Characterization of isoflavone composition in soy-based nutritional supplements via ultra performance liquid chromatography.

The specific isoflavone composition of nutritional supplements is commonly not-labeled, although the stated amounts are strongly dependent on the present isoflavone conjugates. Hence, 11 soy-based dietary supplements were characterized via a newly established ultra performance liquid chromatography (UPLC) method, on both their native conjugated isoflavone spectra, as well as on quantitative amounts derived as total aglycones after enzymatic hydrolysis utilizing Helix pomatia juice. Capitalizing on sub-2 microm particles, the established RP-UPLC technique facilitated efficient chromatographic separation of all 12 soy intrinsic isoflavone forms within 10 min.

Evaluation of different expression systems for the heterologous expression of pyranose 2-oxidase from Trametes multicolor in E. coli.

The heterologous production of the industrially relevant fungal enzyme pyranose 2-oxidase in the prokaryotic host E. coli was investigated using 3 different expression systems, i.e.

Introducing antigen-binding sites in structural loops of immunoglobulin constant domains: Fc fragments with engineered HER2/neu-binding sites and antibody properties.

Yeast surface display libraries of human IgG1 Fc regions were prepared in which loop sequences at the C-terminal tip of the CH3 domain were randomized. A high percentage of these library members bound to soluble CD64 and Protein A indicating that the randomization step did not grossly interfere with the overall structure of the displayed Fc. Sorting these libraries by FACS for binders against HER2/neu yielded antigen-specific Fc binders (Fcab; Fc antigen binding) of which one was affinity matured, resulting in Fcab clone H10-03-6 which showed >10-fold improvement in antigen-binding activity versus the parental clone.

Harnessing ecosystem models and multi-criteria decision analysis for the support of forest management.

The decision-making environment in forest management (FM) has changed drastically during the last decades. Forest management planning is facing increasing complexity due to a widening portfolio of forest goods and services, a societal demand for a rational, transparent decision process and rising uncertainties concerning future environmental conditions (e.g.

Risk factors for lameness in freestall-housed dairy cows across two breeds, farming systems, and countries.

Lameness poses a considerable problem in modern dairy farming. Several new developments (e.g.

A new ultra-pressure liquid chromatography method for the determination of biogenic amines in cheese.

A fast and reliable ultra-performance liquid chromatography (UPLC) method for the determination of biogenic amines (ethanolamine, methylamine, agmatine, histamine, dimethylamine, ethylamine, octopamine, pyrrolidine, dopamine, isopropylamine, propylamine, tyramine, putrescine, butylamine, cadaverine, tryptamine, 2-phenylethylamine, 3-methylbutylamine, spermidine, spermine) in cheese was established. After pre-column derivatization with 6-aminoquinolyl-N-hydroxy-succinimidyl carbamate (AQC), 20 primary and secondary biogenic amines were separated on an Acquity UPLC column (BEH C(18), 1.7 microm; 2.

Starch as a sugar reservoir for nematode-induced syncytia.

The plant parasitic nematode Heterodera schachtii invades the roots of Arabidopsis thaliana to induce nematode feeding structures in the central cylinder. During nematode development, the parasites feed exclusively from these structures. Thus, high sugar import and specific sugar processing of the affected plant cells is crucial for nematode development.

Diversity and activity of sugar transporters in nematode-induced root syncytia.

The plant-parasitic nematode Heterodera schachtii stimulates plant root cells to form syncytial feeding structures which synthesize all nutrients required for successful nematode development. Cellular re-arrangements and modified metabolism of the syncytia are accompanied by massive intra- and intercellular solute allocations. In this study the expression of all genes annotated as sugar transporters in the Arabidopsis Membrane Protein Library was investigated by Affymetrix gene chip analysis in young and fully developed syncytia compared with non-infected Arabidopsis thaliana roots.

Risk factors for lameness in cubicle housed Austrian Simmental dairy cows.

Austrian dairy farming is characterised by predominant use of Simmental cows on small-scale farms. Our aim was to identify lameness risk factors related to housing and management in cubicle housed Austrian dairy cows. Furthermore, we used animal-based parameters (ABP) as integrated measures of cubicle quality and feeding management.

Approaches to assess biocover performance on landfills.

Methane emissions from active or closed landfills can be reduced by means of methane oxidation enhanced in properly designed landfill covers, known as "biocovers". Biocovers usually consist of a coarse gas distribution layer to balance gas fluxes placed beneath an appropriate substrate layer. The application of such covers implies use of measurement methods and evaluation approaches, both during the planning stage and throughout the operation of biocovers in order to demonstrate their efficiency.

Design of a village breeding programme for a llama population in the High Andes of Bolivia.

No national breeding programme for llamas is in place in Bolivia. Initiatives for genetic improvement are rarely found and are usually carried out by NGOs working in rural development or improvement of livestock production or research stations. Farmers in the Province of Ayopaya in the District of Cochabamba have formed a breeders' organization with the aim of improving fibre production.

High-level expression of recombinant beta-galactosidases in Lactobacillus plantarum and Lactobacillus sakei using a Sakacin P-based expression system.

This work presents the cloning and expression of the genes encoding heterodimeric beta-galactosidases from Lactobacillus reuteri L103, Lactobacillus acidophilus R22, Lactobacillus plantarum WCFS1, and Lactobacillus sakei Lb790. These enzymes consist of two subunits of approximately 73 and 35 kDa, which are encoded by two overlapping genes, lacL and lacM, respectively. We have cloned these genes into the lactobacillal expression vectors pSIP403 and pSIP409, which are based on the sakacin P operon of L.

