Hybrid Aryl-Ether-Ketone and Hyperbranched Epoxy Networks

In this dissertation, relationships between chemical structures, cure kinetics and network architectures are correlated to bulk mechanical properties for novel, hybrid epoxy-amine networks. The work is split into two primary sections: the first is the synthesis and characterization of multifunctional glassy networks based on aryl-ether-ketone diamine curatives, while the second is based on the synthesis and characterization of hyperbranched epoxy polymers and their resulting networks. Three aryl-ether-ketone (AEK) diamines of increasing molecular weights were synthesized and used to cure 4,4’-tetraglycidylether of diaminodiphenylmethane (TGDDM); the resulting networks were compared to 4,4’-diaminodiphenyl sulfone cured TGDDM. Architectural differences were created by varying cure profiles, and characteristics such as sub-Tg motions and free volume were altered to study bulk properties such as thermal stability, glass transition, mechanical properties, and moisture resistance. Additional analysis coupled molecular dynamics simulations and free volume data to relate AEK molecular-level characteristics to bulk properties. Simulation showed that enhanced chain packing and conformational freedom provided networks with similar hole free volume characteristics but created increased fractional free volume with increase in Mc. Activation energies of the two sub-Tg relaxations correlated with the free volume findings. Conformational freedom was also related to mechanical properties, which was related to enhanced secondary interactions. Additionally, AEK diamines were copolymerized with DDS to form blended, hybrid networks with TGDDM. Blended networks offered further insight into the effects of chemical composition and Mc effects by varying AEK concentration. It was found that low concentrations of AEK-diamines could significantly alter DDS-cured networks. The second section of this work involved the blending of an epoxide-functional hyperbranched polymer (HBE) into a glassy epoxy network. It was found that the network-level incorporation and high concentration of secondary interactions allowed simultaneous improvements in modulus and toughness. Further thermal and mechanical improvements were found by incorporation of POSS-units onto the hyperbranched polymer. The bulk property improvements were found to correlate with the multiscale dispersion of POSS. Finally, POSS-HBE carbon-fiber composites were fabricated and tested. Composite properties were related to those of the matrix material

Medication Safety in Emergency Medical Services: Approaching an Evidence-Based Method of Verification to Reduce Errors

Lack of verification is often cited as a root cause of medication errors; however, medication errors occur in spite of conventional verification practices and it appears that human factors engineering (HFE) can inform the design of a more effective method. To this end, an HFE-driven process was designed and implemented in an urban, Midwestern emergency medical service agency. Medication error data were collected over a 54-month period, 27 months before and after implementation. A decrease in the average monthly error rate was realized for all medications administered (49.0%) during the post-intervention time period. The average monthly error rate for fentanyl, a commonly administered analgesic, demonstrated a 71.1% error rate decrease. This study is the first to evaluate the effectiveness of a team-based cross-check process for medication verification to prevent errors in the prehospital setting

Evidence for the involvement of lipid rafts localized at the ER-mitochondria associated membranes in autophagosome formation

Mitochondria-associated membranes (MAMs) are subdomains of the endoplasmic reticulum (ER) that interact with mitochondria. This membrane scrambling between ER and mitochondria appears to play a critical role in the earliest steps of autophagy. Recently, lipid microdomains, i.e. lipid rafts, have been identified as further actors of the autophagic process. In the present work, a series of biochemical and molecular analyses has been carried out in human fibroblasts with the specific aim of characterizing lipid rafts in MAMs and to decipher their possible implication in the autophagosome formation. In fact, the presence of lipid microdomains in MAMs has been detected and, in these structures, a molecular interaction of the ganglioside GD3, a paradigmatic “brick” of lipid rafts, with core-initiator proteins of autophagy, such as AMBRA1 and WIPI1, was revealed. This association seems thus to take place in the early phases of autophagic process in which MAMs have been hypothesized to play a key role. The functional activity of GD3 was suggested by the experiments carried out by knocking down ST8SIA1 gene expression, i.e., the synthase that leads to the ganglioside formation. This experimental condition results in fact in the impairment of the ER-mitochondria crosstalk and the subsequent hindering of autophagosome nucleation. We thus hypothesize that MAM raft-like microdomains could be pivotal in the initial organelle scrambling activity that finally leads to the formation of autophagosome. Introduction The interaction of the endoplasmic reticulum (ER) with mito- chondria occurs via certain subdomains of the ER, named mitochondria-associated membranes (MAMs), which allow membrane “scrambling” between these organelles and contrib- utes to the complex series of ER functions.1-3 Indeed, several regions of close apposition between the ER and mitochondria were detected by studies carried out several years ago.4,5 How- ever, since these studies provided only ultrastructural observa- tions, these reports remained neglected for a long time. In particular, while morphological evidence of the physical juxta- position between ER and mitochondria was described since 1959,6 it was experimentally proven only 30 y later. In fact, ana- lyzing ER fractions copurified with mitochondria in velocity sedimentation assays, mainly from rat liver cells, it was observed that mitochondria can tightly be associated with ele- ments of the ER and that the communication and intermixing between ER and mitochondria can be mediated by MAMs.7-12 These works also showed that these cosedimenting fractions were enriched in enzymes responsible for the synthesis of lipids. These findings suggested that MAMs could act as sites

