Cytotoxic Effects of Loperamide Hydrochloride on Canine Cancer Cells

Loperamide is a peripheral opiate agonist that can cause apoptosis and G2/M arrest in human cancer cell lines and may sensitize cells to chemotherapy. The objectives of this study were to investigate the effects of loperamide on viability, apoptosis and cell cycle kinetics in canine cancer cells and to establish whether the drug sensitizes cells to doxorubicin. Cell viability was assessed using Alamar Blue. Cell death and cell cycle were studied using flow cytometry with 7-Aminoactinomycin-D (7-AAD) and propidium iodide (PI), respectively. Loperamide decreased cell viability in a dose-dependent fashion and was most effective against canine osteosarcoma cells. In all cell lines, it induced a dose and time dependent apoptosis and resulted in accumulation in G0/G1. When co-incubated with doxorubicin, loperamide induced a synergistic cell kill in canine carcinoma cells. Investigation is warranted into the role of loperamide in the treatment of canine cancer.     

US strategy for forest management adaptation to climate change: building a framework for decision making

Context

Recent policy changes in the USA direct agencies managing federal forests to analyze the potential effects of climate change on forest productivity, water resource protection, wildlife habitat, biodiversity, and other values.

Aims

This paper describes methods developed to (1) assess current risks, vulnerabilities, and gaps in knowledge; (2) engage internal agency resources and external partners in the development of options and solutions; and (3) manage forest resources for resilience, not just in terms of natural ecosystems but in affected human communities as well.

Methods

We describe an approach designed to characterize certain climate change effects on forests, and estimate the effectiveness of response options ranging from resistance to a realignment of management objectives.

Results

Field testing on a 6,300 km² area of conifer forest in the northwestern USA shows this decision model to be useful and cost-effective in identifying the highest sensitivities relating to vegetation management, biological diversity, water resources and forest transportation systems, and building consensus for adaptive strategies and actions.

Conclusions

Results suggest that this approach is an effective means for guiding management decisions to adapt to the effects of climate change, and provides an empirical basis for setting budgetary and management priorities.     

Feasibility and repeatability of cold and mechanical quantitative sensory testing in normal dogs

Feasibility and inter-session repeatability of cold and mechanical quantitative sensory testing (QST) were assessed in 24 normal dogs. Cold thermal latencies were evaluated using a thermal probe (0 °C) applied to three pelvic limb sites. Mechanical thresholds were measured using an electronic von Frey anesthesiometer (EVF) and a blunt-probed pressure algometer (PA) applied to the dorsal aspect of the metatarsus. All QST trials were performed with dogs in lateral recumbency. Collection of cold QST data was easy (feasible) in 19/24 (79%) dogs. However, only 18.4%, 18.9% and 13.2% of cold QST trials elicited a response at the medial tibia, third digital pad and plantar metatarsal regions, respectively. Collection of mechanical QST data was easy (feasible) in 20/24 (83%) dogs for both EVF and PA.At consecutive sampling times, approximately 2 weeks apart, the average EVF sensory thresholds were 414 ± 186 g and 379 ± 166 g, respectively, and the average PA sensory thresholds were 1089 ± 414 g and 1028 ± 331 g, respectively. There was no significant difference in inter-session or inter-limb threshold values for either mechanical QST device. The cold QST protocol in this study was achievable, but did not provide consistently quantifiable results. Both mechanical QST devices tested provided repeatable, reliable sensory threshold measurements in normal, client-owned dogs. These findings contribute to the validation of the EVF and PA as tools to obtain repeated QST data over time in dogs to assess somatosensory processing changes.     

Does Biochar Improve Establishment of Tree Seedlings in Saline Sodic Soils?

Reforestation of saline sodic soil is increasingly undertaken as a means of reclaiming otherwise unproductive agricultural land. Currently, restoration of degraded land is limited to species with high tolerances of salinity. Biochar application has the potential to improve physical, biological and chemical properties of these soils to allow establishment of a wider range of plants. In a glasshouse trial, we applied biochar made from Acacia pycnantha (5 Mg ha⁻¹) or no biochar to either a low (EC_e 4·75 dS m⁻¹, ESP 6·9), a moderate (EC_e 27·6 dS m⁻¹, ESP 29·3) or a high (EC_e 49·4 dS m⁻¹, ESP 45·1) saline sodic soil. The regional common reforestation species Eucalyptus viminalis and Acacia mearnsii were planted as tubestock in to the soils. Early establishment indicators, including growth, plant condition and nutrition, were assessed at the end of a simulated growing season, 108 days after biochar application. Application of biochar increased height, and decreased root : shoot and the concentration of Mn, N and S in plants of E. viminalis when grown in the highly saline sodic soil. Biochar application increased the concentration of B in leaves of E. viminalis and increased the concentration of P, K and S in leaves of A. mearnsii when grown in the low saline sodic soil. The results confirm that there is potential for biochar to assist in reforestation of saline sodic soils. Copyright © 2015 John Wiley & Sons, Ltd.     

