Lipid-based nanovesicles for nanomedicine

Molecular self-assembly has enabled the fabrication of biologically inspired, advanced nanostructures as lipid-based nanovesicles (L-NVs). The oldest L-NVs, liposomes, have been widely proposed as potential candidates for drug delivery, diagnostic and/or theranostic applications and some liposome-based drug products have already stepped from the lab-bench to the market. This success is attributed to their ability to encapsulate both hydrophobic and/or hydrophilic molecules, efficiently carry and protect them within the body and finally deliver them at the target site. These positive features are also coupled with high biocompatibility. However, liposomes still present some unsolved drawbacks, such as poor colloidal stability, short shelf-life, restricted and expensive conditions of preparation because of the inherent nature of their fundamental constituents (phospholipids). The new tools available in the self-assembly of controlled molecules have significantly advanced the field of L-NV design and synthesis, and nonliposomal L-NVs have been recently developed; this new generation of nanovesicles can represent a paradigm shift in nanomedicine: they may complement liposomes, showing their advantages and overcoming most of their drawbacks. Clearly, being still young, their rocky way to the clinic first and then to the market has just started and it is still long, but they have all the potentialities to reach their objective target. The purpose of this review is to first present the large plethora of L-NVs available, focusing on this new generation of non-liposomal L-NVs and showing their similarities and differences with respect to their ancestors (liposomes). Since the overspread of a nanomaterial to the market is also strongly dependent on the availability of technological-scale preparation methods, we will also extensively review the current approaches exploited for L-NV production. The most cutting-edge approaches based on compressed fluid (CF) technologies will be highlighted here since they show the potential to represent a game-change in the production of L-NVs, favouring their step from the bench to the market. Finally, we will briefly discuss L-NV applications in nanomedicine, looking also for their future perspectives.This work was financially supported by MINECO (DGI), Spain, grants BE-WELL CTQ2013-40480-R, TERARMET (RTC-2014-2207-1), QUATFORFRAG (RTC-2014-2183-5) and UNDERLIPIDS (RTC-2015-3303-1), and by AGAUR, Generalitat de Catalunya, “Grant 2014-SGR-17“. N.G. acknowledges the European Commission (EC) (FP7-PEOPLE-2013-Initial Training Networks (ITN) ‘‘NANO2FUN’’ Project No. 607721) for their Postdoctoral contract. The authors appreciate support from LIPOCELL project financed by CIBER-BBN and Praxis Pharmaceuticals and also acknowledge the financial support from Instituto de Salud Carlos III, through “Acciones CIBER”. The Networking Research Center on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN) is an initiative funded by the VI National R&D&I Plan 2008–2011, Iniciativa Ingenio 2010, Consolider Program, CIBER Actions and financed by the Instituto de Salud Carlos III with assistance from the European Regional Development Fund.Peer reviewe

Prevalence of intestinal parasitosis in patients afferent to the Hospital of Desio (Monza e Brianza), Italy

From may 2008 to June 2010 we have analyzed faecal samples of 2306 subjects: in the subjects was performed the coproparassitological standard examination and in 523 of them was performed scotch test. Of the 2306 subjects examined, 101 resulted positive, 59 to the coproparasitological exam, 43 to scotch test, and 1 to both of them (Taenia spp.). In three cases we have found positivity for two different parassitological species. Pathogenic parasites were found in 77 cases, 61 autoctones and 16 immigrates. The helminths found were Enterobius vermicularis, the prevalent, and Taenia spp., Ascaris lumbricoides, Diphillobothrium latum, Trichuris Trichiura, while the Protozoa were Giardia duodenalis and Entamoeba histolytica/dispar. Non pathogenic species were found in 27 cases, 15 among autoctones and 12 among immigrates: in detail Entamoeba coli, Endolimax nana, Blastocystis hominis. The total prevalence of parassitosis is low (4,2%), 20,0% in immigrate subjects and 3,4% in native subjects. Our results show that the 0-10 age group have an higher prevalence for Enterobius vermicularis. Some considerations are related for improving the complete or more exhaustive diagnosis concerning the intestinal parasitosis

Highly Conserved Non-Coding Sequences and the 18q Critical Region for Short Stature: A Common Mechanism of Disease?

