Wheel Slip Control Applied to an Electric Tractor for Improving Tractive Efficiency and Reducing Energy Consumption.

This work presents an automatic slip control solution applied to a two-wheel-drive (2WD) electric tractor. Considering that the slip can be maintained within a specific range that depends on the type of soil, it is possible to increase the tractive efficiency of the electric vehicle (EV). The control system can be easily designed considering only the longitudinal dynamics of the tractor while using simple proportional-integral (PI) controllers to drive the inverters associated with the rear wheels. The introduced solution is tested on an experimental electric tractor prototype traveling on firm soil considering case studies in which the slip control is enabled and disabled. The acquired results demonstrate that the slip control allows for obtaining a more stable performance and reduced energy consumption.

Effect of Residual Stresses on Fatigue Crack Growth: A Numerical Study Based on Cumulative Plastic Strain at the Crack Tip.

Residual stresses affect the fatigue behavior, given that compressive stresses delay the phenomenon, while tensile stresses accelerate it. However, the mechanisms behind the effect of residual stresses are not totally understood. A numerical study is developed here to understand the effect of thermal residual stresses (TRSs) on fatigue crack growth (FCG). The crack driving force was assumed to be the cumulative plastic strain at the crack tip. The heating of a region ahead of the crack tip produced elastic compressive TRS, which were 69% of material's yield stress. Alternatively, plastic deformation was produced by severe cooling followed by heating to generate compressive residual stresses. The crack propagation in the compressive residual stress field produced a decrease in the FCG rate. On the other hand, without the contact of crack flanks, the TRS showed no effect on FCG. Therefore, the TRSs only affect FCG by changing the crack closure level.

Data Processing to Probe the Cellular Hydrogen Peroxide Landscape.

Hydrogen peroxide (H2O2) regulates signaling pathways by modulating the activity of redox-sensitive proteins denominated redox switches. The magnitude of the transient variations in localized H2O2 pools during signaling events and how these variations impact redox switches present in the cell remain elusive. A canonical model with two chemical reactions comprising the oxidation/reduction cycle of a redox switch is described. The model is dimensionless with respect to the redox switch concentration. Thus, the time-series data required to apply the equations deduced is the percentage of oxidation of a redox switch, avoiding the application of absolute concentrations that are often difficult to measure experimentally. Here, we describe detailed protocols for the processing of experimental data with the canonical model to probe the absolute concentrations of H2O2 found in the vicinity of redox switches and probes, as well as the kinetic parameters that describe the reduction and oxidation of redox switches. The protocols are an analytical tool that helps to depict the cellular hydrogen peroxide signaling landscape, giving new insights on H2O2 signaling mechanisms, and hold the potential to be a framework for a future redox kinetomics analytical platform.

Emerging Nitric Oxide and Hydrogen Sulfide Releasing Carriers for Skin Wound Healing Therapy.

Nitric oxide (NO) and hydrogen sulfide (H2 S) have been recognized as important signalling molecules involved in multiple physiological functions, including wound healing. Their exogenous delivery has been established as a new route for therapies, being the topical application the nearest to commercialization. Nevertheless, the gaseous nature of these therapeutic agents and their toxicity at high levels imply additional challenges in the design of effective delivery systems, including the tailoring of their morphology and surface chemistry to get controllable release kinetics and suitable lifetimes. This review highlights the increasing interest in the use of these gases in wound healing applications by presenting the various potential strategies in which NO and/or H2 S are the main therapeutic agents, with focus on their conceptual design, release behaviour and therapeutic performance. These strategies comprise the application of several types of nanoparticles, polymers, porous materials, and composites as new releasing carriers of NO and H2 S, with characteristics that will facilitate the application of these molecules in the clinical practice.

Fatigue Crack Growth in Metallic Materials.

Mechanical components and structures are submitted to cyclic loads in different applications; therefore, they must be designed to withstand fatigue [...].

Chitosan Biocomposites for the Adsorption and Release of H2S.

