Development of a new method for the complete extraction of carotenoids from cereals with special reference to durum wheat (Triticum durum Desf.)

Due to the growing interest in the role of carotenoids in human health, their qualitative and quantitative analysis in foods is becoming more and more important. High-performance liquid chromatography has become the method of choice for the determination of these phytochemicals. A crucial step prior to the chromatographic separation is the quantitative extraction from the food matrix which was proven to be impeded in durum wheat. To optimize the extraction procedure, several factors with influence on extractability of carotenoids were investigated. Finally, it was shown that soaking of samples in water for 5 min prior to extraction with organic solvents had the strongest impact on extraction yield and led to the most rapid and gentle method. Contents of carotenoids in the extracts of several durum wheat and corn samples were doubled by soaking in water before extracting with methanol/tetrahydrofuran (1/1, v/v). In light of these findings, literature data on contents of carotenoids in cereal grains have to be viewed critically regarding the extraction procedures employed.     

Tissue Doppler Imaging and Two-dimensional Speckle Tracking of Left Ventricular Function in Horses Affected with Recurrent Airway Obstruction before and after Clenbuterol Treatment

Cardiac effects of the β₂-adrenergic agent clenbuterol have been the focus of many studies, but effects on myocardial velocities and myocardial deformation parameters have not yet been evaluated in horses affected with recurrent airway obstruction (RAO) using tissue Doppler imaging (TDI) and two-dimensional speckle tracking (2DST). In our study, 7 horses affected by RAO were treated over 14 days with clenbuterol, 0.8 μg/kg every 12 hours. Standard echocardiographic, TDI (pulsed wave and color TDI), and 2DST examinations were performed before and after the treatment period. Myocardial function was recorded in the right parasternal short-axis view. Percent of fractional shortening and two-dimensional echocardiography (2DE) measurements did not show any significant changes after 2 weeks of treatment. Early diastolic velocity, E, increased significantly after clenbuterol in the left ventricular free wall (LVFW; P = .001). The E/late diastolic velocity (A) quotient (P = .003) and the isovolumetric contractility (P = .035) also increased significantly after treatment. Time parameters, particularly the time interval between the Q-wave in the echocardiograph and atrial release, the time of diastole and Tei index (parameter of global ventricular function), decreased significantly after clenbuterol administration in the LVFW (P = .014/P = .028/P = .015, respectively). The 2DE speckle tracking revealed a significant increase of the early diastolic systolic strain rate (P = .01) in the LVFW after therapy. In conclusion, 2 weeks of treatment with clenbuterol at a dosage of 0.8 μg/kg every 12 hours led to improved cardiac function in severely RAO-affected horses. This could be a sign of myocardial restoration (re-remodeling) after therapy.     

In silico prediction of Gallibacterium anatis pan-immunogens

The Gram-negative bacterium Gallibacterium anatis is a major cause of salpingitis and peritonitis in commercial egg-layers, leading to reduced egg production and increased mortality. Unfortunately, widespread multidrug resistance and antigenic diversity makes it difficult to control infections and novel prevention strategies are urgently needed. In this study, a pan-genomic reverse vaccinology (RV) approach was used to identify potential vaccine candidates. Firstly, the genomes of 10 selected Gallibacterium strains were analyzed and proteins selected on the following criteria; predicted surface-exposure or secretion, none or one transmembrane helix (TMH), and presence in six or more of the 10 genomes. In total, 42 proteins were selected. The genes encoding 27 of these proteins were successfully cloned in Escherichia coli and the proteins expressed and purified. To reduce the number of vaccine candidates for in vivo testing, each of the purified recombinant proteins was screened by ELISA for their ability to elicit a significant serological response with serum from chickens that had been infected with G. anatis. Additionally, an in silico prediction of the protective potential was carried out based on a protein property prediction method. Of the 27 proteins, two novel putative immunogens were identified; Gab_1309 and Gab_2312. Moreover, three previously characterized virulence factors; GtxA, FlfA and Gab_2156, were identified. Thus, by combining the pan-genomic RV approach with subsequent in vitro and in silico screening, we have narrowed down the pan-proteome of G. anatis to five vaccine candidates. Importantly, preliminary immunization trials indicated an in vivo protective potential of GtxA-N, FlfA and Gab_1309.

