Pegylated-liposomes increase the efficacy of Idelalisib in lymphoma B-cells

New drugs and technologies are continuously developed to improve the efficacy and minimize the critical side effects of cancer treatments. The present investigation focuses on the development of a liposomal formulation for Idelalisib, a small-molecule kinase inhibitor approved for the treatment of lymphoid malignancies. Idelalisib is a potent and selective antitumor agent, but it is not indicated nor recommended for first-line treatment due to fatal and serious toxicities. Herein, liposomes are proposed as a delivery tool to improve the therapeutic profile of Idelalisib. Specifically, PEGylated liposomes were prepared, and their physicochemical and technological features were investigated. Light-scattering spectroscopy and cryo-transmission electron microscopy revealed nanosized unilamellar vesicles, which were proved to be stable in storage and in simulated biological fluids. The cytotoxicity of the liposome formulation was investigated in a human non-Hodgkin's lymphoma B cell line. Idelalisib was able to induce death of tumor cells if delivered by the nanocarrier system at increased efficacy. These findings suggest that combining Idelalisib and nanotechnologies may be a powerful strategy to increase the antitumor efficacy of the drug

Self-recognition and Ca2+-dependent carbohydrate–carbohydrate cell adhesion provide clues to the Cambrian explosion

Author Posting. © The Authors, 2009. This is the author's version of the work. It is posted here by permission of Oxford University Press for personal use, not for redistribution. The definitive version was published in Molecular Biology and Evolution 26 (2009): 2551-2561, doi:10.1093/molbev/msp170.The Cambrian explosion of life was a relatively short period ca. 540 million years ago that marked a generalized acceleration in the evolution of most animal phyla, but the trigger of this key biological event remains elusive. Sponges are the oldest extant Precambrian metazoan phylum and thus a valid model to study factors that could have unleashed the rise of multicellular animals. One such factor is the advent of self/non-self recognition systems, which would be evolutionarily beneficial to organisms to prevent germ cell parasitism or the introduction of deleterious mutations resulting from fusion with genetically different individuals. However, the molecules responsible for allorecognition probably evolved gradually before the Cambrian period, and some other (external) factor remains to be identified as the missing triggering event. Sponge cells associate through calcium-dependent, multivalent carbohydrate-carbohydrate interactions of the g200 glycan found on extracellular proteoglycans. Single molecule force spectroscopy analysis of g200-g200 binding indicates that calcium affects the lifetime (+Ca/-Ca: 680 s/3 s) and bond reaction length (+Ca/-Ca: 3.47 Å/2.27 Å). Calculation of mean g200 dissociation times in low and high calcium within the theoretical framework of a cooperative binding model indicates the non-linear and divergent characteristics leading to either disaggregated cells or stable multicellular assemblies, respectively. This fundamental phenomenon can explain a switch from weak to strong adhesion between primitive metazoan cells caused by the well documented rise in ocean calcium levels at the end of Precambrian time. We propose that stronger cell adhesion allowed the integrity of genetically uniform animals composed only of “self” cells, facilitating genetic constitutions to remain within the metazoan individual and be passed down inheritance lines. The Cambrian explosion might have been triggered by the coincidence in time of primitive animals endowed with self/non-self recognition, and of a surge in sea water calcium that increased the binding forces between their calcium-dependent cell adhesion molecules.D.A. and A.K. acknowledge financial support from the Collaborative Research Center SFB 613 from the Deutsche Forschungsgemeinschaft (DFG), and X.F.-B. acknowledges financial support from grants BIO2002-00128, BIO2005-01591, and CSD2006-00012 from the Ministerio de Ciencia y Tecnología, Spain, which included Fondo Europeo de Desarrollo Regional funds, and from grant 2005SGR-00037 from the Generalitat de Catalunya, Spain

Loading of beclomethasone in liposomes and hyalurosomes improved with mucin as effective approach to counteract the oxidative stress generated by cigarette smoke extract

