Synthesis and resistance to in vitro proteolysis of transglutaminase cross-linked phaseolin, the major storage protein from Phaseolus vulgaris

The ability of phaseolin to act as an acyl donor and acceptor substrate of transglutaminase was studied by using an enzyme isolated from Streptoverticillium mobarense. Phaseolin, a trimeric storage protein from Phaseolus vulgaris L., was shown to possess both glutamine and lysine residues reactive for the enzyme. The extent of transglutaminase-catalyzed cross-linking has been studied in function of both incubation time and enzyme concentration. Native- and SDS-PAGE demonstrated that phaseolin is intra- and intermolecularly cross-linked by transglutaminase and gives rise to different polymers as well as to modified forms of the protein having a similar molecular weight but lower Stokes radius if compared to unmodified phaseolin. Cross-linked phaseolin was found to be more resistant to proteolytic cleavage than the unmodified counterpart, as demonstrated by in vitro trypsin and pepsin digestion experiments. This behavior could suggest novel possible uses of the transglutaminase-modified phaseolin.     

Mixotrophy in a Local Strain of Nannochloropsis granulata for Renewable High-Value Biomass Production on the West Coast of Sweden

A local strain of Nannochloropsis granulata (Ng) has been reported as the most productive microalgal strain in terms of both biomass yield and lipid content when cultivated in photobioreactors that simulate the light and temperature conditions during the summer on the west coast of Sweden. To further increase the biomass and the biotechnological potential of this strain in these conditions, mixotrophic growth (i.e., the simultaneous use of photosynthesis and respiration) with glycerol as an external carbon source was investigated in this study and compared with phototrophic growth that made use of air enriched with 1–2% CO₂. The addition of either glycerol or CO₂-enriched air stimulated the growth of Ng and theproduction of high-value long-chain polyunsaturated fatty acids (EPA) as well as the carotenoid canthaxanthin. Bioassays in human prostate cell lines indicated the highest antitumoral activity for Ng extracts and fractions from mixotrophic conditions. Metabolomics detected betaine lipids specifically in the bioactive fractions, suggesting their involvement in the observed antitumoral effect. Genes related to autophagy were found to be upregulated by the most bioactive fraction, suggesting a possible therapeutic target against prostate cancer progression. Taken together, our results suggest that the local Ng strain can be cultivated mixotrophically in summer conditions on the west coast of Sweden for the production of high-value biomass containing antiproliferative compounds, carotenoids, and EPA.     

Assessing wild bees in perennial bioenergy landscapes: effects of bioenergy crop composition,landscape configuration,and bioenergy crop area

Context

Wild bee populations are currently under threat, which has led to recent efforts to increase pollinator habitat in North America. Simultaneously, U.S. federal energy policies are beginning to encourage perennial bioenergy cropping (PBC) systems, which have the potential to support native bees.

Objectives

Our objective was to explore the potentially interactive effects of crop composition, total PBC area, and PBC patches in different landscape configurations.

Methods

Using a spatially-explicit modeling approach, the Lonsdorf model, we simulated the impacts of three perennial bioenergy crops (PBC: willow, switchgrass, and prairie), three scenarios with different total PBC area (11.7, 23.5 and 28.8% of agricultural land converted to PBC) and two types of landscape configurations (PBC in clustered landscape patterns that represent realistic future configurations or in dispersed neutral landscape models) on a nest abundance index in an Illinois landscape.

Results

Our modeling results suggest that crop composition and PBC area are particularly important for bee nest abundance, whereas landscape configuration is associated with bee nest abundance at the local scale but less so at the regional scale.

Conclusions

Strategies to enhance wild bee habitat should therefore emphasize the crop composition and amount of PBC.

     

Unipodal stance influences radiographic evaluation of foot balance in horses

Lifting the contralateral forelimb (unipodal stance) is often used as a method of restraint in horses. This experimental one group pretest, posttest study was conducted to evaluate the effects of unipodal stance on quantitative radiographic parameters in equine forefeet. Seven nonlame horses were randomly selected. Lateromedial (LM) and dorsopalmar (DP) projections were acquired for both forefeet, squarely placed on blocks, using two X‐ray generators. Radiographs of each foot were acquired first in a bipodal stance, immediately followed by the same radiographic projections obtained in a unipodal stance. The following measurements were recorded for each stance: distal interphalangeal joint (DIPJ) space width on both projections; mediolateral joint balance as the difference between lateral and medial DIPJ space widths on DP projections; extensor process‐to‐middle phalangeal condyle distance; and deep digital flexor tendon angle on LM projections. A matched pairs design and Student's t‐test with a 95% confidence level were used to test for statistical significance. Compared to a bipodal stance, lateral DIPJ space width was significantly reduced on unipodal DP views, whereas mediolateral joint imbalance and to a lesser extent medial DIPJ space width were significantly increased. On unipodal LM views, there was a significant higher degree of DIPJ flexion. These findings suggest that stance should be carefully taken into consideration when measuring radiographic parameters in equine forefeet, especially if assessing foot balance and conformation, as unipodal stance significantly affects the mediolateral balance of the DIPJ on DP radiographs and significantly alters the phalangeal axis on LM radiographs.     

