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An under-explored target for SARS-CoV-2 is non-structural protein 14 (Nsp14), a crucial enzyme for viral replication that catalyzes the methylation ofN7-guanosine of the viral RNA at 5′-end; this enables the virus to evade the host immune response by mimicking the eukaryotic post-transcriptional modification mechanism. We sought new inhibitors of the S-adenosyl methionine (SAM)-dependent methyltransferase (MTase) activity of Nsp14 with three large library docking strategies. First, up to 1.1 billion make-on-demand (“tangible”) lead-like molecules were docked against the enzyme’s SAM site, seeking reversible inhibitors. On de novo synthesis and testing, three inhibitors emerged with IC50values ranging from 6 to 43 μM, each with novel chemotypes. Structure-guided optimization andin vitrocharacterization supported their non-covalent mechanism. In a second strategy, docking a library of 16 million tangible fragments revealed nine new inhibitors with IC50values ranging from 12 to 341 μM and ligand efficiencies from 0.29 to 0.42. In a third strategy, a newly created library of 25 million tangible, virtual electrophiles were docked to covalently modify Cys387 in the SAM binding site. Seven inhibitors emerged with IC50values ranging from 3.2 to 39 μM, the most potent being a reversible aldehyde. Initial optimization of a second series yielded a 7 μM acrylamide inhibitor. Three inhibitors characteristic of the new series were tested for selectivity against 30 human protein and RNA MTases, with one showing partial selectivity and one showing high selectivity. Overall, 32 inhibitors encompassing eleven chemotypes had IC50values 50values

Severe acute respiratory coronavirus 2 (SARS-CoV-2) infection causes neurological disease in some patients suggesting that infection can affect both the peripheral and central nervous system (PNS and CNS, respectively). It is not clear whether the outcome of SARS-CoV-2 infection of PNS and CNS neurons is similar, and which are the key factors that cause neurological disease: SARS-CoV-2 infection or the subsequent immune response. Here, we addressed these questions by infecting human induced-pluripotent stem cell-derived CNS and PNS neurons with the β strain of SARS-CoV-2. Our results show that SARS-CoV-2 infects PNS neurons more efficiently than CNS neurons, despite lower expression levels of angiotensin converting enzyme 2. Infected PNS neurons produced interferon λ1, several interferon stimulated genes and proinflammatory cytokines. They also displayed neurodegenerative-like alterations, as indicated by increased levels of sterile alpha and Toll/interleukin receptor motif-containing protein 1, amyloid precursor protein and α-synuclein and lower levels of nicotinamide mononucleotide adenylyltransferase 2 and β-III-tubulin. Interestingly, blockade of the Janus kinase and signal transducer and activator of transcription pathway by Ruxolitinib did not increase SARS-CoV-2 infection, but reduced neurodegeneration, suggesting that an exacerbated neuronal innate immune response contributes to pathogenesis in the PNS.

Chikungunya virus (CHIKV) is an arthritogenic reemerging virus replicating in plasma membrane-derived compartments termed "spherules." Here, we identify the human transmembrane protein CD81 as host factor required for CHIKV replication. Ablation of CD81 results in decreased CHIKV permissiveness, while overexpression enhances infection. CD81 is dispensable for virus uptake but critically required for viral genome replication. Likewise, murine CD81 is crucial for CHIKV permissiveness and is expressed in target cells such as dermal fibroblasts, muscle and liver cells. Whereas related alphaviruses, including Ross River virus (RRV), Semliki Forest virus (SFV), Sindbis virus (SINV) and Venezuelan equine encephalitis virus (VEEV), also depend on CD81 for infection, RNA viruses from other families, such as coronaviruses, replicate independently of CD81. Strikingly, the replication-enhancing function of CD81 is linked to cholesterol binding. These results define a mechanism exploited by alphaviruses to hijack the membrane microdomain-modeling protein CD81 for virus replication through interaction with cholesterol. IMPORTANCE: In this study, we discover the tetraspanin CD81 as a host factor for the globally emerging chikungunya virus and related alphaviruses. We show that CD81 promotes replication of viral genomes in human and mouse cells, while virus entry into cells is independent of CD81. This provides novel insights into how alphaviruses hijack host proteins to complete their life cycle. Alphaviruses replicate at distinct sites of the plasma membrane, which are enriched in cholesterol. We found that the cholesterol-binding ability of CD81 is important for its function as an alphavirus host factor. This discovery thus broadens our understanding of the alphavirus replication process and the use of host factors to reprogram cells into virus replication factories.

