The Productivity of Coastal Meadows in Finland

The coastal meadows of Finland have gained a new interest as a summer pasture for cattle. These habitats have great historical, aesthetic and biological value (Pessa & Anttila, 2000). Typical features of the coastal meadows are the varying vegetation zones and wet, sometimes waterlogged, soils. The meadows are important nesting and feeding habitats for many water birds. When grazing ceases, reeds, trees and shrubs take over and the area loses its openness. Lately the amount of grasslands and pastures has drastically declined all over Europe. In Finland, the area of semi-natural biotopes has decreased to 1% of what it had been at the beginning of the twentieth century (Pitkänen & Tiainen, 2001). The goal of this study was to determine the yield and nutritional value of grass herbage in the meadows

Polarization relaxation in thin-film relaxors compared to that in ferroelectrics

Epitaxial thin films of relaxor PbMg1/3Nb2/3O3 and PbSc0.5Nb0.5O3, and ferroelectric PbZr0.65Ti0.35O3, Pb0.955La0.045Zr0.65Ti0.35O3, and Ba0.4Sr0.6TiO3 were prepared, and their dielectric properties were studied in a broad range of the measurement conditions. In the ferroelectric state, the presence and the change of configuration of the domains determined both the dynamic dielectric nonlinearity and the polarization hysteresis. In thin-film relaxors, the orientation of the randomly interacting dipoles in a random field was responsible for the dynamic dielectric nonlinearity, while the observed hysteresis was suggested to arise due to connection between the applied field and the relaxation times of both the dipoles and the internal field. In thin-film (Ba,Sr)TiO3, the high-temperature dielectric hysteresis was found to be relaxorlike.Comment: 20 pages, 10 figures, submitted to Phys. Rev. B 17.03.200

Effect of ripening temperature on the chemical composition of lingonberries (Vaccinium vitis-idaea L.) of northern and southern origin

Lingonberries (Vaccinium vitis-idaea L.) from two locations, northern (69◦N, 18◦E) and southern (59◦N, 10◦E) Norway, were grown under controlled conditions in a phytotron at two temperatures (9 and 15 ◦C) to study the effects of the ripening temperature and origin on the chemical composition of the berries. The concentrations of phenolic compounds, sugars, and organic acids as well as the profile of volatile organic compounds (VOCs) were determined using chromatographic and mass spectrometric methods. Five anthocyanins, eleven flavonols, eight cinnamic acid derivatives, three flavan-3-ols, three sugars, three organic acids, and 77 VOCs were identified, of which 40 VOCs had not previously been reported in lingonberries. Berries from both locations, were found to have higher contents of anthocyanins and cinnamic acid derivatives when ripened at lower temperature (9 ◦C), compared to the higher temperature (15 ◦C). Lingonberries of northern origin had a different VOC profile and higher contents of anthocyanins and organic acids than berries originating from the south. Lingonberries from the northern location also had higher proportions of cyanidin-3-O-glucoside and cyanidin-3-O-arabinoside than lingonberries from the southern location. The results show that the composition of lingonberries is influenced by both the environment and the origin of the plants, with phenolic compounds mainly influenced by the growth temperature and VOCs mainly influenced by plant origin

The Coordinated Action of MYB Activators and Repressors Controls Proanthocyanidin and Anthocyanin Biosynthesis in Vaccinium

