Multiplexed-Based Assessment of DNA Damage Response to Chemotherapies Using Cell Imaging Cytometry

The current methods for measuring the DNA damage response (DDR) are relatively labor-intensive and usually based on Western blotting, flow cytometry, and/or confocal immunofluorescence analyses. They require many cells and are often limited to the assessment of a single or few proteins. Here, we used the Celigo^® image cytometer to evaluate the cell response to DNA-damaging agents based on a panel of biomarkers associated with the main DDR signaling pathways. We investigated the cytostatic or/and the cytotoxic effects of these drugs using simultaneous propidium iodide and calcein-AM staining. We also describe new dedicated multiplexed protocols to investigate the qualitative (phosphorylation) or the quantitative changes of eleven DDR markers (H2AX, DNA-PKcs, ATR, ATM, CHK1, CHK2, 53BP1, NBS1, RAD51, P53, P21). The results of our study clearly show the advantage of using this methodology because the multiplexed-based evaluation of these markers can be performed in a single experiment using the standard 384-well plate format. The analyses of multiple DDR markers together with the cell cycle status provide valuable insights into the mechanism of action of investigational drugs that induce DNA damage in a time- and cost-effective manner due to the low amounts of antibodies and reagents required.     

Electromagnetic Devices Applications of EBG for Systems Communications

This paper presents the study and applications of EBG (Electromagnetic Band Gap)/PBG (Photonic Band Gap) substrates, of planar microstrip structures, including quadrature directional couplers, rat-race (ring coupler) and impedance transformers. Through specific programs developed in FORTRAN PowerStation, the frequencies and couplings for each structure are obtained. Also are used the PACMO—Computer Aided Design in Microwave program developed by H.C.C. Fernandes. Results of the dimensions and coupling of these devices, varying the frequency band (cellular communication and Wimax systems) and permittivity of the substrate, are obtained, comparing the results of conventional materials with EBG/PBG materials in the s and p polarizations.     

Genomic and Immunological Characterization of Hypermucoviscous Carbapenem-Resistant Klebsiella pneumoniae ST25 Isolates from Northwest Argentina

In recent years, an increase in the prevalence hypermucoviscous carbapenem-resistant Klebsiella pneumoniae with sequence type 25 (ST25) was detected in hospitals of Tucuman (Northwest Argentina). In this work, the virulence and the innate immune response to two K. pneumoniae ST25 strains (LABACER 01 and LABACER 27) were evaluated in a murine model after a respiratory challenge. In addition, comparative genomics was performed with K. pneumoniae LABACER01 and LABACER27 to analyze genes associated with virulence. Both LABACER01 and LABACER27 were detected in the lungs of infected mice two days after the nasal challenge, with LABACER01 counts significantly higher than those of LABACER27. Only LABACER01 was detected in hemocultures. Lactate dehydrogenase (LDH) and albumin levels in bronchoalveolar lavage (BAL) samples were significantly higher in mice challenged with LABACER01 than in LABACER27-infected animals, indicating greater lung tissue damage. Both strains increased the levels of neutrophils, macrophages, TNF-α, IL-1β, IL-6, KC, MCP-1, IFN-γ, and IL-17 in the respiratory tract and blood, with the effect of LABACER01 more marked than that of LABACER27. In contrast, LABACER27 induced higher levels of IL-10 in the respiratory tract than LABACER01. Genomic analysis revealed that K. pneumoniae LABACER01 and LABACER27 possess virulence factors found in other strains that have been shown to be hypervirulent, including genes required for enterobactin (entABCDEF) and salmochelin (iroDE) biosynthesis. In both strains, the genes of toxin–antitoxin systems, as well as regulators of the expression of virulence factors and adhesion genes were also detected. Studies on the genetic potential of multiresistant K. pneumoniae strains as well as their cellular and molecular interactions with the host are of fundamental importance to assess the association of certain virulence factors with the intensity of the inflammatory response. In this sense, this work explored the virulence profile based on genomic and in vivo studies of hypermucoviscous carbapenem-resistant K. pneumoniae ST25 strains, expanding the knowledge of the biology of the emerging ST25 clone in Argentina.     

AES for multiscale localization modeling in granular media

This work presents a multiscale strong discontinuity approach to tackle key challenges in modeling localization behavior in granular media: accommodation of discontinuities in the kinematic fields, and direct linkage to the underlying grain-scale information. Assumed enhanced strain (AES) concepts are borrowed to enhance elements for post-localization analysis, but are reformulated within a recently-proposed hierarchical multiscale computational framework. Unlike classical AES methods, where material properties are usually constants or assumed to evolve with some arbitrary phenomenological laws, this framework provides a bridge to extract evolutions of key material parameters, such as friction and dilatancy, based on grain scale computational or experimental data. More importantly, the phenomenological softening modulus typically used in AES methods is no longer required. Numerical examples of plane strain compression tests are presented to illustrate the applicability of this method and to analyze its numerical performance.     

Physical and mechanical characterization of phyllites and metagreywackes in central Portugal

This work defines the physical and mechanical characteristics of phyllites and metagreywackes belonging to a schist–greywacke complex in central Portugal. The index properties, point load strength, uniaxial compressive strength, slake durability and Schmidt rebound hardness were determined. In general, the metagreywackes gave a higher strength than phyllites with the same weathering degree. Possible correlations and estimation models were established and compared with the equations obtained by different researchers.     

