63 research outputs found

    Comparative Study of Immunogenicity of Split, Intradermal and MF59-adjuvanted Influenza Vaccines in Elderly Institutionalized Subjects

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    Abstract The reduced immunogenicity and effectiveness of influenza vaccines in subjects presenting high risk of influenza-related complications, hospitalization and death, led the innovative drive to search for new strategies to implement the immune response elicited by influenza vaccines including addition of adjuvants, and use of alternative routes of antigen delivery.In this study we evaluated and compared the immune antibody response induced in 252 elderly volunteers living in nursing homes after immunization with three different 2012-2013 seasonal trivalent inactivated influenza vaccines: a conventional split vaccine (n=26), and two potentiated vaccines (a subunit vaccine adjuvanted with MF59 (n=137) or a split vaccine administered intradermally (n=89)), specially licensed for elderly people. Haemagglutination inhibiting (HI) antibody titers were assessed in blood samples collected before and one month after vaccination.The results were evaluated as increase in HI titers found comparing pre- and post-vaccination sera and according to the Committee for Medicinal Products for Human Use (CHMP) criteria for approval of influenza vaccines in the elderly. Significant antibody increases and fulfillment of all the three CHMP requirements were observed against A/H3N2 and B antigens following immunization with the two potentiated vaccines. After immunization with conventional vaccine responses were lower against A/H3N2 and equivalent against the B antigen. The two potentiated vaccines induced significant antibody increases against A/H1N1 antigen, however, only one of the CHMP criteria was reached. The HI antibody increases after conventional vaccine were significant only for the geometric mean titer and none of the CHMP criteria was fulfilled. The antibody responses induced by the two potentiated vaccines against the three vaccine antigens wereequivalent although post-vaccination titers against the B antigen tended to be higher in subjects vaccinated with intradermal vaccine than in individuals receiving MF59-adjuvanted vaccine. In conclusion the use of MF59 adjuvant and intradermal vaccination appear to be appropriate strategies to address the challenge of declining immune response in the elderly after influenza vaccination

    N-Glycans mutations rule oligomeric assembly and functional expression of P2X3 receptor for extracellular ATP

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    N-Glycosylation affects the function of ion channels at the level of multisubunit assembly, protein trafficking, ligand binding and channel opening. Like the majority of membrane proteins, ionotropic P2X receptors for extracellular ATP are glycosylated in their extracellular moiety. Here, we used site-directed mutagenesis to the four predicted N-glycosylation sites of P2X3 receptor (Asn139, Asn170, Asn194 and Asn290) and performed comparative analysis of the role of N-glycans on protein stability, plasma membrane delivery, trimer formation and inward currents. We have found that in transiently transfected HEK293 cells, Asn170 is apparently the most important site for receptor stability, since its mutation causes a primary loss in protein content and indirect failure in membrane expression, oligomeric association and inward current responses. Even stronger effects are obtained when mutating Thr172 in the same glycosylation consensus. Asn194 and Asn290 are the most dispensable, since even their simultaneous mutation does not affect any tested receptor feature. All double mutants containing Asn170 mutation or the Asn139/Asn290 double mutant are instead almost unable to assemble into a functional trimeric structure. The main emerging finding is that the inability to assemble into trimers might account for the impaired function in P2X3 mutants where residue Asn170 is replaced. These results improve our knowledge about the role of N-glycosylation in proper folding and oligomeric association of P2X3 recepto

    The P2Y4 receptor forms homo-oligomeric complexes in several CNS and PNS neuronal cells

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    It is well established that several cell surface receptors interact with each other to form dimers and oligomers, which are essential for their activation. Since little is known about the quaternary structure of P2Y receptors, in the present work, we investigated the expression of the G-protein-coupled P2Y4 subunit as monomeric or higher-order complex protein. We examined both endogenously expressed P2Y4 subtype with the aid of specific anti-P2Y4 antiserum, and heterologously transfected P2Y4-tagged receptors with the use of antitag antibodies. In both cases, we found the P2Y4 receptor displaying molecular masses corresponding to monomeric, dimeric and oligomeric structures. Experiments performed in the absence of reducing agents demonstrated that there is a strict correlation among the multiple protein bands and that the multimeric forms are at least partially assembled by disulphide bonds. The direct demonstration of P2Y4 homodimerisation comes instead from co–transfection and differential co–immunoprecipitation experiments, with the use of differently tagged P2Y4 receptors and antitag antibodies. The structural propensity of the P2Y4 protein to form homo-oligomers may open the possibility of a novel regulatory mechanism of physiopathological functions for this and additional P2Y receptors

    Bollettino Sismico Italiano: Analisys of Early Aftershocks of the 2016 MW 6.0 Amatrice, MW 5.9 Visso and MW 6.5 Norcia earthquakes in Central Italy

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    The Amatrice-Visso-Norcia seismic sequence is the most important of the last 30 years in Italy. The seismic sequence started on 24 August, 2016 and still is ongoing in central Apennines. At the end of February 2017 more than 57,000 events were located, 80,000 events up to the end of September 2017 (Fig. 1). The mainshocks of the sequence occurred on 24 August 2016 (Mw 6.0 and Mw 5.4), 26 October 2016 (Mw 5.4 and Mw 5.9), 30 October 2016 (Mw 6.5), 18 January 2017 (four earthquakes Mw≄ 5.0). In this seismic sequence, all the waveforms recorded by temporary stations deployed by the SISMIKO emergency group (stations T12**; Moretti et al., 2016) where available in real- time at the surveillance room of INGV. Because of the high level of seismicity and the dense seismic network installed in the region, more than 150 events per day were located at the end of February 2017; still 60 events per day were located up to the end of August 2017.The Amatrice-Visso-Norcia is the most important seismic sequence since 2015, the time when the analysis procedures of the BSI group (Bollettino Sismico Italiano) were revised (Nardi et al., 2015). BSI is now available every four months on the web: bulletins contain revised earthquakes (location and magnitude) with ML≄ 1.5, quasi-real time revision of ML≄ 3.5 earthquakes and phase arrivals from waveforms recorded on seismic stations available from the European Integrated Data Archive (EIDA), (Mazza et al., 2012). These last procedures allow the integration of signals from temporary seismic stations (Moretti et al., 2014) installed by the emergency group SISMIKO (Moretti and Sismiko working group, 2016), even when they are not in real time transmission, if they are rapidly archived in EIDA, together with real time signals from the seismic stations of the permanent INGV network. The analysis strategy of the BSI group for the Amatrice -Visso - Norcia seismic sequence (AVN.s.s in the following) was to select the earthquakes located in the box with min/max latitude: 42.2/43.2 - and min/max longitude: 12.4/14.1 to prepare a special volume of BSI on the seismic sequence.PublishedTrieste, Italy1SR. TERREMOTI - Servizi e ricerca per la Societ