Background. Isolated growth hormone deficiency (IGHD) and multiple pituitary hormone deficiency (MPHD) are heterogeneous disorders with several different etiologies and they are responsible for most cases of short stature. Mutations in different genes have been identified but still many patients did not present mutations in any of the known genes. Chromosomal rearrangements may also be involved in short stature and, among others, deletions of 18q23 defined a critical region for the disorder. No gene was yet identified. Methodology/Principal Findings. We now report a balanced translocation X;18 in a patient presenting a breakpoint in 18q23 that was surprisingly mapped about 500 Kb distal from the short stature critical region. It separated from the flanking SALL3 gene a region enriched in highly conserved non-coding elements (HCNE) that appeared to be regulatory sequences, active as enhancers or silencers during embryonic development. Conclusion. We propose that, during pituitary development, the 18q rearrangement may alter expression of 18q genes or of X chromosome genes that are translocated next to the HCNEs. Alteration of expression of developmentally regulated genes by translocation of HCNEs may represent a common mechanism for disorders associated to isolated chromosomal rearrangements. © 2008 Rizzolio et al

Evaluation of an automated method for urinocolture screening

Introduction: Urinary tract infections are one of the most common diseases found in medical practice and are diagnosed with traditional methods of cultivation on plates. In this study we evaluated an automated instrumentation for screening of the urinocultures that can provide results quickly and guarantee traceability. The comparison of results obtained with automatic and plate methods is reported. Methods: 316 urine samples including midstream urine, urine catheter and urine bag have been analyzed by Alfred 60 (Alifax) through light scattering technology that measures the replication of the bacteria. Simultaneously, the samples were sown on agar plates CPS3,Agar Cled, Mc Conkey Agar. Results:A total of 316 samples were analyzed by the automated method, 190 resulted negative, all confirmed by culture, while 126 were found positive. 82 cases were confirmed positive in culture plate, 65 with significant isolation of bacteria and 17 with polymicrobial flora with a significant charge. 44 cases were negative in culture plate but positive for the automated method. Conclusions: The absence of false negative results at low charges can represent a starting point to introduce an automated method for urinocolture screening

Prevalence of intestinal parasitosis in patients afferent to the Hospital of Desio (Monza e Brianza), Italy

From may 2008 to June 2010 we have analyzed faecal samples of 2306 subjects: in the subjects was performed the coproparassitological standard examination and in 523 of them was performed scotch test. Of the 2306 subjects examined, 101 resulted positive, 59 to the coproparasitological exam, 43 to scotch test, and 1 to both of them (Taenia spp.). In three cases we have found positivity for two different parassitological species. Pathogenic parasites were found in 77 cases, 61 autoctones and 16 immigrates. The helminths found were Enterobius vermicularis, the prevalent, and Taenia spp., Ascaris lumbricoides, Diphillobothrium latum, Trichuris Trichiura, while the Protozoa were Giardia duodenalis and Entamoeba histolytica/dispar. Non pathogenic species were found in 27 cases, 15 among autoctones and 12 among immigrates: in detail Entamoeba coli, Endolimax nana, Blastocystis hominis. The total prevalence of parassitosis is low (4,2%), 20,0% in immigrate subjects and 3,4% in native subjects. Our results show that the 0-10 age group have an higher prevalence for Enterobius vermicularis. Some considerations are related for improving the complete or more exhaustive diagnosis concerning the intestinal parasitosis