Cellular compartmentalization of proteins and protein complex formation allow cells to tightly control biological processes. Therefore, understanding the subcellular localization and interactions of a specific protein is crucial to uncover its biological function. The advent of proximity labeling (PL) has reshaped cellular proteomics in infection biology. PL utilizes a genetically modified enzyme that generates a "labeling cloud" by covalently labeling proteins in close proximity to the enzyme. Fusion of a PL enzyme to a specific antibody or a "bait" protein of interest in combination with affinity enrichment mass spectrometry (AE-MS) enables the isolation and identification of the cellular proximity proteome, or proxisome. This powerful methodology has been paramount for the mapping of membrane or membraneless organelles as well as for the understanding of hard-to-purify protein complexes, such as those of transmembrane proteins. Unsurprisingly, more and more infection biology research groups have recognized the potential of PL for the identification of host-pathogen interactions. In this chapter, we introduce the enzymes commonly used for PL labeling as well as recent promising advancements and summarize the major achievements in organelle mapping and nucleic acid PL. Moreover, we comprehensively describe the research on host-pathogen interactions using PL, giving special attention to studies in the field of virology.

The HCV replication cycle is tightly associated with host lipid metabolism: Lipoprotein receptors SR-B1 and LDLr promote entry of HCV, replication is associated with the formation of lipid-rich membranous organelles and infectious particle assembly highjacks the very-low-density lipoprotein (VLDL) secretory pathway. Hence, medications that interfere with the lipid metabolism of the cell, such as statins, may affect HCV infection. Here, we study the interplay between lipoprotein receptors, lipid homeostasis, and HCV infection by genetic and pharmacological interventions. We found that individual ablation of the lipoprotein receptors SR-B1 and LDLr did not drastically affect HCV entry, replication, or infection, but double lipoprotein receptor knock-outs significantly reduced HCV infection. Furthermore, we could show that this effect was neither due to altered expression of additional HCV entry factors nor caused by changes in cellular cholesterol content. Strikingly, whereas lipid-lowering drugs such as simvastatin or fenofibrate did not affect HCV entry or infection of immortalized hepatoma cells expressing SR-B1 and/or LDLr or primary human hepatocytes, ablation of these receptors rendered cells more susceptible to these drugs. Finally, we observed no significant differences between statin users and control groups with regards to HCV viral load in a cohort of HCV infected patients before and during HCV antiviral treatment. Interestingly, statin treatment, which blocks the mevalonate pathway leading to decreased cholesterol levels, was associated with mild but appreciable lower levels of liver damage markers before HCV therapy. Overall, our findings confirm the role of lipid homeostasis in HCV infection and highlight the importance of the mevalonate pathway in the HCV replication cycle.

The SARS-CoV-2 coronavirus has led to a pandemic with millions of people affected. The present study finds prostaglandin E2 (PGE2) blood levels elevated in COVID-19 patients with positive correlation with disease severity. SARS-CoV-2 induces PGE2 generation and secretion in infected lung epithelial cells by upregulating cyclo-oxygenase (COX)-2 and reducing the PG-degrading enzyme 15-hydroxyprostaglandin-dehydrogenase. Also living human-lung-precision-slices infected with SARS-CoV-2 display upregulated COX-2. PGE2 in serum of COVID-19 patients lowers the expression of Paired-Box-Protein-Pax-5 (PAX5), a master regulator of B-cell survival, proliferation and differentiation, in both human and mouse pre-B-cells, while the PGE2 inhibitor taxifolin directly reduces SARS-CoV-2-induced PGE2 production and attenuates viral replication. Risk-factors for severe disease courses, i.e. older age, male sex and air pollution are associated with higher PGE2 production and lower PAX5 expression in pre-B-cells. Since PGE2 acts broadly immunosuppressive its elevation might reduce the early anti-viral defense and its inhibition may therefore reduce severe disease courses.

