Cardiac valves are dynamic structures, exhibiting a highly specialized architecture consisting of cells and extracellular matrix with a relevant proteoglycan and glycosaminoglycan content, collagen and elastic fibers. Biological valve substitutes are obtained from xenogenic cardiac and pericardial tissues. To overcome the limits of such non viable substitutes, tissue engineering approaches emerged to create cell repopulated decellularized scaffolds. This study was performed to determine the glycosaminoglycans content, distribution, and disaccharides composition in porcine aortic and pulmonary valves and in pericardium before and after a detergent-based decellularization procedure. The fine structural characteristics of galactosaminoglycans chondroitin sulfate and dermatan sulfate were examined by FACE. Furthermore, the mechanical properties of decellularized pericardium and its propensity to be repopulated by in vitro seeded fibroblasts were investigated. Results show that galactosaminoglycans and hyaluronan are differently distributed between pericardium and valves and within heart valves themselves before and after decellularization. The distribution of glycosaminoglycans is also dependent from the vascular district and topographic localization. The decellularization protocol adopted resulted in a relevant but not selective depletion of galactosaminoglycans. As a whole, data suggest that both decellularized porcine heart valves and bovine pericardium represent promising materials bearing the potential for future development of tissue engineered heart valve scaffolds.

Apolipoproteins are very heterogeneous protein family, implicated in plasma lipoprotein structural stabilization, lipid metabolism, inflammation, or immunity. Obtaining detailed information on apolipoprotein composition and structure may contribute to elucidating lipoprotein roles in atherogenesis and to developing new therapeutic strategies for the treatment of lipoprotein-associated disorders. This study aimed at developing a comprehensive method for characterizing the apolipoprotein component of plasma VLDL, LDL, and HDL fractions from patients undergoing carotid endarterectomy, by means of two-dimensional electrophoresis (2-DE) coupled with Mass Spectrometry analysis, useful for identifying potential markers of plaque presence and vulnerability. The adopted method allowed obtaining reproducible 2-DE maps of exchangeable apolipoproteins from VLDL, LDL, and HDL. Twenty-three protein isoforms were identified by peptide mass fingerprinting analysis. Differential proteomic analysis allowed for identifying increased levels of acute-phase serum amyloid A protein (AP SAA) in all lipoprotein fractions, especially in LDL from atherosclerotic patients. Results have been confirmed by western blotting analysis on each lipoprotein fraction using apo AI levels for data normalization. The higher levels of AP SAA found in patients suggest a role of LDL as AP SAA carrier into the subendothelial space of artery wall, where AP SAA accumulates and may exert noxious effects.

The hemoglobin of 2048 newborn babies from Sardinia was analyzed by isoelectricfocusing and polyacrylamide gel electrohoresis in order to determine the level of Hb F-Sardinia (with ^Aγ^T) and the A_γ chain. Hb F-Sardinia values of 15.5 ± 2.6% were present in the ^Aγ heterozgote whereas 30.7 ± 5.2% were present in the homozygote. The ^A_γ^T gene frequency was 0.17. Most of the babies tested showed the normal ^G_γ level either in the absence of the ^A_γ^T anomaly (69.6 ± 4.1%), or in the presence of the anomaly in both the heterazygous state (70.9 ± 4.8%) and the homozygous state (71.1 ± 3.4%). Similar values were shown in nine homozygotes for β-thalassentia discovered during the screening. Nine newborn babies. (0.44%) showed particularly low ^G_γ levels (38.3 ± 6.8%) whereas 18 newborn babies showed high ^G_γ levels (83.9 ± 2.6%). The frequencies of the anomalies (0.0022 for the low ^G_γ and 0.0044 for the high ^G_γ were the lowest observed in Caucasian and other ethnic groups. Data suggest the presence of abnormal γ globin gene arrangements in the Sardinian population.