The impact of recombinant fusion-hydrophobin coated surfaces on E. coli and natural mixed culture biofilm formation

The impact of increased surface hydrophobicity on biofilms regarding retardation, repulsion, or attraction was studied with hydrophobin modified glass substrata. Recombinantly produced fungal hydrophobins forming self-assembled monolayers were used as the surface coating. The adsorption dynamics of hydrophobins were analysed with a quartz crystal microbalance which showed the surface coating to be rapid and stable. The change of surface wettability was determined by water contact angle measurements and demonstrated an increase in hydrophobicity in range of 60–62°. The homogeneity of the monolayers was demonstrated by immunofluorescence microscopy. Atomic force microscopy was applied to visualise the uniform texture of the coated materials. The hydrophobin coatings had no impact on different biofilms in terms of spatial distribution, cell numbers, and population composition. In consequence, hydrophobicity might not represent an important parameter for biofilm formation. Nevertheless, recombinant hydrophobins are suitable for large scale surface modification and functionalization with bioactive molecules.     

Humidity-driven changes in growth rate, photosynthetic capacity, hydraulic properties and other functional traits in silver birch (Betula pendula)

A study was performed on saplings of silver birch (Betula pendula Roth) growing at the free air humidity manipulation site, which was established to investigate the effect of increased air humidity on tree performance and canopy functioning. The aim of the experiment was to simulate the impact of the increasing atmospheric humidity on forest ecosystems predicted for northern Europe. Artificially elevated relative humidity (RH), which causes transpirational flux to decrease, diminished nutrient supply to the foliage; leaf nitrogen content, phosphorus content and P:N ratio decreased. The changes in leaf nutritional status brought about a considerable decline in both photosynthetic capacity (A _max, V _cmax, J _max) and tree growth rate. The manipulation induced diverse changes in tree hydraulic architecture and other functional traits. Different segments of the soil-to-leaf water transport pathway responded differently: leaf hydraulic conductance (K _L) decreased, while hydraulic conductance of root systems (K _R) and leaf-specific conductivity of stem-wood increased in response to elevated RH. Humidification caused the Huber values of stems to increase, thus reflecting changes in allocation patterns; relatively more resources were allocated to vascular tissue and less to foliage. The elevated RH induced substantial changes in specific leaf area (increased), branch- (decreased) and stem-wood density (decreased). The observed responses suggest that the expected climate-change-induced increase in the growth rate of trees at northern latitudes (boreal areas) due to the earlier start of the growing season in spring or higher carbon assimilation rate could be smaller or null if temperature rise is accompanied by a rise in atmospheric absolute humidity.     

Bace1 and Neuregulin‐1 cooperate to control formation and maintenance of muscle spindles

The protease β‐secretase 1 (Bace1) was identified through its critical role in production of amyloid‐β peptides (Aβ), the major component of amyloid plaques in Alzheimer's disease. Bace1 is considered a promising target for the treatment of this pathology, but processes additional substrates, among them Neuregulin‐1 (Nrg1). Our biochemical analysis indicates that Bace1 processes the Ig‐containing β1 Nrg1 (IgNrg1β1) isoform. We find that a graded reduction in IgNrg1 signal strength in vivo results in increasingly severe deficits in formation and maturation of muscle spindles, a proprioceptive organ critical for muscle coordination. Further, we show that Bace1 is required for formation and maturation of the muscle spindle. Finally, pharmacological inhibition and conditional mutagenesis in adult animals demonstrate that Bace1 and Nrg1 are essential to sustain muscle spindles and to maintain motor coordination. Our results assign to Bace1 a role in the control of coordinated movement through its regulation of muscle spindle physiology, and implicate IgNrg1‐dependent processing as a molecular mechanism.     

