Comparative triplex tandem mass spectrometry assays of lysosomal enzyme activities in dried blood spots using fast liquid chromatography: application to newborn screening of Pompe, Fabry, and Hurler diseases

We report a comparative study of triplex tandem mass spectrometry (MS/MS) based assays of lysosomal enzymes in dried blood spots for the early detection of Pompe, Fabry, and Hurler diseases in newborns. Four methods have been evaluated that differed in sample handling and the equipment used. A newly developed method uses assay quenching with acetonitrile to precipitate blood proteins followed by analysis on an LC-electrospray/MS/MS system capable of multiple consecutive sample injections on two parallel chromatographic columns. This method requires 1.5 min per a triplex analysis of enzyme products and internal standards, which matches the throughput of the previously reported flow injection method. LC separation reduces matrix effects and allows for more facile sample workup. The new LC-based method showed figures of merit that were superior to those of the currently used method based on liquid-liquid extraction into ethyl acetate and flow injection into the mass spectrometer. The other methods we investigated for comprehensive comparison involved liquid-liquid extraction into ethyl acetate followed by LC-ESI-MS/MS and acetonitrile quenching followed by direct flow injection. Both methods using acetonitrile quenching were found to be robust and provide good quality data while requiring fewer liquid transfer steps and less disposable material and labor than did the extraction methods. The individual merits of the new methods are discussed to present an evaluated alternative approach to high-throughput analysis in newborn screening laboratories.     

Justification and refinements of model B3 for concrete creep and shrinkage 2. Updating and theoretical basis

Following statistical evaluation in part 1, this part deals with the improvement of prediction by updating one or two parameters of the model on the basis of short term tests and theoretical derivation of some formulae. The updating of model parameters is particularly important for high strength concretes and other special concretes containing various admixtures, superplasticizers, water-reducing agents and pozzolanic materials. For the updating of shrinkage prediction, a new method is presented in which the shrinkage half-time is calibrated by simultaneous measurements of water loss. This approach circumvents the ill-posedness of the shrinkage extrapolation problem. Theoretical justifications of various aspects of the model are given and a new formula for the additional creep due to drying (or stress-induced shrinkage) is derived. The new model should allow a more realistic assessment of the creep and shrinkage effects in concrete structures, which significantly affect durability and long term serviceability of civil engineering infrastructure.     

Addendum to “Why Did the World Trade Center Collapse?—Simple Analysis”

The addendum presents the responses to several questions on a preliminary version (which arrived too late for publication as part of the paper). The questions deal with the aircraft impact at a higher floor of the World Trade Center towers on September 11, 2001 damage to the upper part of the collapsing tower, weakness of connections, plastic cushioning of vertical impact, estimation of the equivalent mass, and the collapse of the adjacent lower building.     

Drying creep of concrete: constitutive model and new experiments separating its mechanisms

A new experimental method which allows the direct separation of the components of drying creep due to microcracking and stress-induced shrinkage is developed, demonstrated and validated. The basic idea is to compare the curvature creep of beams subjected to the same bending moment but very different axial forces. The results confirm that drying creep has two different sources: microcracking and stress-induced shrinkage. The latter increases continuously, whereas the former first increases and then decreases. The test results are fitted using a finite element model. The results validate the present model for drying creep. The microcracking is described by an established model, and the free (unrestrained) shrinkage of a material element is shown to depend approximately linearly on the humidity drop.     

Confinement-shear lattice CSL model for fracture propagation in concrete

A previously developed lattice model is improved and then applied to simulations of mixed-mode crack propagation in concrete. The concrete meso-structure is simulated by a three-dimensional lattice system connecting nodes which represent the centers of aggregate particles. These nodes are generated randomly according to the given grain size distribution. Only coarse aggregates are taken into account. Three-dimensional Delaunay triangulation is used to determine the lattice connections. The effective cross-section areas of connecting struts are defined by performing a three-dimensional domain tessellation partly similar to Voronoi tessellation. The deformations of each link connecting two adjacent aggregate pieces are defined in the classical manner of Zubelewicz and Ba?ant in which rigid body kinematics is assumed to characterize the displacement and rotation vectors at the lattice nodes. Each strut connecting adjacent particles can transmit both axial and shear forces. The adopted constitutive law simulates fracture, friction and cohesion at the meso-level. The behavior in tension and shear is made dependent on the transversal confining strain, which is computed assuming a linear displacement field within each tetrahedron of Delaunay triangulation, and neglecting the effect of the particle rotations. A mid-point explicit scheme is used to integrate the governing equations of the problems. General procedures to handle the boundary conditions and to couple the lattice mesh to the usual elastic finite element mesh are also formulated. Numerical simulations of mixed-mode fracture test data are used to demonstrate that the model is capable of accurately predicting complex crack paths and the corresponding load-deflection responses observed in experiments.     

