Utility of P19 Gene-Silencing Suppressor for High Level Expression of Recombinant Human Therapeutic Proteins in Plant Cells

Background: The potential of plants, as a safe and eukaryotic system, is considered in the production of recombinant therapeutic human protein today; but the expression level of heterologous proteins is limited by the post-transcriptional gene silencing (PTGS) response in this new technology. The use of viral suppressors of gene silencing can prevent PTGS and improve transient expression level of foreign proteins. In this study, we investigated the effect of p19 silencing suppressor on recombinant human nerve growth factor expression in Nicotiana benthamiana. Materials and Methods: The p19 coding region was inserted in the pCAMBIA using NcoI and BstEII recognition sites. Also, the cloned synthesized recombinant human NGF (rhNGF) fragment was cloned directly into PVX vector by ClaI and SalI restriction enzymes. The co-agroinfiltration of rhNGF with p19 viral suppressor of gene silencing was evaluated by dot-blot and SDS-PAGE. The amount of expressed rhNGF protein was calculated by AlphaEaseFC software. Results: Co-agroinfiltration of hNGF with P19 suppressor showed about forty-fold increase (8% total soluble protein (TSP)) when compared to the absence of P19 suppressor (0.2%TSP). Conclusion: The results presented here confirmed that the use of P19 gene silencing suppressor derived from tomato bushy stunt virus (TBSV) could efficiently increase the transient expression of recombinant proteins in Nicotiana benthamiana manifold

The Effect of Lateral Wedge Insole on Mediolateral Static Balance in Patients with Mild to Moderate Knee Osteoarthritis

Background and Objectives: Balance has reported to be impaired in patients with knee osteoarthritis. Lateral wedged insoles, have shown positive effect on parameters like knee adduction moment and pain. As a result, the factors that have been shown to reduce the parameters associated with knee osteoarthritis are anticipated to be effective in improving balance. The aim of this study was to investigate the immediate and four-week effects of lateral wedged insoles on static balance in patients with mild and moderate osteoarthritis of knee medial compartment.  Methods: 18 patients (mean age=53±6.07) and body mass index (26/1±2.1 kg/m2) with painful mild and moderate knee osteoarthritis according to the Kellgren-Lawrence grading were recruited. Static standing balance was assessed while standing for 60 seconds on a force plate in three different footwear and two vision conditions: barefoot, wearing shoes without lateral wedged insoles and wearing shoes with lateral wedged insole in open and closed eye conditions. Pain intensity and static standing balance in the mediolateral direction (through the displacement and velocity of center of pressure) were investigated immediately and 4 weeks later.  Results: Lateral wedged insoles had no immediate effect on the center of pressure mediolateral displacement (P=0.70) and velocity (P=0.08). Using the lateral wedged insoles for 4 weeks resulted in significant improvement of pain (P <0.001) in mediolateral center of pressure displacement (P <0.001) and velocity (P <0.001).  Conclusion: No significant immediate effect was seen on the average displacement and velocity of the center of pressure in the mediolateral direction while wearing lateral wedged insoles. Measured parameters improved after 4 weeks. Effect of lateral wedged insoles as a non-invasive treatment can be considered for improving the postural balance in those with knee osteoarthritis