The search for H2S donors has been increasing due to the multiple therapeutic effects of the gas. However, the use of nanoporous materials has not been investigated despite their potential. Zeolites and activated carbons are known as good gas adsorbents and their modification with chitosan may increase the material biocompatibility and simultaneously its release time in aqueous solution, thus making them good H2S donors. Herein, we modified with chitosan a series of A zeolites (3A, 4A and 5A) with different pore sizes and an activated carbon obtained from glycerin. The amount of H2S adsorbed was evaluated by a volumetric method and their release capacity in aqueous solution was measured. These studies aimed to verify which of the materials had appropriate H2S adsorption/release properties to be considered a potential H2S donor. Additionally, cytotoxicity assays using HeLa cells were performed. Considering the obtained results, the chitosan composite with the A zeolite with the larger pore opening was the most promising material to be used as a H2S donor so a further cytotoxicity assay using H2S loaded was conducted and no toxicity was observed.

FCG Modelling Considering the Combined Effects of Cyclic Plastic Deformation and Growth of Micro-Voids.

Fatigue is one of the most prevalent mechanisms of failure. Thus, the evaluation of the fatigue crack growth process is fundamental in engineering applications subjected to cyclic loads. The fatigue crack growth rate is usually accessed through the da/dN-ΔK curves, which have some well-known limitations. In this study a numerical model that uses the cyclic plastic strain at the crack tip to predict da/dN was coupled with the Gurson-Tvergaard-Needleman (GTN) damage model. The crack propagation process occurs, by node release, when the cumulative plastic strain reaches a critical value. The GTN model is used to account for the material degradation due to the growth of micro-voids process, which affects fatigue crack growth. Predictions with GTN are compared with the ones obtained without this ductile fracture model. Crack closure was studied in order to justify the lower values of da/dN obtained in the model with GTN, when compared with the results without GTN, for lower ΔK values. Finally, the accuracy of both variants of the numerical model is accessed through the comparison with experimental results.

Revisiting Classical Issues of Fatigue Crack Growth Using a Non-Linear Approach.

Fatigue crack growth (FCG) has been studied for decades; however, several aspects are still objects of controversy. The objective here is to discuss different issues, using a numerical approach based on crack tip plastic strain, assuming that FCG is driven by crack tip deformation. ΔK was found to control cyclic plastic deformation at the crack tip, while Kmax has no effect. Therefore, alternative mechanisms are required to justify models based on ΔK and Kmax. The analysis of crack tip plastic deformation also showed that there is crack tip damage below crack closure. Therefore, the definition of an effective load range ΔKeff = Kmax - Kopen is not correct, because the portion of load range below opening also contributes to FCG. Below crack closure, damage occurs during unloading while during loading the crack tip deformation is elastic. However, if the maximum load is decreased below the elastic limit, which corresponds to the transition between elastic and elasto-plastic regimes, there is no crack tip damage. Additionally, a significant effect of the crack ligament on crack closure was found in tests with different crack lengths and the same ΔK. Finally, the analysis of FCG after an overload with and without contact of crack flanks showed that the typical variation of da/dN observed is linked to crack closure variations, while the residual stresses ahead of crack tip are not affected by the contact of crack flanks.

A Comparison of Different Approaches to Quantify Nitric Oxide Release from NO-Releasing Materials in Relevant Biological Media.

The development of solid materials that deliver nitric oxide (NO) are of interest for several therapeutic applications. Nevertheless, due to NO's reactive nature, rapid diffusion and short half-life, reporting their NO delivery characteristics is rather complex. The full knowledge of this parameter is fundamental to discuss the therapeutic utility of these materials, and thus, the NO quantification strategy must be carefully considered according to the NO-releasing scaffold type, to the expected NO-releasing amounts and to the medium of quantification. In this work, we explore and discuss three different ways of quantifying the release of NO in different biological fluids: haemoglobin assay, Griess assay and NO electrochemical detection. For these measurements, different porous materials, namely zeolites and titanosilicates were used as models for NO-releasing platforms. The oxyhaemoglobin assay offers great sensitivity (nanomolar levels), but it is only possible to monitor the NO release while oxyhaemoglobin is not fully converted. On the other hand, Griess assay has low sensitivity in complex biological media, namely in blood, and interferences with media make NO measurements questionable. Nevertheless, this method can measure micromolar amounts of NO and may be useful for an initial screening for long-term release performance. The electrochemical sensor enabled real-time measurements in a variety of biological settings. However, measured NO is critically low in oxygenated and complex media, giving transient signals, which makes long-term quantification impossible. Despite the disadvantages of each method, the combination of all the results provided a more comprehensive NO release profile for these materials, which will help to determine which formulations are most promising for specific therapeutic applications. This study highlights the importance of using appropriate NO quantification tools to provide accurate reports.