Pressure-responsive, surfactant-free CO2‑based nanostructured fluids

Microemulsions are extensively used in advanced material and chemical processing. However, considerable amounts of surfactant are needed for their formulation, which is a drawback due to both economic and ecological reasons. Here, we describe the nanostructuration of recently discovered surfactant-free, carbon dioxide (CO2)-based microemulsion-like systems in a water/organic-solvent/CO2 pressurized ternary mixture. “Water-rich” nanodomains embedded into a “water-depleted” matrix have been observed and characterized by the combination of Raman spectroscopy, molecular dynamics simulations, and small-angle neutron scattering. These single-phase fluids show a reversible, pressure-responsive nanostructuration; the “water-rich” nanodomains at a given pressure can be instantaneously degraded/expanded by increasing/decreasing the pressure, resulting in a reversible, rapid, and homogeneous mixing/demixing of their content. This pressure-triggered responsiveness, together with other inherent features of these fluids, such as the absence of any contaminant in the ternary mixture (e.g., surfactant), their spontaneous formation, and their solvation capability (enabling the dissolution of both hydrophobic and hydrophilic molecules), make them appealing complex fluid systems to be used in molecular material processing and in chemical engineering.N.G. acknowledges the European Commission (EC) (FP7-PEOPLE-2013-Initial Training Networks (ITN) “‘NANO2FUN’” project no. 607721) for her Postdoctoral contract. A.S.B. thanks funding from the European Research Council under ERC starting grant agreement no. 637654. The authors appreciate the economical support from DGI, MINECO, Spain (Grant MAT2016-80826-R and “Severo Ochoa” Programme for Centres of Excellence in R&D (SEV- 2015-0496)), and the Instituto de Salud Carlos III, through “Acciones CIBER”. The ICTS “Nanbiosis”, more specifically U6 Unit, is also acknowledged since some of the studies here reported have been performed there. Neutron beam time at the Institut Laue-Langevin (ILL) Grenoble, France, is acknowledged together with Dr. Thomas Sottmann, who has kindly provided the high-pressure cell used for SANS measurements. The authors acknowledge the valuable support of Julian Jonathan Schuster from FAU with respect to the processing of Raman spectra and the valuable help of David Bowyer during experiments at ILL.We acknowledge support by the CSIC Open Access Publication Initiative through its Unit of Information Resources for Research (URICI).Peer reviewe

Highly Stable and Red‐Emitting Nanovesicles Incorporating Lipophilic Diketopyrrolopyrroles for Cell Imaging

Diketopyrrolopyrroles (DPPs) have recently attracted much interest as very bright and photostable red‐emitting molecules. However, their tendency to form nonfluorescent aggregates in water through the aggregation‐caused quenching (ACQ) effect is a major issue that limits their application under the microscope. Herein, two DPP molecules have been incorporated into the membrane of highly stable and water‐soluble quatsomes (QS; nanovesicles composed of surfactants and sterols), which allow their nanostructuration in water and, at the same time, limits the ACQ effect. The obtained fluorescent organic nanoparticles showed superior structural homogeneity, along with long‐term colloidal and optical stability. A thorough one‐ (1P) and two‐photon (2P) fluorescence characterization revealed the promising photophysical features of these fluorescent nanovesicles, which showed a high 1P and 2P brightness. Finally, the fluorescent QSs were used for the in vitro bioimaging of Saos‐2 osteosarcoma cell lines; this demonstrates their potential as nanomaterials for bioimaging applications.This work was financially supported by the DGI, Spain (Grant MOTHER; MAT2016‐80826‐R); AGAUR, Generalitat de Catalunya (Grant 2017 SGR 918); and the Networking Research Center on Bioengineering, Biomaterials, and Nanomedicine (CIBER‐BBN). Part of the characterization work has been performed by the ICTS “NANBIOSIS”, at the Biomaterial Processing and Nanostructuring Unit #6 of the CIBER in Bioengineering, Biomaterials & Nanomedicine (CIBER‐BBN). D.B. and A.A. acknowledge the European Commission (EC) FP7‐PEOPLE‐2013‐Initial Training Networks (ITN) “NANO2FUN” project no. 607721 for their predoctoral contracts. This work was also supported by the Ministero dell'Istruzione, dell'Università e della Ricerca (MIUR; Progetto PRIN 2012 prot. 2012A4Z2RY and Project PON02_00563_3316357 (PON MAAT)). Financial support from the Fonds National Suisse de la Recherche Scientifique (no. 200020‐165890) and the University of Geneva is also acknowledged.Peer reviewe