Several cationic amphiphilic drugs (CADs) have been found to inhibit cell entry of filoviruses and other enveloped viruses. Structurally unrelated CADs may have antiviral activity, yet the underlying common mechanism and structure-activity relationship are incompletely understood. We aimed to understand how widespread antiviral activity is among CADs and which structural and physico-chemical properties are linked to entry inhibition. We measured inhibition of Marburg virus pseudoparticle (MARVpp) cell entry by 45 heterogeneous and mostly FDA-approved CADs and cytotoxicity in EA.hy926 cells. We analyzed correlation of antiviral activity with four chemical properties: pKa, hydrophobicity (octanol/water partitioning coefficient; ClogP), molecular weight, and distance between the basic group and hydrophobic ring structures. Additionally, we quantified drug-induced phospholipidosis (DIPL) of a CAD subset by flow cytometry. Structurally similar compounds (derivatives) and those with similar chemical properties but unrelated structures (analogues) to those of strong inhibitors were obtained by two in silico similarity search approaches and tested for antiviral activity. Overall, 11 out of 45 (24%) CADs inhibited MARVpp by 40% or more. The strongest antiviral compounds were dronedarone, triparanol, and quinacrine. Structure-activity relationship studies revealed highly significant correlations between antiviral activity, hydrophobicity (ClogP > 4), and DIPL. Moreover, pKa and intramolecular distance between hydrophobic and hydrophilic moieties correlated with antiviral activity but to a lesser extent. We also showed that in contrast to analogues, derivatives had antiviral activity similar to that of the seed compound dronedarone. Overall, one-quarter of CADs inhibit MARVpp entry in vitro, and antiviral activity of CADs mostly relies on their hydrophobicity yet is promoted by the individual structure.

Clinical data of patients suffering from COVID-19 indicates that statin therapy, used to treat hypercholesterolemia, is associated with a better disease outcome. Whether statins directly affect virus replication or influence the clinical outcome through modulation of immune responses is unknown. We therefore investigated the effect of statins on SARS-CoV-2 infection in human lung cells and found that only fluvastatin inhibited low and high pathogenic coronaviruses in vitro and ex vivo in a dose-dependent manner. Quantitative proteomics revealed that fluvastatin and other tested statins modulated the cholesterol synthesis pathway without altering innate antiviral immune responses in infected lung epithelial cells. However, fluvastatin treatment specifically downregulated proteins that modulate protein translation and viral replication. Collectively, these results support the notion that statin therapy poses no additional risk to individuals exposed to SARS-CoV-2 and that fluvastatin has a moderate beneficial effect on SARS-CoV-2 infection of human lung cells., Graphical abstract, Drugs; Virology

This paper is concerned with the application of the strip yield models implemented in NASGRO to estimate fatigue crack growth under random loading. The two different strip yield model options (constant constraint-loss (CCL) and variable constraint-loss (VCL)) implemented in the software are considered and compared. In addition, three crack grow rate relations, obtained from constant amplitude data, the NASGRO material database and the literature, were also considered and compared. The capacity of the models to estimate the fatigue life and variability is analyzed by comparing simulated results with experimental data of fatigue crack growth under different stationary Gaussian random load processes on compact, C(T), specimens of aluminum alloy 2024-T351. The analysis performed showed that both model options provide very similar good fatigue life predictions. The estimated fatigue lives are within ±13% of the test lives with the best option. The variability of the results due to the randomness of the load is also analyzed. In this case, the variable constraint-loss option provides better estimation than the constant constraint-loss option.