An in vitro evaluation of browser and grazer fermentation efficiency and microbiota using European moose spring and summer foods

Evolutionary morphological and physiological differences between browsers and grazers contribute to species‐specific digestion efficiency of food resources. Rumen microbial community structure of browsers is supposedly adapted to characteristic nutrient composition of the diet source. If this assumption is correct, domesticated ruminants, or grazers, are poor model animals for assessing the nutritional value of food consumed by browsing game species. In this study, typical spring and summer foods of the European moose (Alces alces) were combined with rumen fluid collected from both dairy cows (Bos taurus) and from moose, with the aim of comparing fermentation efficiency and microbial community composition. The nutritional value of the food resources was characterized by chemical analysis and advanced in vitro measurements. The study also addressed whether or not feed evaluation based on in vitro techniques with cattle rumen fluid as inoculum could be a practical alternative when evaluating the nutritional value of plants consumed by wild browsers. Our results suggest that the fermentation characteristics of moose spring and summer food are partly host‐specific and related to the contribution of the bacterial phyla Firmicutes and Bacteriodetes to the rumen microbial community. Host‐specific adaptations of the ruminal microbial community structure could be explained from the evolutionary adaptations related to feeding habitats and morphophysiological differences between browsers and grazers. However, the observed overall differences in microbial community structure could not be related to ruminal digestion parameters measured in vitro. The in vitro evaluation of digestion efficiency reveals that equal amounts of methane were produced across all feed samples regardless of whether the ruminal fluid was from moose or dairy cow. The results of this study suggested that the nutritional value of browsers' spring and summer food can be predicted using rumen fluid from domesticated grazers as inoculum in in vitro assessments of extent of digestion when excluding samples of the white water lily root, but not of fermentation characteristics as indicated by the proportions of individual fermentation fatty acids to the total of volatile fatty acids.     

Pluripotent Stem Cells for Uncovering the Role of Mitochondria in Human Brain Function and Dysfunction

Mitochondrial dysfunctions are a known pathogenetic mechanism of a number of neurological and psychiatric disorders. At the same time, mutations in genes encoding for components of the mitochondrial respiratory chain cause mitochondrial diseases, which commonly exhibit neurological symptoms. Mitochondria are therefore critical for the functionality of the human nervous system. The importance of mitochondria stems from their key roles in cellular metabolism, calcium handling, redox and protein homeostasis, and overall cellular homeostasis through their dynamic network. Here, we describe how the use of pluripotent stem cells (PSCs) may help in addressing the physiological and pathological relevance of mitochondria for the human nervous system. PSCs allow the generation of patient-derived neurons and glia and the identification of gene-specific and mutation-specific cellular phenotypes via genome engineering approaches. We discuss the recent advances in PSC-based modeling of brain diseases and the current challenges of the field. We anticipate that the careful use of PSCs will improve our understanding of the impact of mitochondria in neurological and psychiatric disorders and the search for effective therapeutic avenues.     

IgG glycosylation and DNA methylation are interconnected with smoking

Background

Glycosylation is one of the most common post-translation modifications with large influences on protein structure and function. The effector function of immunoglobulin G (IgG) alters between pro- and anti-inflammatory, based on its glycosylation. IgG glycan synthesis is highly complex and dynamic.

Characteristics of oviposition behaviour of the pollen beetle, Meligethes aeneus on four different host plants

Oviposition behaviour of Meligethes aeneus F. (Coleoptera: Nitidulidae) is characterised and quantified on four different plant species. Six behavioural components are identified: W—walking, WA—walking with abdomen on surface, R—resting, B—biting, AOH—placing abdomen over the bite hole and OVI—oviposition. Comparison of host acceptance behaviours on Brassica napus L., Brassica juncea (L.) Czern, Brassica nigra (L.) Koch and Sinapis alba L. showed that S. alba was accepted as a host only after a long exposure to the plant. Behaviour on the Brassica species was similar, however on B. nigra beetles spent a high proportion of time actually ovipositing. We conclude that important cues for oviposition are located both on the bud surface and inside the bud.     