Numerical Study on the Variability of Plastic CTOD.

This paper presents a numerical study on the influence of material parameters and loading variability in the plastic crack tip opening displacement (CTOD) results. For this purpose, AA7050-T6 was selected as reference material and a middle-cracked tension specimen geometry was considered. The studied input parameters were the Young's modulus, Poisson's ratio, isotropic and kinematic hardening parameters and the maximum and minimum applied loads. The variability of the input parameters follows a Gaussian distribution. First, screening design-of-experiments were performed to identify the most influential parameters. Two types of screening designs were considered: one-factor-at-a-time and fractional factorial designs. Three analysis criteria were adopted, based on: main effect, index of influence and analysis of variance. Afterwards, metamodels were constructed to establish relationships between the most influential parameters and the plastic crack tip opening displacement (CTOD) range, based on two types of designs: Face-Centered Central Composite Design and Box-Behnken design. Finally, the metamodels were validated, enabling the expeditious evaluation of the variability in the plastic CTOD range; in addition, the variability in the fatigue crack growth rate was also evaluated.

Tuning Cellular Biological Functions Through the Controlled Release of NO from a Porous Ti-MOF.

Materials for the controlled release of nitric oxide (NO) are of interest for therapeutic applications. However, to date, many suffer from toxicity and stability issues, as well as poor performance. Herein, we propose a new NO adsorption/release mechanism through the formation of nitrites on the skeleton of a titanium-based metal-organic framework (MOF) that we named MIP-177, featuring a suitable set of properties for such an application: (i) high NO storage capacity (3 µmol mg-1solid ), (ii) excellent biocompatibility at therapeutic relevant concentrations (no cytotoxicity at 90 µg mL-1 for wound healing) due to its high stability in biological media (<9 % degradation in 72 hours) and (iii) slow NO release in biological media (≈2 hours for 90 % release). The prospective application of MIP-177 is demonstrated through NO-driven control of mitochondrial respiration in cells and stimulation of cell migration, paving the way for the design of new NO delivery systems for wound healing therapy.

Human Aquaporin-5 Facilitates Hydrogen Peroxide Permeation Affecting Adaption to Oxidative Stress and Cancer Cell Migration.

Reactive oxygen species (ROS), including H2O2, contribute to oxidative stress and may cause cancer initiation and progression. However, at low concentrations, H2O2 can regulate signaling pathways modulating cell growth, differentiation, and migration. A few mammalian aquaporins (AQPs) facilitate H2O2 diffusion across membranes and participate in tumorigenesis. AQP3 and AQP5 are strongly expressed in cancer tissues and AQP3-mediated H2O2 transport has been related to breast cancer cell migration, but studies with human AQP5 are lacking. Here, we report that, in addition to its established water permeation capacity, human AQP5 facilitates transmembrane H2O2 diffusion and modulates cell growth of AQP5-transformed yeast cells in response to oxidative stress. Mutagenesis studies revealed that residue His173 located in the selective filter is crucial for AQP5 permeability, and interactions with phosphorylated Ser183 may regulate permeation through pore blockage. Moreover, in human pancreatic cancer cells, the measured AQP5-mediated H2O2 influx rate indicates the presence of a highly efficient peroxiporin activity. Cell migration was similarly suppressed by AQP3 or AQP5 gene silencing and could be recovered by external oxidative stimuli. Altogether, these results unveiled a major role for AQP5 in dynamic fine-tuning of the intracellular H2O2 concentration, and consequently in activating signaling networks related to cell survival and cancer progression, highlighting AQP5 as a promising drug target for cancer therapies.

New generation of nitric oxide-releasing porous materials: Assessment of their potential to regulate biological functions.

