Intramammary infusion of a live culture for treatment of bovine mastitis: effect of live lactococci on the mammary immune response

In the accompanying article, we demonstrated that a live culture of Lactococcus lactis compares favourably with antibiotics for treatment of bovine mastitis in two initial field trials. In an effort to explain the mechanism involved, this study investigated the effect of culture administration on the local immune response. In this respect we initially observed that infusion of the live culture Lactococcus lactis stimulated substantial recruitment of polymorphonucleocytes (PMN) and lymphocytes to the udder. For instance, in one assay, quarters infused with the probiotic experienced a dramatic increase (approximately 20,000-fold) in neutrophils over the first 48-h period from an average value of 83.6 cells/ml pre-treatment to 1.78 x 106 cells/ml 48 h post-infusion. Levels of the acute phase proteins haptaglobin and milk amyloid A were also elevated significantly in comparison with controls following infusion of the culture. The results of flow cytometric assays also demonstrated that while infusion of a live lactococcal culture led to an enhanced recruitment of PMN to the udder (from 1.85 x 104 cells/ml pre-infusion to 1.45 x 106 cells/ml 24 h post-infusion) cell-free supernatant from the same culture was not able to do so, indicating that live Lc. lactis can specifically trigger the mammary immune response to elicit PMN accumulation. These results suggest that the mechanism responsible for this probiotic treatment of mastitis is associated with stimulation of the host intramammary immune system.     

Comparison of a time- and a speed-based traffic light assistance system

Traffic light assistance systems (TLASs) can be infrastructure based or on-board, and in the latter case they can inform the driver about the time remaining to green, or about the recommended speed for a smooth passage at green. A speed-based and a time-based on-board system prototype was compared against each other and against a baseline without any assistance system. Using a within-subjects design, 18 participants drove in a fixed-base simulator along a suburban road with signalised intersections, where the delay to green was set to zero (allowing a passage at the current speed), “half-speed” (requiring a clear speed reduction) and “stop” (requiring a substantial speed reduction). Driving behaviour, visual attention distribution and acceptance were evaluated. Both support systems improved driving efficiency and comfort over baseline, with the time-based system achieving higher scores in general. Both systems attracted a substantial amount of visual attention in the current setting; however, single-glance durations were below 1 s, and the number of glances forward were equal in the time-based condition compared to baseline, but lower in the speed-based condition. No red or amber light violations were registered in baseline, while some occurred with any of the systems. Acceptance for both systems was high, with higher scores for the time-based prototype. Overall, an on-board TLAS with a countdown timer to green has the potential to increase efficiency and comfort without strong indications for attention disruption, but the risk for increased red/amber light violations has to be addressed. Improved system design as a way to mitigate potential issues is discussed.     

Characteristics of and selection criteria for support materials for cell immobilization in wastewater treatment

For treatment of wastewater with immobilized cells, support materials need to meet the following criteria: insoluble, not biodegradable, high mechanical stability, high diffusivity, simple immobilization procedure, high biomass retention, minimal attachment of other organisms and preferably a low cost price. In order to compare which support materials are the most suitable, characteristics of several natural and synthetic materials have been determined. For this, both literature and experimental data were used. The immobilization procedures of natural gel materials, like alginate and carrageenan, are mild and cells grow well in these supports. Furthermore, the effective diffusion coefficients of substrates are close to those in water. These supports, however, appeared to be soluble, biodegradable and liable to abrasion. Synthetic gels, on the contrary, have better mechanical properties, but mostly lower substrate diffusion coefficients. Immobilization conditions are less mild resulting in low biomass retention. For application of entrapped nitrifying cells in wastewater-treatment systems synthetic gels, however, are promising.     

Composite Nanostructures of TiO2 and ZnO for Water Splitting Application: Atomic Layer Deposition Growth and Density Functional Theory Investigation

The commercialization of solar fuel devices requires the development of novel engineered photoelectrodes for water splitting applications which are based on redundant, cheap, and environmentally friendly materials. In the current study, a combination of titanium dioxide (TiO₂) and zinc oxide (ZnO) onto nanotextured silicon is utilized for a composite electrode with the aim to overcome the individual shortcomings of the respective materials. The properties of conformal coverage of TiO₂ and ZnO layers are designed on the atomic scale by the atomic layer deposition technique. The resulting photoanode shows not only promising stability but also nine times higher photocurrents than an equivalent photoanode with a pure TiO₂ encapsulation onto the nanostructured silicon. Density functional theory calculations indicate that segregation of TiO₂ at the ZnO surfaces is favorable and leads to the stabilization of the ZnO layers in water environments. In conclusion, the novel designed composite material constitutes a promising base for a stable and effective photoanode for the water oxidation reaction.     

Involvement of GPR17 in Neuronal Fibre Outgrowth

Characterization of new pharmacological targets is a promising approach in research of neurorepair mechanisms. The G protein-coupled receptor 17 (GPR17) has recently been proposed as an interesting pharmacological target, e.g., in neuroregenerative processes. Using the well-established ex vivo model of organotypic slice co-cultures of the mesocortical dopaminergic system (prefrontal cortex (PFC) and substantia nigra/ventral tegmental area (SN/VTA) complex), the influence of GPR17 ligands on neurite outgrowth from SN/VTA to the PFC was investigated. The growth-promoting effects of Montelukast (MTK; GPR17- and cysteinyl-leukotriene receptor antagonist), the glial cell line-derived neurotrophic factor (GDNF) and of two potent, selective GPR17 agonists (PSB-16484 and PSB-16282) were characterized. Treatment with MTK resulted in a significant increase in mean neurite density, comparable with the effects of GDNF. The combination of MTK and GPR17 agonist PSB-16484 significantly inhibited neuronal growth. qPCR studies revealed an MTK-induced elevated mRNA-expression of genes relevant for neuronal growth. Immunofluorescence labelling showed a marked expression of GPR17 on NG2-positive glia. Western blot and RT-qPCR analysis of untreated cultures suggest a time-dependent, injury-induced stimulation of GPR17. In conclusion, MTK was identified as a stimulator of neurite fibre outgrowth, mediating its effects through GPR17, highlighting GPR17 as an interesting therapeutic target in neuronal regeneration.     