You like it,you learn it: affectivity and learning in competitive social role play gaming

Despite the alleged ability of digital game-based learning (DGBL) to foster positive affect and in turn improve learning, the link between affectivity and learning has not been sufficiently investigated in this field. Regarding learning from team-based games with competitive elements, even less is known about the relationship between competitiveness (as a dispositional trait) and induced positive affect. In this media comparison study with between-subject design, participants (N?=?325; high school and college students) learned about the EU’s policy agenda by means of a debate-based method delivered through one of three educational media: a) through a social role-playing game with competitive elements played on computers, b) through a very similar game played without computers and c) through a non-game workshop. Unlike many previous DGBL studies, this study used participant randomization and strived to address the teacher effect and the length of exposure effect, while also using the same learning materials and a very similar educational method for all three treatments. Both games induced comparatively higher generalized positive affect and flow. Participants also learned more with the games. Positive affect, but not flow, mediated the influence of educational media on learning gains. Participants’ competitiveness was partly related to positive affect and experiencing flow but unrelated to learning gains. These outcomes held both when the game was played using computers, as well as without them. The study indicates that the ability of an educational intervention to instigate positive affect is an important feature that should be considered by educational designers.     

Electric field driven addressing of ATPS droplets in microfluidic chips

The possibility of controlled droplet motion (droplet addressing) mediated by DC electric field in aqueous two-phase systems (ATPS) is here reported for the first time. Three ATPS of polyethylene glycol (PEG)/salt type, namely PEG/phosphate, PEG/sulphate, and PEG/carbonate, were selected for this study. We observed fast motion of salty droplets dispersed in PEG continuous phase induced by electric field of relative low strength. Hence, three fluidic systems with separated electrode chambers for the evaluation of electrophoretic mobilities and for addressing experiments were fabricated. Electrophoretic mobilities of salty droplets always exceeded the value of \(1\times 10^{-7}\, \hbox {m}^2\hbox {V}^{-1}\hbox {s}^{-1}\), which is about by one magnitude higher value than those typically measured in water–oil droplet systems. The electrophoretic mobilities in systems with free surface are the same or even smaller than in closed microfluidic structures, which is accounted mainly to the fact that a significant part of salty droplets is exposed to air and does not contribute to droplet forcing. Series of addressing and merging experiments in a microfluidic chip shows that DC electric field can be used as a powerful tool for smart manipulation of droplets in microfluidic systems with PEG/salt ATPS.     

Size-Effect Testing of Cohesive Fracture Parameters and Nonuniqueness of Work-of-Fracture Method

The cohesive crack model has been widely accepted as the best compromise for the analysis of fracture of concrete and other quasibrittle materials. The softening stress-separation law of this model is now believed to be best described as a bilinear curve characterized by four parameters: the initial and total fracture energies Gf and GF, the tensile strength ft′, and the knee-point ordinate σ1. The classical work-of-fracture test of a notched beam of one size can deliver a clear result only for GF. Here it is shown computationally that the same complete load-deflection curve can be closely approximated with stress-separation curves in which the ft′ values differ by 77% and Gf values by 68%. It follows that the work-of-fracture test alone cannot provide an unambiguous basis for quasibrittle fracture analysis. It is found, however, that if this test is supplemented by size-effect testing, all four cohesive crack model parameters can be precisely identified and the fracture analysis of structures becomes unambiguous. It is shown computationally that size-effect tests do not suffice for determining GF and ft′, which indicates that they provide a sufficient basis for computing neither the postpeak softening of fracturing structures nor the peak loads of a very large structure. However, if the size-effect tests are supplemented by one complete softening load-deflection curve of a notched specimen, an unambiguous calculation of peak loads and postpeak response of structures becomes possible. To this end, the notched specimen tests must be conducted in a certain size range, whose optimum is here established by extending a previous analysis. Combination of the work-of-fracture and size-effect testing could be avoided only if the ratios GF/Gf and σ1/ft′ were known a priori, but unfortunately their estimates are far too uncertain.     

Practical prediction of cyclic humidity effect in creep and shrinkage of concrete

The previously published BP Model for prediction of concrete creep and shrinkage is generalized to cover the increase of creep caused by the cyclic component of environmental relative humidity. A simple formula, which is partly empirical, partly based on the diffusion theory, is proposed and is calibrated by comparisons with test data from the literature.     

Temperature Effect on Concrete Creep Modeled by Microprestress-Solidification Theory

The previously developed microprestress-solidification theory for concrete creep and shrinkage is generalized for the effect of temperature (not exceeding 100°C). The solidification model separates the viscoelasticity of the solid constituent, the cement gel, from the chemical aging of material caused by solidification of cement and characterized by the growth of volume fraction of hydration products. This permits considering the viscoelastic constituent as non-aging. The temperature dependence of the rates of creep and of volume growth is characterized by two transformed time variables based on the activation energies of hydration and creep. The concept of microprestress achieves a grand unification of theory in which the long-term aging and all transient hygrothermal effects simply become different consequences of one and the same physical phenomenon. The microprestress, which is independent of the applied load, is initially produced by incompatible volume changes in the microstructure during hydration, and later builds up when changes of moisture content and temperature create a thermodynamic imbalance between the chemical potentials of vapor and adsorbed water in the nanopores of cement gel. As recently shown, this simultaneously captures two basic effects: First, the creep decreases with increasing age at loading after the growth of the volume fraction of hydrated cement has ceased; and, second, the drying creep, i.e., the transient creep increases due to drying (Pickett effect) which overpowers the effect of steady-state moisture content (i.e., less moisture—less creep). Now it is demonstrated that the microprestress buildup and relaxation also captures a third effect: The transitional thermal creep, i.e., the transient creep increase due to temperature change. For computations, an efficient (exponential-type) integration algorithm is developed. Finite element simulations, in which the apparent creep due to microcracking is taken into account separately, are used to identify the constitutive parameters and a satisfactory agreement with typical test data is achieved.