Nitric oxide (NO) presents innumerable biological roles, and its exogenous supplementation for therapeutic purposes has become a necessity. Some nanoporous materials proved to be potential vehicles for NO with high storage capacity. However, there is still a lack of information about their efficiency to release controlled NO and if they are biocompatible and biologically stable. In this work, we address this knowledge gap starting by evaluating the NO release and stability under biological conditions and their toxicity with primary keratinocyte cells. Titanosilicates (ETS-4 and ETS-10 types) and clay-based materials were the materials under study, which have shown in previous studies suitable NO gas adsorption/release rates. ETS-4 proved to be the most promising material, combining good biocompatibility at 180 µg/mL, stability and slower NO release. ETS-10 and ETAS-10 showed the best biocompatibility at the same concentration and, in the case of clay-based materials, CoOS is the least toxic of those tested and the one that releases the highest NO amount. The potentiality of these new NO donors to regulate biological functions was assessed next by controlling the mitochondrial respiration and the cell migration. NO-loaded ETS-4 regulates O2 consumption and cell migration in a dose-dependent manner. For cell migration, a biphasic effect was observed in a narrow range of ETS-4 concentration, with a stimulatory effect becoming inhibitory just by doubling ETS-4 concentration. For the other materials, no effective regulation was achieved, which highlights the relevance of the new assessment presented in this work for nanoporous NO carriers that will pave the way for further developments.

Measuring intracellular concentration of hydrogen peroxide with the use of genetically encoded H2O2 biosensor HyPer.

In this study, we propose a method for quantification of average hydrogen peroxide concentration within a living cell that is based on the use of genetically encoded H2O2 biosensor HyPer. The method utilizes flow cytometric measurements of HyPer fluorescence in H2O2-exposed cells to analyze the biosensor oxidation kinetics. Fitting the experimental curves with kinetic equations allows determining the rate constants of HyPer oxidation/reduction which are used further for the calculation of peroxide concentrations in the cells of interest both in the presence and absence of external H2O2. Applying this method to K562 cells, we have estimated the gradient as about 390-fold between the extracellular and intracellular level of exogenous H2O2 in cells exposed to the micromole doses of peroxide, as well as the average basal level of H2O2 in the cytosol of undisturbed cells ( [Formula: see text] ). The method can be extended to other H2O2-sensitive redox probes or to procedures in which, rather than adding external peroxide, intracellular production of peroxide is triggered, providing a tool to quantitate not only basal average H2O2 concentrations but also the concentration of peroxide build up in the vicinity of redox probes.

Using in vivo oxidation status of one- and two-component redox relays to determine H2O2 levels linked to signaling and toxicity.

BACKGROUND: Hydrogen peroxide (H2O2) is generated as a by-product of metabolic reactions during oxygen use by aerobic organisms, and can be toxic or participate in signaling processes. Cells, therefore, need to be able to sense and respond to H2O2 in an appropriate manner. This is often accomplished through thiol switches: Cysteine residues in proteins that can act as sensors, and which are both scarce and finely tuned. Bacteria and eukaryotes use different types of such sensors-either a one-component (OxyR) or two-component (Pap1-Tpx1) redox relay, respectively. However, the biological significance of these two different signaling modes is not fully understood, and the concentrations and peroxides driving those types of redox cascades have not been determined, nor the intracellular H2O2 levels linked to toxicity. Here we elucidate the characteristics, rates, and dynamic ranges of both systems. RESULTS: By comparing the activation of both systems in fission yeast, and applying mathematical equations to the experimental data, we estimate the toxic threshold of intracellular H2O2 able to halt aerobic growth, and the temporal gradients of extracellular to intracellular peroxides. By calculating both the oxidation rates of OxyR and Tpx1 by peroxides, and their reduction rates by the cellular redoxin systems, we propose that, while Tpx1 is a sensor and an efficient H2O2 scavenger because it displays fast oxidation and reduction rates, OxyR is strictly a H2O2 sensor, since its reduction kinetics are significantly slower than its oxidation by peroxides, and therefore, it remains oxidized long enough to execute its transcriptional role. We also show that these two paradigmatic H2O2-sensing models are biologically similar at pre-toxic peroxide levels, but display strikingly different activation behaviors at toxic doses. CONCLUSIONS: Both Tpx1 and OxyR contain thiol switches, with very high reactivity towards peroxides. Nevertheless, the fast reduction of Tpx1 defines it as a scavenger, and this efficient recycling dramatically changes the Tpx1-Pap1 response to H2O2 and connects H2O2 sensing to the redox state of the cell. In contrast, OxyR is a true H2O2 sensor but not a scavenger, being partially insulated from the cellular electron donor capacity.