Monitoring Climate Sensitivity Shifts in Tree-Rings of Eastern Boreal North America Using Model-Data Comparison

The growth of high-latitude temperature-limited boreal forest ecosystems is projected to become more constrained by soil water availability with continued warming. The purpose of this study was to document ongoing shifts in tree growth sensitivity to the evolving local climate in unmanaged black spruce (Picea mariana (Miller) B.S.P.) forests of eastern boreal North America (49°N–52°N, 58°W–82°W) using a comparative study of field and modeled data. We investigated growth relationships to climate (gridded monthly data) from observed (50 site tree-ring width chronologies) and simulated growth data (stand-level forest growth model) over 1908–2013. No clear strengthening of moisture control over tree growth in recent decades was detected. Despite climate warming, photosynthesis (main driver of the forest growth model) and xylem production (main driver of radial growth) have remained temperature-limited. Analyses revealed, however, a weakening of the influence of growing season temperature on growth during the mid- to late twentieth century in the observed data, particularly in high-latitude (> 51.5°N) mountainous sites. This shift was absent from simulated data, which resulted in clear model-data desynchronization. Thorough investigations revealed that desynchronization was mostly linked to the quality of climate data, with precipitation data being of particular concern. The scarce network of weather stations over eastern boreal North America (> 51.5°N) affects the accuracy of estimated local climate variability and critically limits our ability to detect climate change effects on high-latitude ecosystems, especially at high altitudinal sites. Climate estimates from remote sensing could help address some of these issues in the future.     

Myogenic differentiation of primary myoblasts and mesenchymal stromal cells under serum-free conditions on PCL-collagen I-nanoscaffolds

Background

The creation of functional skeletal muscle via tissue engineering holds great promise without sacrificing healthy donor tissue. Different cell types have been investigated regarding their myogenic differentiation potential under the influence of various media supplemented with growth factors. Yet, most cell cultures include the use of animal sera, which raises safety concerns and might lead to variances in results. Electrospun nanoscaffolds represent suitable matrices for tissue engineering of skeletal muscle, combining both biocompatibility and stability.We therefore aimed to develop a serum-free myogenic differentiation medium for the co-culture of primary myoblasts (Mb) and mesenchymal stromal cells derived from the bone marrow (BMSC) and adipose tissue (ADSC) on electrospun poly-ε-caprolacton (PCL)-collagen I-nanofibers.

Results

Rat Mb were co-cultured with rat BMSC (BMSC/Mb) or ADSC (ADSC/Mb) two-dimensionally (2D) as monolayers or three-dimensionally (3D) on aligned PCL-collagen I-nanofibers. Differentiation media contained either AIM V, AIM V and Ultroser® G, DMEM/Ham’s F12 and Ultroser® G, or donor horse serum (DHS) as a conventional differentiation medium. In 2D co-culture groups, highest upregulation of myogenic markers could be induced by serum-free medium containing DMEM/Ham’s F12 and Ultroser® G (group 3) after 7 days. Alpha actinin skeletal muscle 2 (ACTN2) was upregulated 3.3-fold for ADSC/Mb and 1.7-fold for BMSC/Mb after myogenic induction by group 3 serum-free medium when compared to stimulation with DHS. Myogenin (MYOG) was upregulated 5.2-fold in ADSC/Mb and 2.1-fold in BMSC/Mb. On PCL-collagen I-nanoscaffolds, ADSC showed a higher cell viability compared to BMSC in co-culture with Mb. Myosin heavy chain 2, ACTN2, and MYOG as late myogenic markers, showed higher gene expression after long term stimulation with DHS compared to serum-free stimulation, especially in BMSC/Mb co-cultures. Immunocytochemical staining with myosin heavy chain verified the presence of a contractile apparatus under both serum free and standard differentiation conditions.

Conclusions

In this study, we were able to myogenically differentiate mesenchymal stromal cells with myoblasts on PCL-collagen I-nanoscaffolds in a serum-free medium. Our results show that this setting can be used for skeletal muscle tissue engineering, applicable to future clinical applications since no xenogenous substances were used.