X-ray Fluorescence Uptake Measurement of Functionalized Gold Nanoparticles in Tumor Cell Microsamples

Quantitative cellular in vitro nanoparticle uptake measurements are possible with a large number of different techniques, however, all have their respective restrictions. Here, we demonstrate the application of synchrotron-based X-ray fluorescence imaging (XFI) on prostate tumor cells, which have internalized differently functionalized gold nanoparticles. Total nanoparticle uptake on the order of a few hundred picograms could be conveniently observed with microsamples consisting of only a few hundreds of cells. A comparison with mass spectroscopy quantification is provided, experimental results are both supported and sensitivity limits of this XFI approach extrapolated by Monte-Carlo simulations, yielding a minimum detectable nanoparticle mass of just 5 pg. This study demonstrates the high sensitivity level of XFI, allowing non-destructive uptake measurements with very small microsamples within just seconds of irradiation time.     

Hemi- and Homozygous Loss-of-Function Mutations in DSG2 (Desmoglein-2) Cause Recessive Arrhythmogenic Cardiomyopathy with an Early Onset

About 50% of patients with arrhythmogenic cardiomyopathy (ACM) carry a pathogenic or likely pathogenic mutation in the desmosomal genes. However, there is a significant number of patients without positive familial anamnesis. Therefore, the molecular reasons for ACM in these patients are frequently unknown and a genetic contribution might be underestimated. Here, we used a next-generation sequencing (NGS) approach and in addition single nucleotide polymor-phism (SNP) arrays for the genetic analysis of two independent index patients without familial medical history. Of note, this genetic strategy revealed a homozygous splice site mutation (DSG2–c.378+1G>T) in the first patient and a nonsense mutation (DSG2–p.L772X) in combination with a large deletion in DSG2 in the second one. In conclusion, a recessive inheritance pattern is likely for both cases, which might contribute to the hidden medical history in both families. This is the first report about these novel loss-of-function mutations in DSG2 that have not been previously identi-fied. Therefore, we suggest performing deep genetic analyses using NGS in combination with SNP arrays also for ACM index patients without obvious familial medical history. In the future, this finding might has relevance for the genetic counseling of similar cases.     

An Engineered sgsh Mutant Zebrafish Recapitulates Molecular and Behavioural Pathobiology of Sanfilippo Syndrome A/MPS IIIA

Mucopolysaccharidosis IIIA (MPS IIIA, Sanfilippo syndrome type A), a paediatric neurological lysosomal storage disease, is caused by impaired function of the enzyme N-sulfoglucosamine sulfohydrolase (SGSH) resulting in impaired catabolism of heparan sulfate glycosaminoglycan (HS GAG) and its accumulation in tissues. MPS IIIA represents a significant proportion of childhood dementias. This condition generally leads to patient death in the teenage years, yet no effective therapy exists for MPS IIIA and a complete understanding of the mechanisms of MPS IIIA pathogenesis is lacking. Here, we employ targeted CRISPR/Cas9 mutagenesis to generate a model of MPS IIIA in the zebrafish, a model organism with strong genetic tractability and amenity for high-throughput screening. The sgsh^Δex5−6 zebrafish mutant exhibits a complete absence of Sgsh enzymatic activity, leading to progressive accumulation of HS degradation products with age. sgsh^Δex5−6 zebrafish faithfully recapitulate diverse CNS-specific features of MPS IIIA, including neuronal lysosomal overabundance, complex behavioural phenotypes, and profound, lifelong neuroinflammation. We further demonstrate that neuroinflammation in sgsh^Δex5−6 zebrafish is largely dependent on interleukin-1β and can be attenuated via the pharmacological inhibition of Caspase-1, which partially rescues behavioural abnormalities in sgsh^Δex5−6 mutant larvae in a context-dependent manner. We expect the sgsh^Δex5−6 zebrafish mutant to be a valuable resource in gaining a better understanding of MPS IIIA pathobiology towards the development of timely and effective therapeutic interventions.     

Numerical characterization and experimental verification of an in-plane MEMS-actuator with thin-film aluminum heater

In this paper, a novel concept of a thermo-mechanical MEMS actuator using aluminum thin-film heaters on a thermal oxide for electrical insulation is presented. The actuator is part of an universal tensile testing platform for thermo-mechanical material characterization of one dimensional materials on a micro- and nano-scopic scale under different environmental conditions, as varying temperatures, pressure, moisture or even vacuum and is realised in BDRIE technology. It is shown, that the actuator concept fulfills the requirements for the use in a tensile loading stage along with heterogeneously integrated nanofunctional elements, following a specimen centered approach in line with bottom-up self-assembly processes. Simulation and experiment agree very well in the thermal and mechanical domain and allow subsequent optimisation of the actuator performance.     

