93 research outputs found

    Revealing the mechanical and microstructural performance of multiphase steels during tensile, forming and flanging operations

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    The mechanical performance of Dual Phase (DP) and Complex Phase (CP) steels was investigated by SEM analysis, tensile testing, Forming Limit Curve investigation and flange formability testing. The alloys of interest were Dual Phase (DP) untempered, Dual Phase (DP) tempered and Complex Phase (CP) steels. Phase content analysis showed that the distribution of the ferrite and martensite phases was the same for the two DP alloys, but the grain size and condition (tempered/untempered) for the martensite islands was much different in the two alloys. In the tempered DP steel, the smaller grain size for the martensite and the tempering process resulted in increased elongation, more formability and ability to form a flange (flangeability). In CP steels the soft ferrite phase is replaced by harder bainite, yielding a bainitic-martensitic microstructure. Bainite reduced the total elongation of the alloy during tensile testing, reduced the formability (especially under plane strain conditions) of the alloy but improved the flangeability of the alloy. Under flanging conditions, CP steels deformed to higher strains, at tighter radii with minimum springback. Microstructural inspections at the outer radius of the flanged specimens revealed that in CP steels bainite deforms similarly to martensite, therefore the strain partitioning is smaller in CP steels in comparison to DP steels. Plastic deformation in CP steels upon flanging occurs with the formation of strong slip bands in both martensite and bainite. In contrast, the martensite and ferrite grains in DP steels deform quite differently leading to strong strain localisations. Void nucleation and cracking occurred at the martensite islands or within the soft ferrite phase next to the martensite islands. In CP steels no voids or damage was observed within the matrix. A special case study was done with a thicker and stronger alloy, a Martensitic 1400 steel to reveal the flangeability limits for advanced high strength steels. Neither cracks nor damage were observed visually on the flanged specimens. However SEM observations at the outer radius of the flanged samples revealed significant void growth at inclusion sites and cracks nucleating within the matrix adjacent to the inclusions.Publisher Statement: This is an open access article under the CC BY license (http://creativecommons.org/licenses/BY/4.0/)</p

    Multiscale characterisation of the mechanical properties of austenitic stainless steel joints

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    A multiscale investigation was pursued in order to obtain the strain distribution and evolution during tensile testing both at the macro- and micro-scale for a diffusion bonded 316L stainless steel. The samples were designed for the purpose to demonstrate that the bond line properties were equal or better than the parent material in a sample geometry that was extracted from a larger component. The macroscopic stress-strain curves were coupled to the strain distributions using a camera-based 2D – Digital Image Correlation system. Results showed significant amount of plastic deformation predominantly concentrated in shear bands which were extended over a large region, crossing through the joint area. Yet it was not possible to be certain whether the joint has shown significant plastic deformation. In order to obtain the joints’ mechanical response in more detail, in situ micromechanical testing was conducted in the SEM chamber that allowed areas of 1x1 mm2 and 50x50 mm2 to be investigated. The size of the welded region was rather small to be accurately captured from the camera based DIC system. Therefore a microscale investigation was pursued where the samples were tested within an SEM chamber. Low magnification SEM imaging was utilised in order to cover a viewing area of 1 mm×1 mm while high magnification SEM imaging was employed to provide evidence of the occurrence of plastic deformation within the joint, at an area of just 50 μm×50 μm. The strain evolution over the microstructural level, within the joint and at the base material was obtained. The local strains were highly non-homogeneous through the whole test. Final failure occurred approximately 0.2 mm away from the joint. Large local strains were measured within the joint region, while SEM imaging showed that plastic deformation occurs via the formation of strong slip bands, followed by the activation of additional slip systems upon further plastic deformation which end up in additional slip bands to form on the surface. Plastic deformation occurred by slip and twinning mechanisms. Upon necking, significant out of plane deformations and slip deformation mechanisms were observed which suggested that plastic deformation was also happening at the last stages of damage evolution for the specific alloy. This was also evident from the large difference between the 600 MPa UTS stress value and the low stress values before final failure (which in many cases was below 30 MPa)

    Structural basis of nuclear import of flap endonuclease 1 (FEN1)

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    Flap endonuclease 1 (FEN1) is a member of the nuclease family and is structurally conserved from bacteriophages to humans. This protein is involved in multiple DNA-processing pathways, including Okazaki fragment maturation, stalled replication-fork rescue, telomere maintenance, long-patch base-excision repair and apoptotic DNA fragmentation. FEN1 has three functional motifs that are responsible for its nuclease, PCNA-interaction and nuclear localization activities, respectively. It has been shown that the C-terminal nuclear localization sequence (NLS) facilitates nuclear localization of the enzyme during the S phase of the cell cycle and in response to DNA damage. To determine the structural basis of the recognition of FEN1 by the nuclear import receptor importin alpha, the crystal structure of the complex of importin alpha with a peptide corresponding to the FEN1 NLS was solved. Structural studies confirmed the binding of the FEN1 NLS as a classical bipartite NLS; however, in contrast to the previously proposed (KRKX8KKK367)-K-354 sequence, it is the (354)KRX(10)KKAK(369) sequence that binds to importin alpha. This result explains the incomplete inhibition of localization that was observed on mutating residues (KKK367)-K-365. Acidic and polar residues in the X-10 linker region close to the basic clusters play an important role in binding to importin alpha. These results suggest that the basic residues in the N-terminal basic cluster of bipartite NLSs may play roles that are more critical than those of the many basic residues in the C-terminal basic cluster

    Damage in dual phase steel DP1000 investigated using digital image correlation and microstructure simulation

