An efficient Foxtail mosaic virus vector system with reduced environmental risk

<p>Abstract</p> <p>Background</p> <p>Plant viral vectors offer high-yield expression of pharmaceutical and commercially important proteins with a minimum of cost and preparation time. The use of <it>Agrobacterium tumefaciens </it>has been introduced to deliver the viral vector as a transgene to each plant cell via a simple, nonsterile infiltration technique called "agroinoculation". With agroinoculation, a full length, systemically moving virus is no longer necessary for excellent protein yield, since the viral transgene is transcribed and replicates in every infiltrated cell. Viral genes may therefore be deleted to decrease the potential for accidental spread and persistence of the viral vector in the environment.</p> <p>Results</p> <p>In this study, both the coat protein (CP) and triple gene block (TGB) genetic segments were eliminated from <it>Foxtail mosaic virus </it>to create the "FECT" vector series, comprising a deletion of 29% of the genome. This viral vector is highly crippled and expresses little or no marker gene within the inoculated leaf. However, when co-agroinoculated with a silencing suppressor (p19 or HcPro), FECT expressed GFP at 40% total soluble protein in the tobacco host, <it>Nicotiana benthamiana</it>. The modified FoMV vector retained the full-length replicase ORF, the TGB1 subgenomic RNA leader sequence and either 0, 22 or 40 bases of TGB1 ORF (in vectors FECT0, FECT22 and FECT40, respectively). As well as <it>N. benthamiana</it>, infection of legumes was demonstrated. Despite many attempts, expression of GFP via syringe agroinoculation of various grass species was very low, reflecting the low <it>Agrobacterium</it>-mediated transformation rate of monocots.</p> <p>Conclusions</p> <p>The FECT/40 vector expresses foreign genes at a very high level, and yet has a greatly reduced biohazard potential. It can form no virions and can effectively replicate only in a plant with suppressed silencing.</p

RANDOMIZED COMPARISON OF TWO APPROACHES TO WARFARIN DOSAGE IN CARDIOVASCULAR HOSPITAL SECTION

Aim. To compare two approaches to warfarin dosage formulation: the standard and with the clinical algorithm.Material and methods. As an approach to warfarin dosage, we have used the method that included clinical properties of the patients. Into the group of the studied approach (intervention group), we randomized 31 person, into the standard approach group (controls) — 29 persons with a variety of indications for vitamin K antagonists treatment. Target diapason of International Normalized Ratio (INR) for all the patients was 2,0 to 3,0.Results. A stable target INR in both groups was reached during hospitalization just in a small part of the patients: 19,4% in intervention group and 17,2% in controls. The patients from intervention group reached stable INR by 6,8 days in average, that is almost two times faster than controls, who had their target INR by 12,4 day in average (p&lt;0,05). Part of the INR values higher than 4,0 in intervention group was 3,6%, in control group — 18,2% (p&lt;0,05). Thromboembolic events (strokes, pulmonary embolism repeats) has not developed in both groups. There was one bleeding episode in each of the groups.Conclusion. Opportunities for targeting INR, not dependent on dosing regimen, are quite restricted in inpatient settings. The patients having reached target INR, usage of clinical algorithm of warfarin dosage helped to achieve these values much faster than in standard approach. Prevalence of excessive hypocoagulation while using the algorithm was lesser than in standard group. Key words: