L-Type Bovine Spongiform Encephalopathy in Genetically Susceptible and Resistant Sheep: Changes in Prion Strain or Phenotypic Plasticity of the Disease-Associated Prion Protein?

Background. Sheep with prion protein (PrP) gene polymorphisms QQ171 and RQ171 were shown to be susceptible to the prion causing L-type bovine spongiform encephalopathy (L-BSE), although RQ171 sheep specifically propagated a distinctive prion molecular phenotype in their brains, characterized by a high molecular mass proteaseresistant PrP fragment (HMM PrPres), distinct from L-BSE in QQ171 sheep. Methods. The resulting infectious and biological properties of QQ171 and RQ171 ovine L-BSE prions were investigated in transgenic mice expressing either bovine or ovine PrP. Results. In both mouse lines, ovine L-BSE transmitted similarly to cattle-derived L-BSE, with respect to survival periods, histopathology, and biochemical features of PrPres in the brain, as well as splenotropism, clearly differing from ovine classic BSE or from scrapie strain CH1641. Nevertheless and unexpectedly, HMM PrPres was found in the spleen of ovine PrP transgenic mice infected with L-BSE from RQ171 sheep at first passage, reminiscent, in lymphoid tissues only, of the distinct PrPres features found in RQ171 sheep brains. Conclusions. The L-BSE agent differs from both ovine classic BSE or CH1641 scrapie maintaining its specific strain properties after passage in sheep, although striking PrPres molecular changes could be found in RQ171 sheep and in the spleen of ovine PrP transgenic mice

Synthesis of Copper-Based Nanostructures in Liquid Environments by Means of a Non-equilibrium Atmospheric Pressure Nanopulsed Plasma Jet

The influence of the liquid composition on the chemical and morphological properties of copper-based nanostructures synthesized by a non-equilibrium atmospheric plasma treatment is investigated and discussed. The synthesis approach is simple and environmentally friendly, employs a non-equilibrium nanopulsed atmospheric pressure plasma jet as a contactless cathode and a Cu foil as immersed anode. The process was studied using four distinct electrolyte solutions composed of distilled water and either NaCl + NaOH, NaCl only or NaOH only at two different concentrations, without the addition of any copper salts. CuO crystalline structures with limited impurities (e.g. Cu and Cu(OH)2phases) were produced from NaCl + NaOH containing solutions, mainly CuO and CuCl2structures were synthesized in the electrolyte solution containing only NaCl and no synthesis occurred in solutions containing only NaOH. Both aggregated and dispersed nanostructures were produced in the NaCl + NaOH and NaCl containing solutions. Reaction pathways leading to the formation of the nanostructures are proposed and discussed

Plasma-activated Ringer's Lactate Solution Displays a Selective Cytotoxic Effect on Ovarian Cancer Cells

Epithelial Ovarian Cancer (EOC) is one of the leading causes of cancer-related deaths among women and is characterized by the diffusion of nodules or plaques from the ovary to the peritoneal surfaces. Conventional therapeutic options cannot eradicate the disease and show low efficacy against resistant tumor subclones. The treatment of liquids via cold atmospheric pressure plasma enables the production of plasma-activated liquids (PALs) containing reactive oxygen and nitrogen species (RONS) with selective anticancer activity. Thus, the delivery of RONS to cancer tissues by intraperitoneal washing with PALs might be an innovative strategy for the treatment of EOC. In this work, plasma-activated Ringer's Lactate solution (PA-RL) was produced by exposing a liquid substrate to a multiwire plasma source. Subsequently, PA-RL dilutions are used for the treatment of EOC, non-cancer and fibroblast cell lines, revealing a selectivity of PA-RL, which induces a significantly higher cytotoxic effect in EOC with respect to non-cancer cells

A rapid and reliable detection procedure of Atlantic trout introgression at the diagnostic lactate dehydrogenase chain-1 gene

