Protease-activated receptor-2 expression in IgA nephropathy: a potential role in the pathogenesis of interstitial fibrosis

An increasing body of evidence suggests that proteases may play a key role in the pathogenesis of tissue fibrosis. Protease-activated receptor-2 (PAR-2) is cleaved and activated by trypsin-like proteolytic enzymes, including tryptase and activated coagulation factor X (FXa). Both these soluble mediators have been demonstrated, directly or indirectly, at the interstitial level in progressive renal diseases, including IgA nephropathy (IgAN). PAR-2 mRNA and protein levels were investigated by RT-PCR and immunohistochemistry, respectively, in 17 biopsies from IgAN patients and 10 normal kidneys. PAR-2 expression was also evaluated, by RT-PCR and western blotting, in cultured human mesangial and proximal tubular cells. Finally, gene expression of plasminogen activator inhibitor-1 (PAI-1) and TGF-beta, two powerful fibrogenic factors, was evaluated in FXa-, trypsin-, and PAR-2 activating peptide-stimulated human proximal tubular cells by Northern blot. In normal kidneys, PAR-2 gene expression was barely detectable, whereas in IgAN biopsies the mRNA levels for this protease receptor were strikingly increased and directly correlated with the extent of interstitial fibrosis. Immunohistochemical staining demonstrated that PAR-2 protein expression in IgAN biopsies was mainly localized in the proximal tubuli and within the interstitial infiltrate. Proximal tubular cells in culture expressed PAR-2. Activation of this receptor by FXa in tubular cells induced a striking increase in intracellular calcium concentration. In addition, incubation of both cell lines with trypsin, FXa, or PAR-2 activating peptide caused a marked upregulation of PAI-1 gene expression that was not counterbalanced by an increased expression of plasminogen activators. Finally, PAR-2 activation induced a significant upregulation of TGF-beta gene and protein expression in both mesangial and tubular cells. On the basis of our data, we can suggest that PAR-2 expressed by renal resident cells and activated by either mast cell tryptase or FXa may induce extracellular matrix deposition modifying the PAI-1/PA balance and inducing TGF-beta expression. These molecular mechanisms may underlie interstitial fibrosis in IgAN.     

Carbonic anhydrase inhibitors: Hypoxia-activatable sulfonamides incorporating disulfide bonds that target the tumor-associated isoform IX

An approach for designing bioreductive, hypoxia-activatable carbonic anhydrase (CA, EC 4.2.1.1) inhibitors targeting the tumor-associated isoforms is reported. Sulfonamides incorporating 3,3'-dithiodipropionamide/2,2'-dithiodibenzamido moieties were prepared and reduced enzymatically/chemically in conditions present in hypoxic tumors, leading to thiols. The X-ray crystal structure of the most promising compound, 4-(2-mercaptophenylcarboxamido)benzenesulfonamide, which as disulfide showed a K(I) against hCA IX of 653 nM (in reduced form of 9.1 nM), in adduct with hCA II showed the inhibitor making favorable interactions with Gln92, Val121, Phe131, Leu198, Thr199, Thr200, Pro201, and Pro202, whereas the sulfamoyl moiety was coordinated to the Zn2+ ion. The same interactions were preserved in the adduct with hCA IX, but in addition, a hydrogen bond between the SH moiety of the inhibitor and the amide nitrogen of Gln67 was evidenced, which may explain the almost 2 times more effective inhibition of the tumor-associated isozyme over the cytosolic isoform.     

Localization of primary sensory afferents in the prothoracic ganglion of the wood ant (Formica lugubris Zett.): a combined light and electron microscopic study of secondary degeneration

Secondary degeneration of foreleg afferents was traced in the prothoracic ganglion of the common wood ant (Formica lugubris Zett) with the aid of phase contrast and electron microscopy. The legs were amputated at the coxal level in 43 workers; 1–35 days were allowed for degeneration. Localization of degenerated sensory nerves in the entering nerve root and within the neuropil of the ganglion was opumal after 4-10 days. Afferent fibers were found predominantly in the ipsilateral half of the ganglion. However, a few fibers seemed to cross the midline and to end in the medial portion of the contralateral side. No degenerations could be located within the cranial and caudal connectives. The significance of the findings with respect to the problem of functional localization pattern in the neuropil of an insect ganglion is discussed.     

Comparison of active zones in spinal cord of anesthetized and unanesthetized rats: a high voltage electron microscope study.

High voltage electronmicroscopy was applied to the study of presynaptic nerve terminals and specifically the arrangement and staining properties of synaptic vesicles in and around the active zone. The zinc iodideosmium tetroxide technique was used for the impregnation of neuropil derived from the spinal cord of anesthetized (40 mg/kg pentobarbital sodium) and unanesthetized rats. Under optimal conditions of section thickness and tilt angles stereoscopic views of a paracrystalline lattice arrangement of synaptic vesicles could be ascertained. The ZIO affinity was significantly higher in synaptic vesicles of the anesthetized as compared with the unanesthetized preparations.     

Considerations about our approach to obstetric psychoprophylaxis

Usually the term "obstetric psychoprophylaxis" refers to a specific method or technique. We prefer to consider it as a procedure that involves on one side the woman, the child and its family, and on the other the services entitled to give pre- and post-natal assistance. In order to realize this, a reformation of our methodological parameters and a critical analysis of the results obtained are required. In the courses of obstetric psychoprophylaxis that are held in the Department of Obstetrics and Gynaecology of the University of Padua we take into consideration the following themes: - Methodological approach - Professional training of the staff - Significance of psychosocial culture in the management of the pregnancy by the health services.