Induction of tetraploidy in meristematically active seeds of Japanese barberry (Berberis thunbergii var. atropurpurea) through exposure to colchicine and oryzalin

Japanese barberry (Berberis thunbergii DC) is an invasive shrub, widely naturalized across the United States, whose numerous cultivars remain an important horticultural commodity. Maintaining this crop for the future necessitates the development of sterile clones. Exposure to the mitotic inhibitors colchicine and oryzalin is a traditional method for inducing tetraploidy in breeding lines as a precursor to creating sterile genotypes. Treatments utilized pre-germinated B. t. var. atropurpurea seeds with emerged radicles. Seeds were immersed in aqueous solutions of colchicine (.02%, .05%, .1% and .2%) and oryzalin (.002%, .005%, .01% and .02%) dissolved in 1% DMSO for 6, 12 and 24 h durations. Seedling ploidy level was determined via flow cytometry following 6 and 52 weeks of growth in the greenhouse. Both anti-mitotic chemicals proved effective at inducing tetraploidy and produced comparable efficiency rates. The survival rate of treated seeds decreased in response to both increased mitotic inhibitor concentration and longer exposure duration. While exposure to oryzalin produced greater seed mortality than colchicine, most seedlings that survived had altered ploidy levels. The most efficient oryzalin concentration was 0.002% with a rating of 28%, while the most efficient colchicine concentrations were in the range from 0.05% to 0.2%. Duration of exposure to mitotic inhibitor was not a significant factor over the range from 6 to 24 h. Reversion of tetraploid plants to the diploid state occurred at a low frequency following a dormancy period. Some tetraploid seedlings derived from exposure to both chemicals displayed foliar abnormalities including irregular leaf margins and mottled lamina. The primary advantage of colchicine was low seedling toxicity, while oryzalin was notable for its ability to induce tetraploidy at low concentrations.     

Application of unmanned aerial systems for high throughput phenotyping of large wheat breeding nurseries

Background

Low cost unmanned aerial systems (UAS) have great potential for rapid proximal measurements of plants in agriculture. In the context of plant breeding and genetics, current approaches for phenotyping a large number of breeding lines under field conditions require substantial investments in time, cost, and labor. For field-based high-throughput phenotyping (HTP), UAS platforms can provide high-resolution measurements for small plot research, while enabling the rapid assessment of tens-of-thousands of field plots. The objective of this study was to complete a baseline assessment of the utility of UAS in assessment field trials as commonly implemented in wheat breeding programs. We developed a semi-automated image-processing pipeline to extract plot level data from UAS imagery. The image dataset was processed using a photogrammetric pipeline based on image orientation and radiometric calibration to produce orthomosaic images. We also examined the relationships between vegetation indices (VIs) extracted from high spatial resolution multispectral imagery collected with two different UAS systems (eBee Ag carrying MultiSpec 4C camera, and IRIS+ quadcopter carrying modified NIR Canon S100) and ground truth spectral data from hand-held spectroradiometer.

Results

We found good correlation between the VIs obtained from UAS platforms and ground-truth measurements and observed high broad-sense heritability for VIs. We determined radiometric calibration methods developed for satellite imagery significantly improved the precision of VIs from the UAS. We observed VIs extracted from calibrated images of Canon S100 had a significantly higher correlation to the spectroradiometer (r = 0.76) than VIs from the MultiSpec 4C camera (r = 0.64). Their correlation to spectroradiometer readings was as high as or higher than repeated measurements with the spectroradiometer per se.

Conclusion

The approaches described here for UAS imaging and extraction of proximal sensing data enable collection of HTP measurements on the scale and with the precision needed for powerful selection tools in plant breeding. Low-cost UAS platforms have great potential for use as a selection tool in plant breeding programs. In the scope of tools development, the pipeline developed in this study can be effectively employed for other UAS and also other crops planted in breeding nurseries.

     

Comparisons of Mayan forest management, restoration, and conservation

Numerous communities associated with at least five distinct ethnic Mayan groups in southern Mexico and Central America continue to rely upon forested areas as integral components of their agricultural systems. They carefully manage these areas so that forests provide food, raw materials, and animals. Management practices include removing and planting of woody and herbaceous species, apiculture, and seed harvest. Mayan agroforestry systems in geographically and ecologically distinct areas of Mesoamerica were evaluated to better understand traditional agroforestry system components and how indigenous Mayan agroforestry could be a part of regional forest conservation and restoration. Systems were within Mexican land grant areas (ejidos) or on contested land. Although these systems rely upon different woody species and management techniques, common among them are: (1) the use of multi-stage and successional pathways with forest as a part of the larger system, (2) species that are believed by traditional ecological knowledge (TEK) to accelerate forest regeneration - more than 30 tree species are recognized and managed as potential facilitators of forest regeneration and (3) direct human consumption of forest products at all stages of regeneration.     

Induction of tumors in mice by genomic hypomethylation

Genome-wide DNA hypomethylation occurs in many human cancers, but whether this epigenetic change is a cause or consequence of tumorigenesis has been unclear. To explore this phenomenon, we generated mice carrying a hypomorphic DNA methyltransferase 1 (Dnmt1) allele, which reduces Dnmt1 expression to 10% of wild-type levels and results in substantial genome-wide hypomethylation in all tissues. The mutant mice were runted at birth, and at 4 to 8 months of age they developed aggressive T cell lymphomas that displayed a high frequency of chromosome 15 trisomy. These results indicate that DNA hypomethylation plays a causal role in tumor formation, possibly by promoting chromosomal instability.