Systemic distribution and elimination of plain and with Cy3.5 functionalized poly(vinyl alcohol) coated superparamagnetic maghemite nanoparticles after intraarticular injection in sheep in vivo

PVA coated and fluorescent dye (Cy3.5) functionalized vinyl alcohol/vinyl amine copolymer coated superparamagnetic iron oxide nanoparticles (SPION) were evaluated for systemic distribution and elimination after intraarticular injection in sheep. Observation was done at 3, 24, 72, and 120 hours after injection using light microscopy, fluorescent microscopy, and confocal microscopy. No pathologic influence of SPION on the tissue harvested could be seen. A significantly increased iron content could be identified in the kidneys, lymph nodes, and spleen after injection of SPION. No particles were detected in the liver, the urinary, and the gall bladder. No positive fluorescent signal could be attributed to SPION throughout the organs. Our results indicated that the iron component of the SPION is possible to be incorporated into the physiologic iron metabolism after reabsorption in the proximal tubule system of the kidney and that concentration levels of Cy3.5 are too low to be detected throughout the body.     

Partially reversible adsorption of annexin A1 on POPC/POPS bilayers investigated by QCM measurements, SFM, and DMC simulations

The kinetics of annexin A1 binding to solid-supported lipid bilayers consisting of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC)/1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoserine (POPS; 4:1) has been investigated as a function of the calcium ion concentration in the bulk phase. Quartz crystal microbalance measurements in conjunction with scanning force microscopy, fluorescence microscopy, and computer simulations indicate that at a given Ca2+ concentration annexin A1 adsorbs irreversibly on membrane domains enriched in POPS. By contrast, annexin A1 adsorbs reversibly on the POPC-enriched phase, which is composed of single POPS molecules embedded within a POPC matrix. The overall area occupied by the POPS-enriched phase is controlled by the CaCl2 concentration. Monte Carlo simulations suggest that the area of the POPS-enriched phase increases by a factor of 7 when the Ca2+ concentration is changed from 0.01 to 1 mM.     

Polyaza crown ethers as non-nucleosidic building blocks in DNA conjugates: synthesis and remarkable stabilization of dsDNA

The synthesis of amphiphilic polyaza crown ether monomers X (benzyl-substituted), Y (palmityl-substituted) and Z (cholesteryl-substituted) and their incorporation into oligonucleotides are described. Their effects on thermal duplex stability were investigated by UV melting curve analysis in different alkaline metal buffer solutions. Thermal-denaturation experiments showed remarkable stabilization of dsDNA by polyaza crown ether monomers when incorporated in opposite positions. The series of polyaza crown ether monomers (X, Y and Z) with different overall lipophilicities showed a trend of increased stability of the corresponding dsDNA with increasing lipophilicity of the polyaza crown ether monomer. Multiple incorporations of benzyl-substituted polyaza crown ether monomer X as dangling ends on both sides of dsDNA resulted in strongly increased stability of the corresponding duplex.     

Saccharomyces cerevisiae S288C genome annotation: a working hypothesis

The S. cerevisiae genome is the most well-characterized eukaryotic genome and one of the simplest in terms of identifying open reading frames (ORFs), yet its primary annotation has been updated continually in the decade since its initial release in 1996 (Goffeau et al., 1996). The Saccharomyces Genome Database (SGD; www.yeastgenome.org) (Hirschman et al., 2006), the community-designated repository for this reference genome, strives to ensure that the S. cerevisiae annotation is as accurate and useful as possible. At SGD, the S. cerevisiae genome sequence and annotation are treated as a working hypothesis, which must be repeatedly tested and refined. In this paper, in celebration of the tenth anniversary of the completion of the S. cerevisiae genome sequence, we discuss the ways in which the S. cerevisiae sequence and annotation have changed, consider the multiple sources of experimental and comparative data on which these changes are based, and describe our methods for evaluating, incorporating and documenting these new data.     

Co-deposition and densification of SiC with MgO for biomedical applications

This article focuses on the electrophoretic co-deposition of SiC and MgO, which has not been previously reported. The EPD of SiC has been widely investigated, whilst the EPD of SiC with sintering additives is usually not taken into account since every compound added to the suspension seriously affects the zeta-potential and the conductivity, the two main parameters that should be optimized to achieve a good deposit. We comprehensively observed the effects of the individual compounds on the colloidal behaviour of a suspension suitable for co-deposition to achieve good homogeneity and the highest possible green density. The obtained green densities reached up to 1.92 g/cm³, which correspond to 60% of theoretical density, whilst after sintering in open air at 1350 °C the densities reached 2.33 g/cm³. SEM and TEM analyses revealed that the microstructure is composed of SiC grains embedded in a SiO₂ matrix, whilst XRD confirmed that even though the sintering caused a partial oxidation and the appearance of an amorphous phase, the prevailing crystalline phase is still β-SiC. In the presence of MgO, SiO₂ also appears in the crystalline form as cristobalite.