European contribution to the study of ROS: A summary of the findings and prospects for the future from the COST action BM1203 (EU-ROS).

The European Cooperation in Science and Technology (COST) provides an ideal framework to establish multi-disciplinary research networks. COST Action BM1203 (EU-ROS) represents a consortium of researchers from different disciplines who are dedicated to providing new insights and tools for better understanding redox biology and medicine and, in the long run, to finding new therapeutic strategies to target dysregulated redox processes in various diseases. This report highlights the major achievements of EU-ROS as well as research updates and new perspectives arising from its members. The EU-ROS consortium comprised more than 140 active members who worked together for four years on the topics briefly described below. The formation of reactive oxygen and nitrogen species (RONS) is an established hallmark of our aerobic environment and metabolism but RONS also act as messengers via redox regulation of essential cellular processes. The fact that many diseases have been found to be associated with oxidative stress established the theory of oxidative stress as a trigger of diseases that can be corrected by antioxidant therapy. However, while experimental studies support this thesis, clinical studies still generate controversial results, due to complex pathophysiology of oxidative stress in humans. For future improvement of antioxidant therapy and better understanding of redox-associated disease progression detailed knowledge on the sources and targets of RONS formation and discrimination of their detrimental or beneficial roles is required. In order to advance this important area of biology and medicine, highly synergistic approaches combining a variety of diverse and contrasting disciplines are needed.

Incidência de infecções primárias na corrente sanguínea em uma UTI neonatal / Incidence of primary bloodstream infections in a neonatal intensive care UNIT

As infecções primárias da corrente sanguínea são umas das principais causas de morbimortalidade intra-hospitalar. Além da grande responsabilidade sobre as mortes perinatais, estas são as principais infecções em Unidade de Terapia Intensiva Neonatal (UTI neonatal) e estão associadas a altos custos hospitalares, representando assim um grave problema de saúde pública. O objetivo é identificar a incidência de Infecções Primárias da Corrente Sanguínea (IPCS) em uma UTI neonatal de uma unidade hospitalar do Estado de Santa Catarina, Trata-se de um estudo descritivo retrospectivo, de abordagem quantitativa, tendo por base os dados coletados pela Comissão de Controle de Infeções relacionadas à assistência à saúde (CCIH) . Esta caracteriza as infecções com base nos Critérios Diagnósticos de Infecções Relacionadas à Assistência à Saúde da Agência Nacional de Vigilância Sanitária (ANVISA) as IPCS ocorridas na UTI neonatal nos anos de 2014 a 2015. Ocorreram neste período um total de 30 casos, sendo que em ambos os anos a maioria dos casos de IPCS ocorreram em pacientes do sexo feminino e com mais de 1.500g de peso ao nascer. A caraterização das IPCS laboratorial foi positiva em 88,5% e dentre as bactérias isoladas, apenas 4 (44,44%) no ano de 2014 foram classificadas como multirresistentes e não foram encontradas estas no ano de 2015. As bactérias mais comumente isoladas foram Staphylococcus aureus (41,6%) e Staphylococcus coagulase negativa (37,5%). Sendo que o desfecho dos casos foi predominantemente alta hospitalar.

The primary bloodstream infections are a major cause of in-hospital mortality. Besides the great responsibility on perinatal deaths, these are the main infections in neonatal ICUs and are associated with high hospital costs, thus representing a serious public health problem. The aim of this article is to analyze the incidence of Primary Blood Stream Infections in the Intensive Care Unit of a hospital in the state of Santa Catarina, as well as identify the most prevalent bacteria in IPCS and its sensitivity to antibiotics. This is a retrospective descriptive study with a quantitative approach, based on data collected by the Commission of Hospital Infection Control (CCIH) of Tereza Ramos Hospital (HTR), which characterized the IPCS occurred in the Neonatal Intensive Care Unit of the same hospital in the years 2014 and 2015. Occurred in this period a total of 30 cases, and in both years the majority of cases of IPCS occurred in female patients and over 1.500g and the predominant outcome was discharged. Blood cultures were positive in 88.5% and among the bacteria isolated, only 4 (44.44%) in 2014 were classified as multirresistentes. As most commonly isolated bacteria were Staphylococcus aureus (41.6%) and staphylococcus coagulase negative (37.5%). It is understood finally that the characteristics of the patients in our ICU IPCS follows specific pattern and increasing values, showing that are required targeted and effective measures to reduce the number of infections at our institution.

