Tailoring dry microparticles for pulmonary drug delivery: ultrasonic spray freeze-drying with mannitol and salbutamol sulphate

Spray freeze-drying has emerged as a valid alternative to traditional spray drying to produce therapeutic dry microparticles. In particular, the spherical shape and high porosity of spray freeze-dried microparticles make them suitable for pulmonary drug delivery through dry powder inhalers. However, an appropriate particle size and fine particle fraction are required to guarantee lung deposition. This study used ultrasonic spray freeze-drying to generate dry microparticles composed of mannitol either alone or added with the bronchodilator salbutamol sulphate. The influence of the solid concentration and the feed flow rate on the particle size, morphology, surface area, porosity, and crystallinity was investigated. Growing particle size was observed, increasing the concentration and feed flow rate. Similarly, the addition of the drug led to a larger particle size and surface area. The in vitro simulation of drug deposition highlighted the dependence of the aerodynamic properties on the solid concentration and feed flow rate. Due to the lower density and particle geometric size, the highest fine particle fraction (26%) and smallest mass median aerodynamic diameter (4.4 μm) were reached at the lowest solid concentration and feed flow rate

Lipid-based nanoparticles for delivery of vaccine adjuvants and antigens : toward multicomponent vaccines

Despite the many advances that have occurred in the field of vaccine adjuvants, there are still unmet needs that may enable the development of vaccines suitable for more challenging pathogens (e.g., HIV and tuberculosis) and for cancer vaccines. Liposomes have already been shown to be highly effective as adjuvant/delivery systems due to their versatility and likely will find further uses in this space. The broad potential of lipid-based delivery systems is highlighted by the recent approval of COVID-19 vaccines comprising lipid nanoparticles with encapsulated mRNA. This review provides an overview of the different approaches that can be evaluated for the design of lipid-based vaccine adjuvant/delivery systems for protein, carbohydrate, and nucleic acid-based antigens and how these strategies might be combined to develop multicomponent vaccines

Città di villeggiatura a Santa Cesarea Terme (LE)-2009

Progetto di masterplan per una piccola città di villegiatura a Santa Cesarea Terme (Le)CONCEPT DEL PIANO 1 Città di villeggiatura e commistione funzionale (Il programma funzionale) 2 Elementi di fondazione: “infrastrutture leggere” (Il viale e il tratturo) 3 Ricostruzione del paesaggio: strategia delle tipologie di vegetazione (Il paesaggio) 4 Integrazione nella rete delle strade esistenti (Le infrastrutture) 5 Intreccio tra spazi urbani e spazi della campagna (Gli spazi ed edifici pubblici/comuni) 6 Tracciati esistenti e nuovi tracciati regolatori (I muri, i recinti, le colture e le regole di impianto) / Uso variabile della densità (Le densità alta, media, bassa) 7 Gradualità dei microclimi (Il nuovo ambiente) 8 Nuovo equilibrio energetico (Le risorse rinnovabili

Synthesis of protein conjugates adsorbed on cationic liposomes surface

The well-known Toll like receptor 9 (TLR9) agonist CpG ODN has shown promising results as vaccine adjuvant in preclinical and clinical studies, however its in vivo stability and potential systemic toxicity remain a concern. In an effort to overcome these issues, different strategies have been explored including conjugation of CpG ODN with proteins or encapsulation/adsorption of CpG ODN into/onto liposomes. Although these methods have resulted in enhanced immunopotency compared to co-administration of free CpG ODN and antigen, we believe that this effect could be further improved. Here, we designed a novel delivery system of CpG ODN based on its conjugation to serve as anchor for liposomes. Thiol-maleimide chemistry was utilised to covalently ligate model protein with the CpG ODN TLR9 agonist. Due to its negative charge, the protein conjugate readily electrostatically bound cationic liposomes composed of 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), cholesterol and dimethyldioctadecylammonium bromide (DDA) in a very high degree. The novel cationic liposomes-protein conjugate complex shared similar vesicle characteristics (size and charge) compared to free liposomes. The conjugation of CpG ODN to protein in conjunction with adsorption on cationic liposomes, could promote co-delivery leading to the induction of immune response at low antigen and CpG ODN doses. • The CpG ODN Toll-like receptor (TLR) 9 agonist was conjugated to protein antigens via thiol-maleimide chemistry. • Due to their negative charge, protein conjugates readily electrostatically bound cationic liposomes composed of 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), cholesterol and dimethyldioctadecylammonium bromide (DDA) resulting to the design of novel cationic liposomes-protein conjugate complexes. • The method is suited for the liposomal delivery of a variety of adjuvant-protein conjugates