Vaccinium berries are regarded as “superfoods” owing to their high concentrations of anthocyanins, flavonoid metabolites that provide pigmentation and positively affect human health. Anthocyanin localization differs between the fruit of cultivated highbush blueberry (V. corymbosum) and wild bilberry (V. myrtillus), with the latter having deep red flesh coloration. Analysis of comparative transcriptomics across a developmental series of blueberry and bilberry fruit skin and flesh identified candidate anthocyanin regulators responsible for this distinction. This included multiple activator and repressor transcription factors (TFs) that correlated strongly with anthocyanin production and had minimal expression in blueberry (non-pigmented) flesh. R2R3 MYB TFs appeared key to the presence and absence of anthocyanin-based pigmentation; MYBA1 and MYBPA1.1 co-activated the pathway while MYBC2.1 repressed it. Transient overexpression of MYBA1 in Nicotiana benthamiana strongly induced anthocyanins, but this was substantially reduced when co-infiltrated with MYBC2.1. Co-infiltration of MYBC2.1 with MYBA1 also reduced activation of DFR and UFGT, key anthocyanin biosynthesis genes, in promoter activation studies. We demonstrated that these TFs operate within a regulatory hierarchy where MYBA1 activated the promoters of MYBC2.1 and bHLH2. Stable overexpression of VcMYBA1 in blueberry elevated anthocyanin content in transgenic plants, indicating that MYBA1 is sufficient to upregulate the TF module and activate the pathway. Our findings identify TF activators and repressors that are hierarchically regulated by SG6 MYBA1, and fine-tune anthocyanin production in Vaccinium. The lack of this TF module in blueberry flesh results in an absence of anthocyanins.publishedVersio

Low prevalence of zoonotic multidrug-resistant bacteria in veterinarians in a country with prudent use of antimicrobials in animals

The occurrence of multidrug-resistant zoonotic bacteria in animals has been increasing worldwide. Working in close contact with livestock increases the risk of carriage of these bacteria. We investigated the occurrence of extended-spectrum beta-lactamase (ESBL) and plasmidic AmpC beta-lactamase producing Enterobacteriaceae (ESBL/pAmpC-PE) and livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) in Finnish veterinarians (n = 320). In addition to microbiological samples, background information was collected. Bacterial whole genome sequencing was performed to deduce sequence types (STs), spa types and resistance genes of the isolates. In total, 3.0% (9/297) of the veterinarians carried ESBL producing Escherichia coli, with one ESBL producing E. coli isolate producing also AmpC. Seven different STs, sequences of several different plasmid groups as well as several different bla(ESBL/pAmpC )genes existed in different combinations. No carbapenemase or colistin resistance genes were detected. MRSA was detected in 0.3% (1/320) of the samples. The strain belonged to LA-MRSA clonal complex (CC) 398 (ST398, spa type 011, lacking Panton-Valentine leukocidin genes). In conclusion, this study shows low carriage of multidrug-resistant zoonotic bacteria in Finnish veterinarians. However, finding LA-MRSA for the first time in a sample from a veterinarian in a country with prudent use of animal antimicrobials and regarding the recent rise of LA-MRSA on Finnish pig farms, a strong recommendation to protect people working in close contact with animals carrying LA-MRSA CC398 is given. Further studies are needed to explain why the prevalence of LA-MRSA in veterinarians is lower in Finland than in other European countries.Peer reviewe

Novel Information on the Epitope of an Inverse Agonist Monoclonal Antibody Provides Insight into the Structure of the TSH Receptor

The TSH receptor (TSHR) comprises an extracellular leucine-rich domain (LRD) linked by a hinge region to the transmembrane domain (TMD). Insight into the orientation of these components to each other is required for understanding how ligands activate the receptor. We previously identified residue E251 at the LRD-hinge junction as contributing to coupling TSH binding with receptor activation. However, a single residue cannot stabilize the LRD-hinge unit. Therefore, based on the LRD crystal structure we selected for study four other potential LRD-hinge interface charged residues. Alanine substitutions of individual residues K244, E247, K250 and R255 (as well as previously known E251A) did not affect TSH binding or function. However, the cumulative mutation of these residues in varying permutations, primarily K250A and R255A when associated with E251A, partially uncoupled TSH binding and function. These data suggest that these three residues, spatially very close to each other at the LRD base, interact with the hinge region. Unexpectedly and most important, monoclonal antibody CS-17, a TSHR inverse agonist whose epitope straddles the LRD-hinge, was found to interact with residues K244 and E247 at the base of the convex LRD surface. These observations, together with the functional data, exclude residues K244 and E247 from the TSHR LRD-hinge interface. Further, for CS-17 accessibility to K244 and E247, the concave surface of the TSHR LRD must be tilted forwards towards the hinge region and plasma membrane. Overall, these data provide insight into the mechanism by which ligands either activate the TSHR or suppress its constitutive activity