Background & Aims The lipid-binding protein, SEC14L2, is crucial for the efficient viral replication of clinical hepatitis C virus (HCV) isolates in cell culture. Given the role of SEC14L2 in HCV replication, we aimed to study a large number of HCV positive sera carrying genotypes 1–4, to identify viral factors associated with efficient replication in culture. Additionally, we investigated whether 13 single nucleotide polymorphisms (SNPs) of SEC14L2 have an impact on RNA replication of naturally occurring HCV isolates. Methods We generated Huh-7.5 cell lines overexpressing SEC14L2 or 13 coding SNPs and tested 73 different HCV positive sera for in vitro replication. Furthermore, we genotyped a cohort of 262 patients with chronic HCV for the common SNP (rs757660) and investigated its effect on the clinical phenotype. Results HCV isolates from genotype 1, 2, 3 and 4 replicate in Huh-7.5 cells overexpressing SEC14L2. Interestingly, only subgenomic replicons from genotypes 1 and 3 showed enhanced replication whereas genotypes 2 and 4 remained unaffected. Furthermore, replication was independent of viral load. Importantly, all tested SNPs supported HCV RNA replication in vitro, while 1 SNP was associated with decreased SEC1L2 expression and viral RNA. All SNPs exhibited comparable cellular cholesterol and vitamin E abundance in naive Huh-7.5 cells. Conclusions This large screen of natural HCV isolates of 4 genotypes underscores the relevance of SEC14L2 as an in vitro HCV host factor. Additionally, SEC14L2 variants appear to recapitulate the wild-type enhancement of HCV replication. Variant rs191341134 showed a decreased effect due to lowered stability, whereas variant rs757660, a high prevalence mutant, showed a similar phenotype to the wild-type. Lay summary Until the year 2015, consistent replication of patient-derived isolates of hepatitis C virus (HCV) in an in vitro model remained a limitation in HCV research. In 2015 a group of authors identified a protein named SEC14L2 that enabled the replication of HCV isolates in cell culture. We performed a large screen encompassing 73 isolates of 4 different HCV genotypes. Additionally, we replaced the natural SEC14L2 with 13 different mutants to test if the protein variation significantly altered its HCV replication enhancing functions. We showed that different genotypes of HCV react differently to the presence of this protein and the variants of the protein mimic the behavior of the wild-type.

The crack initiation and propagation behaviour of St52-3N steel is investigated under biaxial conditions. Short crack behaviour is studied and pearlite bands are found to modify the crack growth rate. The behaviour under proportional and non-proportional (90°) loading is studied with different critical plane models; Wang–Brown, Fatemi–Socie, Smith–Watson–Topper, Liu I and Liu II. Smith–Watson–Topper and Liu I are found to best predict the crack orientation under proportional loading and Fatemi–Socie and Liu II under non-proportional loading. The best predictions in terms of fatigue life are obtained by Fatemi–Socie, both under proportional and non-proportional loads.

This work presents a novel methodology for characterising fatigue cracks under biaxial conditions on a low carbon steel. It allows both short crack and early propagation stages to be studied in tubular specimens. Short crack growth is studied with a long-distance microscope acquiring images of the bare metal surface. Results showed oscillations in crack growth rate due to microstructure. Early propagation stage is studied with high magnification Digital Image Correlation (DIC) technique for measuring displacement and strain crack-tip fields. By applying micro-speckle pattern on the metal surface it is possible to achieve high magnification for DIC technique. Ultra-fine black and white speckles were created by electro-spray technique. The validity of this novel technique is demonstrated by direct comparison with extensometer measurements, under combined tension–compression and torsion conditions. It was also possible to estimate satisfactorily the mixed-mode stress intensity factor.

