Targeted Expression to Liver of an antimiR-33 Sponge as a Gene Therapy Strategy against Hypercholesterolemia: In Vitro Study.

Atherosclerosis is the leading cause of cardiovascular diseases in Mexico and worldwide. The membrane transporters ABCA1 and ABCG1 are involved in the reverse transport of cholesterol and stimulate the HDL synthesis in hepatocytes, therefore the deficiency of these transporters promotes the acceleration of atherosclerosis. MicroRNA-33 (miR-33) plays an important role in lipid metabolism and exerts a negative regulation on the transporters ABCA1 and ABCG1. It is known that by inhibiting the function of miR-33 with antisense RNA, HDL levels increase and atherogenic risk decreases. Therefore, in this work, a genetic construct, pPEPCK-antimiR-33-IRES2-EGFP, containing a specific antimiR-33 sponge with two binding sites for miR-33 governed under the PEPCK promoter was designed, constructed, and characterized, the identity of which was confirmed by enzymatic restriction, PCR, and sequencing. Hep G2 and Hek 293 FT cell lines, as well as a mouse hepatocyte primary cell culture were transfected with this plasmid construction showing expression specificity of the PEPCK promoter in hepatic cells. An analysis of the relative expression of miR-33 target messengers showed that the antimiR-33 sponge indirectly induces the expression of its target messengers (ABCA1 and ABCG1). This strategy could open new specific therapeutic options for hypercholesterolemia and atherosclerosis, by blocking the miR-33 specifically in hepatocytes.

Immunobiotic and Paraprobiotic Potential Effect of Lactobacillus casei in a Systemic Toxoplasmosis Murine Model.

One of the main characteristics of probiotics is their ability to stimulate and modulate the immune response regardless of their viability. Lactobacillus casei (Lc) can stimulate local and systemic immunity, in addition to the activation of macrophages at sites distant from the intestine. Activated macrophages limit the replication of intracellular protozoa, such as Toxoplasma gondii, through the production of nitric oxide. The present study aimed to evaluate the protection generated by treatment with viable and non-viable Lc in the murine systemic toxoplasmosis model. CD1 male mice were treated with viable Lc (immunobiotic) and non-viable Lc (paraprobiotic), infected with tachyzoites of Toxoplasma gondii RH strain. The reduction of the parasitic load, activation of peritoneal macrophages, inflammatory cytokines, and cell populations was evaluated at 7 days post-infection, in addition to the survival. The immunobiotic and paraprobiotic reduced the parasitic load, but only the immunobiotic increased the activation of peritoneal macrophages, and the production of interferon-gamma (IFN-γ), tumor necrosis factor (TNF), and interleukin-6 (IL-6) while the paraprobiotic increased the production of monocyte chemoattractant protein-1 (MCP-1) and T CD4+CD44+ lymphocytes. Viable and non-viable Lc increases survival but does not prevent the death of animals. The results provide evidence about the remote immunological stimulation of viable and non-viable Lc in an in vivo parasitic model.

Gold nanoparticles with chitosan, N-acylated chitosan, and chitosan oligosaccharide as DNA carriers.

Currently, gold nanoparticles have found applications in engineering and medical sciences, taking advantage from their properties and characteristics. Surface plasmon resonance, for instance, is one of the main features for optical applications and other physical properties, like high density, that represents the key for cellular uptake. Among other applications, in the medical field, some diseases may be treated by using gene therapy, including monogenetic or polygenetic disorders and infections. Gene adding, suppression, or substitution is one of the many options for genetic manipulation. This work explores an alternative non-viral method for gene transfer by using gold nanoparticles functionalized with organic polymers; two routes of synthesis were used: one of them with sodium borohydride as reducing agent and the other one with chitosan oligosaccharide as reducing and stabilizing agent. Gold nanoparticles conjugated with chitosan, acylated chitosan and chitosan oligosaccharide, were used to evaluate transfection efficiency of plasmid DNA into cell culture (HEK-293). Physical and chemical properties of gold nanocomposites were characterized by using UV-Vis Spectroscopy, ξ-potential, and transmission electron microscopy. Furthermore, the interaction between gold nanoparticles and plasmid DNA was demonstrated by using agarose gel electrophoresis. Transfection tests were performed and evaluated by ß-galactosidase activity and green fluorescence protein expression. The percentage of transfection obtained with chitosan, acylated chitosan, and chitosan oligosaccharide were of 27%, 33%, and 60% respectively.

Thermal diffusivity measurement for urchin-like gold nanofluids with different solvents, sizes and concentrations/shapes.

The thermal properties of nanofluids are an especially interesting research topic because of the variety of potential applications, which range from bio-utilities to next-generation heat-transfer fluids. In this study, photopyroelectric calorimetry for measuring the thermal diffusivity of urchin-like colloidal gold nanofluids as a function of particle size, concentration and shape in water, ethanol and ethylene glycol is reported. Urchin-like gold nanoparticles were synthesised in the presence of hydroquinone through seed-mediated growth with homogeneous shape and size ranging from 55 to 115 nm. The optical response, size and morphology of these nanoparticles were characterised using UV-visible spectroscopy and transmission electron microscopy. The thermal diffusivity of these nanofluids decreased as the size of the nanoparticles increased, and the enhancement depended on the thermal diffusivity of the solvent. The opposite effect (increase in thermal diffusivity) was observed when the nanoparticle concentration was increased. These effects were more evident for urchin-like gold nanofluids than for the corresponding spherical gold nanofluids.