Quantitative biology of hydrogen peroxide signaling.

Hydrogen peroxide (H2O2) controls signaling pathways in cells by oxidative modulation of the activity of redox sensitive proteins denominated redox switches. Here, quantitative biology concepts are applied to review how H2O2 fulfills a key role in information transmission. Equations described lay the foundation of H2O2 signaling, give new insights on H2O2 signaling mechanisms, and help to learn new information from common redox signaling experiments. A key characteristic of H2O2 signaling is that the ratio between reduction and oxidation of redox switches determines the range of H2O2 concentrations to which they respond. Thus, a redox switch with low H2O2-dependent oxidability and slow reduction rate responds to the same range of H2O2 concentrations as a redox switch with high H2O2-dependent oxidability, but that is rapidly reduced. Yet, in the first case the response time is slow while in the second case is rapid. H2O2 sensing and transmission of information can be done directly or by complex mechanisms in which oxidation is relayed between proteins before oxidizing the final regulatory redox target. In spite of being a very simple molecule, H2O2 has a key role in cellular signaling, with the reliability of the information transmitted depending on the inherent chemical reactivity of redox switches, on the presence of localized H2O2 pools, and on the molecular recognition between redox switches and their partners.

Vitamin B3 metal-organic frameworks as potential delivery vehicles for therapeutic nitric oxide.

The synthesis and structural characterization of two isostructural metal (M=Ni, Co) 3D framework structure that integrate vitamin B3 building blocks with NO delivery capabilities and low toxicity is presented. The compounds with a formula [M2(µ2-H2O)(µ-vitamin B3)4]·2H2O contain two crystallographic distinct divalent metal centres connected by a bridging water and carboxylate group from vitamin B3. The porous compounds have the capability of storing and releasing nitric oxide (NO) in a slow and reversible manner, with released amounts of 2.6 and 2.0µmol NOmgsolid-1, on the Ni and Co compound, respectively. The NO release followed a convenient slow release kinetic profile in both gas and liquid phases. Haemoglobin tests demonstrated that NO is released to the medium in a biologically active form, thus suitable to trigger the desired response in biological systems. The toxicity of the samples with and without loaded NO was evaluated from cytotoxicity tests in HeLa and HEKn cells, showing low toxicity of the compounds at concentrations below 180µgcm-3. The overall results indicate that these bio based MOFs are of interest for therapeutic applications related with NO delivery. STATEMENT OF SIGNIFICANCE.

Rat Aquaporin-5 Is pH-Gated Induced by Phosphorylation and Is Implicated in Oxidative Stress.

Aquaporin-5 (AQP5) is a membrane water channel widely distributed in human tissues that was found up-regulated in different tumors and considered implicated in carcinogenesis in different organs and systems. Despite its wide distribution pattern and physiological importance, AQP5 short-term regulation was not reported and mechanisms underlying its involvement in cancer are not well defined. In this work, we expressed rat AQP5 in yeast and investigated mechanisms of gating, as well as AQP5's ability to facilitate H2O2 plasma membrane diffusion. We found that AQP5 can be gated by extracellular pH in a phosphorylation-dependent manner, with higher activity at physiological pH 7.4. Moreover, similar to other mammalian AQPs, AQP5 is able to increase extracellular H2O2 influx and to affect oxidative cell response with dual effects: whereas in acute oxidative stress conditions AQP5 induces an initial higher sensitivity, in chronic stress AQP5 expressing cells show improved cell survival and resistance. Our findings support the involvement of AQP5 in oxidative stress and suggest AQP5 modulation by phosphorylation as a novel tool for therapeutics.