This work uses the strip yield model implemented in NASGRO software to estimate fatigue life under random loading. Simulated results were compared with experimental data previously obtained by the authors using different random loading processes in Al2024-T351. Test data under constant amplitude loading from different authors have been considered in order to characterize the material behaviour and fit the model parameters. The two different strip yield model implemented in NASGRO software were considered. The ratio of simulated to experimental fatigue lives was between 0.71 and 1.52 considering all options and between 0.87 and 1.12 with the best option. Ministerio de Economía y Competitividad DPI2012-33382 Junta de Andalucía TEP-3244

In this paper a methodology for evaluating crack initiation under biaxial conditions is presented. The methodology consists of evaluating the crack length automatically with digital processing of highmagnification images of the crack. The methodology was applied to study five different strain conditions on a low carbon ferritic-pearlitic steel specimen with tubular shape. A hole of 150 μm diameter was drilled to enforce the crack to initiate at a particular spot. Different combinations of axial and torsional strains were analysed during the initiation stage of the crack. The setup employed allowed detection of the crack to within 6 μm from the edge of the hole on average and monitoring of the crack during early stages. Fatigue crack propagation curves clearly showed oscillations due to microstructure. It was also observed that these oscillations decreased as the torsional component of the strain was increased. Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech. Junta de Andalucía through Proyectos de Excelencia grant reference TEP-3244 and of Spanish Ministerio de Economía y Competitividad through grant reference DPI2012-33382

Until April 2007 the MAGIC telescope used a 300 MSamples/s FADC system to sample the shaped PMT signals produced by the captured Cherenkov photons of air showers. Different algorithms to reconstruct the signal from the read-out samples (extractors) have been implemented and are described and compared. Criteria based on the obtained charge and time resolution/bias are defined and used to judge the different extractors, by applying them to calibration, cosmic and pedestal signals. The achievable charge and time resolution have been derived as functions of the incident number of photo-electrons., Accepted for publication in NIM A

Filoviruses infect a wide range of cell types with the exception of lymphocytes. The intracellular proteins cathepsin B and L, two-pore channel 1 and 2, and bona fide receptor Niemann&ndash, Pick Disease C1 (NPC1) are essential for the endosomal phase of cell entry. However, earlier steps of filoviral infection remain poorly characterized. Numerous plasma membrane proteins have been implicated in attachment but it is still unclear which ones are sufficient for productive entry. To define a minimal set of host factors required for filoviral glycoprotein-driven cell entry, we screened twelve cell lines and identified the nonlymphocytic cell line SH-SY5Y to be specifically resistant to filovirus infection. Heterokaryons of SH-SY5Y cells fused to susceptible cells were susceptible to filoviruses, indicating that SH-SY5Y cells do not express a restriction factor but lack an enabling factor critical for filovirus entry. However, all tested cell lines expressed functional intracellular factors. Global gene expression profiling of known cell surface entry factors and protein expression levels of analyzed attachment factors did not reveal any correlation between susceptibility and expression of a specific host factor. Using binding assays with recombinant filovirus glycoprotein, we identified cell attachment as the step impaired in filovirus entry in SH-SY5Y cells. Individual overexpression of attachment factors T-cell immunoglobulin and mucin domain 1 (TIM-1), Axl, Mer, or dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin (DC-SIGN) rendered SH-SY5Y cells susceptible to filovirus glycoprotein-driven transduction. Our study reveals that a lack of attachment factors limits filovirus entry and provides direct experimental support for a model of filoviral cell attachment where host factor usage at the cell surface is highly promiscuous.

The paper describes the different methods, used in the MAGIC experiment, to unfold experimental energy distributions of cosmic ray particles (gamma-rays). Questions and problems related to the unfolding are discussed. Various procedures are proposed which can help to make the unfolding robust and reliable. The different methods and procedures are implemented in the MAGIC software and are used in most of the analyses., Comment: Submitted to NIM A

