentially the same. The MRI activity scores correlated well with both (99m)Tc-DTPA and Tc-depreotide UA. The mean (99m)Tc-DTPA UA value of 25 inactive GO orbits was (6.55 +/- 1.4) x 10(-6) ID x cm3 (where ID is the injected dose); the values of the active GO orbits (12 orbits with an MRI score of 1 and five orbits with an MRI score of 2) were significantly higher: (8.87 +/- 1.63) x 10(-6) and (10.36 +/- 1.60) x 10(-6) ID x cm3, respectively. Similar differences were observed for the (99m)Tc-depreotide UA values: the averages in the inactive and active groups with MRI scores of 1 and 2 were (5.23 +/- 1.23) x 10(-6), (7.69+/-2.24) x 10 and (10.92 +/- 3.85) x 10(-6) ID x cm3, respectively. The (99m)Tc-DTPA accumulation pattern in the orbital region of active GO patients was similar to that of the more specific (99m)Tc-depreotide. There was a good correlation (r = 0.71, P<0.001) between the UA values of the two radiopharmaceuticals, but CAS did not correlate with either of the UA values. CONCLUSIONS: (99m)Tc-DTPA eye SPET is a potentially useful method, allowing rapid imaging at an acceptable cost. It provides essential supplementary information to traditional CAS in assessing disease activity in Graves' orbitopathy.

Activated protein C resistance in anterior ischaemic optic neuropathy.

PURPOSE: Protein C is a major component of the natural anticoagulant pathway. Resistance of coagulation factor V (FV) to activated protein C (APC), mostly due to FV Leiden mutation, is the most common cause of inherited thrombophilia. The aim of this retrospective study was to determine the prevalence of APC resistance and Leiden mutation in patients with non-arteritic anterior ischaemic optic neuropathy (NAION). METHODS: A total of 25 patients with NAION were examined between 1997 and 2002. The patients were screened for APC resistance and FV Leiden mutation as well as for acquired risk factors of vascular disease such as diabetes mellitus, hypercholesterolaemia, hypertonia and ischaemic heart disease. A control group of subjects without ocular vascular disease and with homogenous distribution of the same risk factors was used for comparison. RESULTS: Six of the 25 patients (24%) with NAION had APC resistance due to the heterozygous Leiden mutation of FV. The frequency of the same genetic mutation in the control group was only 5.9%. Odds ratio calculations showed that patients with the Leiden mutation were at a significantly higher risk of NAION than control patients (p < or = 0.021). CONCLUSION: The high frequency of Leiden mutation in NAION suggests a pathogenic role of the mutation in the disease.

Using morphologic parameters of extraocular muscles for diagnosis and follow-up of Graves' ophthalmopathy: diameters, areas, or volumes?

OBJECTIVE: The objective of this study was to find the most appropriate, accurate, and convenient muscle parameter that can be used as a substitute for volume in monitoring the effectiveness of therapy of patients with Graves' ophthalmopathy. SUBJECTS AND METHODS: The four rectus muscles in 110 orbits (35 patients and 20 control subjects) were evaluated with MR imaging. The diameter at the largest extent of the muscle belly, as well as the long and the short diameters and the cross-sectional areas in a preselected coronal scan, were measured for each muscle and were compared with the corresponding muscle volume measured on contiguous T1-weighted transverse slices. RESULTS: The measured coronal area correlated well with the volume of the superior (r = 0.694, p < 0.0001) and inferior (r = 0.783, p < 0.0001) recti, and the largest transverse diameter showed strong correlation with the volume of the lateral (r = 0.868, p < 0.0001) and medial (r = 0.869, p < 0.0001) recti. For the latter muscle, the coronal area also exhibited a good correlation with the volume (r = 0.838, p < 0.0001). Coronal cross-sectional areas can be well estimated by measuring both the short and long coronal muscle diameters (r values were between 0.914 and 0.966; p < 0.0001). CONCLUSION: In Graves' ophthalmopathy, the volume of three of the rectus muscles can be well estimated by simple measurements on a single coronal slice, and the largest transverse diameter of the lateral rectus is suitable for the same purpose.