Caracterização reológica de suspensões cimentícias mistas com cales ou filitos

   Filitos são tipos de rochas metamórficas que vêm sendo utilizados ultimamente como insumos em composições de argamassa de revestimento, em substituição à cal. A melhora da trabalhabilidade e da plasticidade dos materiais cimentícios, principais motivos para a utilização das cales, são as características obtidas também com a utilização dos filitos. Este trabalho tem como objetivo avaliar as características de suspensões cimentícias formuladas com cal ou filito, a partir de ensaios de reometria rotacional para a avaliação das propriedades das suspensões durante o fluxo, e oscilatória para a avaliação da cinética de consolidação. Os resultados mostraram que, apesar de as características físicas, químicas e mineralógicas dos pós serem muito diferentes, as cales hidratadas do tipo I (CHI) não se diferenciaram dos filitos durante o fluxo, mas durante o endurecimento foram observadas consideráveis alterações na cinética de aglomeração

RYK promotes the stemness of glioblastoma cells via the WNT/ \u3b2-catenin pathway

Glioblastoma multiforme (GBM) is characterized by a strong self-renewal potential and a poor differentiation state. Since receptor-like tyrosine kinase (RYK) activates the WNT/\u3b2-catenin pathway essential for cancer stem cell maintenance, we evaluated its contribution in conferring stemness to GBM cells. Here, we report that Ryk (related-to-receptor tyrosine kinase), an atypical tyrosine kinase receptor, is upregulated in samples from GBM patients as well as in GSCs. Ryk overexpression confers stemness properties to GBM cells through the modulation of the canonical Wnt signaling and by promoting the activation of pluripotency-related transcription factor circuitry and neurosphere formation ability. In contrast, siRNA-mediated knockdown of Ryk expression suppresses this stem-like phenotype. Rescue experiments reveal that stemness-promoting activity of Ryk is attributable, at least in part, to \u3b2-catenin stabilization. Furthermore, Ryk overexpression improves cell motility and anchorage independent cell growth. Taken together, our findings demonstrate that Ryk promotes stem cell-like and tumorigenic features to glioma cells its essential for the maintenance of GSCs and could be a target of novel therapies

Sugar-Protein Connectivity Impacts on the Immunogenicity of Site-Selective Salmonella O-Antigen Glycoconjugate Vaccines

A series of glycoconjugates with defined connectivity were synthesized to investigate the impact of coupling Salmonella typhimurium O-antigen to different amino acids of CRM197 protein carrier. In particular, two novel methods for site-selective glycan conjugation were developed to obtain conjugates with single attachment site on the protein, based on chemical modification of a disulfide bond and pH-controlled transglutaminase-catalyzed modification of lysine, respectively. Importantly, conjugation at the C186-201 bond resulted in significantly higher anti O-antigen bactericidal antibody titers than coupling to K37/39, and in comparable titers to conjugates bearing a larger number of saccharides. This study demonstrates that the conjugation site plays a role in determining the immunogenicity in mice and one single attachment point may be sufficient to induce high levels of bactericidal antibodies

Aspetti metodologici del monitoraggio di inquinanti aerodiffusi persistenti mediante “MOSS-BAGS” di nuova generazione

I muschi, grazie alla loro elevata efficienza nell’accumulo di particolato atmosferico, sono considerati degli ottimi biomonitor. Una tecnica oramai diffusa prevede l’esposizione del materiale all’interno di reticelle, formando le cosiddette “moss-bags”, che possono essere distribuite sul territorio secondo schemi espositivi ottimali. Tra i principali limiti di questa tecnica si ricorda che il muschio viene raccolto in natura, e ciò ha un certo impatto ambientale, mentre l’operazione di preparazione delle bags presenta aspetti “artigianali” che rappresentano un serio handicap per la sua diffusione. Per ovviare a questi problemi un team di ricercatori europei ha sviluppato il progetto FP7 MOSSCLONE, che ha tra gli obiettivi anche quelli di (1) individuare e caratterizzare un muschio particolarmente performante per clonarlo, coltivarlo in condizioni controllate, e quindi usarlo come materiale di uso standard, dopo un processo di devitalizzazione; (2) ottimizzare le moss-bags per forma, taglia, ampiezza delle maglie della rete di contenimento e quantità di muschio, proponendo un prototipo di nuova concezione da sottoporre a brevetto internazionale; (3) standardizzare le condizioni di esposizione, in termini di altezza e durata di esposizione; (4) confrontare i dati ottenuti con le moss-bags di nuova generazione con i dati ottenuti con i metodi chimicofisici tradizionali. In questa sede si presentano i risultati del primo anno e mezzo di attività, in particolare quelli relativi ai punti (2) e (3), basati sull’esposizione di più di 250 bags di vecchia e nuova generazione per periodi di diversa durata, da 3 a un massimo di 12 settimane, in sette siti campani con diverso uso del suolo; ciò in attesa di avere a disposizione i dati relativi ad altre esposizioni condotte in due regioni europee climaticamente molto diverse, la Galizia e l’Austria orientale. Viene inoltre illustrato in prima nazionale il nuovo prototipo

