Dynamic light scattering and X-ray photoelectron spectroscopy characterization of PEGylated polymer nanocarriers: internal structure and surface properties.

In this work, nanospheres and nanocapsules are precipitated in confined impinging jet mixers through solvent displacement and characterized. Acetone and water are used as the solvent and antisolvent, respectively, together with polymethoxypolyethylene glycol cyanoacrylate-co-hexadecylcyanoacrylate and Miglyol as the copolymer and oil, respectively. Characterization is performed with dynamic light scattering, with electrophoretic measurements, and for the first time with X-ray photoelectron spectroscopy. Results show that the presence of polyethylene glycol chains seems to be more pronounced on the surface of nanospheres than on that of nanocapsules. The thickness of the copolymer layer in nanocapsules ranges from 1 to 10 nm, depending on the value of the oil:copolymer mass ratio. Fast dilution is confirmed to have a positive effect in suppressing aggregation but can induce further copolymer precipitation.

miRNA signatures in sera of patients with active pulmonary tuberculosis.

Several studies showed that assessing levels of specific circulating microRNAs (miRNAs) is a non-invasive, rapid, and accurate method for diagnosing diseases or detecting alterations in physiological conditions. We aimed to identify a serum miRNA signature to be used for the diagnosis of tuberculosis (TB). To account for variations due to the genetic makeup, we enrolled adults from two study settings in Europe and Africa. The following categories of subjects were considered: healthy (H), active pulmonary TB (PTB), active pulmonary TB, HIV co-infected (PTB/HIV), latent TB infection (LTBI), other pulmonary infections (OPI), and active extra-pulmonary TB (EPTB). Sera from 10 subjects of the same category were pooled and, after total RNA extraction, screened for miRNA levels by TaqMan low-density arrays. After identification of "relevant miRNAs", we refined the serum miRNA signature discriminating between H and PTB on individual subjects. Signatures were analyzed for their diagnostic performances using a multivariate logistic model and a Relevance Vector Machine (RVM) model. A leave-one-out-cross-validation (LOOCV) approach was adopted for assessing how both models could perform in practice. The analysis on pooled specimens identified selected miRNAs as discriminatory for the categories analyzed. On individual serum samples, we showed that 15 miRNAs serve as signature for H and PTB categories with a diagnostic accuracy of 82% (CI 70.2-90.0), and 77% (CI 64.2-85.9) in a RVM and a logistic classification model, respectively. Considering the different ethnicity, by selecting the specific signature for the European group (10 miRNAs) the diagnostic accuracy increased up to 83% (CI 68.1-92.1), and 81% (65.0-90.3), respectively. The African-specific signature (12 miRNAs) increased the diagnostic accuracy up to 95% (CI 76.4-99.1), and 100% (83.9-100.0), respectively. Serum miRNA signatures represent an interesting source of biomarkers for TB disease with the potential to discriminate between PTB and LTBI, but also among the other categories.

Production of PEGylated nanocapsules through solvent displacement in confined impinging jet mixers.

The growth of importance of nanocapsules (and other particulate systems) in different fields requires fast and reproducible methods for their production. Confined impinging jet mixers were successfully used for the production of nanospheres and are now tested for the first time for the production of nanocapsules. This work focuses on the understanding of formation mechanisms and on the quantification of the effect of the most important operating parameters involved in their production. Solvent displacement is employed here for the assembly of the nanocapsules by using a PEGylated derivative of cyanoacrylate as copolymer. A comparison with nanospheres obtained under the same operating conditions is also reported. Results show that the oil-to-copolymer mass ratio (MR) is the main factor affecting the final size distribution and that small nanocapsules are obtained only at low oil-to-copolymer MR. The effect of mixing is significant, proving that mixing of solvent and antisolvent also affects the final size distribution; this depends mainly on the inlet jet velocity, but the size of the mixer is also important. The Reynolds number may be useful to take this into account for geometrically similar systems. Quenching by dilution allows to stabilize the nanocapsules, evidencing the role of aggregation and ripening.