Susceptibility to and transmission of H5N1 and H7N1 highly pathogenic avian influenza viruses in bank voles (Myodes glareolus)

The study of influenza type A (IA) infections in wild mammals populations is a critical gap in our knowledge of how IA viruses evolve in novel hosts that could be in close contact with avian reservoir species and other wild animals. The aim of this study was to evaluate the susceptibility to infection, the nasal shedding and the transmissibility of the H7N1 and H5N1 highly pathogenic avian influenza (HPAI) viruses in the bank vole (Myodes glareolus), a wild rodent common throughout Europe and Asia. Two out of 24 H5N1-infected voles displayed evident respiratory distress, while H7N1-infected voles remained asymptomatic. Viable virus was isolated from nasal washes collected from animals infected with both HPAI viruses, and extra-pulmonary infection was confirmed in both experimental groups. Histopathological lesions were evident in the respiratory tract of infected animals, although immunohistochemistry positivity was only detected in lungs and trachea of two H7N1-infected voles. Both HPAI viruses were transmitted by direct contact, and seroconversion was confirmed in 50% and 12.5% of the asymptomatic sentinels in the H7N1 and H5N1 groups, respectively. Interestingly, viable virus was isolated from lungs and nasal washes collected from contact sentinels of both groups. The present study demonstrated that two non-rodent adapted HPAI viruses caused asymptomatic infection in bank voles, which shed high amounts of the viruses and were able to infect contact voles. Further investigations are needed to determine whether bank voles could be involved as silent hosts in the transmission of HPAI viruses to other mammals and domestic poultry.     

Definitive‐intent intensity‐modulated radiation therapy provides similar outcomes to those previously published for definitive‐intent three‐dimensional conformal radiation therapy in dogs with primary brain tumors: A multi‐institutional retrospective study

Radiotherapy with or without surgery is a common choice for brain tumors in dogs. Although numerous studies have evaluated use of three‐dimensional conformal radiotherapy, reports of definitive‐intent, IMRT for canine intracranial tumors are lacking. Intensity‐modulated radiation therapy has the benefit of decreasing dose to nearby organs at risk and may aid in reducing toxicity. However, increasing dose conformity with IMRT calls for accurate target delineation and daily patient positioning, in order to decrease the risk of a geographic miss. To determine survival outcome and toxicity, we performed a multi‐institutional retrospective observational study evaluating dogs with brain tumors treated with IMRT. Fifty‐two dogs treated with fractionated, definitive‐intent IMRT at four academic radiotherapy facilities were included. All dogs presented with neurologic signs and were diagnosed via MRI. Presumed radiological diagnoses included 37 meningiomas, 12 gliomas, and one peripheral nerve sheath tumor. One dog had two presumed meningiomas and one dog had either a glioma or meningioma. All dogs were treated in the macroscopic disease setting and were prescribed a total dose of 45‐50 Gy (2.25‐2.5 Gy per fraction in 18‐20 daily fractions). Median survival time for all patients, including seven cases treated with a second course of therapy was 18.1 months (95% confidence of interval 12.3‐26.6 months). As previously described for brain tumors, increasing severity of neurologic signs at diagnosis was associated with a worse outcome. Intensity‐modulated radiation therapy was well tolerated with few reported acute, acute delayed, or late side effects.     

ULTRASONOGRAPHIC CHARACTERISTICS OF THE CISTERNA CHYLI IN EIGHT DOGS AND FOUR CATS

Ultrasonography of the cisterna chyli has been used in humans to diagnose increased lymphatic flow or lymph flow obstruction and to guide percutaneous embolization of the thoracic duct via the cisterna chyli. The aim of this study was to describe the ultrasonographic characteristics of the dorsal portion of cisterna chyli in dogs and cats with chylous ascites or chylothorax and in a group of healthy dogs and cats. The aorta and the cranial mesenteric artery were used as anatomic landmarks. Ultrasonography was performed before and 2 h after a fatty meal in healthy dogs and cats. The visualized structure was confirmed to be a dilated cisterna chyli at necropsy in a dog with chylous ascites. The confirmed or presumed cisterna chyli was consistently detected using ultrasonography in nonfasted healthy animals and clinically affected animals and appeared as an anechoic tubular structure, without detectable flow, at the right dorsolateral aspect of the aorta. It had a similar ultrasonographic appearance in patients with chyloabdomen and in nonfasted healthy dogs and cats. There was considerable overlap in diameters of the cisterna chyli for affected and healthy animals. The shape and size of the cisterna chyli in an individual animal were variable during the same ultrasound examination and between different examinations. This study demonstrated the appearance of the presumed dorsal portion of the cisterna chyli by ultrasonography and might provide useful preliminary data for further studies into the feasibility of ultrasound‐guided injections or aspirations of the cisterna chyli in dogs and cats.