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During inflammation, endothelial cells (EC) are the first elements to be exposed to mediators circulating in the bloodstream. EC react with finely tuned responses mediated by different pathways, including the Phosphoinositide (PI) signal transduction system. The PI pathway contributes to a variety of cell functions, including hormone secretion, neurotransmitter signal transduction, cell growth, membrane trafficking, ion channel activity, cytoskeleton regulation, cell cycle control, apoptosis, embryonic development, organogenesis, and cell/tissue polarity. The Phosphoinositide-specific phospholipase C (PI-PLC) enzymes contribute to the regulation of the spatio-temporal balance of molecules belonging to the PI system. Thirteen mammalian PI-PLC enzymes have been identified, divided into six sub-families on the basis of amino acid sequence, domain structure and mechanism of recruitment. Isoforms within sub-families share sequence similarity, common domain organization, and general regulatory mechanism. The expression of PI-PLCs is strictly tissue specific, and evidences suggest that it varies under different conditions, such as tumor progression or cell activation. We obtained the complete panel of expression of PI-PLC isoforms in human umbilical vein endothelial cells (HUVEC), a widely used experimental model for EC. Then, we analyzed the mRNA concentration of PI-PLCs in LPS treated HUVEC by using the multiliquid bioanalyzer methodology after 3, 6, 24 48 and 72 hours from LPS administration. Marked differences in the expression of most PI-PLC codifying genes were evident., Italian Journal of Anatomy and Embryology, Vol 118, No 2 (Supplement) 2013

Much recent experimental evidence suggests that inflammatory reactions are involved in aneurysm formation and progression [1]. Vessel wall weakening has been attributed to the infiltration of a variety of cells, which leads to the upregulation of multiple cytokines [2]. A growing interest is currently focused in the definition of cytokines associated with the final stage of the disease that precedes the rupture. In the present study full aortic segments were collected from patients with thoracic aortic aneurysms, according to the declaration of Helsinki. Control aorta tissue from organ donors was included as reference. Microtome sections of paraffin embedded samples were analyzed for the presence of IL-17 and IL-23 positivity. Aortic sections were also stained with hematoxylin and eosin and Verhoeff-Van Gieson for elastin. Immunohistochemistry detected IL-17 and IL-23 positive cells at the level of adventitia and muscle cell layer. Our data reinforce previous results showing that IL-17 plays a central role in the promotion of vascular inflammation. The identification of main cytokines released during inflammatory processes underlying aneurysm formation and progression may lead to the set up of pharmaceutic strategies that may prove effective for the stabilization of aortic aneurysms., Italian Journal of Anatomy and Embryology, Vol 118, No 2 (Supplement) 2013

The rationale of our study was to test the potential of the autologous elements of peripheral white blood cells and platelets to promote neoangiogenesis, thereby overcoming hypoxemia, the major obstacle to the regeneration of the tissue. An highly innovative technique, based on an autologous leuco-platelet preparation, applied by direct infiltration to the ulcer was employed to treat 65 patients with ulcers of different etiologies. Hyaluronic acid was used as a scaffold, supplemented with autologous cells. Patients were followed up until complete healing of the lesions. Morphometric analysis was perfomed on histologic sections to determine the presence of neoangiogenesis. The wound healing was obtained in all cases after 2 until 22 months with this treatment (median time is 11.4 months). More than half of the patients recovered completely within the first 12 months. Morphometric analysis on histologic sections determined the presence of an abundant neovasculature in the close proximity of the infiltrations of leuco-platelet concentrate. Our data show that autologous leucoplatelet preparation layered onto ischemic ulcers may improve neoangiogenesis leading to wound healing. These findings suggest that this cell-based therapy may be a useful tool for the treatment for intractable skin ulcers resulting from diabetes, ischemia and collagen diseases., Italian Journal of Anatomy and Embryology, Vol 118, No 2 (Supplement) 2013

Our data suggest that MBP interacts with astroglial cells, priming their activation, as proven by HLA class II modulation and HO-1 induction. MBP binding seems to be mediated by LRP,an endocytosis-mediating receptor. Such an interaction is followed by an increase of intracellular ROS content which, possibly, is counteracted by HO-1 induction. Many reports outlined the expression of HO-1 in multiple sclerosis and EAE, but its role in these diseases is still unclear (Chakrabarty et al. 2003), even if a growing body of evidence suggests that overexpression of HO-1 may protect tissues from immune-mediated injury either through prevention of oxidative damage or via a local immunomodulatory influence on infiltrating inflammatory cells (Kapturczak et al., 2004).