Improving active Mealy machine learning for protocol conformance testing

Using a well-known industrial case study from the verification literature, the bounded retransmission protocol, we show how active learning can be used to establish the correctness of protocol implementation I relative to a given reference implementation R. Using active learning, we learn a model M _R of reference implementation R, which serves as input for a model-based testing tool that checks conformance of implementation I to M _R . In addition, we also explore an alternative approach in which we learn a model M _I of implementation I, which is compared to model M _R using an equivalence checker. Our work uses a unique combination of software tools for model construction (Uppaal), active learning (LearnLib, Tomte), model-based testing (JTorX, TorXakis) and verification (CADP, MRMC). We show how these tools can be used for learning models of and revealing errors in implementations, present the new notion of a conformance oracle, and demonstrate how conformance oracles can be used to speed up conformance checking.     

Nature-Inspired Meta-Heuristics on Modern GPUs: State of the Art and Brief Survey of Selected Algorithms

Graphic processing units (GPUs) emerged recently as an exciting new hardware environment for a truly parallel implementation and execution of Nature and Bio-inspired Algorithms with excellent price-to-power ratio. In contrast to common multicore CPUs that contain up to tens of independent cores, the GPUs represent a massively parallel single-instruction multiple-data devices that can nowadays reach peak performance of hundreds and thousands of giga floating-point operations per second. Nature and Bio-inspired Algorithms implement parallel optimization strategies in which a single candidate solution, a group of candidate solutions (population), or multiple populations seek for optimal solution or set of solutions of given problem. Genetic algorithms (GA) constitute a family of traditional and very well-known nature-inspired populational meta-heuristic algorithms that have proved its usefulness on a plethora of tasks through the years. Differential evolution (DE) is another efficient populational meta-heuristic algorithm for real-parameter optimization. Particle swarm optimization (PSO) can be seen as nature-inspired multiagent method in which the interaction of simple independent agents yields intelligent collective behavior. Simulated annealing (SA) is global optimization algorithm which combines statistical mechanics and combinatorial optimization with inspiration in metallurgy. This survey provides a brief overview of the latest state-of-the-art research on the design, implementation, and applications of parallel GA, DE, PSO, and SA-based methods on the GPUs.     

Development of high‐definition aqueous polyvinylpyrrolidone photoresists for cathode ray tubes

Aqueous photoresists that are capable of higher resolutions than are currently employed by the cathode ray tube (CRT) industry were developed by combining photoactivators, which activate at shorter UV wavelengths with polyvinylpyrrolidone (PVP). Two photoactivators were synthesized: 4,4′‐diazido‐2,2′‐biphenyl disodium disulfonate (DABP), which has a maximum absorbance at 264 nm, and 4,4′‐diazido‐2,2′‐biphenylethane disodium disulfonate (DABPE), which has a maximum absorbance at 258 nm. The PVP/DABP and PVP/DABPE photoresists successfully imaged a pattern with resolution as small as 4.4 μm with photoactivator concentrations greater than 20% of the PVP concentration. Addition of silane and emulgen greatly improved the performance of the photoresists with more uniform coatings of thicknesses of up to 1.4 μm. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 102: 1637–1644, 2006     

New aspects in cationization of lignocellulose materials. IX. Flame retardancy effect of modification with nitrogen and sulfur containing groups

Lignocellulose materials were modified with 3-chlor-2-hydroxypropyltrimethylammoniumchloride, 1,3-bis(3-chlor-2-hydroxypropyl)imidazoliumhydrogensulphate, and 2-chlorethyl-sodiumsulphonate and the flame-retardant properties of materials obtained were studied. The flame-retardant effect was proved using the limited oxygen index (LOI) method and values up to 32.8 vol % of oxygen were determined. The LOI values were higher when the modification was done without using NaOH for activation, but in that case the alkylating groups were not chemically bonded to the material. The NaOH activation was necessary to obtain material with flame-retardant properties stable against washing. No synergistic effects were observed when the material was modified with both nitrogen- and sulfur-containing groups.     

Dynamic mechanical behavior of chlorosubstituted derivatives of poly(ethyl methacrylate) and poly(ethyl acrylate)

A freely oscillating torsional pendulum was used in the investigation of the influence of trichloroethyl, tetrachloroethyl, trichloromethoxyethyl, and trichloroethoxyethyl side groups on the molecular mobility in the glassy state and on the glass transition temperature of poly(meth)acrylates. All the polymers under study, which may be used as fire retardants, exhibit a simple relaxation behavior. While the parameters of the low-temperature and secondary relaxation process in the glassy state are not noticeably affected by the substituents used, the glass transition temperature T_g, increases with rising polarity and volume of side chains. The increase is larger in the series of polyacrylates, so that differences in the softening temperatures of polymethacrylates and polyacrylates having the same side chains decrease considerably with growing substitution.     

Experiments on the suspension of spheres in inclined tubes—II. Suspension by air

Data are presented on the suspension of single spheres by air, in inclined tubes ranging in diameter from 0·97 to 5·13 cm. The angle of inclination of the tubes to the horizontal axis varied from 0 to 90°. The data, which relate the pressure drop and suspending velocity to the geometric and physical parameters of the system, are compared with data generated previously using water as the suspending fluid. Dimensionless expressions are presented which correlate both sets of data for pressure drop and suspending velocity in terms of sphere/tube diameter ratio, tube diameter, density of solid and fluid and angle of inclination of the tube.