In this work beclomethasone dipropionate was loaded into liposomes and hyalurosomes modified with mucin to improve the ability of the payload to counteract the oxidative stress and involved damages caused by cigarette smoke in the airway. The vesicles were prepared by dispersing all components in the appropriate vehicle and sonicating them, thus avoiding the use of organic solvents. Unilamellar and bilamellar vesicles small in size (~117 nm), homogeneously dispersed (polydispersity index lower than 0.22) and negatively charged (~−11 mV), were obtained. Moreover, these vesicle dispersions were stable for five months at room temperature (~25◦C). In vitro studies performed using the Next Generation Impactor confirmed the suitability of the formulations to be nebulized as they were capable of reaching the last stages of the impactor that mimic the deeper airways, thus improving the deposition of beclomethasone in the target site. Further, biocompatibility studies performed by using 16HBE bronchial epithelial cells confirmed the high biocompatibility and safety of all the vesicles. Among the tested formulations, only mucin-hyalurosomes were capable of effectively counteracting the production of reactive oxygen species (ROS) induced by cigarette smoke extract, suggesting that this formulation may represent a promising tool to reduce the damaging effects of cigarette smoke in the lung tissues, thus reducing the pathogenesis of cigarette smoke-associated diseases such as chronic obstructive pulmonary disease, emphysema, and cancer

Potential therapeutic effect of curcumin loaded hyalurosomes against inflammatory and oxidative processes involved in the pathogenesis of rheumatoid arthritis : the use of fibroblast-like synovial cells cultured in synovial fluid

In the present work curcumin loaded hyalurosomes were proposed as innovative systems for the treatment of rheumatoid arthritis. Vesicles were prepared using a one-step and environmentally friendly method. Aiming at finding the most suitable formulation in terms of size, surface charge and stability on storage, an extensive pre-formulation study was performed using different type and amount of phospholipids. Curcumin loaded vesicles prepared with 180 mg/ml of Phospholipon 90G (P90G) and immobilized with sodium hyaluronate (2 mg/ml) were selected because of their small size (189 nm), homogeneous dispersion (PI 0.24), negative charge (−35 mV), suitable ability to incorporate high amount of curcumin (E% ∼88%) and great stability on storage. The in vitro study using fibroblast-like synovial cells cultured in synovial fluid, demonstrated the ability of these vesicles to downregulate the production of anti-apoptotic proteins IAP1 and IAP2 and stimulate the production of IL-10, while the production of IL-6 and IL-15 and reactive oxygen species was reduced, confirming their suitability in counteracting pathogenesis of rheumatoid arthritis

Copolimeri ad innesto PLGA-g-PVP per la veicolazione e il rilascio controllato di farmaci antimalariali

I copolimeri PLGA-g-PVP, aventi una catena di PLGA centrale ad alto peso molecolare e segmenti laterali oligomerici di PVP sono state recentemente sintetizzati tramite un processo sintetico a singolo step. La reazione consiste in una polimerizzazione a trasferimento di catena del N-vinilpirrolidone (N-VP) a partire da PLGA fuso, in presenza di un iniziatore radicalico quale \ue8 l\u2019AIBN (azobisisobutirronitrile). Diverse percentuali di PVP sono state innestati sulla catena principale, in modo da variare l\u2019idrofilia del copolimero. Lo scopo di questo lavoro \ue8 stato quello di formulare tali copolimeri PLGA-g-PVP in presenza di farmaci idrofobici, quali l\u2019artemisinina e la curcumina, sotto forma di nanocapsule da utilizzare come sospensioni antimalariche

A novel aminohydroxy sulfonamide formulated in PEGylated liposomes with potential antitumor activity

