Differential response to hepatic differentiation stimuli of amniotic epithelial cells isolated from four regions of the amniotic membrane

Human Amniotic Epithelial Cells (hAEC) isolated from term placenta are a promising source for regenerative medicine. However, it has long been debated whether the hAEC population consists of heterogeneous or homogeneous cells. In a previous study, we investigated the characteristics of hAEC isolated from four different regions of the amniotic membrane finding significant heterogeneity. The aim of this study was to evaluate the hepatic differentiation capability of hAEC isolated from these four regions. Human term placentae were collected after caesarean section and hAEC were isolated from four regions of the amniotic membrane (R1-R4, according to their relative distance from the umbilical cord) and treated in hepatic differentiation conditions for 14 days. hAEC-derived hepatocyte-like cells showed marked differences in the expression of hepatic markers: R4 showed higher levels of Albumin and Hepatocyte Nuclear Factor (HNF) 4α whereas R1 expressed higher Cytochrome P450 enzymes, both at the gene and protein level. These preliminary results suggest that hAEC isolated from R1 and R4 of the amniotic membrane are more prone to hepatic differentiation. Therefore, the use of hAEC from a specific region of the amniotic membrane should be taken into consideration as it could have an impact on the outcome of therapeutic applications.     

The role of carrier ATPases and neuromediators in neurological disorders

Physiological and biochemical processes which take place in the nervous system at stroke and neurotrauma are reviewed, and the experience of using low doses of steroid derivatives with piridamole in the treatment of central nervous system (CNS) disorders is summarized. ATPases (including Na,K-ATPase) are reported to play an important role in CNS functioning, the correlation between Na,K-ATPase activity and the extent of CNS injury is revealed. The use of NMR-spectroscopy method for investigation of brain and spinal cord condition in vivo is suggested.     

Tyrosine mutations within the alpha platelet-derived growth factor receptor kinase insert domain abrogate receptor-associated phosphatidylinositol-3 kinase activity without affecting mitogenic or chemotactic signal transduction.

A phosphatidylinositol-3 (PI-3) kinase activity of unknown biological function associates with tyrosine kinase-containing proteins, including a number of growth factor receptors after ligand stimulation. In the beta platelet-derived growth factor (beta PDGF) receptor, phosphorylation of a specific tyrosine residue within the kinase insert domain was required for its interaction with this enzyme. We show that substitutions of phenylalanine for tyrosine residue 731 or 742 within the kinase insert domain of the alpha PDGF receptor do not impair PDGF-induced tyrosine phosphorylation of the receptor or of an in vivo substrate, phospholipase C-gamma. Moreover, phosphatidylinositol turnover in response to ligand stimulation is unaffected. However, both lesions markedly impair receptor association with PI-3 kinase. Antiphosphotyrosine antibody-recoverable PI-3 kinase was also dramatically reduced in PDGF-stimulated cells expressing either mutant receptor. Since neither mutation abolished PDGF-induced mitogenesis or chemotaxis, we conclude that alpha PDGF receptor-associated PI-3 kinase activity is not required for either of these major PDGF signalling functions.     

Conclusive diagnosis of hepatic and pancreatic malignancies by fine needle aspiration

A study of the diagnosis of hepatic and pancreatic malignancies by fine needle aspiration (FNA) was made, based on 221 aspirates obtained from 209 patients with histologic or clinical confirmation: 159 with hepatic and 50 with pancreatic lesions. The values of sensitivity, specificity and predictivity for positive FNA results were, respectively, 0.84, 0.96 and 1.0 for the liver and 0.76, 1.0 and 1.0 for the pancreas. The composition of the case material showed an incidence of malignant tumors of the liver and pancreas of 84% and 60%, respectively (among which the primary malignancies were 39% and 48%), while nonneoplastic lesions had incidences of 14% and 40%. However, conclusive FNA diagnoses of the histologic type of the primary and the site of origin of metastatic tumors were made in 60% of the hepatic lesions but in only 9% of the pancreatic lesions. Primary hepatocellular carcinoma was diagnosed by FNA of the liver in 95% of the cases; FNA specifically diagnosed 42% of intrahepatic bile duct carcinomas and 40% of hepatic metastases. These findings correlate with the unique cytologic features of primary hepatocellular carcinoma of intrahepatic rather nonspecific morphology of carcinoma of intrahepatic and extrahepatic origin, as well as of pancreatic ductal origin.     

Enzyme-linked immunosorbent assay (ELISA) for the identification of mosquito bloodmeals

An ELISA for the identification of mosquito bloodmeals (MBM) is described. The technique makes use of commercial antisera and allows to investigate at least three different animal sources on the same MBM. The efficiency of identification is 100% for up to 20 hrs-digested MBM.     

Rifampicin-resistant bacteriophage PBS2 infection and RNA polymerase in Bacillus subtilis

Bacteriophage PBS2 replication is unaffected by rifampicin and other rifamycin derivatives, which are potent inhibitors of Bacillus subtilis RNA synthesis. Extracts of gently-lysed infected cells contain a DNA-dependent RNA polymerase activity which is specific for uracil-containing PBS2 DNA. The PBS2-induced RNA polymerase is insensitive to rifamycin derivatives which inhibit the host's RNA polymerase.