Production and characterisation of monoclonal antibodies to cell wall components of the flax rust fungus

Monoclonal antibodies have been raised against an haustorium-enriched sample prepared from flax leaves infected with the biotrophic flax rust pathogen Melampsora lini. The monoclonal antibodies were produced following conventional and co-immunisation procedures and the range of antibody specificities was compared. The preparation used as immunogen for the conventional protocol was a crude isolate of haustoria consisting of approx. 65% fungal haustoria, the other components being mainly mesophyll cells or cell wall and chloroplast fragments. Following hybridoma production, 40% of positive cell lines produced antibodies that reacted with haustoria and other fungal cells, but 60% bound to plant cells in the infected leaves. For the co-immunisation protocol, the preparation used for immunisation consisted of the crude isolate of haustoria mixed with serum raised against an haustorium-depleted leaf homogenate. In two fusions, 92-94% of the antibodies reacted with fungal cells, including 3 cell lines that localised specifically to the cell wall of haustoria. Only 6-8% of the antibodies produced via co-immunisation reacted with plant cells. The antigens targeted by the three haustorium-specific monoclonal antibodies are incorporated into the wall at early stages of haustorium development, remain in the wall throughout haustorium maturation, and are present in both compatible and incompatible interactions. The epitopes recognised by the monoclonal antibodies are oligosaccharide in nature and the antigens are highly resistant to extraction from the wall. These results highlight the value of the co-immunisation protocol for the production of monoclonal antibodies to specific components in an impure preparation and provide direct evidence for molecular differentiation within the wall of the haustorium of M. lini.     

Effects of forest floor planting and stock type on growth and root emergence of Pinus contorta seedlings in a cold northern cutblock

A two-year field trial was conducted to determine the growth response, and root emergence pattern of interior lodgepole pine (Pinus contorta Dougl. var. latifolia Engelm.) seedlings in response to container type and forest floor removal. Seedlings were grown in Styroblocks^TM, Copperblocks^TM, or AirBlocks^TM, and were planted directly into the undisturbed forest floor or into manually prepared planting spots where the forest floor had been scraped away to expose the mineral soil. Seedlings planted into scalped planting spots exhibited marginally but significantly (7%) greater above-ground growth rates (seedling stem volume); whereas seedlings planted into the forest floor produced significantly more (11%) new roots. There were no differences in above- or below-ground biomass. Seedlings grown in Copperblock^TM containers produced a higher proportion of roots near the top of the plug when tested at lifting, however this pattern was not observed in the field. Given that scalping is more costly than forest floor planting, and that the increased shoot growth was relatively small, we recommend that forest floor planting be considered as an alternative to manual spot scalping for sites, such as the site tested here: those with cold, but well-drained soils and where competition from other plants is not a serious problem.     

Orientation of asymmetric stem cell division by the APC tumor suppressor and centrosome

Stem cell self-renewal can be specified by local signals from the surrounding microenvironment, or niche. However, the relation between the niche and the mechanisms that ensure the correct balance between stem cell self-renewal and differentiation is poorly understood. Here, we show that dividing Drosophila male germline stem cells use intracellular mechanisms involving centrosome function and cortically localized Adenomatous Polyposis Coli tumor suppressor protein to orient mitotic spindles perpendicular to the niche, ensuring a reliably asymmetric outcome in which one daughter cell remains in the niche and self-renews stem cell identity, whereas the other, displaced away, initiates differentiation.     

Stabilities of daidzin,glycitin, genistin,and generation of derivatives during heating

The soy isoflavones daidzin, glycitin, and genistin were purified from defatted soy flour using preparative-scale reverse-phase HPLC. The stabilities of the three isoflavones at different heating temperatures were investigated. Daidzin, glycitin, and genistin were lost at a rate of 26, 27, and 27% of their original concentration, respectively, after 3 min at 185 degrees C. At 215 degrees C, decreases of daidzin, glycitin, and genistin were 65, 98, and 74% after 3 min and 91, 99, and 94% after 15 min, respectively. The order of the thermal stabilities, from lowest to highest, was glycitin, genistin, and daidzin. Acetyl daidzin and acetyl genistin, daidzein, glycitein, and genistein were produced during heating at temperatures above 135 degrees C. The rate of binding of an acetyl group to form acetyl daidzin and acetyl genistin from daidzin and genistin was higher than the rate of loss of a glucoside group to form daidzein and genistein. However, acetyl daidzin and acetyl genistin decreased sharply at temperatures above 200 degrees C, while daidzein, glycitein, and genistein were relatively stable over 30 min. The stability of daidzein was higher than that of glycitein or genistein.     

The first reported case of rabbit hemorrhagic disease in Canada

In March 2011, rabbit hemorrhagic disease (RHD) was suspected in a 1-year-old male neutered lop-eared rabbit that had acute onset liver failure. Gross pathology, histopathology, immunohistochemistry, partial nucleic acid sequencing and phylogenetic analysis of the major capsid protein (VP60) and animal inoculation studies all supported this diagnosis making it the first confirmed case of RHD in Canada.     