The blazar Markarian 501 (Mrk 501) was observed at energies above 0.10 TeV with the MAGIC telescope from May through July 2005. The high sensitivity of the instrument enabled the determination of the flux and spectrum of the source on a night-by-night basis. Throughout our observational campaign, the flux from Mrk 501 was found to vary by an order of magnitude. Intra-night flux variability with flux-doubling times down to 2 minutes was observed during the two most active nights, namely June 30 and July 9. These are the fastest flux variations ever observed in Mrk 501. The ~20-minute long flare of July 9 showed an indication of a 4 +/- 1 min time delay between the peaks of F(1.2 TeV), which may indicate a progressive acceleration of electrons in the emitting plasma blob. The flux variability was quantified for several energy ranges, and found to increase with the energy of the gamma-ray photons. The spectra hardened significantly with increasing flux, and during the two most active nights, a spectral peak was clearly detected at 0.43 +/- 0.06 TeV and 0.25 +/- 0.07 TeV, respectively for June 30 and July 9. There is no evidence of such spectral feature for the other nights at energies down to 0.10 TeV, thus suggesting that the spectral peak is correlated with the source luminosity. These observed characteristics could be accommodated in a Synchrotron-Self-Compton (SSC) framework in which the increase in gamma-ray flux is produced by a freshly injected (high energy) electron population.

During its first data cycle, between 2005 and the beginning of 2006, the fast repositioning system of the MAGIC telescope allowed the observation of nine different gamma-ray bursts as possible sources of very high energy gamma-rays. These observations were triggered by alerts from Swift, HETE-2, and INTEGRAL; they started as quickly as possible after the alerts and lasted for several minutes, with an energy threshold varying between 80 and 200 GeV, depending on the zenith angle of the burst. No evidence for gamma signals was found, and upper limits for the flux were derived for all events using the standard analysis chain of MAGIC. For the bursts with measured redshifts, the upper limits are compatible with a power-law extrapolation, when the intrinsic fluxes are evaluated taking into account the attenuation due to the scattering in the metagalactic radiation field.

Many mechanical components are subjected to multiaxial fatigue. These conditions are typically coming from external loads, the geometry of the component and/or residual stresses. However the majority of experimental data available in the literature are focused on the simpler uni-axial fatigue problem. The present work describes a series of experimental tests conducted to characterise in a comprehensive way the multiaxial behaviour of a ST52-3N structural steel. First, the monotonic properties of the steel were obtained experimentally. Then cyclic properties were also measured both in the longitudinal and torsional axes. Finally another series of tests were carried out to study the multiaxial response of the material. Both in-phase (proportional) and out-of-phase (non proportional) loadings were employed, thus providing a complete database for improving current models which describe the multiaxial behaviour of materials.

There are a number of theories available to model biaxial fatigue problems. Among these, the critical plane models try to predict not only the fatigue life but also the orientation of the crack or failure plane. This work attempt to analyse the biaxial tension-torsion problem from a strain-based analysis focused on plane failures predictions. Fatigue life and critical plane predictions are carried out applying multiaxial models proposed by Brown-Miller, Fatemi-Socie and Smith-Watson-Topper. The theoretical results are compared with constant amplitude test data, in-phase and out-of-phase (90˚) in a structural steel. The results suggest that Smith-Watson-Topper model predictions of fracture plane do not match the real fracture plane. The critical fracture plane predictions of the Brown-Miller and Fatemi-Socie models do not match in most cases with experimental observations either, but considering the second plane of the maximum shear strain amplitude, the predictions of the fracture plane and fatigue life improve substantially.

r Fatigue crack closure is strongly influenced by the plastic wake developed because of the action of precedent cycles. It is related to the load sequence and is an additional difficulty for its determination. Finite-element method has been an alternative for the study of crack closure, commonly in constant-amplitude loading. However, such analyses are complex and computationally expensive, the length of the wake simulated being a critical parameter. In this paper, a comprehensive study of the effect of the plastic wake in fatigue crack closure is made. It is structured in two parts: one regarding the correct numerical modelling process and the other focussed on the influence of the wake under moderate variable-amplitude loading. After a brief description of the numerical methodology, results for different wake lengths are reported. The plastic wake is modelled with a new approach in which the final crack tip is fixed. Information regarding potential error is provided. Second, the load sequence effect is discussed. Crack closure is obtained for several load ratios, with wakes developed with different stress ratios. This permits the analysis of the influence of the load sequence over the crack-driving force compared with constant-amplitude loading.

