Correction to the pathogenic alternative splicing, caused by the common GNB3 c.825C>T allele, using a novel, antisense morpholino

The very common GNB3 c.825C>T polymorphism (rs5443), is present in approximately half of all human chromosomes. Significantly the presence of the GNB3 825T allele has been strongly associated, with predisposition to essential hypertension. Paradoxically the presence of the GNB3 825T allele, in exon 10, introduces a pathogenic alternative RNA splice site into the middle of exon 9. To attempt to correct this pathogenic aberrant splicing, we therefore bioinformatically designed, using a Gene Tools® algorithm, a GNB3 specific, antisense morpholino. It was hoped that this morpholino would behave in vitro as either a potential “ splice blocker and/or exon skipper, to both bind and inhibit/reduce the aberrant splicing of the GNB3, 825T allele. On transfecting a human lymphoblast cell line homozygous for the 825T allele, with this antisense morpholino, we encouragingly observed both a significant reduction (from ~58% to ~5%) in the production of the aberrant smaller GNB3 transcript, and a subsequent increase in the normal GNB3 transcript (from ~42% to ~95%). Our results demonstrate the potential use of a GNB3 specific antisense morpholino, as a pharmacogenetic therapy for essential hypertension.

Optimization of physical and optical properties of biodegradable edible films based on pea starch and guar gum

The influence of process variables (pea starch, guar gum and glycerol) on the viscosity (V), solubility (SOL), moisture content (MC), transparency (TR), Hunter parameters (L, a, and b), total color difference (ΔE), yellowness index (YI), and whiteness index (WI) of the pea starch based edible films was studied using three factors with three level Box–Behnken response surface design. The individual linear effect of pea starch, guar and glycerol was significant (p < 0.05) on all the responses. However, a value was only significantly (p < 0.05) affected by pea starch and guar gum in a positive and negative linear term, respectively. The effect of interaction of starch × glycerol was also significant (p < 0.05) on TR of edible films. Interaction between independent variables starch × guar gum had a significant impact on the b and YI values. The quadratic regression coefficient of pea starch showed a significant effect (p < 0.05) on V, MC, L, b, ΔE, YI, and WI; glycerol level on ΔE and WI; and guar gum on ΔE and SOL value. The results were analyzed by Pareto analysis of variance (ANOVA) and the second order polynomial models were developed from the experimental design with reliable and satisfactory fit with the corresponding experimental data and high coefficient of determination (R2) values (>0.93). Three-dimensional response surface plots were established to investigate the relationship between process variables and the responses. The optimized conditions with the goal of maximizing TR and minimizing SOL, YI and MC were 2.5 g pea starch, 25% glycerol and 0.3 g guar gum. Results revealed that pea starch/guar gum edible films with appropriate physical and optical characteristics can be effectively produced and successfully applied in the food packaging industry.