Cancer ranks as a leading cause of death worldwide, with liver cancer being one of the most commonly diagnosed. Currently, several molecules are being studied in order to find new therapeutic options that can reduce cancer recurrence rate and increase patient survival. This study proposes PEGylated liposomes for the delivery of a newly synthesized aminohydroxy sulfonamide, BupM-NH2, which has shown dose-dependent cytotoxicity towards hepatic tumor cells. The prepared PEG-liposomes were nanosized, spherical, and unilamellar, as shown by light scattering data and cryo-TEM micrographs. The physical stability of the PEG-liposomes was preserved when tested in simulated body fluids. Similar results were found for the storage stability evaluation. Furthermore, the PEG-liposomes efficiently entrapped BupM-NH2 and modulated its release. The antitumor activity of BupM-NH2 in PEG-liposomes was assessed in vitro in hepatocarcinoma cells. The viability assay showed that PEG-liposomes were able to control the release of BupM-NH2 over time and induce the death of HepG2 cells at a concentration about two-times lower than that required by free BupM-NH2 (IC50: 33.31 vs. 57.05 μM). Furthermore, the liposomal formulation showed a less cytotoxic effect against non-cancerous cell line (IHH) compared to the free molecule (IC50 > 200 vs. 106.9 μM), encouraging further investigation to confirm its effective and safe use

The prion-like RNA-processing protein HNRPDL forms inherently toxic amyloid-like inclusion bodies in bacteria

BACKGROUND: The formation of protein inclusions is connected to the onset of many human diseases. Human RNA binding proteins containing intrinsically disordered regions with an amino acid composition resembling those of yeast prion domains, like TDP-43 or FUS, are being found to aggregate in different neurodegenerative disorders. The structure of the intracellular inclusions formed by these proteins is still unclear and whether these deposits have an amyloid nature or not is a matter of debate. Recently, the aggregation of TDP-43 has been modelled in bacteria, showing that TDP-43 inclusion bodies (IBs) are amorphous but intrinsically neurotoxic. This observation raises the question of whether it is indeed the lack of an ordered structure in these human prion-like protein aggregates the underlying cause of their toxicity in different pathological states. RESULTS: Here we characterize the IBs formed by the human prion-like RNA-processing protein HNRPDL. HNRPDL is linked to the development of limb-girdle muscular dystrophy 1G and shares domain architecture with TDP-43. We show that HNRPDL IBs display characteristic amyloid hallmarks, since these aggregates bind to amyloid dyes in vitro and inside the cell, they are enriched in intermolecular β-sheet conformation and contain inner amyloid-like fibrillar structure. In addition, despite their ordered structure, HNRPDL IBs are highly neurotoxic. CONCLUSIONS: Our results suggest that at least some of the disorders caused by the aggregation of human prion-like proteins would rely on the formation of classical amyloid assemblies rather than being caused by amorphous aggregates. They also illustrate the power of microbial cell factories to model amyloid aggregation. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12934-015-0284-7) contains supplementary material, which is available to authorized users

Amyloidogenic Regions and Interaction Surfaces Overlap in Globular Proteins Related to Conformational Diseases

Protein aggregation underlies a wide range of human disorders. The polypeptides involved in these pathologies might be intrinsically unstructured or display a defined 3D-structure. Little is known about how globular proteins aggregate into toxic assemblies under physiological conditions, where they display an initially folded conformation. Protein aggregation is, however, always initiated by the establishment of anomalous protein-protein interactions. Therefore, in the present work, we have explored the extent to which protein interaction surfaces and aggregation-prone regions overlap in globular proteins associated with conformational diseases. Computational analysis of the native complexes formed by these proteins shows that aggregation-prone regions do frequently overlap with protein interfaces. The spatial coincidence of interaction sites and aggregating regions suggests that the formation of functional complexes and the aggregation of their individual subunits might compete in the cell. Accordingly, single mutations affecting complex interface or stability usually result in the formation of toxic aggregates. It is suggested that the stabilization of existing interfaces in multimeric proteins or the formation of new complexes in monomeric polypeptides might become effective strategies to prevent disease-linked aggregation of globular proteins

Caratterizzazione di alcuni siti della rete accelerometrica nazionale al fine di individuare la risposta sismica locale

