Electron microscopy analysis of skin biopsies in CADASIL disease.

Cerebral Autosomal Dominant Arteriopathy with Subcortical Infarcts and Leukoencephalopathy (CADASIL) is an inherited vascular disorder, non-amyloid and non-atherosclerotic, affecting predominantly the central nervous system. We examined samples of skin biopsies from six patients (men, 43-52-year-old), admitted for treatment in the Neurology Clinic regarding the presence of partial motor impairment on upper and lower right limbs, facial asymmetry and phrasing impairment (three of the patients); These three patients had family history remarkable for early-onset strokes: mother and two brothers deceased by early strokes (40-50-year-old). Skin biopsy samples were fixed in glutaraldehyde and post-fixed in osmium tetroxyde. After dehydration, tissue samples were embedded in Epon. Ultrathin sections were mounted on copper grids and stained with uranyl acetate and lead citrate as usual and examined with a transmission electron microscope Phillips CM100. In all cases ultrastructural study showed granular osmiophilic material (GOM) in extracellular locations, between degenerating smooth muscle cells in dermal arteries or in their indentations. Deposits of GOM varied in size and electron density. Degeneration and loss of smooth muscle cells (SMCs) leads to abnormal enlargement of the space between these cells Ultrastructural analysis in three cases showed chromatin condensation and peripheral aggregation of nuclear material suggesting cells entry to apoptosis. These aspects and the marked destruction of the vascular wall were correlated with MRI findings and the severity of clinical manifestations at these patients. Our study showed that findings of GOM deposits, degeneration and loss of SMCs (probably by apoptosis), cell adhesion elements disturbance are characteristic for CADASIL disease and sufficient for diagnose of certainty. Moreover, electron microscopy analysis of skin biopsies is a useful tool for a differential diagnosis and can be considered as first choice method.

Two cases of Marfan syndrome.

The early identification of the Marfan Syndrome should be essential for the prevention of the aortic dilatation and dissection, but the wide phenotypic expression of the disorder makes the clinical diagnosis very difficult. The aim of this study is to emphasise the necessity of a widely applicable method of morpho-clinical diagnosis. The diagnosis was confirmed on surgical biopsies from ascending aortic wall and aortic cusps, using routine morphological techniques. The macroscopical examination revealed a thin ascending aortic wall, presenting a transversal intimal tear, without secondary dissecting hematoma, and an aortic dilated annulus, producing aortic regurgitation. Microscopically, the diagnosis was supported by the identification of cystic medical necrosis, a specific degenerative lesion in the Marfan Syndrome, in patients with or without family history of aortic aneurysms. This study defines the morpho-clinical changes of the disease, emphasising the necessity of a permanent monitorization of the patient after surgery, due to the risk of late complications.

The risk factors of fibrosis in nonalcoholic steatohepatitis.

UNLABELLED: Nonalcoholic steatohepatitis (NASH) can be a severe progressive liver disease leading to the development of cirrhosis. The aim of this study was to assess the risk factors of fibrosis in NASH. METHODS: The clinical, biochemical and histological features of 80 patients with NASH admitted at University Hospital from 1998 to 2000, were analyzed. The patients had no alcoholic, viral, autoimmune, drug induced or genetic liver disease. The fibrosis and necroinflammatory activity was associated with clinico-biochemical parameters and with oxidative stress. The clinical and biochemical parameters consist of: age, sex, body mass index (BMI), serum glucose, tryglycerides, alanine aminotransferase (ALT) and serum iron study (transferring saturation and ferritin). Liver iron overload was assessed by Pearls'staining and graded in four classes. The oxidative stress was evaluated by the levels of serum malon-dyaldehide (MDA) and serum total gluthatione (GSI4). RESULTS: Septal fibrosis was present in 30 patients (27%) including cirrhosis in 4 patients (5%). Age > 45 years, B.M.I. >30 Kg/m2, serum tryglycerides >180 mg/dl, hyperglycemia > 220 mg/dl, serum ALT > 3N, increased hepatic iron and transferrin saturation percentage were independently associated with sepal fibrosis. Linear regression analysis showed that increased hepatic iron had the greatest association with the increase of lipid peroxidation (MDA > 250 nanomol/dl) and the decrease of serum gluthatione (< 40 micromol/dl). CONCLUSIONS: The clinical and biochemical parameters which consist of age, BMI, serum ALT, glucose and tryglycerides, Perls' grade and the serum index of oxidative stress (MDA and GSH) are independent risk factors for fibrosis in NASH raising therapeutic implications for the management of these patients.