Electronic supplementary material

The online version of this article (doi:10.1186/s13567-014-0080-0) contains supplementary material, which is available to authorized users.     

MAGNETIC RESONANCE IMAGING CONTRAST ENHANCEMENT OF EXTRA‐OCULAR MUSCLES IN DOGS WITH NO CLINICAL EVIDENCE OF ORBITAL DISEASE

Enhancement of extra‐ocular muscles has been reported in cases of orbital pathology in both veterinary and medical magnetic resonance imaging. We have also observed this finding in the absence of orbital disease. The purpose of this retrospective study was to describe extra‐ocular muscle contrast enhancement characteristics in a group of dogs with no known orbital disease. Magnetic resonance images (MRI) from dogs with no clinical evidence of orbital disease and a reportedly normal MRI study were retrieved and reviewed. Contrast enhancement percentages of the medial, lateral, ventral, and dorsal rectus muscles were calculated based on signal‐to‐noise ratios that were in turn determined from hand‐traced regions of interest in precontrast, immediate postcontrast and 10‐min postcontrast scans. Comparison measurements were made in the pterygoid muscle. Contrast enhancement of the extra‐ocular muscles was observed in all patients (median contrast enhancement percentage 45.0%) and was greater than that of pterygoid muscle (median contrast enhancement percentage 22.7%). Enhancement of the extra‐ocular muscles persisted 10 min after contrast administration (median contrast enhancement percentage 43.4%). Findings indicated that MRI contrast enhancement of extra‐ocular muscles is likely normal in dogs.     

Simple and Rapid Quality Control of Sulfated Glycans by a Fluorescence Sensor Assay—Exemplarily Developed for the Sulfated Polysaccharides from Red Algae Delesseria sanguinea

Sulfated polysaccharides (SP) from algae are of great interest due to their manifold biological activities. Obstacles to commercial (especially medical) application include considerable variability and complex chemical composition making the analysis and the quality control challenging. The aim of this study was to evaluate a simple microplate assay for screening the quality of SP. It is based on the fluorescence intensity (FI) increase of the sensor molecule Polymer-H by SP and was originally developed for direct quantification of SP. Exemplarily, 65 SP batches isolated from the red alga Delesseria sanguinea (D.s.-SP) and several other algae polysaccharides were investigated. Their FI increase in the Polymer-H assay was compared with other analytical parameters. By testing just one concentration of a D.s.-SP sample, quality deviations from the reference D.s.-SP and thus both batch-to-batch variability and stability can be detected. Further, structurally distinct SP showed to differ in their concentration-dependent FI profiles. By using corresponding reference compounds, the Polymer-H assay is therefore applicable as identification assay with high negative predictability. In conclusion, the Polymer-H assay showed to represent not only a simple method for quantification, but also for characterization identification and differentiation of SP of marine origin.     

Questioning the contemporary forest planning paradigm: making use of local knowledge

The forest planning system of large Swedish forest owners follows a three step procedure: long-term, medium-term, and short-term planning. The system is sequential and hierarchical in the sense that longer-term plans form the framework for shorter-term plans, and that top-level management prepares the long range plans and the lower management levels develop plans with successively shorter horizons. Studies indicate that this approach does not fully use existing knowledge within the organization. Problems associated with the top-down approach are also recognized in the general literature on organization and management. A proposal for a bottom-up approach is developed that aim at the use of local level knowledge to enhance accuracy and applicability of the forest plans. After top-level management has issued some fundamental planning directives, medium-term planning is conducted by the districts. Then the district plans are consolidated at the top-level for coordination and revision. A simulated planning process provides an illustration of the approach. The Heureka system is used here to optimize harvests and road costs with a mixed integer programming model of the problem, spanning 10 years with three seasons per year. The importance of detailed local knowledge to the outcome of planning is indicated, and needs for continued decision support systems development is discussed.