Le indagini geotecniche finalizzate alla stima della risposta sismica locale si limitano molto spesso ai primi 30 m di profondità, valore che è diventato uno standard per la classificazione delle caratteristiche di un sito. Negli anni ’90 Borcherdt (1994) e Martin e Dobry (1994) suggerirono 30 m come la profondità standard di indagine per la verifica delle strutture. Boore et al. (1993, 1994, 1997) e Boore e Joyner (1997) basarono le regressioni per il calcolo delle leggi predittive del moto del suolo sullo stesso parametro. Nel 1997 negli Stati Uniti il National Earthquake Hazards Reduction Program (NEHRP) nella stesura delle norme tecniche per le costruzioni in zona sismica (FEMA, 1997) utilizza per la prima volta il parametro Vs30 come indice per la classificazione dei suoli, con lo scopo di definirne l’amplificazione. Le norme tecniche per le costruzioni in zona sismica della comunità Europea, EC8 (ENV, 1998) ente da dati provenienti dagli Stati Uniti occidentali e, utilizzando dati provenienti dalla stessa regione, Wald & Mori (2000) segnalano che le VS,30 non sono molto ben correlate con l’entità dell’amplificazione, in quanto esiste una forte dispersione dei dati. La figura 1.1 mostra il rapporto tra le amplificazioni, mediate sull’intervallo di frequenza compreso tra 3-5 Hz. raccomandano lo stesso parametro per suddividere i terreni, anche se le classi differiscono in parte dalla classificazione NEHRP. Infine, anche in Italia, le Norme Tecniche per le Costruzioni (Normative Tecniche per le Costruzioni, Gazzetta Ufficiale del 14/01/2008) adottano la stessa suddivisione dei terreni adottata dall’EC8.L’attendibilità della velocità delle onde di taglio nei primi 30 m (VS,30) come estimatore della risposta sismica di un sito, in termini di frequenza e amplificazione, è tuttavia molto discussa.Innanzitutto il parametro è stato ricavato unicamente da dati provenienti dagli Stati Uniti occidentali e, utilizzando dati provenienti dalla stessa regione, Wald & Mori (2000) segnalano che le Vs30 non sono molto ben correlate con l’entità dell’amplificazione, in quanto esiste una forte dispersione dei dati. La figura 1.1 mostra il rapporto tra le amplificazioni, mediate sull’intervallo di frequenza compreso tra 3-5 Hz. I valori risultano effettivamente molto dispersi, ma questo risultato può essere spiegato col fatto che non tutte le classi di sito hanno frequenza di risonanza compreso in questo intervallo di frequenza. Perciò per alcuni siti la media è stata calcolata nell’intorno della frequenza di risonanza (sulle amplificazioni massime), mentre per altri è stata calcolata sulle armoniche superiori, che hanno ampiezze minori. Lavori eseguiti con dati provenienti da altre regioni sottolineano come le Vs30 non siano buoni estimatori per la predizione di amplificazioni in bacini profondi (Park & Hashash, 2004), per la stima delle amplificazioni in altre regioni (Stewart et al., 2003) o in presenza di inversioni di velocità (Di Giacomo et al., 2005). Uno studio recente, eseguito su dati giapponesi (Zhao et al., 2006) si è evitato l’uso della Vs30 perché strati spessi di terreno rigido posti sopra il substrato roccioso amplificano il moto di lungo periodo, mentre gli strati sottili e soffici tendono ad amplificare il moto di corto periodo: ciò significa che la VS,30 non può rappresentare il periodo predominante del sito, dato che si basa solo sugli strati superficiali. Secondo Mucciarelli e Gallipoli (2006) il confronto tra l’amplificazione sismica al sito e la Vs30 mostra che quest’ultimo parametro non è adeguato per spiegare gli effetti di sito osservati in Italia a causa delle situazioni geologiche particolari che sono diffuse nel nostro paese. La figura 1.2 mostra la distribuzione dell’ampiezza rispetto alla classe di sito, in cui si vede che le classi sono mal discriminate e le mediane delle classi A e B (indicate dalla linea nera) sono uguali. È però necessario notare che questo grafico è stato costruito utilizzando le ampiezze ricavate col metodo dei rapporti spettrali H/V, ma in letteratura (Bard, 1999) è dimostrato che tali rapporti spettrali permettono di stimare la frequenza di risonanza, ma falliscono nella stima del valore di amplificazione. In particolare la Vs30 sottostima gli effetti locali ai siti con inversione di velocità e li sovrastima in siti con bacini profondi. La Vs30 sembra fornire dei buoni risultati solo in siti che abbiano un profilo di velocità monotono, crescente con la profondità e un forte contrasto di impedenza nella prima decina di metri. Questo studio si propone di verificare l’attendibilità della velocità delle onde di taglio valutate nei primi 30 m come estimatore della risposta sismica di un sito. Per questo scopo sono state selezionate 45 stazioni della Rete Accelerometrica Nazionale, di cui si conoscono i profili stratigrafici e i profili di velocità delle onde di taglio e di compressione. Inoltre sono state raccolte le registrazioni strong motion relative ai terremoti registrati da queste stazioni. Gli effetti di sito sono stati valutati in due modi: · Le registrazioni sono state utilizzate per calcolare i rapporti spettrali H/V per ricavare la frequenza fondamentale propria di ciascun sito (f0) e il relativo valore di amplificazione; · I profili di velocità delle onde di taglio sono serviti per ricavare il modello teorico monodimensionale per il calcolo della funzione di trasferimento del sito, eseguito per mezzo del modello proposto da Haskell e Thomson (Haskell, 1953, Thomson 1950), da cui ricavare la f0 e l’amplificazione. I valori ottenuti con i due metodi sono stati poi confrontati per verificare la congruenza dei risultati. I profili di velocità hanno permesso di classificare le stazioni utilizzando la velocità media delle onde di taglio nei primi 30 m (Vs30), secondo la normativa italiana. I risultati ottenuti dalla valutazione della risposta di ciascun sito, espressi in termini di frequenza fondamentale e amplificazione, sono stati correlati con la rispettiva classe di sito per verificare l’attendibilità del parametro delle Vs30 come estimatore degli effetti di sito