Tissue distribution of the laminin β1 and β2 chain during embryonic and fetal human development

Laminins are the major glycoproteins present in all basement membranes. Previously, we showed that perlecan is present during human development. Although an overview of mRNA-expression of the laminin β1 and β2 chains in various developing fetal organs is already available, a systematic localization of the laminin β1 and β2 chains on the protein level during embryonic and fetal human development is missing. Therefore, we studied the immunohistochemical expression and tissue distribution of the laminin β1 and β2 chains in various developing embryonic and fetal human organs between gestational weeks 8 and 12. The laminin β1 chain was ubiquitously expressed in the basement membrane zones of the brain, ganglia, blood vessels, liver, kidney, skin, pancreas, intestine, heart and skeletal system. Furthermore, the laminin β2 chain was present in the basement membrane zones of the brain, ganglia, skin, heart and skeletal system. The findings of this study support and expand upon the theory that these two laminin chains are important during human development

Structure-Based Discovery of A2A Adenosine Receptor Ligands

The recent determination of X-ray structures of pharmacologically relevant GPCRs has made these targets accessible to structure-based ligand discovery. Here we explore whether novel chemotypes may be discovered for the A(2A) adenosine receptor, based on complementarity to its recently determined structure. The A(2A) adenosine receptor signals in the periphery and the CNS, with agonists explored as anti-inflammatory drugs and antagonists explored for neurodegenerative diseases. We used molecular docking to screen a 1.4 million compound database against the X-ray structure computationally and tested 20 high-ranking, previously unknown molecules experimentally. Of these 35% showed substantial activity with affinities between 200 nM and 9 microM. For the most potent of these new inhibitors, over 50-fold specificity was observed for the A(2A) versus the related A(1) and A(3) subtypes. These high hit rates and affinities at least partly reflect the bias of commercial libraries toward GPCR-like chemotypes, an issue that we attempt to investigate quantitatively. Despite this bias, many of the most potent new ligands were novel, dissimilar from known ligands, providing new lead structures for modulation of this medically important target

Stability of the Neurotensin Receptor NTS1 Free in Detergent Solution and Immobilized to Affinity Resin

Purification of recombinant membrane receptors is commonly achieved by use of an affinity tag followed by an additional chromatography step if required. This second step may exploit specific receptor properties such as ligand binding. However, the effects of multiple purification steps on protein yield and integrity are often poorly documented. We have previously reported a robust two-step purification procedure for the recombinant rat neurotensin receptor NTS1 to give milligram quantities of functional receptor protein. First, histidine-tagged receptors are enriched by immobilized metal affinity chromatography using Ni-NTA resin. Second, remaining contaminants in the Ni-NTA column eluate are removed by use of a subsequent neurotensin column yielding pure NTS1. Whilst the neurotensin column eluate contained functional receptor protein, we observed in the neurotensin column flow-through misfolded NTS1.To investigate the origin of the misfolded receptors, we estimated the amount of functional and misfolded NTS1 at each purification step by radio-ligand binding, densitometry of Coomassie stained SDS-gels, and protein content determination. First, we observed that correctly folded NTS1 suffers damage by exposure to detergent and various buffer compositions as seen by the loss of [(3)H]neurotensin binding over time. Second, exposure to the neurotensin affinity resin generated additional misfolded receptor protein.Our data point towards two ways by which misfolded NTS1 may be generated: Damage by exposure to buffer components and by close contact of the receptor to the neurotensin affinity resin. Because NTS1 in detergent solution is stabilized by neurotensin, we speculate that the occurrence of aggregated receptor after contact with the neurotensin resin is the consequence of perturbations in the detergent belt surrounding the NTS1 transmembrane core. Both effects reduce the yield of functional receptor protein