Recessive loss of function PIGN alleles, including an intragenic deletion with founder effect in La Réunion Island, in patients with Fryns syndrome.

Fryns syndrome (FS) is a multiple malformations syndrome with major features of congenital diaphragmatic hernia, pulmonary hypoplasia, craniofacial dysmorphic features, distal digit hypoplasia, and a range of other lower frequency malformations. FS is typically lethal in the fetal or neonatal period. Inheritance is presumed autosomal recessive. Although no major genetic cause has been identified for FS, biallelic truncating variants in PIGN, encoding a component of the glycosylphosphatidylinositol (GPI)-anchor biosynthesis pathway, have been identified in a limited number of cases with a phenotype compatible with FS. Biallelic variants in PIGN, typically missense or compound missense with truncating, also cause multiple congenital anomalies-hypotonia-seizures syndrome 1 (MCAHS1). Here we report six further patients with FS with or without congenital diaphragmatic hernia and recessive loss of function PIGN alleles, including an intragenic deletion with a likely founder effect in La Réunion and other Indian Ocean islands. Our results support the hypothesis that a spectrum of phenotypic severity is associated with recessive PIGN variants, ranging from FS at the extreme end, caused by complete loss of function, to MCAHS1, in which some residual PIGN function may remain. Our data add FS resulting from PIGN variants to the catalog of inherited GPI deficiencies caused by the disruption of the GPI-anchor biosynthesis pathway.

Nanoparticulate delivery systems for alkyl gallates: Influence of the elaboration process on particle characteristics, drug encapsulation and in-vitro release.

PLA nanoparticles loaded with n-alkyl gallates (AGs) were prepared either by nanoprecipitation (NP) or by O/W emulsion/solvent evaporation (E/SE). A nonionic hydrophobically modified polysaccharide was used for surface coverage and for ensuring colloidal stability. Different parameters were systematically assessed to enhance the drug incorporation, with the aim of obtaining monomodal and narrow particle size distributions. The nanoparticles were characterized by 1H NMR, transmission electron microscopy (TEM) and laser light scattering granulometry. The colloidal stability of suspensions was evaluated after incubation in NaCl solutions and was maintained up to 1M NaCl. The mean particle diameter and the width of size distribution were found very similar for both processes (slightly lower diameters when using E/SE) with various drug loadings. The amount of encapsulated AG by E/SE was about twice that encapsulated by NP. The in-vitro release of AG was evaluated under sink conditions and no burst effect was observed. Release curves were successfully modeled using the Fick diffusion model with a constant diffusion coefficient and assuming non-swellable particles. Diffusion coefficients of AG loaded in nanoparticles prepared by NP were higher than those found in nanoparticles elaborated by E/SE.

Amphiphilic Polysaccharides Acting both as Stabilizers and Surface Modifiers during Emulsification in Microfluidic Flow-Focusing Junction.

A continuous emulsion/solvent diffusion process was designed for the preparation of polysaccharide-covered poly(d,l-lactide) (PLA) microparticles. The emulsification step was carried out in a flow-focusing junction where ethyl acetate containing dissolved PLA was dispersed into an aqueous solution of hydrophobically modified dextran. It was demonstrated that poly(dimethylsiloxane) devices could be used for oil-in-water emulsion preparation provided that the microfluidic devices were preconditioned by simply circulating the aqueous phase containing the amphiphilic polysaccharide during a sufficient time (30 h). The adsorption of the polymers at the surface of the channel walls permitted the wetting by the aqueous phase with a hydrophilic character maintained at least throughout 2 months. The preconditioning time was significantly reduced by pretreating the microfluidic device with piranha solution and KOH solution during 15 min each before the circulation of the aqueous solution of dextran derivative. Dextran-covered PLA microparticle aqueous suspensions were produced with well-controlled size distribution. The suspensions could be lyophilized and reconstituted by retrieving the initial size distribution without adding any cryoprotectant. The reported procedure was used for preparing octyl gallate-loaded PLA microparticles.