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In perennial ryegrass breeding programmes, dry‐matter yield (DMY) of individual plots is monitored destructively at the different cuts or derived from non‐destructive canopy height measurements using devices like rising plate meters (RPM). These approaches both have constraints. Destructive sampling implies low temporal resolution, restraining the study of dry‐matter accumulation rates, while RPM measurements are influenced by the canopy structure and limit intra‐field variability identification. We present a phenotyping methodology, based on the use of an affordable RGB camera mounted on an unmanned aerial vehicle (UAV), to monitor the spatial and temporal evolution of canopy height and to estimate DMY. Weekly flights were carried out from April to October above a field comprising a diverse set of accessions. To test the capacity of UAV imagery to estimate canopy height, 8 ground control points and 28 artificial height references were placed at different locations. Accurate flights with an RMSE as low as 0.94 cm were achieved. In addition, canopy height was recorded using an RPM and destructive biomass samples were collected. Different models (linear, multiple linear, principal components, partial least squares regression and random forest) were used to predict DMY, and their performance was evaluated. The best estimations were obtained by combining variables including canopy height, vegetation indices and environmental data in a multiple linear regression (R2 = .81). All models built using UAV data obtained a lower RMSE than the one using RPM data. The approach presented is a possibility for breeders to incorporate new information in their selection process. [ABSTRACT FROM AUTHOR]

ABSTRACT BACKGROUND Multimodality imaging such as proton magnetic resonance spectroscopy (MRS) and positron emission tomography (PET) have provided information specific to the underlying mechanisms of many brain diseases, including multiple sclerosis (MS). PURPOSE To determine the structural and metabolic characterization of two particular variants of MS, namely tumefactive MS and Balo's concentric sclerosis (BCS). METHODS Conventional MR imaging, diffusion and perfusion MR, MR spectroscopy and PET imaging with F-18 fluorodeoxyglucose (FDG) and F-18 fluoromethylcholine (FCho) were performed. RESULTS In a case with pathologically proven tumefactive MS, magnetic resonance imaging (MRI) showed a pseudotumoral lesion with incomplete ring enhancement, peripheral diffusion restriction, and high choline and lactate peaks on MRS. On follow-up, the lesion showed significant growth. In a case of BCS, MRI showed an onion-like lesion without contrast enhancement or diffusion restriction, and only a moderate increase in choline on MRS. The lesion remained stable on follow-up. On PET, there was no uptake of F-18 FDG in either type of MS lesion. Conversely, uptake of F-18 FCho was moderate in tumefactive MS, whereas no F-18 FCho uptake was noted in the lesion with, on MRI, typical features of BCS. CONCLUSIONS Our findings illustrate that metabolic features may differ between variants of MS possibly signifying different disease activity. [ABSTRACT FROM AUTHOR]

Low-density lipoprotein receptor-related protein (LRP) contributes to factor VIII (FVIII) catabolism in vivo. Besides LRP, FVIII also interacts with very low-density lipoprotein receptor (VLDLR) in vitro. We investigated the physiological role of VLDLR in FVIII catabolism, using knockout mouse models for VLDLR and LRP, alone and in combination. VLDLR−/− mice displayed normal plasma FVIII, whereas VLDLR−/− LRP− double-knockout mice had slightly increased FVIII compared with LRP-deficient mice. Remarkably, VLDLR deficiency slightly accelerated FVIII clearance. Adenovirus-mediated overexpression of VLDLR did not lower plasma FVIII in LRP-deficient mice. We conclude that VLDLR does not act in concert with LRP in FVIII clearance in vivo. [ABSTRACT FROM AUTHOR]

The functional defect caused by substitution of Arg527 (→ Trp) and Arg531 (→ Gly, His) in factor VIII (FVIII), was explored by employing FVIII derived from patient plasma and recombinant FVIII variants. Mutation of these residues is associated with mild haemophilia A. For both FVIII-R527W and FVIII-R531H, activity was lower than antigen, indicating a functional defect for both variants. In contrast to FVIII-R527W, the amount of FVIII-R531H heterodimer present in plasma was reduced compared to heavy and light chain levels. Factor X (FX) activation experiments employing recombinant FVIII-R531G revealed that the activated FVIII-R531G heterotrimer was less stable than normal FVIIIa, apparently due to rapid dissociation of the A2 domain. These findings suggest that Arg531 is involved in maintaining the stability of both the heterodimer and the activated FVIII heterotrimer. Recombinant FVIII-R527W displayed reduced stimulation of FX activation, suggesting a defect in interaction with factor IXa (FIXa). The contribution of Arg527 in the interaction with FIXa was supported by the observation that FVIII-derived synthetic peptide Tyr511-Leu530 was able to inhibit FX activation and that this inhibition could be overcome by addition of increasing concentrations of FIXa. Furthermore, in the three-dimensional FVIII model residues Val517-Arg527 are located near the FIXa binding site Ser558-Gln565. Therefore we propose that Arg527 is part of an extended FIXa binding site, comprising residues Ser558-Gln565 and Val517-Arg527. [ABSTRACT FROM AUTHOR]

Activation and limited proteolysis of factor VIII have been investigated with respect to the role of the heavy-chain region LysT13-Arg740. The kinetics of factor VIII activation have been analyzed in a system consisting of human factor VIII, factor IXa, factor X, phospholipids, and thrombin or factor Xa. Plasma-derived factor VIII is activated by thrombin with a second-order rate constant of 3.3±0.3×106 M-1 s-1, which proved to be slightly higher than for activation by factor Xa. The second-order rate constant of activation by thrombin of plasma-derived factor VIII in the presence of a monoclonal antibody against the sequence Lys713-Arg740 is markedly reduced. The same result was obtained for activation by thrombin and factor Xa of factor VIII with a deletion including the sequence Lys713-Arg740, des-(713–1637) factor VIII. This suggests that the region Lys713-Arg740 promotes factor VIII activation by both thrombin and factor Xa. Since factor VIII activation is associated with proteolysis, cleavage of factor VIII heavy and light chains was analyzed quantitatively. These studies indicated that heavy-chain cleavage of des-(713–1637)-factor VIII is similar to that of plasma-derived factor VIII. In contrast, cleavage of the light chain of des-(713–1637)-factor VIII is clearly reduced. Furthermore, the second-order rate constant (0.2±0.1×106 M-1 s-1) of des-(713–1637)-factor VIII light-chain cleavage by thrombin was reduced tenfold compared with that of plasma-derived factor VIII. Proteolysis by factor Xa yielded similar results. The rate of des-(713–1637)-factor VIII light chain cleavage by thrombin is similar to that of isolated light-chain, but isolated light-chain is cleaved by factor Xa 20-fold more efficiently than the light chain in des (713–1637)-factor VIII. We conclude that activation of factor VIII by both thrombin and factor Xa is closely associated with light-chain cleavage. Furthermore, within the factor VIII heterodimer, the heavy-chain sequence Lys713-Arg740 promotes both activation and light-chain proteolysis. [ABSTRACT FROM AUTHOR]