The effect of chondrocyte cryopreservation on cartilage engineering

Chondrocytes were collected from the stifle joints of four pigs to study the effect of cryopreservation on the chondrogenic potential of chondrocytes. Half of the cells were cryopreserved for 3months. Polyglycolic acid scaffolds were cultured with fresh or cryopreserved chondrocytes for 4weeks. Cell morphology and the quality of engineered tissue were evaluated by scanning electron microscopy, histopathology and biochemical methods. More cells attached to scaffolds at 48h when fresh chondrocytes were seeded. At 4weeks, the numbers of cells, DNA and collagen II were greater in constructs engineered by fresh cells. However, the collagen II/DNA ratio did not differ between the two groups. More matrix was identified on a scanning electron microscope and by histopathology in the fresh group. Cartilage engineered with cryopreserved chondrocytes may contain less matrix and fewer cells. These findings most likely resulted from a lack of cell attachment on the matrix secondary to cryopreservation. Future studies are needed to further evaluate the mechanism by which cryopreservation may affect chondrocyte attachment.     

Doxorubicin area under the curve is an important predictor of neutropenia in dogs with naturally occurring cancers

Doxorubicin (DOX) area‐under‐the‐curve (AUC) was calculated for 40 dogs with spontaneously occurring cancers using a previously validated limited‐sampling approach. All dogs were administered a dose of 30 mg/m² by intravenous infusion and serum samples were collected at 5, 45 and 60 minutes post‐infusion. DOX and its major metabolite, doxorubicinol (doxol), were quantified in serum samples using high‐performance liquid chromatography tandem‐mass spectrometry. Wide interpatient variability was observed in the predicted DOX AUC with a coefficient of variation of 34%. A significant relationship was found between DOX AUC and absolute white blood cell count (P = 0.003), absolute neutrophil count (ANC; P = 0.002) and surviving fraction of neutrophils (P = 0.03) approximately 1 week after dosing (nadir). No changes in other hematologic parameters (red blood cells, platelets, lymphocytes, haemoglobin) were found to correlate with DOX AUC. The absolute dose (mg) and the dose per unit body weight (mg/kg) were not significantly correlated with nadir ANC. No relationships were found between maximum serum doxol concentration and myelosuppression. Baseline ANC was also significantly correlated to nadir ANC and a model was constructed using baseline ANC and DOX AUC that significantly described the nadir ANC. These findings demonstrate the important relationship between systemic DOX exposure and degree of neutropenia in dogs, and suggest a potential for individualized, pharmacokinetically‐guided DOX dosing in dogs.     

A decision support system to simulate and compare silvicultural scenarios for pure even-aged larch stands

In sustained forest management, it is particularly useful to test the adequacy of various silvicultural scenarios, but decision-making is also becoming increasingly complex because forest managers have to simultaneously meet several different objectives in response to society demands. In order to help forest managers make appropriate choices in silvicultural systems, we propose a SDSS (Silvicultural Decision Support System) that applies to pure and even-aged larch stands (Larix sp.) in lowland areas where site conditions are similar to those encountered in Southern Belgium. Its main purpose is (i) to predict the influence of silvicultural treatments on stand evolution and (ii) assist forest managers in comparing different scenarios with respect to predefined goals. This SDSS consists in three modules designed to elaborate silvicultural scenarios involving (i) stand growth prediction and thinning simulation; (ii) assessment of indicators selected to define the scenarios referring to wood production, financial, technico-economic and ecological components, tree stability and wood quality; and (iii) comparison of the scenarios on the basis of the set of previously assessed indicators (multi-criteria analysis). User-friendly “MGC_Larch” software helps managers to formulate, evaluate and compare different silvicultural scenarios for larch.     