S-layer stabilized lipid membranes (Review).

The present review focuses on a unique bio-molecular construction kit based on surface-layer (S-layer) proteins as building blocks and patterning elements, but also major classes of biological molecules such as lipids, membrane-active peptides and membrane proteins, and glycans for the design of functional supported lipid membranes. The biomimetic approach copying the supramolecular building principle of most archaeal cell envelopes merely composed of a plasma membrane and a closely associated S-layer lattice has resulted in robust and fluid lipid membranes. Most importantly, S-layer supported lipid membranes spanning an aperture or generated on solid and porous substrates constitute highly interesting model membranes for the reconstitution of responsive transmembrane proteins and membrane-active peptides.

The water framework directive -can we reach the target?

The WFD is an overarching piece of legislation that aims to harmonize existing European water policy; since 2000 it requires managing the river basins so that the quality and quantity of water does not affect the ecological services and to promote sustainable water use of any specific water body. Nevertheless the goals of other directives, such as drinking water, bathing water and urban wastewater treatment (UWWT), are not yet harmonised mainly concerning microbiological and PS/PSR/PHS contamination. Great challenges are due to emerging contaminants especially the technical improvement and harmonisation for risk identification and risk assessment: for biological effects and for chemical analysis and, finally integrate this knowledge to preserve the "good status".

Sucrose supply to nematode-induced syncytia depends on the apoplasmic and symplasmic pathways.

The plant parasitic nematode Heterodera schachtii induces syncytial feeding structures in the roots of host plants. Nematode-induced syncytia become strong sink tissues in the plant solute circulation system as the parasites start withdrawing nutrients. In the present work, the expression pattern of the phloem-specific sucrose transporter AtSUC4 (also described as AtSUT4) is analysed in syncytia induced by H.

Comparison of production systems and selection criteria of Ankole cattle by breeders in Burundi, Rwanda, Tanzania and Uganda.

A survey in Burundi, Rwanda, Tanzania and Uganda was conducted in order to determine the different production systems under which Ankole cattle are currently kept. Additionally, selection criteria of livestock keepers were documented. In Burundi, Rwanda and parts of Uganda, livestock keepers are sedentary and herds are small, whereas in the other areas Ankole cattle are kept in large herds, some of them still under a (semi-)nomadic system.

Individual-based assessment of population structure and admixture in Austrian, Croatian and German draught horses.

All over Europe, the number of draught horses has decreased drastically during the last 50 years. As a prerequisite for efficient management decisions, we analysed the conservation status in Austrian (Noriker Carinthia - NC, Noriker Salzburg - NS), Croatian (Croatian Coldblood - C, Posavina horse - P) and German (Altmaerkisch Coldblood - A, Black Forest horse - BF, Mecklenburg Coldblood - M, Rhenish German Draught horse - R, Saxon Thuringa Coldblood - ST, Schleswig Draught horse - Sch, South German Coldblood - SG) draught horses (434) using multilocus genotypic information from 30 (effectively 27) microsatellite loci. Populations located in areas with less intensive agricultural production (C, NC, NS, P and SG) had greater diversity within the population and estimated effective population size than A, BF, Sch, M, R and ST populations.

Modelling the contribution of arbuscular mycorrhizal fungi to plant phosphate uptake.

In this paper, we investigate the role of arbuscular mycorrhizal fungi in plant phosphorus nutrition. We develop a mathematical model which quantitatively assesses the contribution of external fungal hyphae to plant phosphate uptake. We derive an equation for solute uptake by a growing fungal mycelium which we couple with a model for root uptake.

Engineering of Pichia pastoris for improved production of antibody fragments.

The methylotrophic yeast Pichia pastoris has been used for the expression of many proteins, including antibody fragments. However, limitations became obvious especially when secreting heterodimeric Fab fragments. Up-to-date, antibody fragments have only been expressed under control of the strong inducible alcohol oxidase 1 (AOX1) promoter, which may stress the cells by excessive transcription.

Simultaneous determination of type A-& B-trichothecenes and zearalenone in cereals by High Performance Liquid Chromatography - Tandem Mass Spectrometry.

A rapid quantitative method for the simultaneous determination of the majorFusarium mycotoxins nivalenol, deoxynivalenol, fusarenon-X, 3-acetyl-deoxynivalenol, 15-acetyl-deoxynivalenol, diacetoxyscirpenol, HT-2 toxin, T-2 toxin and zearalenone in maize and wheat was developed. Raw extracts (acetonitrile/water 84/16) are cleaned-up with MycoSep(®) columns., Chromatographic separation and end determination is carried out by HPLC-APCI-MS/MS.

Ancestral gene fusion in cellobiose dehydrogenases reflects a specific evolution of GMC oxidoreductases in fungi.

Cellobiose dehydrogenases (CDHs) are extracellular hemoflavoenzymes that are thought to be involved in the degradation of two of the most abundant biopolymers in the biosphere, cellulose and lignin. To date, these enzymes, consisting of a cytochrome domain and a flavin domain, have been detected and sequenced exclusively in the kingdom of fungi. Independent phylogenetic analyses of two distinct domains of CDH genes reveal that they evolved in parallel as fused genes.

Characterisation of cellobiose dehydrogenases from the white-rot fungi Trametes pubescens and Trametes villosa.

Cellobiose dehydrogenase (CDH) is an extracellular haemoflavoenzyme that is produced by a number of wood-degrading and phytopathogenic fungi and it has a proposed role in the early events of lignocellulose degradation and wood colonisation. In the presence of a suitable electron acceptor, e.g.