The Marine Fungal Metabolite,AD0157, Inhibits Angiogenesis by Targeting the Akt Signaling Pathway

In the course of a screening program for the inhibitors of angiogenesis from marine sources, AD0157, a pyrrolidinedione fungal metabolite, was selected for its angiosupressive properties. AD0157 inhibited the growth of endothelial and tumor cells in culture in the micromolar range. Our results show that subtoxic doses of this compound inhibit certain functions of endothelial cells, namely, differentiation, migration and proteolytic capability. Inhibition of the mentioned essential steps of in vitro angiogenesis is in agreement with the observed antiangiogenic activity, substantiated by using two in vivo angiogenesis models, the chorioallantoic membrane and the zebrafish embryo neovascularization assays, and by the ex vivo mouse aortic ring assay. Our data indicate that AD0157 induces apoptosis in endothelial cells through chromatin condensation, DNA fragmentation, increases in the subG1 peak and caspase activation. The data shown here altogether indicate for the first time that AD0157 displays antiangiogenic effects, both in vitro and in vivo, that are exerted partly by targeting the Akt signaling pathway in activated endothelial cells. The fact that these effects are carried out at lower concentrations than those required for other inhibitors of angiogenesis makes AD0157 a new promising drug candidate for further evaluation in the treatment of cancer and other angiogenesis-related pathologies.     

Differentiating between the supply of N to wheat from above and belowground residues of preceding crops of pea and canola

Belowground N comprises an important component of total residue N, but the N from this pool is often not included in studies that use ¹⁵N approaches to trace residue-derived N in succeeding crops. The objective of this greenhouse study was to differentiate between N supplied from aboveground and belowground crop residues, including rhizodeposits, of pea and canola to wheat using ¹⁵N labeling. A cross ¹⁵N-labeling approach was used in which wheat was grown on ¹⁵N-labeled belowground residues and non-labeled aboveground residues and vice versa. On average, the amount of N in wheat derived from belowground residues was almost twice the amount from aboveground residues. The higher input of both aboveground and belowground residue biomass and hence residue N from canola compared to pea tended to increase the residue-derived N in wheat from canola (6.4 %) relative to pea (4.7 %). However, differences in the percent recovery of ¹⁵N based on the amount of residue-¹⁵N initially applied revealed that a higher proportion of belowground residue N from pea (13.4 %) was recovered in wheat compared to the corresponding aboveground residue N from pea (8.8 %), and both belowground and aboveground residue N from canola (6.5 and 7.3 %, respectively). The total supply of N to wheat from preceding pea and canola crops was relatively low, likely due to wide C/N ratios. This study demonstrates the importance of belowground residue N to the supply of N to succeeding crops as well as differences between species.     

Evaluation of barcoded magnetic bead–based immunoassays for the simultaneous detection of feline leukemia virus antigen and antibody against feline immunodeficiency virus

Testing platforms that leverage automation, require minimal sample volume, and enable various tests to be performed simultaneously on a single sample have the potential to improve workflow and efficiency in veterinary diagnostic laboratories. We evaluated a barcoded magnetic bead (BMB) technology using established immunoassays for detection of feline leukemia virus (FeLV) p27 antigen and antibody against feline immunodeficiency virus (FIV). Analytical sensitivity, limit of blank, and limit of detection were used to establish a functional sensitivity of 1.00 ng/mL of inactivated FeLV antigen and 35.7 ng/mL of anti-FIV monoclonal antibody. Common interferents, such as hemoglobin, lipid, and bilirubin, were not found to interfere with the performance of the assay. Intra- and inter-assay CVs were <13% for both assays using manufactured samples. Using a set of 116 feline samples, the diagnostic accuracy of our multiplex assay was 100% compared to reference assays. Performance in a convenience set of 1,000 feline samples submitted to a commercial diagnostic laboratory revealed a proportion of positive results of 1.3% for FeLV and 3.7% for FIV. BMB technology should enable rapid screening of samples for various markers in a single immunoassay well.     

Epithelial‐to‐mesenchymal transition: immunohistochemical investigation of related molecules in canine cutaneous epithelial tumours