The atmospheric Cherenkov telescope MAGIC for ground-based gamma-ray astronomy is operating since late 2003 on the Canary island of La Palma. Its 17 m diameter mirror is composed of 964 square all-aluminum mirrors of ∼ 0.5 m side, making up a parabola of 236 m 2 area. Each mirror is composed of a sandwich of two thin aluminum layers interspaced by a honeycomb structure that ensures rigidity, high temperature conductivity and low weight. The surface of each raw blank is diamond milled to provide high reflectivity and a slightly different focal length to fit the overall parabolic shape of the reflector. We report about the stability and performance of the surface exposed to the atmosphere for over 3 years. For the construction of the clone of the first telescope, dubbed MAGIC II, major improvements of the design and performance of the reflective surface were required. Given the good experience with aluminum mirrors, a similar assembly was tested, but the area was increased to 1 m 2 , which allowed to skip the inter-alignment of four mirrors within a panel and to reduce substantially the weight. The increased rigidity of the mirror unit resulted in an improved focussing quality. In addition, a second class of mirrors will be installed in the outermost part of the reflector, namely glass mirrors obtained by cold-slumping replica technique. Details on the construction of both type MAGIC II new mirrors and the 17 m reflector will be presented.

The fatigue life of metallic materials is strongly influenced by crack closure effects. Finite element (FE) methods allow the study of crack closure with great detail and can provide valuable information about phenomena occurring in the bulk of the material. In this work the distribution of stresses through the thickness of a cracked specimen has been studied using 3D FE simulations. It was found that the transition between the interior of the specimen (plane strain) and the surface (plane stress) differs from that predicted by 2D plane stress models. In addition, an attempt is presented to experimentally validate the results at the surface level. For this purpose full-field image correlation technique was utilized. This allowed direct comparison between the displacement field predicted by the numerical simulations and the experimental results measured by digital image correlation.

Plasticity-induced crack closure is an observed phenomenon during fatigue crack growth. However, accurate determination of fatigue crack closure has been a complex task for years. It has been approached by means of experimental and numerical methods. The finite element method (FEM) has been the principal numerical tool employed. In this paper the results of a broad study of fatigue crack closure in plane stress and plane strain by means of FEM are presented. The effect of three principal factors has been analysed in depth, the maximum load, the crack length and the stress ratio. It has been found that the results are independent of maximum load and the crack length, and there exists a direct influence of the stress ratio. This relation has been numerically correlated and compared with experimental results. Differences have also been established between opening and closure points and between the different criteria employed to compute crack closure.

We report on very high energy gamma-observations with the MAGIC Telescope of the pulsar PSR B1951+32 and its associated nebula, CTB 80. Our data constrain the cutoff energy of the pulsar to be less than 32 GeV, assuming the pulsed gamma-ray emission to be exponentially cut off. The upper limit on the flux of pulsed gamma-ray emission above 75 GeV is 4.3*10^-11 photons cm^-2 sec^-1, and the upper limit on the flux of steady emission above 140 GeV is 1.5*10^-11 photons cm^-2 sec^-1. We discuss our results in the framework of recent model predictions and other studies.

This paper focuses on the study of the plastic zone in fatigue crack closure based on the results obtained by means of 3D Finite Element Analysis (FEA). These results show the crack behavior through the thickness. The plastic zone is visualized and quantified. It does not correspond to the classical shape. The plastic zone in the interior surface is similar to those obtained in 2D plane strain conditions and a reduced effect of closure is observed. However, close to the external surface, 2D plane stress results are not reproduced, the plastic zone size is smaller and an important change is observed. This transition is developed in a thin external portion of the specimen and it can only be captured if a fine mesh of the thickness is done.

