Superswelling Microneedle Arrays for Dermal Interstitial Fluid (Prote)Omics

The noninvasive sampling of dermal interstitial fluid (ISF) for the monitoring of clinical biomarkers is a greatly appealing area of research. The identification of molecular biomarkers in biological fluids has been accelerated with -omics analyses but remains limited in ISF because of its time-consuming and complex extraction process. Here, the generation of microneedle (MN) patches made of superabsorbent acrylate-based hydrogels for the rapid sampling of dermal ISF is described to explore its proteome. In depth, iterative optimization allows the identification of novel acrylate-based compositions with the required chemical, mechanical, and biocompatibility properties allowing proteomic analysis of the extracted ISF for the first time after sampling with swelling MNs. The generated MN arrays show no cytotoxic effect, successfully cross the stratum corneum, and can collect up to 6 µL of dermal ISF in 10 min in vivo. Proteomics lead to the detection of 176 clinically relevant biomarkers in the collected samples validating the use of ISF as a relevant bodily fluid for disease monitoring and diagnostic. Importantly, it is discovered that extraction fingerprint is strongly dependent on the MNs chemistry, and thus specific biomarkers could be selectively extracted by tuning the composition of the patch, making the system versatile and specific.     

Workers’ rest allowance and smoothing of the workload in assembly lines

Ergonomic aspects have a crucial role in manual assembly systems. They impact on the workers’ health, final product quality and productivity. For these reasons, there is the necessity to integrate them into the assembly line balancing phase as, whereas, only time and cost variables are considered. In this study, human energy expenditures are considered as ergonomic aspects and we integrate them, for the first time, into the assembly line balancing problem type 2 through the rest allowance evaluation. We consider as an objective function the minimization of the smoothness index. Firstly, a new optimal method based on mixed integer linear programming and a new linearization methodology are proposed. Then, a heuristic approach is introduced. To complete the study, a computational experimentation is presented to validate the mathematical model and to compare the methodologies proposed in terms of computational time, complexity and solution. Additionally, we provide a detailed analysis of the impact that rest allowance evaluation can have on productivity comparing the results obtained, taking into account the rest allowance integration before, during and after the assembly balancing process.     

A self‐scalable load injection service

Load testing of applications is an important and costly activity for software provider companies. Classical solutions are very difficult to set up statically, and their cost is prohibitive in terms of both human and hardware resources. Virtualized cloud computing platforms provide new opportunities for stressing an application's scalability, by providing a large range of flexible and less expensive (pay‐per‐use model) computation units. On the basis of these advantages, load testing solutions could be provided on demand in the cloud. This paper describes a Benchmark‐as‐a‐Service solution that automatically scales the load injection platform and facilitates its setup according to load profiles. Our approach is based on: (i) virtualization of the benchmarking platform to create self‐scaling injectors; (ii) online calibration to characterize the injector's capacity and impact on the benched application; and (iii) a provisioning solution to appropriately scale the load injection platform ahead of time. We also report experiments on a benchmark illustrating the benefits of this system in terms of cost and resource reductions. Copyright © 2013 John Wiley & Sons, Ltd.     

Patterning of the chick forebrain anlage by the prechordal plate

We analysed the role of the prechordal plate in forebrain development of chick embryos in vivo. After transplantation to uncommitted ectoderm a prechordal plate induces an ectopic, dorsoventrally patterned, forebrain-like vesicle. Grafting laterally under the anterior neural plate causes ventralization of the lateral side of the forebrain, as indicated by a second expression domain of the homeobox gene NKX2.1. Such a lateral ventralization cannot be induced by the secreted factor Sonic Hedgehog alone, as this is only able to distort the ventral forebrain medially. Removal of the prechordal plate does not reduce the rostrocaudal extent of the anterior neural tube, but leads to significant narrowing and cyclopia. Excision of the head process results in the caudal expansion of the NKX2.1 expression in the ventral part of the anterior neural tube, while PAX6 expression in the dorsal part remains unchanged. We suggest that there are three essential steps in early forebrain patterning, which culminate in the ventralization of the forebrain. First, anterior neuralization occurs at the primitive streak stage, when BMP-4-antagonizing factors emanate from the node and spread in a planar fashion to induce anterior neural ectoderm. Second, the anterior translocation of organizer-derived cells shifts the source of neuralizing factors anteriorly, where the relative concentration of BMP-4-antagonists is thus elevated, and the medial part of the prospective forebrain becomes competent to respond to ventralizing factors. Third, the forebrain anlage is ventralized by signals including Sonic Hedgehog, thereby creating a new identity, the prospective hypothalamus, which splits the eye anlage into two lateral domains.     