Changes in membrane lipids drive increased endocytosis following Fas ligation

Once activated, some surface receptors promote membrane movements that open new portals of endocytosis, in part to facilitate the internalization of their activated complexes. The prototypic death receptor Fas (CD95/Apo1) promotes a wave of enhanced endocytosis that induces a transient intermixing of endosomes with mitochondria in cells that require mitochondria to amplify death signaling. This initiates a global alteration in membrane traffic that originates from changes in key membrane lipids occurring in the endoplasmic reticulum (ER). We have focused the current study on specific lipid changes occurring early after Fas ligation. We analyzed the interaction between endosomes and mitochondria in Jurkat T cells by nanospray-Time-of-flight (ToF) Mass Spectrometry. Immediately after Fas ligation, we found a transient wave of lipid changes that drives a subpopulation of early endosomes to merge with mitochondria. The earliest event appears to be a decrease of phosphatidylcholine (PC), linked to a metabolic switch enhancing phosphatidylinositol (PI) and phosphoinositides, which are crucial for the formation of vacuolar membranes and endocytosis. Lipid changes occur independently of caspase activation and appear to be exacerbated by caspase inhibition. Conversely, inhibition or compensation of PC deficiency attenuates endocytosis, endosome-mitochondria mixing and the induction of cell death. Deficiency of receptor interacting protein, RIP, also limits the specific changes in membrane lipids that are induced by Fas activation, with parallel reduction of endocytosis. Thus, Fas activation rapidly changes the interconversion of PC and PI, which then drives enhanced endocytosis, thus likely propagating death signaling from the cell surface to mitochondria and other organelles

Human complement factor H. Two factor H proteins are derived from alternatively spliced transcripts

The human complement factor H is an important component in the control of the alternative pathway of complement activation. We have previously shown that at least three factor H homologous mRNA species of 4.3 kb, 1.8 kb and 1.4 kb in length are constitutively expressed in human liver. In addition, several factor Hrelated proteins have been detected in human sera using antibodies directed against the classical human factor H glycoprotein of 150 kDa.The structure of the additional polypeptides has not been shown so far. Circumstantial evidence suggests that the 1.8-kb mRNA might encode the 43-kDa factor H-like polypeptide. Here we report the isolation, characterization and eukaryotic expression of the first full-length cDNA representing the major 4.3-kb mRNA and the 1.8-kb mRNA of human factor H. We show that the 4.3-kb transcript encodes the 150-kDa-factor H glycoprotein and the 1.8-kb mRNA the 43-kDa factorH polypeptide. The identity of the two cDNA in a region of 1400 nucleotides suggests that the two factor H-related transcripts are derived from one gene by a process of alternative splicing

Autophagy generates citrullinated peptides in human synoviocytes: a possible trigger for anti-citrullinated peptide antibodies

OBJECTIVES: Autophagy may represent a functional processing event that creates a substrate for autoreactivity. In particular, autophagy may play a role in the pathogenesis of RA, since autophagy is a key cellular event involved in the generation of citrullinated peptides, with consequent breakage of tolerance. Thus, in RA, autophagy may be the common feature in several situations (including smoking, joint injury and infection) that may drive the adaptive responses to citrullinated self-proteins. The aim of this study was the analysis, in vitro, of the role of autophagy in the generation of citrullinated peptides and, in vivo, of the relationship between autophagy and the production of anti-CCP antibodies (Abs). METHODS: For autophagy induction, fibroblast-like synoviocytes, primary fibroblasts and monocytes were stimulated with tunicamycin or rapamycin. Peptidyl arginine deiminase activity was tested by enzyme-linked immunosorbent assay, and protein citrullination was evaluated by western blotting. The main citrullinated RA candidate antigens, vimentin, α-enolase and filaggrin, were demonstrated by immunoprecipitation. The relationship between autophagy and anti-CCP Abs was analysed in 30 early-active RA patients. RESULTS: Our results demonstrated in vitro a role for autophagy in the citrullination process. Cells treated with tunicamycin or rapamycin showed peptidyl arginine deiminase 4 activation, with consequent protein citrullination. Immunoblotting and immunoprecipitation experiments, using specific Abs, identified the main citrullinated proteins: vimentin, α-enolase and filaggrin. In vivo, a significant association between levels of autophagy and anti-CCP Abs was observed in treatment-naïve early-active RA patients. CONCLUSION: These findings support the view that the processing of proteins in autophagy generates citrullinated peptides recognized by the immune system in RA