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    Microstructure failure mechanisms and void nucleation in dual-phase (DP) steels during deformation have been studied using a combination of in situ tensile testing in a scanning electron microscope (SEM), digital image correlation (DIC) and finite element (FE) modelling. SEM images acquired during in situ tests were used to follow the evolution of damage within the microstructure of a DP1000 steel. From these images, strain maps were generated using DIC and used as boundary conditions for a FE model to investigate the stress state of martensite and ferrite before the onset of the martensite phase cracking. Based on the simulation results, a maximum principal stress of about 1700 MPa has been estimated for crack initiation in the martensite of the investigated DP1000 steel. The SEM image observations in combination with the FE analyses provide new insights for the development of physically-based damage models for DP-steels

    Deformation-induced microstructural banding in TRIP steels

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    Microstructure inhomogeneities can strongly influence the mechanical properties of advanced high-strength steels in a detrimental manner. This study of a transformation-induced plasticity (TRIP) steel investigates the effect of pre-existing contiguous grain boundary networks (CGBNs) of hard second-phases and shows how these develop into bands during tensile testing using in situ observations in conjunction with digital image correlation (DIC). The bands form by the lateral contraction of the soft ferrite matrix, which rotates and displaces the CGBNs of second-phases and the individual features within them to become aligned with the loading direction. The more extensive pre-existing CGBNs that were before the deformation already aligned with the loading direction are the most critical microstructural feature for damage initiation and propagation. They induce micro-void formation between the hard second-phases along them, which coalesce and develop into long macroscopic fissures. The hard phases, retained austenite and martensite, were not differentiated as it was found that the individual phases do not play a role in the formation of these bands. It is suggested that minimizing the presence of CGBNs of hard second-phases in the initial microstructure will increase the formability

    Potent Inhibition of HIV-1 Replication by a Tat Mutant

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    Herein we describe a mutant of the two-exon HIV-1 Tat protein, termed Nullbasic, that potently inhibits multiple steps of the HIV-1 replication cycle. Nullbasic was created by replacing the entire arginine-rich basic domain of wild type Tat with glycine/alanine residues. Like similarly mutated one-exon Tat mutants, Nullbasic exhibited transdominant negative effects on Tat-dependent transactivation. However, unlike previously reported mutants, we discovered that Nullbasic also strongly suppressed the expression of unspliced and singly-spliced viral mRNA, an activity likely caused by redistribution and thus functional inhibition of HIV-1 Rev. Furthermore, HIV-1 virion particles produced by cells expressing Nullbasic had severely reduced infectivity, a defect attributable to a reduced ability of the virions to undergo reverse transcription. Combination of these inhibitory effects on transactivation, Rev-dependent mRNA transport and reverse transcription meant that permissive cells constitutively expressing Nullbasic were highly resistant to a spreading infection by HIV-1. Nullbasic and its activities thus provide potential insights into the development of potent antiviral therapeutics that target multiple stages of HIV-1 infection

    Human PAPS Synthase Isoforms Are Dynamically Regulated Enzymes with Access to Nucleus and Cytoplasm

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    In higher eukaryotes, PAPS synthases are the only enzymes producing the essential sulphate-donor 3′-phospho-adenosine-5′-phosphosulphate (PAPS). Recently, PAPS synthases have been associated with several genetic diseases and retroviral infection. To improve our understanding of their pathobiological functions, we analysed the intracellular localisation of the two human PAPS synthases, PAPSS1 and PAPSS2. For both enzymes, we observed pronounced heterogeneity in their subcellular localisation. PAPSS1 was predominantly nuclear, whereas PAPSS2 localised mainly within the cytoplasm. Treatment with the nuclear export inhibitor leptomycin B had little effect on their localisation. However, a mutagenesis screen revealed an Arg-Arg motif at the kinase interface exhibiting export activity. Notably, both isoforms contain a conserved N-terminal basic Lys-Lys-Xaa-Lys motif indispensable for their nuclear localisation. This nuclear localisation signal was more efficient in PAPSS1 than in PAPSS2. The activities of the identified localisation signals were confirmed by microinjection studies. Collectively, we describe unusual localisation signals of both PAPS synthase isoforms, mobile enzymes capable of executing their function in the cytoplasm as well as in the nucleus

    Association of Tat with Promoters of PTEN and PP2A Subunits Is Key to Transcriptional Activation of Apoptotic Pathways in HIV-Infected CD4+ T Cells

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    Apoptosis in HIV-1-infected CD4+ primary T cells is triggered by the alteration of the PI3K and p53 pathways, which converge on the FOXO3a transcriptional activator. Tat alone can cause activation of FOXO3a and of its proapoptotic target genes. To understand how Tat affects this pathway, we carried out ChIP-Chip experiments with Tat. Tat associates with the promoters of PTEN and two PP2A subunit genes, but not with the FOXO3a promoter. PTEN and PP2A encode phosphatases, whose levels and activity are increased when Tat is expressed. They counteract phosphorylation of Akt1 and FOXO3a, and so activate transcriptional activity of FOXO3a. FOXO3a promotes increased transcription of Egr-1, which can further stimulate the transcription of PTEN, thereby reinforcing the pathway that leads to FOXO3a transcriptional activation. RNAi experiments support the role of PTEN and PP2A in the initiation of the Tat-mediated cascade, which is critical to apoptosis. The increased accumulation of PTEN and PP2A subunit mRNAs during Tat expression is more likely to be the result of increased transcription initiation and not relief of promoter-proximal pausing of RNAPII. The Tat-PTEN and -PP2A promoter interactions provide a mechanistic explanation of Tat-mediated apoptosis in CD4+ T cells
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