Analysis of factor V in zebrafish demonstrates minimal levels needed for early hemostasis

In humans, coagulation factor V (FV) deficiency is a rare, clinically heterogeneous bleeding disorder, suggesting that genetic modifiers may contribute to disease expressivity. Zebrafish possess many distinct advantages including high fecundity, optical clarity, external development, and homology with the mammalian hemostatic system, features that make it ideal for genetic studies. Our aim was to study the role of FV in zebrafish through targeted mutagenesis and apply the model to the study of human F5 variants. CRISPR-mediated genome editing of the zebrafish f5 locus was performed, generating mutants homozygous for a 49 base pair deletion in exon 4. Thrombus formation secondary to vascular endothelial injury was absent in f52/2 mutant embryos and larvae. Despite this severe hemostatic defect, homozygous mutants survived before succumbing to severe hemorrhage in adulthood. Human F5 variants of uncertain significance from patients with FV deficiency were evaluated, and the causative mutations identified and stratified by their ability to restore thrombus formation in larvae. Analysis of these novel mutations demonstrates variable residual FV function, with minimal activity being required to restore hemostasis in response to laser-induced endothelial injury. This in vivo evaluation may be beneficial for patients whose factor activity levels lack correlation with bleeding symptomatology, although limitations exist. Furthermore, homozygous mutant embryos tolerate what is a severe and lethal defect in mammals, suggesting the possibility of species-specific factors enabling survival, and allowing further study not possible in the mouse. Identification of these factors or other genetic modifiers could lead to novel therapeutic modalities

Targeted sequencing to identify novel genetic risk factors for deep vein thrombosis: a study of 734 genes

Essentials Deep vein thrombosis (DVT) has a large unknown genetic component. We sequenced coding areas of 734 hemostasis-related genes in 899 DVT patients and 599 controls. Variants in F5, FGA-FGG, CYP4V2-KLKB1-F11, and ABO were associated with DVT risk. Associations in KLKB1 and F5 suggest a more complex genetic architecture than previously thought. Summary: Background Although several genetic risk factors for deep vein thrombosis (DVT) are known, almost all related to hemostasis, a large genetic component remains unexplained. Objectives To identify novel genetic determinants by using targeted DNA sequencing. Patients/Methods We included 899 DVT patients and 599 controls from three case\u2013control studies (DVT-Milan, Multiple Environmental and Genetic Assessment of risk factors for venous thrombosis [MEGA], and the Thrombophilia, Hypercoagulability and Environmental Risks in Venous Thromboembolism [THE-VTE] study) for sequencing of the coding regions of 734 genes involved in hemostasis or related pathways. We performed single-variant association tests for common variants (minor allele frequency [MAF] 65 1%) and gene-based tests for rare variants (MAF 64 1%), accounting for multiple testing by use of the false discovery rate (FDR). Results Sixty-two of 3617 common variants were associated with DVT risk (FDR 0.2). Conclusions We confirmed associations between DVT and common variants in F5,ABO,FGA\u2013FGG, and CYP4V2\u2013KLKB1\u2013F11, and observed secondary signals in F5 and CYP4V2\u2013KLKB1\u2013F11 that warrant replication and fine-mapping in larger studies

Validation of a spatial-temporal soil water movement and plant water uptake model

Management and irrigation of plants increasingly relies on accurate mathematical models for the movement of water within unsaturated soils. Current models often use values for water content and soil parameters that are averaged over the soil profile. However, many applications require models to more accurately represent the soil–plant–atmosphere continuum, in particular, water movement and saturation within specific parts of the soil profile. In this paper a mathematical model for water uptake by a plant root system from unsaturated soil is presented. The model provides an estimate of the water content level within the soil at different depths, and the uptake of water by the root system. The model was validated using field data, which include hourly water content values at five different soil depths under a grass/herb cover over 1 year, to obtain a fully calibrated system for plant water uptake with respect to climate conditions. When compared quantitatively to a simple water balance model, the proposed model achieves a better fit to the experimental data due to its ability to vary water content with depth. To accurately model the water content in the soil profile, the soil water retention curve and saturated hydraulic conductivity needed to vary with depth

Mathematical modelling of flow in curved, compliant arteries

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How changing root system architecture can help tackle a reduction in soil phosphate (P) levels for better plant P acquisition

The readily available global rock phosphate (P) reserves may run out within the next 50-130 years, causing soils to have a reduced P concentration which will affect plant P uptake. Using a combination of mathematical modelling and experimental data we investigated potential plant-based options for optimising crop P uptake in reduced soil P environments. By varying the P concentration within a well-mixed agricultural soil, for high and low P (35.5 to 12.5 mg l-1 respectively, using Olsen’s P index), we investigated branching distributions within a wheat root system that maximise P uptake. Changing the root branching distribution from linear (evenly spaced branches) to strongly exponential (a greater number of branches at the top of the soil), improves P uptake by 142% for low P soils when root mass is kept constant between simulations. This causes the roots to emerge earlier and mimics topsoil foraging. Manipulating root branching patterns, to maximise P uptake, is not enough on its own to overcome the drop in soil P from high to low P. Further mechanisms have to be considered to fully understand the impact of P reduction on plant development