Radiotherapy isocenters verified by matching to bony landmarks of the canine and feline head differ when localized using volumetric versus planar imaging

The “gold standard” for verification of patient positioning before linear accelerator‐based stereotactic radiation therapy is kilovoltage cone‐beam computed tomography (kV‐CBCT), which is not uniformly available or utilized; planar imaging is sometimes used instead. The primary aim of this study was to determine if the position of the bony skull, when used as a surrogate for isocenter verification, is different when orthogonal megavoltage (MV) portal or kilovoltage (kV/kV) radiographs are used for image guidance, rather than kV‐CBCT. A secondary aim was to determine the influence of intra‐observer variability, body size and skull conformation on positioning, as determined using these three imaging modalities. Dogs and cats receiving radiotherapy of the head were recruited for this prospective analytical study. Planar (MV portal and kV/kV images) and volumetric (kV‐CBCT) images were acquired before treatment, and manually coregistered with reference images. Differences in skull position when matched based on MV portal, kV/kV images and kV‐CBCT were compared. A total of 65 subjects and 148 unique datasets were evaluated. The Wilcoxon rank‐sum test was used to evaluate effects of transitioning between imaging modalities. When comparing magnitude of shifts in MV to kV‐CBCT, MV to kV/kV and kV/kV to kV‐CBCT, there were statistically significant differences. Results were not measurably impacted by body size, skull conformation or interobserver differences. Based on shift magnitude and direction, an isotropic setup margin of at least 1 mm should be incorporated within the planning target volume when MV or kV planar imaging is used for position verification.     

Leptospirosis in beef herds from western Canada: serum antibody titers and vaccination practices

One study described the frequency of pre-breeding vaccination for leptospirosis in 205 cow-calf herds from across western Canada and the prevalence of positive Leptospira antibody titers in unvaccinated, weaned calves from 61 of these herds. The percentages of herds vaccinated for leptospirosis were 13.7% in 2001 and 8.4% in 2002. Of 1539 calves examined, 13 (0.8%) had a positive antibody titer for a Leptospira serovar; the most common serovar detected was hardjo. A second study examined the prevalence of positive Leptospira antibody titers during the summer grazing season in 313 vaccinated and 478 unvaccinated cows from 40 cow-calf herds in southern Saskatchewan. Antibody titers for 7 Leptospira serovars were measured during the grazing season. Of the non-vaccinated cows, 9.6% were positive in the spring for serovar pomona, 6.7% for serovar grippotyphosa, and 6.1% for serovar icterohaemorrhagiae; the corresponding percentages for the fall were 5.5%, 3.0%, and 1.3%, respectively. Of 781 vaccinated and unvaccinated cows that were sampled twice, 11.3% of vaccinated cows and 2.3% of unvaccinated cows had increases in Leptospira antibody titers during the grazing season.     

Extraction of Antioxidants from Wheat Bran Using Conventional Solvent and Microwave-Assisted Methods

Total phenolic and tocopherol contents and free radical scavenging capability of wheat bran extracted using conventional and microwave-assisted solvent extraction methods were studied. Three different solvents (methanol, acetone, and hexane) were used in the conventional solvent extraction. Methanol was the most effective solvent, producing higher extraction yield (4.86%), total phenolic compound content (241.3 μg of catechin equivalent/g of wheat bran), and free radical scavenging capability (0.042 μmol of trolox equivalent/g of wheat bran) than either acetone or hexane. However, there was no significant difference in the total tocopherol contents (13.6–14.8 μg/g of wheat bran) among the three different solvent extraction methods. Microwave-assisted solvent extraction using methanol significantly increased the total phenolic compound content to 467.5 and 489.5 μg of catechin equivalent; total tocopherol content to 18.7 and 19.5 μg; and free radical scavenging capability to 0.064 and 0.072 μmol of trolox equivalent/g of wheat bran at extraction temperatures of 100 and 120°C, respectively. However, extraction yields of conventional methanol solvent and microwave-assisted methanol extractions at different temperatures were not significantly different.     

Stop using dichotomous terms to reference observations of scale-dependent habitat selection

Context

Many studies dichotomize habitat selection into “local” or “landscape” effects, with little explanation regarding what each represents. Ambiguous use of these terms across studies may confound observations of selection operating across different spatiotemporal resolutions and impede synthesis of scale-dependent habitat selection.

Objective

To examine the consistency of use of the terms “local” and “landscape” level habitat selection and evaluate potential implications for conservation science.

Methods

We reviewed 136 multi-level studies observing local and/or landscape effects. From each study, we identified which of Johnson’s (Ecology 61:65–71, 1980) selection orders was observed at the local and landscape scale. We assessed consistency in the selection order observed at each scale.

Results

At the local scale, 54% of studies observed 3rd order selection and 38% of studies observed 4th order selection. At the landscape scale, 64% of studies observed 2nd order selection, while only 8% of studies observed 1st order selection. Overlap also occurred; 2nd order selection was observed at the local scale in 8% of studies, while 3rd and 4th order selection was observed at the landscape scale in 24 and 3% of studies, respectively.

Conclusions

Selection orders observed at local and landscape scales are inconsistent across studies, making comparisons and synthesis difficult. This inconsistency makes the terms “local” and “landscape” meaningless, and may lead to misidentification of limiting factor(s) most important for conservation efforts. Dichotomous terms should no longer be used in reference to levels of selection, but in reference to specific explanatory variables whose characteristic scale(s) best fit the appropriate definition.