Kinetic analysis of the coding properties of O6-methylguanine in DNA: the crucial role of the conformation of the phosphodiester bond

Production by N-nitroso compounds of O6-alkylguanine (O6-alkylG) in DNA directs the misincorporation of thymine during DNA replication, leading to G:C to A:T transition mutations, despite the fact that DNA containing O6-alkylG:T base pairs is less stable than that containing O6-alkylG:C pairs. We have examined the kinetics of incorporation by Klenow fragment (KF) of Escherichia coli DNA polymerase I of thymine (T) and of cytosine (C) opposite O6-MeG in the template DNA strand. Both T and C were incorporated opposite O6-MeG much slower than nucleotides forming regular A:T or G:C base pairs. Using various concentrations of dTTP, dCTP, or their phosphorothioate (Sp)-dNTP alpha S analogues, or a mixture of dTTP and dCTP, the progress of incorporation of a single nucleotide in a single catalytic cycle of a preformed KF-DNA complex was measured (pre-steady-state kinetics). The results were consistent with the kinetic scheme (Kuchta, R. D., Benkovic, P., & Benkovic, S. J. (1988) Biochemistry 27, 6716-6725): (1) binding of dNTP to polymerase-DNA; (2) conformational change in polymerase; (3) formation of phosphodiester between the dNTP and the 3'-OH of the primer; (4) conformational change of polymerase; (5) release of pyrophosphate. The results were analyzed mathematically to identify the steps at which the rate constants differ significantly between the incorporation of T and C. The only significant difference was the 5-fold difference in the rates of formation of the phosphodiester bond (for dTTP, kforward = 3.9 s-1 and kback = 1.9 s-1; for dCTP, kforward = 0.7 s-1 and kback = 0.9 s-1). These pre-steady-state progress curves were biphasic with a rapid initial burst followed by an apparently steady-state rise. Deconvolution of these curves gave direct evidence for the importance of the conformational change after polymerization by showing that the curves represented the sum of the rapid accumulation of the product of step 3 followed by the slow conversion of that to the product of step 5 (because of the rapidity of the release of pyrophosphate there was no significant accumulation of the product of step 4). The equilibrium constants for each step suggest that the greatest change in the Gibbs free energy occurs at the conformational change after polymerization and that while the formation of the phosphodiester bond to T is slightly exothermic, that to C is slightly endothermic.(ABSTRACT TRUNCATED AT 400 WORDS)     

The possibility of new high-transmission,high finesse,Perot-Fabry interferometers for far ir spectroscopy

Finesse as high as 260 and peak transmission close to 98% at frequencies around 30 cm^?1 are predicted at 10 K by coating the Perot-Fabry (PF) mirrors with a suitable thickness of High Tc (HTC) superconductor YBaCuO. In fact, Finesse will be limited by the lack of parallelism and the lack of perfect flatness of the mirrors to values around 100. These predictions are based upon phenomenological models for both the YBaCuO coating and the MgO support, i.e. from direct transmission measurements of MgO and YBaCuO in the Far IR, which had to be fitted by the models.     

Evidence of High Vertical Wave-Number Behavior in a Continuously Stratified Reservoir: Boadella, Spain

High vertical wave-number modes clearly dominate the internal wave field during the stratification period in Boadella reservoir in northeast Spain. In this period, the extraction of hypolimnetic water, due to summer irrigation, brought the surface level down by 6 m in one month and the epilimnetic water progressively occupied the whole water column. The temperature profile, with the exception of a few meters at the surface layer, presented an almost constant temperature gradient of about 0.7°C/m. The period of the main vertical mode is 24 h with an amplitude of around 1 m. Thermistor chain records and meteorological data allow us to deduce that this mode is, at least, a third vertical mode forced by the wind, which normally has a typical periodicity of 24 h. However, when the wind changes direction from south to north, the circulation cells developed due to this forced nonstationary oscillation are destroyed. When this occurs, the Bulk Richardson number is Rib ～ 1. Similar vertical structures as a response to wind forcing should be expected in similar systems, although this has not been reported in the literature.     

Sequential batch reactor for eucalypt kraft pulp effluent treatment with Trametes versicolor

BACKGROUND: A sequential batch reactor (SBR) was used for eucalypt kraft pulp effluent treatment with Trametes versicolor. A 2³ full factorial design and response surface methodology were applied to optimise the batch fermentation conditions. Effluent concentration, culture medium and inoculum age were the factors selected for this study in order to optimise the reduction of chemical oxygen demand (COD). RESULTS: The presence of Trametes Defined Medium (TDM) in the fermentation was required to obtain a significant COD reduction. Experiments in the batch reactor confirmed, in general, the predicted results of optimisation developed from Erlenmeyer batch assays. The T. versicolor culture remained active during 42 days of study in the SBR, providing approximately 80% of COD reduction. CONCLUSION: Trametes versicolor may be considered as very promising for the biological treatment of effluents from kraft pulp mills in an SBR system instead of the activated sludge mixed cultures traditionally used. Copyright © 2008 Society of Chemical Industry