Safety and immunogenicity of capsular polysaccharide-tetanus toxoid conjugate vaccines for group B streptococcal types Ia and Ib

The translocation of spin-labeled analogues of phosphatidylcholine (4-doxylpentanoyl-PC, SL-PC), phosphatidylethanolamine (SL-PE), phosphatidylserine (SL-PS), and sphingomyelin (SL-SM) from the outer to the inner leaflet of the plasma membrane bilayer was investigated in dog kidney MDCK II and human colon Caco-2 cells. Disappearance from the outer leaflet was assayed using back-exchange to serum albumin. Experiments with cells in suspension as well as with polarized cells on filters were performed at reduced temperatures (10 and 20 degreesC) to suppress endocytosis and hydrolysis of spin-labeled lipids. For both epithelial cell lines, a fast ATP-dependent inward movement of the aminophospholipids SL-PS and SL-PE was found, while SL-SM was only slowly internalized without any effect of ATP depletion. The kinetics of redistribution of SL-PC were clearly different between the two cell lines. In MDCK II cells, SL-PC was rapidly internalized in an ATP-dependent and N-ethylmaleimide-sensitive manner and at a rate similar to that of the aminophospholipids. In contrast, in Caco-2 cells the inward movement of SL-PC was much slower than that of the aminophospholipids, did not depend on ATP, and was not N-ethylmaleimide-sensitive. Inhibitor studies indicated that the outward-translocating multidrug resistance P-glycoprotein present in these cells did not affect the kinetics of inward translocation. Internalization was always similar on the apical and basolateral cell surface, suggesting the presence of the same phospholipid translocator(s) on both surface domains of epithelial cells. We propose that Caco-2 cells contain the well-known aminophospholipid translocase, while MDCK II cells contain either two translocases, namely, the aminophospholipid translocase and a phosphatidylcholine-specific translocase, or one translocase of a new type, translocating aminophospholipids as well as phosphatidylcholine.     

Das Biegedrillknickverhalten von Glasträgern aus Verbundglas

The LTB‐behaviour of beams made of laminated glass. In many cases the lateral torsional buckling is governing the design of beams made of laminated glass. Then the question arises of how to consider the composite effect of the PVB‐layers between the glass panels, which always shows a strong time‐ and temperature‐dependant behaviour. Also the combination and sequence of permanent and variable load and temperature phenomena plays an important rule for the prediction of the LTB‐behaviour, as these effects have significant influence on the stability load capacity of laminated glass. In the following paper the most important steps for the solution of a new developed design concept [1] are presented, by which the complex time dependant LTB‐capacities of beams of laminated glass can be verified. On the basis of the experimentally obtained time‐ and temperature dependant stiffness‐behaviour of the interlayer the load capacities of these beams can analytically be predicted. Comparisons with large scale tests and simulations show good agreements with the results. Finally the design procedure is able to take into account also sequence effects due to variable loads as well as temperature.     

Water Supply System Performance for Different Pipe Materials Part II: Sensitivity Analysis to Pressure Variation

In water supply systems there are many situations during normal operation that induce the occurrence of pressure transients, where high pressures are followed by low, sometimes even negative pressures. These transients may cause ruptures in pipes creating thus leaks or opportunities for contaminants to enter the water supply system. Thus severe pressures transients should be avoided or adequately controlled in potable drinking systems. The level of service provided by water distribution systems is an important matter in the water industry of today. However, the measure of the performance of a pipe system network is not a straightforward task. In this study the performance of pressures in two networks (a cast iron network and a polyethylene network) with the same typology was compared. The transient state conditions were induced by different typical hydromechanical devices operation characterised by a sudden pumps trip-off, a leakage occurrence and a closure of an automatic control valve. For the hydraulic simulations, advanced models based on numerical computation for steady and transient state conditions were used. A performance evaluation model was developed to analyse each type of situation since the simulation time period and the concerns regarding the system behaviour can be fairly different.     

Extracellular Matrix Synthesis and Remodeling by Mesenchymal Stromal Cells Is Context-Sensitive

Matrix remodeling could be an important mode of action of multipotent mesenchymal stromal cells (MSC) in extracellular matrix (ECM) disease, but knowledge is limited in this respect. As MSC are well-known to adapt their behavior to their environment, we aimed to investigate if their mode of action would change in response to healthy versus pathologically altered ECM. Human MSC-derived ECM was produced under different culture conditions, including standard culture, culture on Matrigel-coated dishes, and stimulation with the pro-fibrotic transforming growth factor-β1 (TGFβ1). The MSC-ECM was decellularized, characterized by histochemistry, and used as MSC culture substrate reflecting different ECM conditions. MSC were cultured on the different ECM substrates or in control conditions for 2 days. Culture on ECM increased the presence of surface molecules with ECM receptor function in the MSC, demonstrating an interaction between MSC and ECM. In MSC cultured on Matrigel-ECM and TGFβ1-ECM, which displayed a fibrosis-like morphology, gene expression of collagens and decorin, as well as total matrix metalloproteinase (MMP) activity in the supernatant were decreased as compared with control conditions. These results demonstrated that MSC adapt to their ECM environment, which may include pathological adaptations that could compromise therapeutic efficacy.     