The Italian-native Mediterranean brown trout (Salmo ghigii) is a seriously threatened freshwater fish, especially by anthropogenic hybridisation with the domestic strains of Atlantic origin that have been repeatedly released into the wild for angling. A PCR-restriction fragment length polymorphism (RFLP) assay of the diagnostic lactate dehydrogenase chain-1 (LDH-C1) gene sequences has been routinely applied to distinguish exotic from native brown trout lineages and detect Atlantic introgression signals in the Mediterranean wild populations. Here, we used dermal swab DNA obtained from 28 wild trout to improve laboratory procedures to genetically characterise trout samples at the LDH-C1gene through (1) a capillary electrophoresis analysis of the RFLP fragments and (2) the optimisation of a diagnostic single nucleotide polymorphism analysable through mini-sequencing approaches. The developed methods were fully consistent with those obtained through the traditional approach, but their analytical process is almost entirely automated and digitalised, thus improving result readability and accuracy in the detection of alien introgressed traces in wild Mediterranean brown trout populations

Gerstmann-Sträussler-Scheinker disease subtypes efficiently transmit in bank voles as genuine prion diseases.

Gerstmann-Sträussler-Scheinker disease (GSS) is an inherited neurodegenerative disorder associated with mutations in the prion protein gene and accumulation of misfolded PrP with protease-resistant fragments (PrPres) of 6–8 kDa

Hybridosomes from spruce needle homogenate

Introduction: Being of compatible structure with biomembranes, lipid–based nanoparticles are considered as convenient platforms for drug delivery systems. In the proposed work we considered formation of lipid nanovesicles associated with bioactive phytochemicals from spruce needle homogenate (here called hybridosomes). We formed hybridosomes by mixing appropriate amounts of lecithin, supernatant of isolation of extracellular particles from spruce needle homogenate and glycerol. Methods: We visualized hybridosomes by light microscopy and cryogenic transmission electron microscopy and assessed them by flow cytometry, dynamic light scattering, ultraviolet–visual spectroscopy and interferometric microscopy. Results: We found that the particles consisted of a bilayer membrane and a fluid-like interior. Flow cytometry and interferometric light microscopy measurements showed that the majority of the particles were nano-sized. Dynamic light scattering and interferometric light microscopy measurements agreed well with the determined average hydrodynamic radius of the particles Rh (between 140 and 180 nm) while their number densities were in the range between 10^13 and 10^14/mL indicating that hybridosomes present about 2/3 of the mixture, excluding solvent and other small molecules. Discussion: Simple and low-cost preparation method, non-demanding saving process and efficient formation procedure suggest that large scale production of hybridosomes from lipids and spruce needle homogenate is feasible.Small New World 2.0 4-5 September 2023., Graz, Austri

Hybridosomes from spruce needle homogenate

Introduction: Being of compatible structure with biomembranes, lipid–based nanoparticles are considered as convenient platforms for drug delivery systems. In the proposed work we considered formation of lipid nanovesicles associated with bioactive phytochemicals from spruce needle homogenate (here called hybridosomes). We formed hybridosomes by mixing appropriate amounts of lecithin, supernatant of isolation of extracellular particles from spruce needle homogenate and glycerol. Methods: We visualized hybridosomes by light microscopy and cryogenic transmission electron microscopy and assessed them by flow cytometry, dynamic light scattering, ultraviolet–visual spectroscopy and interferometric microscopy. Results: We found that the particles consisted of a bilayer membrane and a fluid-like interior. Flow cytometry and interferometric light microscopy measurements showed that the majority of the particles were nano-sized. Dynamic light scattering and interferometric light microscopy measurements agreed well with the determined average hydrodynamic radius of the particles Rh (between 140 and 180 nm) while their number densities were in the range between 10^13 and 10^14/mL indicating that hybridosomes present about 2/3 of the mixture, excluding solvent and other small molecules. Discussion: Simple and low-cost preparation method, non-demanding saving process and efficient formation procedure suggest that large scale production of hybridosomes from lipids and spruce needle homogenate is feasible.Small New World 2.0 4-5 September 2023., Graz, Austri