Synergistic effects of neurons and astrocytes on the differentiation of brain capillary endothelial cells in culture

Brain capillary endothelial cells form a functional barrier between blood and brain, based on the existence of tight junctions that limit paracellular permeability. Occludin is one of the major transmembrane proteins of tight junctions and its peripheral localization gives indication of tight junction formation. We previously reported that RBE4.B cells (brain capillary endothelial cells), cultured on collagen IV, synthesize occludin and correctly localize it at the cell periphery only when cocultured with neurons. In the present study, we describe a three-cell type-culture system that allowed us to analyze the combined effects of neurons and astrocytes on differentiation of brain capillary endothelial cells in culture. In particular, we found that, in the presence of astrocytes, the neuron-induced synthesis and localization of occludin is precocious as compared to cells cocultured with neurons only

Synthesis and characterisation of novel chemical conjugates based on alpha, beta-polyaspartylhydrazide and beta-cyclodextrins

A new family of supramolecular systems based on a synthetic polyaminoacid and cyclic oligosaccharides such as beta-cyclodextrins (beta-CDs) was synthesised. The pharmaceutical potential of these systems arises from the proper combination between the complexing properties of cyclodextrins and the particular pharmacokinetic profile that can be obtained by using macromolecular conjugates with a biocompatible backbone. Five supramolecular conjugates were synthesised by using alpha,beta-polyaspartylhydrazide (PAHy) as a polymeric component and various amounts of two P-CD derivatives. In particular, by reaction of PAHy with beta-CD monoaldehyde, samples named as A(1), A(2) and A(3), bearing, respectively, 4.0, 7.5 and 10.7 mol% of beta-CDs were obtained. The reaction of PAHy with 6-[aminoethyl(4'-carboxybutanamide)]-beta-CD afforded samples named as B-1 and B-2, bearing, respectively, 1.8 and 2.6 mol% of beta-CDs linked to the polymer. The occurrence of the conjugation reactions as well as the evaluation of the amount of oligosaccharides conjugated to the polymeric backbone were confirmed by FT-IR, H-1 NMR, DSC, SEC analyses and viscosimetric measurements. Molecular weight values obtained by SEC analysis were in good agreement with the theoretical increase of molecular weight of PAHy due to the beta-CD moieties linked to the polymeric backbone. Fluorescence studies on the conjugate A(3) evidenced an interaction of a probe molecule with beta-CDs linked to PAHy greater than that found with beta-CDs alone and even in both cases the formation of a 1:1 host-guest complex occurs

Coinfection of Helicobacter pylori and Epstein-Barr virus in patients with gastric disease in Southern Italy

Helicobacter pylor

MOLECULAR EPIDEMIOLOGY OF MYCOBACTERIUM TUBERCULOSIS COMPLEX IN PALERMO, ITALY

Background: We aimed to investigate the molecular epidemiology of Mycobacterium tuberculosis complex (MTBC) in the province of Palermo, Italy, by characterizing 183 isolates identified in the years 2004-2012. A comparison with 104 MTBC strains identified in the same geographic area in the years 1994-2000 was also carried out. Methods: MTBC isolates were analyzed by spoligotyping and the 24 mycobacterial interspersed repetitive unit (MIRU)-variablenumber tandem-repeat (VNTR) method typing. Susceptibility testing to streptomycin, isoniazid, rifampin and ethambutol was also carried out. Furthermore, the spoligotyping dataset obtained from 104 MTBC isolates identified in the years 1994-2000 in the same geographic area was reanalyzed. Distribution into lineages and clustering of isolates in the two periods was compared. Results: Out of the 177 isolates of MTBC submitted to molecular typing, 108 were from Italian-born and 69 from foreign-born individuals. Eleven different lineages were identified with the most represented being Haarlem, T, LAM and S. Except for the Haarlem lineage, where isolates from foreign-born patients were overrepresented, the distribution of isolates in the families belonging to the Euro-American phylogeographical clone reflected the proportions of the two subpopulations. A total of 27 strains (15.2%) were clustered and three cluster were mixed. 25% of the isolates proved to be resistant to at least one anti-TB drug, with only three isolates being categorized as multidrug resistant (MDR). When MTBC isolates identified in the years 1994-2000 were reanalyzed, lineages T, LAM, Haarlem and S proved also to be predominant. No MTBC isolates belonging to CAM, U, CAS, Turkish and Ural lineages were identified in this set of isolates. Moreover, the proportion of clustered isolates was similar, but all the clustered isolates were from Italian patients. Conclusions: A wide heterogeneity (11 lineages and 35 families-subfamilies) was detected among the MTBC strains isolated in the years 2004-2012. Ten clusters were identified, of which three mixed and four involving newly identified lineages. Compared with isolates from 1994-2000, six new lineages were detected. Comparison between distribution of lineages and the characteristics of isolates in the two consecutive periods depicts rapid and deep changes in the tuberculosis epidemiology in Palermo, Italy

Draft Genome Sequence and Biofilm Production of a Carbapenemase-Producing Klebsiella pneumoniae (KpR405) Sequence Type 405 Strain Isolated in Italy

Rapid identification and characterization of multidrug-resistant Klebsiella pneumoniae strains is essential to diagnose severe infections in patients. In clinical routine practice, K. pneumoniae is frequently identified and characterized for outbreak investigation. Pulsed-field gel electrophoresis or multilocus sequence typing could be used, but, unfortunately, these methods are time-consuming, laborious, expensive, and do not provide any information about the presence of resistance and virulence genes. In recent years, the decreasing cost of next-generation sequencing and its easy use have led to it being considered a useful method, not only for outbreak surveillance but also for rapid identification and evaluation, in a single step, of virulence factors and resistance genes. Carbapenem-resistant strains of K. pneumoniae have become endemic in Italy, and in these strains the ability to form biofilms, communities of bacteria fixed in an extracellular matrix, can defend the pathogen from the host immune response as well as from antibiotics, improving its persistence in epithelial tissues and on medical device surfaces