A technique for creating critical-size defects in the metatarsus of sheep for use in investigation of healing of long-bone defects

OBJECTIVE: To develop a technique for use in investigation of healing of long-bone defects by creation of a critical-size defect in the left metarsal III and IV bone (metatarsus) of sheep. ANIMALS: 18 healthy adult sheep. PROCEDURE: Sheep were allocated to 4 groups (3, 3, 5, and 7 sheep in groups 1 to 4, respectively). An ostectomy with various segmental length-to-diaphyseal diameter ratios (0.5, 1.0, 2.0, and 2.0 for groups 1 to 4, respectively) was performed on the left metatarsus of each sheep. The defect was left empty in sheep of groups 1, 2, and 3, whereas the defect was filled with a massive corticocancellous bone autograft in sheep of group 4. RESULTS: All sheep tolerated the surgical procedure well and were able to use the affected limb the day after surgery. Radiographic and histologic examinations conducted 16 weeks after surgery revealed nonunion in all sheep of groups 1, 2, and 3, whereas consistent bone healing with abundant bone formation was observed in all sheep of group 4. CONCLUSIONS AND CLINICAL RELEVANCE: Analysis of these findings suggests that the sheep metatarsal model is a critical-size defect model with low morbidity. It should allow the assessment of new technologies for bone regeneration in conditions closely mimicking the clinical setting. IMPACT FOR HUMAN MEDICINE: Use of this technique in sheep should be of benefit for the preclinical study of osteoconductive, osteoinductive, or osteogenic biomaterials for use in humans.     

Low-Molecular-Weight Fucoidan Induces Endothelial Cell Migration via the PI3K/AKT Pathway and Modulates the Transcription of Genes Involved in Angiogenesis

Low-molecular-weight fucoidan (LMWF) is a sulfated polysaccharide extracted from brown seaweed that presents antithrombotic and pro-angiogenic properties. However, its mechanism of action is not well-characterized. Here, we studied the effects of LMWF on cell signaling and whole genome expression in human umbilical vein endothelial cells and endothelial colony forming cells. We observed that LMWF and vascular endothelial growth factor had synergistic effects on cell signaling, and more interestingly that LMWF by itself, in the absence of other growth factors, was able to trigger the activation of the PI3K/AKT pathway, which plays a crucial role in angiogenesis and vasculogenesis. We also observed that the effects of LMWF on cell migration were PI3K/AKT-dependent and that LMWF modulated the expression of genes involved at different levels of the neovessel formation process, such as cell migration and cytoskeleton organization, cell mobilization and homing. This provides a better understanding of LMWF’s mechanism of action and confirms that it could be an interesting therapeutic approach for vascular repair.     

Improved effectiveness and efficiency in measuring soil enzymes as universal soil quality indicators using microplate fluorimetry

We tested the relevance of the microplate fluorimetric (F) assay for five enzymes in contrasting land uses, including woodland, grassland, cultivated and contaminated lands, as compared to the standard spectrophotometric (P) method. Enzymatic activity measured by the P method ranged from 0 to 56.04 nmol-pNP g^?1 min^?1 (median = 4) while the F method revealed lower values ranging from 0 to 6.22 nmol-MUB g^?1 dry soil min^?1 (median = 1). The values obtained by the P method were around 8 times higher than those revealed by the F method. However, the F method revealed significant differences in enzyme activity in orchard parcels (land use with low variations in soil properties). We concluded that the F method improves the effectiveness and the efficiency of measuring universal soil quality indicators using enzymes.     

Fabrication and characterization of nanofibers based on poly(lactic acid)/chitosan blends by electrospinning and their functionalization with phospholipase A1

In this work, electrospinning of poly(lactic acid) (PLA), chitosan and their blends has been investigated, and nanofibers with a diameter ranging from 90 nm to 1.9 microns were produced and used as carriers for immobilization of the phospholipase A1. A strong influence of chitosan (CS) and the solvent trifluoroacetic acid (TFA) on the morphology, distribution of the nanofibers diameter and on their hydrophobicity was observed. The yield of phospholipase A1 (PLA1) on non-woven fibers was evaluated using the method of Bradford. Their activities and their reutilisability were assessed titrimetrically using soybean lecithin as substrate. The results showed that the degree of immobilization on the non-woven fibers of pure PLA and mixtures PLA/CS4 and PLA/SC6 are 73, 54, 45 % respectively and can be reused up to 4 cycles without significant loss of enzyme activity. Moreover, a remarkable improvement of the activity of phospholipase A1 on non-woven based on pure PLA fibers was observed, indicating that most of the enzymes were probably in their active form.     

Linking changes in the soil microbial community to C and N dynamics during crop residue decomposition

Crop residues are among the main inputs that allow the organic carbon (C) and nutrients to be maintained in agricultural soil. It is an important management strategy that can improve soil fertility and enhance agricultural productivity. This work aims to evaluate the extent of the changes that may occur in the soil heterotrophic microbial communities involved in organic matter decomposition and C and nitrogen (N) mineralization after the addition of crop residues. Soil microcosm experiments were performed at 28°C for 90 days with the addition of three crop residues with contrasting biochemical qualities: pea (Pisum sativum L.), rapeseed (Brassica napus L.), and wheat (Triticum aestivum L.). Enzyme activities, C and N mineralization, and bacterial and fungal biomasses were monitored, along with the bacterial and fungal community composition, by the high-throughput sequencing of 16S rRNA and ITS genes. The addition of crop residues caused decreases in β-glucosidase and arylamidase activities and simultaneous enhancement of the C mineralization and net N immobilization, which were linked to changes in the soil microbial communities. The addition of crop residues decreased the bacterial and fungal biomasses 90 days after treatment and there were shifts in bacterial and fungal diversity at the phyla, order, and genera levels. Some specific orders and genera were dependent on crop residue type. For example, Chloroflexales, Inquilinus, Rubricoccus, Clitocybe, and Verticillium were identified in soils with pea residues; whereas Thermoanaerobacterales, Thermacetogenum, and Hypoxylon were enriched in soils with rapeseed residues, and Halanaerobiales, Rubrobacter, and Volutella were only present in soils with wheat residues. The findings of this study suggest that soil C and N dynamics in the presence of the crop residues were driven by the selection of specific bacterial and fungal decomposers linked to the biochemical qualities of the crop residues. If crop residue decomposition processes showed specific bacterial and fungal operational taxonomic unit (OTU) signatures, this study also suggests a strong functional redundancy that exists among soil microbial communities.     

Potential of combined biological control agents to cope with <Emphasis Type="Italic">Phytophthora parasitica,</Emphasis> a major pathogen of <Emphasis Type="Italic">Choisya ternata</Emphasis>

The Cook Agronomy Farm of Washington State University is a long-term precision agriculture study site. Since 2000, the farm has been in various three-year no-till rotations with winter wheat, spring wheat, and various rotation crops such as barley, canola, peas, lentils, and chickpea. The spatial distribution of root lesion nematodes (Pratylenchus spp.) and their relationships with soil and terrain variables were studied by collecting soil samples at 127 and 124 geo-referenced points in two 12-ha fields during the spring of 2010 and 2011, respectively. Pratylenchus spp. were detected in more than 85% of the sampled locations in both fields. Nematode densities spatially mapped using ArcGIS software occurred in an aggregated pattern. Previous rotation crops had no effect on the spatial distribution. Classification and regression tree analysis (CART) using soil and terrain variables, and nematode density, explained 61 and 34% of the variability associated with nematode density in 2010 and 2011, respectively. Soil edaphic factors, such as organic matter, were stronger predictors of nematode populations than rotation effects.