Previous experiments have shown that human neoplastic and embryonic glial cell lines synthesize and secrete in culture, alpha 2 macroglobulin (alpha 2M), a broad spectrum proteinase inhibitor present in serum and extracellular fluids. The present study was aimed to investigate the presence of alpha 2M receptors on glial cell membrane, since several non-neural cell types producing alpha 2M also express alpha 2M receptors. By flow cytometric analysis, immunofluorescence and immunoelectronmicroscopy techniques we demonstrate an alpha 2M receptor-related immunoreactivity on the plasma membrane of a human glioma cell line. Ultrastructural experiments reveal a close colocalization of immunoreactivities for alpha 2M and its receptor in clathrin-coated pits and vesicles, structures typically involved in receptor-mediated endocytic pathways.

The proteinase inhibitor set in skeletal muscle is poorly characterized at present. This study was aimed to investigate in mouse skeletal muscle 1) the tissue-associated counterpart, if any, of serum protease inhibitors (which may also play antiproteolytic functions in tissues) and 2) calpastatin, a tissue inhibitor of calcium-activated neutral proteases (calpains). Triton-extracts were prepared from muscle homogenates of mice, which had been perfused extensively with phosphate buffered saline (PBS) (under deep anesthesia) to remove blood inhibitors. Among various inhibitors tested, the following muscle-associated inhibitors were identified by western-blotting: alpha-2-macroglobulin (185, 165, 35 kDa), alpha-1-antitrypsin (52 kDa), inter-alpha-trypsin inhibitor (220, 180 kDa) and calpastatin (70 kDa). Combined light microscope and confocal immunohistochemical experiments revealed that, in all muscles examined (soleus, plantaris, extensor digitorum longus) the above specific immunoreactivities were localized outside the muscle fibers (in periendomysium, blood vessel wall) as well as within them. Inter-alpha-trypsin inhibitor, however, completely lacked the intracellular localization. This wide distribution of proteinase inhibitors suggests that numerous muscular structures may be normally protected from unwanted proteolysis, thus providing an essential background for further studies on pathological models with altered proteolysis (m. dystrophy, denervation atrophy, etc.).

SUMMARY 1. The aim of the present study was to evaluate whether postural blood pressure (BP) change could predict office-daytime BP disparity and the nocturnal BP fall in young, mild essential hypertensives. We investigated 411 males aged between 18 and 45 years with never treated borderline to mild hypertension. BP was measured three times after a 5 min rest in the supine position and thereafter three times after 2 min of standing. The mean of six BP measurements obtained during two visits in the lying position was defined as office BP. 2. Twenty-four hour ambulatory BP monitoring was performed with either the A and D TM-2420 model 7 or the ICR Spacelabs 90207. BP values were averaged for day—and nighttime periods. The nocturnal BP fall was defined as the difference between the average day—and night-time BP. 3. The standing-lying difference was significantly inversely correlated with the office-daytime difference for both systolic blood pressure (SBP) (r = -0.34, P 0.001) and diastolic blood pressure (DBP) (r = -0.24, P 0.001). These correlations did not change when the obese subjects (body mass index > 30 kg/m2) were excluded from the analysis. No significant correlation between standing-lying difference and nocturnal BP fall was found. 4. Our results indicate that white coat hypertension assessed as the office-daytime BP disparity is partially related (in a negative fashion) to the BP reaction to standing. The postural BP change does not predict nocturnal BP fall in young, mild essential hypertensives.

The distribution of inter-alpha-trypsin inhibitor (ITI) and related inhibitors was investigated in normal human tissues and body fluids by using an enzyme-linked immunosorbent assay (ELISA) and a streptavidin-biotin-peroxidase immunohistochemical technique. ITI-related immunoreactivity was localized in different cell types of various organs, such as liver, kidney, testis, gross intestine, cutis and brain. Specific immunoreactivity was also detected in serum, urine and bronchial mucus. This widespread, but not ubiquitous pattern of localization suggests that, in addition to the well known plasmatic role, ITI and/or ITI-related inhibitors may play a number of different physiological roles in various human tissues.