Targeting lipid rafts as a strategy against coronavirus

Lipid rafts are functional membrane microdomains containing sphingolipids, including gangliosides, and cholesterol. These regions are characterized by highly ordered and tightly packed lipid molecules. Several studies revealed that lipid rafts are involved in life cycle of different viruses, including coronaviruses. Among these recently emerged the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). The main receptor for SARS-CoV-2 is represented by the angiotensin-converting enzyme-2 (ACE-2), although it also binds to sialic acids linked to host cell surface gangliosides. A new type of ganglioside-binding domain within the N-terminal portion of the SARS-CoV-2 spike protein was identified. Lipid rafts provide a suitable platform able to concentrate ACE-2 receptor on host cell membranes where they may interact with the spike protein on viral envelope. This review is focused on selective targeting lipid rafts components as a strategy against coronavirus. Indeed, cholesterol-binding agents, including statins or methyl-β-cyclodextrin (MβCD), can affect cholesterol, causing disruption of lipid rafts, consequently impairing coronavirus adhesion and binding. Moreover, these compounds can block downstream key molecules in virus infectivity, reducing the levels of proinflammatory molecules [tumor necrosis factor alpha (TNF-α), interleukin (IL)-6], and/or affecting the autophagic process involved in both viral replication and clearance. Furthermore, cyclodextrins can assemble into complexes with various drugs to form host–guest inclusions and may be used as pharmaceutical excipients of antiviral compounds, such as lopinavir and remdesivir, by improving bioavailability and solubility. In conclusion, the role of lipid rafts-affecting drugs in the process of coronavirus entry into the host cells prompts to introduce a new potential task in the pharmacological approach against coronavirus

Longevity city: Urbanistica e qualità della vita. Un percorso virtuoso per garantire il ben-essere

The present study starts from the idea that the abandonment of small centres in inner areas can be contrasted through sustainable and innovative strategies The regeneration of the depopulating centres is possible only investing in the territorial capital and the endogenous resources assuming at the base a "project of knowledge" of the potential present therein. It means committing to safeguarding the cultural and landscaping heritage by experimenting with new design paradigms as briefly represented in the "ecoborghi" pilot project.     The aim of the research is to restore and enhance the original meaning of small centres through a re-reading of the real conditions and a re-proposal of the potential and opportunities for qualitative development starting from the great historical-cultural heritage in the essential interaction between protection, maintenance and management of historical landscapes. To do this we propose the pilot project ecoborghi, an enhancement project aimed at redeveloping public spaces in historic centres that is an integral part of the Local Landscape Observatory. Longevity city: Urbanistica e qualità della vita. Un percorso virtuoso per garantire il ben-essereSi assume come principio base quello secondo il quale l’abbandono dei piccoli centri può essere contrastato attraverso un percorso virtuoso che garantisca una strategia urbanistica diversa rispetto a quella fin qui attuata. Le considerazioni che seguono sono finalizzate a rendere sostenibile un percorso innovativo a favore delle aree interne e dei centri abbandonati. La rigenerazione dei centri in via di spopolamento è possibile solo se si riesce ad investire sul capitale territoriale e sulle risorse endogene assumendo alla base un “progetto di conoscenza” delle potenzialità presenti. Significa impegnarsi a salvaguardare il patrimonio culturale e paesaggistico sperimentando nuovi paradigmi progettuali per come rappresentati nel progetto pilota “ecoborghi”. Lo scopo della ricerca è di restituire ed esaltare il significato originario dei piccoli centri attraverso una ri-lettura delle reali condizioni e di una riproposizione delle potenzialità e delle opportunità di sviluppo qualitativo a partire dal grande patrimonio storico-culturale nell’imprescindibile interazione tra protezione, manutenzione e gestione dei paesaggi storici. Per fare ciò si propone Il progetto pilota ecoborghi, un progetto di valorizzazione volto a riqualificare spazi pubblici nei centri storici che è parte integrante dell’Osservatorio Locale del Paesaggio.Si assume come principio base quello secondo il quale l’abbandono dei piccoli centri può essere contrastato attraverso un percorso virtuoso che garantisca una strategia urbanistica diversa rispetto a quella fin qui attuata. Le considerazioni che seguono sono finalizzate a rendere sostenibile un percorso innovativo a favore delle aree interne e dei centri abbandonati. La rigenerazione dei centri in via di spopolamento è possibile solo se si riesce ad investire sul capitale territoriale e sulle risorse endogene assumendo alla base un “progetto di conoscenza” delle potenzialità presenti. Significa impegnarsi a salvaguardare il patrimonio culturale e paesaggistico sperimentando nuovi paradigmi progettuali per come rappresentati nel progetto pilota “ecoborghi”. Lo scopo della ricerca è di restituire ed esaltare il significato originario dei piccoli centri attraverso una ri-lettura delle reali condizioni e di una riproposizione delle potenzialità e delle opportunità di sviluppo qualitativo a partire dal grande patrimonio storico-culturale nell’imprescindibile interazione tra protezione, manutenzione e gestione dei paesaggi storici. Per fare ciò si propone Il progetto pilota ecoborghi, un progetto di valorizzazione volto a riqualificare spazi pubblici nei centri storici che è parte integrante dell’Osservatorio Locale del Paesaggio. Longevity city: Urban planning and quality of life. A virtuous path to guarantee well-beingThe present study starts from the idea that the abandonment of small centres in inner areas can be contrasted through sustainable and innovative strategies The regeneration of the depopulating centres is possible only investing in the territorial capital and the endogenous resources assuming at the base a "project of knowledge" of the potential present therein. It means committing to safeguarding the cultural and landscaping heritage by experimenting with new design paradigms as briefly represented in the "ecoborghi" pilot project.     The aim of the research is to restore and enhance the original meaning of small centres through a re-reading of the real conditions and a re-proposal of the potential and opportunities for qualitative development starting from the great historical-cultural heritage in the essential interaction between protection, maintenance and management of historical landscapes. To do this we propose the pilot project ecoborghi, an enhancement project aimed at redeveloping public spaces in historic centres that is an integral part of the Local Landscape Observatory