Elective cancer surgery in COVID-19-free surgical pathways during the SARS-CoV-2 pandemic: An international, multicenter, comparative cohort study

PURPOSE As cancer surgery restarts after the first COVID-19 wave, health care providers urgently require data to determine where elective surgery is best performed. This study aimed to determine whether COVID-19–free surgical pathways were associated with lower postoperative pulmonary complication rates compared with hospitals with no defined pathway. PATIENTS AND METHODS This international, multicenter cohort study included patients who underwent elective surgery for 10 solid cancer types without preoperative suspicion of SARS-CoV-2. Participating hospitals included patients from local emergence of SARS-CoV-2 until April 19, 2020. At the time of surgery, hospitals were defined as having a COVID-19–free surgical pathway (complete segregation of the operating theater, critical care, and inpatient ward areas) or no defined pathway (incomplete or no segregation, areas shared with patients with COVID-19). The primary outcome was 30-day postoperative pulmonary complications (pneumonia, acute respiratory distress syndrome, unexpected ventilation). RESULTS Of 9,171 patients from 447 hospitals in 55 countries, 2,481 were operated on in COVID-19–free surgical pathways. Patients who underwent surgery within COVID-19–free surgical pathways were younger with fewer comorbidities than those in hospitals with no defined pathway but with similar proportions of major surgery. After adjustment, pulmonary complication rates were lower with COVID-19–free surgical pathways (2.2% v 4.9%; adjusted odds ratio [aOR], 0.62; 95% CI, 0.44 to 0.86). This was consistent in sensitivity analyses for low-risk patients (American Society of Anesthesiologists grade 1/2), propensity score–matched models, and patients with negative SARS-CoV-2 preoperative tests. The postoperative SARS-CoV-2 infection rate was also lower in COVID-19–free surgical pathways (2.1% v 3.6%; aOR, 0.53; 95% CI, 0.36 to 0.76). CONCLUSION Within available resources, dedicated COVID-19–free surgical pathways should be established to provide safe elective cancer surgery during current and before future SARS-CoV-2 outbreaks