Tokens vs. Signals: On Conformance between Formal Models of Dataflow and Hardware

Designing hardware often involves several types of modeling and analysis, e.g., in order to check system correctness, to derive performance properties such as throughput, to optimize resource usages (e.g., buffer sizes), and to synthesize parts of a circuit (e.g., control logic). Working directly with low-level hardware models such as finite-state machines (FSMs) to answer such questions is often infeasible, e.g., due to state explosion. Instead, designers often use dataflow models such as SDF and CSDF, which are more abstract than FSMs, and less expensive to use since they come with more efficient analysis algorithms. However, dataflow models are only abstractions of the real hardware, and often omit critical information. This raises the question, when can one say that a certain dataflow model faithfully captures a given piece of hardware? The question is of more than simply academic interest. Indeed, as illustrated in this paper, dataflow-based analysis outcomes may sometimes be defensive (e.g., buffers that are too big) or even incorrect (e.g., buffers that are too small). To answer the question of faithfully capturing hardware using dataflow models, we develop a formal conformance relation between the heterogeneous formalisms of (1) finite-state machines with synchronous semantics, typically used to model synchronous hardware, and (2) asynchronous processes communicating via queues, used as a formal model for dataflow. The conformance relation preserves performance properties such as worst-case throughput and latency.     

Molds and Resists Studies for Nanoimprint Lithography of Electrodes in Low-Voltage Polymer Thin-Film Transistors

A low-cost patterning of electrodes was investigated looking forward to replacing conventional photolithography for the processing of low-operating voltage polymeric thin-film transistors. Hard silicon, etched by sulfur hexafluoride and oxygen gas mixture, and flexible polydimethylsiloxane imprinting molds were studied through atomic force microscopy (AFM) and field emission gun scanning electron microscopy. The higher the concentration of oxygen in reactive ion etching, the lower the etch rate, sidewall angle, and surface roughness. A concentration around 30 % at 100 mTorr, 65 W and 70 sccm was demonstrated as adequate for submicrometric channels, presenting a reduced etch rate of 176 nm/min. Imprinting with positive photoresist AZ1518 was compared to negative SU-8 2002 by optical microscopy and AFM. Conformal results were obtained only with the last resist by hot embossing at 120 °C and 1 kgf/cm² for 2 min, followed by a 10 min post-baking at 100 °C. The patterning procedure was applied to define gold source and drain electrodes on oxide-covered substrates to produce bottom-gate bottom-contact transistors. Poly(3-hexylthiophene) (P3HT) devices were processed on high-κ titanium oxynitride (TiO _x N _y ) deposited by radiofrequency magnetron sputtering over indium tin oxide-covered glass to achieve low-voltage operation. Hole mobility on micrometric imprinted channels may approach amorphous silicon (～0.01 cm²/V s) and, since these devices operated at less than 5 V, they are not only suitable for electronic applications but also as sensors in aqueous media.     

Moisture Sensitive Smart Yarns and Textiles from Self‐Balanced Silk Fiber Muscles

Smart textiles that sense, interact, and adapt to environmental stimuli have provided exciting new opportunities for a variety of applications. However, current advances have largely remained at the research stage due to the high cost, complexity of manufacturing, and uncomfortableness of environment‐sensitive materials. In contrast, natural textile materials are more attractive for smart textiles due to their merits in terms of low cost and comfortability. Here, water fog and humidity‐driven torsional and tensile actuation of thermally set twisted, coiled, plied silk fibers, and weave textiles from these silk fibers are reported. When exposed to water fog, the torsional silk fiber provides a fully reversible torsional stroke of 547° mm^?1. Coiled‐and‐thermoset silk yarns provide a 70% contraction when the relative humidity is changed from 20% to 80%. Such an excellent actuation behavior originates from water absorption‐induced loss of hydrogen bonds within the silk proteins and the associated structural transformation, which are corroborated by atomistic and macroscopic characterization of silk and molecular dynamics simulations. With its large abundance, cost‐effectiveness, and comfortability for wearing, the silk muscles will open up additional possibilities in industrial applications, such as smart textiles and soft robotics.     

Comparison of different separative techniques in the quantitative determination of active compound enclosed in liposomal systems

The suitability of three different separative techniques, dialysis, gel filtration and centrifugation, for determining the percentage of active compound included (PAI) in liposomal systems was assessed. Two model compounds, glucose and vitamin E acetate were encapsulated in dipalmitoylphosphatidylcholine (DPPC), soybean lecithin (SL) and hydrogenated soybean lecithin (HSL) multilamellar vesicles (MLV). Vitamin E acetate PAI values from DPPC MLV liposomes obtained by dialysis, gel filtration and centrifugation, were compared with those determined by differential scanning calorimetry. Glucose PAI values from DPPC MLV liposomes, obtained using the same separative techniques, were compared with that calculated by taking into account the glucose content of the liposome internal aqueous phase on the basis of liposome mean size determined by light scattering.
Vitamin E acetate and glucose PAI values from SL and HSL liposomes were compared with those obtained for DPPC liposomes. Dialysis proved suitable for PAI determination for both lipophilic and hydrophilic compounds, centrifugation was found to be suitable only for the determination of lipophilic compound PAI values while gel filtration using Sephadex G-25M proved inadequate for the determination of PAI values for both lipophilic and hydrophilic compounds in the experimental conditions used in this study.     

Enhancing effect of Cl2 atmosphere on transition aluminas transformation

In the present study we examine the changes in phase composition and microstructure undergone by solids after thermal treatments performed on transition aluminas in chlorine and air atmospheres. The thermal transformation to the stable -Al2O3 was faster for the chlorine-heated samples than for the air-heated ones, and for chlorinated samples single crystals of -Al2O3 were observed. We propose a mechanism by which AlCl3(g) formation allowed vapour mass transport that contributed to the growth of crystals of -Al2O3. Hydroxyls contained in transition aluminas could enhance gaseous transport and could also react with chlorine molecules. The subsequent -Al2O3 crystallization would be assisted by water vapour.