VKORC1 and CYP2C9 polymorphisms related to adverse events in case-control cohort of anticoagulated patients

Vitamin K antagonists (VKAs) are highly effective but have a narrow therapeutic index and require routine monitoring of the INR. The primary aim of pharmacogenetics (PGx) is to optimize patient care, achieving drug treatments that are personalized according to the genetic profile of each patient. The best-characterized genes involved in VKA PGx involve pharmacokinetics (VKORC1) and pharmacodynamics (CYP2C9) of VKA metabolism. The role of these genes in clinical outcomes (bleeding and thrombosis) during oral anticoagulant (OAC) therapy is controversial. The aim of the present study was to evaluate any potential association between genotype VKORC1 and CYP2C9 and adverse events (hemorrhagic and/or thrombotic), during initiation and long-term VKA treatment, in Caucasian patients. Furthermore, we aimed to determine if the concomitant prescription of other selected drugs affected the association between genotype and adverse events.We performed a retrospective, matched case-control study to determine associations between multiple gene variants, drug intake, and any major adverse effects in anticoagulated patients, monitored in 2 Italian anticoagulation clinics.Our results show that anticoagulated patients have a high risk of adverse events if they are carriers of 1 or more genetic polymorphisms in the VKORC1 (rs9923231) and CYP2C9 (rs1799853 and rs1057910) genes.Information on CYP2C9 and VKORC1 variants may be useful to identify individualized oral anticoagulant treatment for each patient, improve management and quality of VKA anticoagulation control, and monitor drug surveillance in pharmacovigilance programs

Advances in the Pathophysiology of Thrombosis in Antiphospholipid Syndrome. Molecular Mechanisms and Signaling through Lipid Rafts

The pathological features of antiphospholipid syndrome (APS) are related to the activity of circulating antiphospholipid antibodies (aPLs) associated with vascular thrombosis and obstetric complications. Indeed, aPLs are not only disease markers, but also play a determining pathogenetic role in APS and exert their effects through the activation of cells and coagulation factors and inflammatory mediators for the materialization of the thromboinflammatory pathogenetic mechanism. Cellular activation in APS necessarily involves the interaction of aPLs with target receptors on the cell membrane, capable of triggering the signal transduction pathway(s). This interaction occurs at specific microdomains of the cell plasma membrane called lipid rafts. In this review, we focus on the key role of lipid rafts as signaling platforms in the pathogenesis of APS, and propose this pathogenetic step as a strategic target of new therapies in order to improve classical anti-thrombotic approaches with "new " immunomodulatory drugs