Discovery of an amylose-free starch mutant in cassava (Manihot esculenta Crantz)

One of the objectives of the cassava-breeding project at CIAT is the identification of clones with special root quality characteristics. A large number of self-pollinations have been made in search of useful recessive traits. During 2006 harvests an S1 plant produced roots that stained brownish-red when treated with an iodine solution, suggesting that it had lower-than-normal levels of amylose in its starch. Colorimetric and DSC measurements indicated low levels (3.4%) and an absence of amylose in the starch, respectively. SDS-PAGE demonstrated the absence of GBSS enzyme in the starch from these roots. Pasting behavior was analyzed with a rapid visco-analyzer and resulted in larger values for peak viscosity, gel breakdown, and setback in the mutant compared with normal cassava starch. Solubility was considerably reduced, while the swelling index and volume fraction of the dispersed phase were higher in the mutant. No change in starch granule size or shape was observed. This is the first report of a natural mutation in cassava that drastically reduces amylose content in root starch.     

Residual toxicity of two insecticides on three field populations of Lygus lineolaris (Hemiptera: Miridae) collected along the St Lawrence valley in eastern Canada

Insecticides are still the single main pest control method employed today by most growers to mitigate damage done by the tarnished plant bug, Lygus lineolaris (Palisot de Beauvois) (Hemiptera: Miridae). In eastern Canada, the complex agricultural ecosystem, which may be described as a mosaic of farmlands dispersed among natural habitats (forest, prairies), allows tarnished plant bug adults to fly and move from sprayed to non-sprayed areas. In 2004 (late August to early September), three populations of L. lineolaris were collected from three mixed vegetation strips adjacent to orchards and vineyards along the St Lawrence valley: the Niagara Peninsula (Ontario), Dunham (Quebec) and La Pocatière (Quebec). Assays were done in the laboratory by confining adults in glass vials coated with dried residues. The estimated LC(50) values for the three populations varied from 11.2 to 16.8 x 10(-5) g L(-1) for azinphos-methyl and from 0.8 to 1.4 x 10(-5) g L(-1) for cypermethrin. In contrast to the Mississippi delta, no tolerance to insecticides was found in the populations collected. Possible explanations for this non-tolerance to insecticides includes a very low selection pressure as a result of the reduced number of insecticide treatments done in the context of the diversified agricultural landscapes encountered in eastern Canada which allow movements of adults from treated to non-treated areas.     

Haemonchus contortus (Nematoda: Trichostrongylidae) infection in lambs elicits an unequivocal Th2 immune response

Selection of resistant animals and host immunization have been proposed as alternative methods for the control of gastrointestinal nematode parasites. However, a better knowledge of the mechanisms involved in protective immunity against these parasites is required for the development of optimal strategies. In this study, 3 month old INRA 401 lambs (n = 81) were allocated into three groups: uninfected control, challenged either once (primary-infected animals) or twice (previously-infected animals) with 10,000 Haemonchus contortus L3. Uninfected control and challenged animals were sequentially sacrificed at 3, 7, 15 and 28 days post challenge. In both challenged groups, a clear Th2-oriented immune response was observed in the abomasal lymph node and mucosa. IL-4 and IL-13 mRNA over-expression, recruitment of eosinophils, mast cells and globule leukocytes and production of specific systemic IgG and mucosal IgA were observed earlier in previously-infected animals than in primary-infected ones. At 28 days post infection, no differences between intensities of these responses were observed between the challenged groups. Worm establishment rates were similar in previously-infected and primary-infected lambs. However, reductions of worm development, female fecundity and fecal egg output were observed in previously-infected sheep. We conclude that H. contortus infection in young INRA 401 lambs induced an unequivocal Th2 immune response resulting in the regulation of worm egg production without affecting their establishment.     

Is the 2000 CDC growth reference appropriate for developing countries?

In 2000, the Centers for Disease Control and Prevention (CDC) produced a revised growth reference. This has already been used in different settings outside the USA. Using data obtained during a nutritional survey in Madagascar, we compare results produced by using both the 2000 CDC and the 1978 National Center for Health Statistics (NCHS)/World Health Organization (WHO) growth references. We show that changing the reference has an important impact on nutritional diagnosis. In particular, the prevalence of wasting is greatly increased. This could generate substantial operational and clinical difficulties. We recommend continued use of the 1978 NCHS/WHO reference until release of the new WHO multi-country growth charts.