Transcathether aortic valve implantation with the new repositionable self-expandable Evolut R versus CoreValve system: A case-matched comparison

Background: Despite promising results following transcatheter aortic valve implantation (TAVI), several relevant challenges still remain. To overcome these issues, new generation devices have been developed. The purpose of the present study was to determine whether TAVI with the new self-expanding repositionable Evolut R offers potential benefits compared to the preceding CoreValve, using propensity matching. Methods: Between June 2007 and November 2015, 2148 consecutive patients undergoing TAVI either CoreValve (n = 1846) or Evolut R (n = 302) were prospectively included in the Italian TAVI ClinicalService® project. For the purpose of our analysis 211 patients treated with the Evolut R were matched to 211 patients treated with the CoreValve. An independent core laboratory reviewed all angiographic procedural data and an independent clinical events committee adjudicated all events. Results: Patients treated with Evolut R experienced higher 1-year overall survival (log rank test p = 0.045) and a significantly lower incidence of major vascular access complications, bleeding events and acute kidney injury compared to patients treated with the CoreValve. Recapture manoeuvres to optimize valve deployment were performed 44 times, allowing a less implantation depth for the Evolut R. As a consequence, the rate of more than mild paravalvular leak and new permanent pacemaker was lower in patients receiving the Evolut R. Conclusion: In this matched comparison of high surgical risk patients undergoing TAVI, the use of Evolut R was associated with a significant survival benefit at 1. year compared with the CoreValve. This was driven by lower incidence of periprocedural complications and higher rates of correct anatomic positioning

Design of a novel vaccine nanotechnology-based delivery system comprising CpGODN-protein conjugate anchored to liposomes

Although the well-known Toll like receptor 9 (TLR9) agonist CpGODN has shown promising results as vaccine adjuvant in preclinical and clinical studies, its in vivo stability and potential systemic toxicity remain a concern. In an effort to circumvent these issues, different strategies have been employed to increase its stability, localise action and reduce dosage. These include conjugation of CpGODN with proteins or encapsulation/adsorption of CpGODN into/onto liposomes, and have resulted in enhanced immunopotency compared to co-administration of free CpGODN and antigen. Here, we designed a novel delivery system of CpGODN based on its conjugation to serve as anchor for liposomes. Thiol-maleimide chemistry was utilised to covalently ligate the Group B Streptococcus (GBS) GBS67 protein antigen with the CpGODN TLR9 agonist. This treatment did not alter protein's ability to be recognised by specific antibodies or the CpGODN to function as a TLR9 agonist. Due to its negative charge, the protein conjugate readily electrostatically bound cationic liposomes composed of 1, 2-distearoyl-sn-glycero-3-phosphocholine (DSPC), cholesterol and dimethyldioctadecylammonium bromide (DDA). The novel cationic liposomes-protein conjugate complex (GBS67-CpGODN+L) shared similar vesicle characteristics (size and charge) compared to free liposomes but exhibited different structure and morphology. Following intramuscular immunisation, GBS67-CpGODN+L formed a vaccine depot at the injection site and induced a remarkable increase of functional immune responses against GBS compared to the simple co-administration of GBS67, CpGODN and liposomes. This work demonstrates that the conjugation of CpGODN to GBS67 in conjunction with adsorption on cationic liposomes, can promote co-delivery leading to the induction of a multifaceted immune response at low antigen and CpGODN doses. Our findings highlight the potential for harnessing the immunostimulatory properties of different adjuvants to develop more effective nanostructure-based vaccine platforms