An 8-fold {beta}{alpha} barrel protein with redundant folding possibilities

Protein sequences containing redundant segments of secondarystructure at both termini have the choice a priori of foldinginto several possible circularly permuted variants of the wild-typetertiary structure. To test this hypothesis the gene of phosphoribosylanthranilate isomerase from yeast, which is a single-domain8-fold ß barrel protein, was modified to produce a10-fold ß homologue in Escherichia coli. It containeda duplicate of the two C-terminal ß units of supersecondarystructure fused to its N-terminus. Most of the protein was recoveredfrom the insoluble fraction of disrupted cells by dissolutionin guanidinium chloride solutions and refolding. Pristine proteinwas purified from the soluble fraction. The purified (ß)₁₀proteins were enzymically almost fully active. Absorbance, fluorescenceand circular dichroism spectra as well as the reversible unfoldingbehaviour of both proteins were also very similar to the propertiesof the original (ß)₈ protein. Digestion with endopeptidasesconverted both the pristine and the refolded (ß)₁₀variant to the same large fragment that had the N-terminal sequenceand mol. wt of the wild-type ß)₈ protein. The datasuggest that the folding of the (ß)₁₀ variant is controlledthermodynamically both in vivo and in vitro.     

A highly parallelized framework for computationally intensive MR data analysis

Object

The goal of this study was to develop a comprehensive magnetic resonance (MR) data analysis framework for handling very large datasets with user-friendly tools for parallelization and to provide an example implementation.

Materials and methods

Commonly used software packages (AFNI, FSL, SPM) were connected via a framework based on the free software environment R, with the possibility of using Nvidia CUDA GPU processing integrated for high-speed linear algebra operations in R. Three hundred single-subject datasets from the 1,000 Functional Connectomes project were used to demonstrate the capabilities of the framework.

Results

A framework for easy implementation of processing pipelines was developed and an R package for the example implementation of Fully Exploratory Network ICA was compiled. Test runs on data from 300 subjects demonstrated the computational advantages of a processing pipeline developed using the framework compared to non-parallelized processing, reducing computation time by a factor of 15.

Conclusion

The feasibility of computationally intensive exploratory analyses allows broader access to the tools for discovery science.     

N-Hydroxysuccinimide-Modified Ethynylphosphonamidates Enable the Synthesis of Configurationally Defined Protein Conjugates

Herein, the application of N-hydroxysuccinimide-modified phosphonamidate building blocks for the incorporation of cysteine-selective ethynylphosphonamidates into lysine residues of proteins, followed by thiol addition with small molecules and proteins, is reported. It is demonstrated that the building blocks significantly lower undesired homo-crosslinking side products that can occur with commonly applied succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC) under physiological pH. The previously demonstrated stability of the phosphonamidate moiety additionally solves the problem of premature maleimide hydrolysis, which can hamper the efficiency of subsequent thiol addition. Furthermore, a method to separate the phosphonamidate enantiomers to be able to synthesize protein conjugates in a defined configuration has been developed. Finally, the building blocks are applied to the construction of functional antibody–drug conjugates, analogously to FDA-approved, SMCC-linked Kadcyla, and to the synthesis of a functional antibody–protein conjugate.     

Effects of aluminum treatment on phosphorus, carbon, and nitrogen distribution in lake sediment: a 31P NMR study

The effects of aluminum (Al) treatment on sediment composition of carbon (C), nitrogen (N) and phosphorus (P) were investigated in sediment representing pre- and post-treatment years in the Danish Lake S?nderby. 31P NMR spectroscopy analysis of EDTA-NaOH extracts revealed six functional P groups. Direct effects of the Al treatment were reflected in the orthophosphate profile revealing increased amounts of Al-P in the sediment layers representing the post-treatment period, as well as changes in organic P groups due to precipitation of phytoplankton and bacteria at the time of Al addition. Furthermore, changes in phytoplankton community structure and lowered production due to the Al treatment resulted in decreased concentrations of sediment organic P groups and total C. Exponential regressions were used to describe the diagenesis of C, N, and P in the sediment. From these regressions, half-life degradation times and C, N, and P burial rates were determined.