Background –  Epithelial‐to‐mesenchymal transition (EMT) is a multistep process, important in tumour invasion and metastasis, characterized by loss of epithelial markers, redistribution of β‐catenin and gain of mesenchymal markers. Hyposthesis/Objectives –  Our aim was to investigate the immunohistochemical aberrant expression of cytokeratin, vimentin, survivin and heat shock protein 72 (Hsp72) in canine cutaneous epithelial tumours, to understand the association of expression of these molecules with features of malignancy and their role in the EMT phenotype. Methods –  Ten canine squamous cell carcinomas (SCCs; one with lymph node metastasis), 30 canine hair follicle tumours (six pilomatricomas, eight infundibular keratinizing acanthomas, six trichoepitheliomas and 10 trichoblastomas) and five normal skin samples were investigated by immunohistochemistry using specific anti‐vimentin, ‐cytokeratin, ‐survivin and ‐Hsp72 antibodies. A semi‐quantitative method was used to analyse the results, as follows: 0 to <5%; ≥5 to <10%; ≥10 to <25%; and ≥25% of positive cells. Immunofluorescence was performed to investigate survivin–vimentin and survivin–Hsp72 colocalization in selected SCCs. Results –  In malignant hair follicle tumours and SCCs, a reduced intensity of cytokeratin and increased survivin and Hsp72 expression were observed. In SCCs, loss of cytokeratin expression and vimentin immunolabelling, suggestive of the EMT phenotype, were evident in <5% of neoplastic cells in the front of tumour invasion. In the same areas, strong nuclear survivin and cytoplasmic Hsp72 staining was evident, often colocalizing. Only a few neoplastic cells in the front of tumour invasion showed vimentin–survivin colocalization. Conclusions and clinical importance –  A possible simultaneous involvement of survivin and Hsp72 in tumour invasion and the multistep process of EMT of cutaneous epithelial tumours of dogs is suggested.     

Prevalence of Bartonella henselae antibodies in serum of cats with and without clinical signs of central nervous system disease

Bartonella henselae is occasionally associated with neurological dysfunction in people and some experimentally infected cats. The purpose of this study was to determine whether B henselae seroprevalence or titer magnitude varies among cats with neurological disease, cats with non-neurological diseases, and healthy cats while controlling for age and flea exposure. There was no difference in B henselae seroprevalence rates between cats with seizures and cats with other neurological diseases. Cats with non-neurological disease and healthy cats were more likely than cats with neurological disease to be seropositive. While the median B henselae antibody titer was greater in cats with seizures than in cats with other neurological disease, the median B henselae antibody titer was also greater in healthy cats than cats with seizures. The results suggest that titer magnitude cannot be used alone to document clinical disease associated with B henselae infection and that presence of B henselae antibodies in serum of cats with neurological disease does not prove the clinical signs are related to B henselae.     

Fecal microbial diversity and putative function in captive western lowland gorillas (Gorilla gorilla gorilla), common chimpanzees (Pan troglodytes), Hamadryas baboons (Papio hamadryas) and binturongs (Arctictis binturong)

Microbial populations in the gastrointestinal tract contribute to host health and nutrition. Although gut microbial ecology is well studied in livestock and domestic animals, little is known of the endogenous populations inhabiting primates or carnivora. We characterized microbial populations in fecal cultures from gorillas (Gorilla gorilla gorilla), common chimpanzees (Pan troglodytes), Hamadryas baboons (Papio hamadryas) and binturongs (Arctictis binturong) to compare the microbiomes associated with different gastrointestinal morphologies and different omnivorous feeding strategies. Each species was fed a distinct standardized diet for 2 weeks prior to fecal collection. All diets were formulated to reflect the species' feeding strategies in situ. Fresh fecal samples were pooled within species and used to inoculate in vitro batch cultures. Acetate, propionate, butyrate and valerate were measured after 24 h of incubation. Eubacterial DNA was extracted from individual fecal samples, pooled, and the cpn60 gene region was amplified and then sequenced to identify the major eubacterial constituents associated with each host species. Short chain fatty acids (P < 0.001) and methane (P < 0.001) were significantly different across species. Eubacterial profiles were consistent with fermentation data and suggest an increase in diversity with dietary fiber.     

Projecting species distributions using fishery-dependent data

Many marine species are shifting their distributions in response to changing ocean conditions, posing significant challenges and risks for fisheries management. Species distribution models (SDMs) are used to project future species distributions in the face of a changing climate. Information to fit SDMs generally comes from two main sources: fishery-independent (scientific surveys) and fishery-dependent (commercial catch) data. A concern with fishery-dependent data is that fishing locations are not independent of the underlying species abundance, potentially biasing predictions of species distributions. However, resources for fishery-independent surveys are increasingly limited; therefore, it is critical we understand the strengths and limitations of SDMs developed from fishery-dependent data. We used a simulation approach to evaluate the potential for fishery-dependent data to inform SDMs and abundance estimates and quantify the bias resulting from different fishery-dependent sampling scenarios in the California Current System (CCS). We then evaluated the ability of the SDMs to project changes in the spatial distribution of species over time and compare the time scale over which model performance degrades between the different sampling scenarios and as a function of climate bias and novelty. Our results show that data generated from fishery-dependent sampling can still result in SDMs with high predictive skill several decades into the future, given specific forms of preferential sampling which result in low climate bias and novelty. Therefore, fishery-dependent data may be able to supplement information from surveys that are reduced or eliminated for budgetary reasons to project species distributions into the future.