The applications of mullite based ceramics cause that along their useful life they are subjected to important mechanical and thermal solicitations. This is the reason why it is very interesting to establish procedures that allow to predict their behaviour in function of the initial condition. For this work the ceramic used was a 2Al2O3:1SiO2 mullite material obtained using a method of chemical processing of kaolinite and aluminium metal wastes. Mullite based ceramic materials with different sintering grade were obtained using various firing conditions. Strength of ceramics was measured in four point bending test. Weibull distribution function was used to characterize statistically the variation of the mechanical strength. A surface flaws mapping was established by scanning electron microscopy and distributions of pore-size, orientation and shape factor were obtained for each sample. A study of strength is done using Weibull´s theory and the surface flaws mapping.Las aplicaciones propias de los materiales cerámicos de mullita propician que a lo largo de su vida útil se vean sometidos a importantes solicitaciones mecánicas y térmicas. Es por ello interesante establecer procedimientos que permitan predecir su comportamiento en función del estado inicial. Para este estudio se utilizó una mullita del tipo 2Al2O3:1SiO2 obtenida usando un método de procesado químico a partir de caolinita y residuos de aluminio. Se fabricaron varias series de dicho material para ser sometidas a distintos ciclos de cocción que proporcionasen materiales con diferente grado de sinterización. La resistencia a la fractura de dichas cerámicas se determinó mediante ensayo a flexión en cuatro puntos. La función de distribución de Weibull se usó para caracterizar estadísticamente la variación de la resistencia mecánica. A continuación se procedió al estudio de su superficie por microscopía electrónica de barrido, para establecer un mapeado de defectos que posibilitase la obtención de la distribución de tamaño de poro, orientación y factor de forma de cada probeta. Finalmente se llevó a cabo un estudio de resistencia usando la teoría de Weibull y el mapeado de defectos superficiales.

This article analyses fatigue crack growth under random loading using experimental results taken from literature on the subject and from growth simulations carried out using the Strip Yield Model. The capacity of the Strip Yield Model in representing the retardation effects produced during the growth process is analysed. The effect of different statistical parameters of the random load process and of the representative histories of the same on the crack growth life and on the variability of the results obtained in different representative tests of the same real load process is likewise studied. Some of the parameters considered are the bandwidth of the random loading process, the number of cycles of the load histories employed and the effect of truncating the histories. It can be seen that under certain conditions, the numerical model employed can quite closely predict some of the behaviours of the crack growth. Likewise, it can also be seen that the bandwidth has a significant effect on the fatigue life, and that the length of the histories employed in tests or simulations has a great effect on the variability of the obtained results. It can also be seen that, although the elimination of the overloads tends to give rise to shorter lives, in certain cases, from a statistical point of view, it may be the origin of non-conservative predictions.

Measuring opening or closure stress is a complex process that influences the low accuracy of obtained data. Finite element models have been one of the available ways to deal with this problem. The difficulty of modelling the whole process of crack growth (due to the great number of cycles implied) as the great complexity of the phenomenon itself (with a high plastic strain concentrated in a small area, with elevated stress gradients) has made the results to be quite varied, being influenced by a great number of modelling parameters. Of those parameters, the minimum size of the element used to mesh the area around the crack tip vicinity presents a great influence on the results. In this work, a detailed analysis of the influence of this parameter in the results in terms of closure or opening stress is presented. The effect that different meshing criteria can have on the result is complex and it has been necessary to reduce the element size around the crack tip to a size that had not been reached before. Procedures and modelling criteria stricter than the ones shown in the current bibliography are proposed. A methodology for the correct interpretation of the results is also established.

Mullite ceramics have been prepared from an aqueous suspension of kaolinite (raw or ground) and aluminum hydroxide. The precursor was coprecipitated in the mixture using hexamethylenediamine (HMDA) or ammonium hydroxide, and a solution of aluminum chloride from acid dissolution of wastes of aluminum metal. The precursor and the resulting materials were characterized and studied by X-ray diffraction, thermal methods, and mechanical strength and porosity measurements. The feasibility of the proposed chemical processing route for mullite preparation was demonstrated, in particular using HMDA as a precipitating agent for aluminum hydroxide instead of ammonium hydroxide, which adversely affects the system reactivity. The use of HMDA, as compared with ammonium hydroxide, and ground kaolinite produces single-phase mullite and enhances the flexural strength (maximum of 49 MPa) of the resultant ceramic porous bodies (porosity ca. 52–45 vol%) fired at 1550–1600°C for 30 min.