319 research outputs found
Property Optimization for TWIP Steels – Effect of Pre-deformation Temperature on Fatigue Properties
The current work investigates the impact of pre-deformation temperatures on the microstructure evolution and the subsequent cyclic stress-strain response of high-manganese steel showing twinning-induced plasticity (TWIP) at room temperature (RT). Deformation at low temperatures increases the hardening rate at low to medium degrees of deformation through concurrent martensitic transformation. In contrast, high temperatures promote dislocation slip. Thus, employing pre-treatments at temperatures below and above RT leads to the evolution of considerably different microstructures. Low-cycle fatigue experiments revealed distinct differences for the pre-treated TWIP steels
Expression of nodal signalling components in cycling human endometrium and in endometrial cancer
<p>Abstract</p> <p>Background</p> <p>The human endometrium is unique in its capacity to remodel constantly throughout adult reproductive life. Although the processes of tissue damage and breakdown in the endometrium have been well studied, little is known of how endometrial regeneration is achieved after menstruation. Nodal, a member of the transforming growth factor-beta superfamily, regulates the processes of pattern formation and differentiation that occur during early embryo development.</p> <p>Methods</p> <p>In this study, the expression of Nodal, Cripto (co-receptor) and Lefty A (antagonist) was examined by RT-PCR and immunohistochemistry across the menstrual cycle and in endometrial carcinomas.</p> <p>Results</p> <p>Nodal and Cripto were found to be expressed at high levels in both stromal and epithelial cells during the proliferative phase of the menstrual cycle. Although immunoreactivity for both proteins in surface and glandular epithelium was maintained at relatively steady-state levels across the cycle, their expression was significantly decreased within the stromal compartment by the mid-secretory phase. Lefty expression, as has previously been reported, was primarily restricted to glandular epithelium and surrounding stroma during the late secretory and menstrual phases. In line with recent studies that have shown that Nodal pathway activity is upregulated in many human cancers, we found that Nodal and Cripto immunoreactivity increased dramatically in the transition from histologic Grade 1 to histologic Grades 2 and 3 endometrial carcinomas. Strikingly, Lefty expression was low or absent in all cancer tissues.</p> <p>Conclusion</p> <p>The expression of Nodal in normal and malignant endometrial cells that lack Lefty strongly supports an important role for this embryonic morphogen in the tissue remodelling events that occur across the menstrual cycle and in tumourogenesis.</p
EPHA3 (EPH receptor A3)
Review on EPHA3 (EPH receptor A3), with data on DNA, on the protein encoded, and where the gene is implicated
Separation of VUV/UV photons and reactive particles in the effluent of a He/O2 atmospheric pressure plasma jet
Cold atmospheric pressure plasmas can be used for treatment of living tissues
or for inactivation of bacteria or biological macromolecules. The treatment is
usually characterized by a combined effect of UV and VUV radiation, reactive
species, and ions. This combination is usually beneficial for the effectiveness
of the treatment but it makes the study of fundamental interaction mechanisms
very difficult. Here we report on an effective separation of VUV/UV photons and
heavy reactive species in the effluent of a micro scale atmospheric pressure
plasma jet (-APPJ). The separation is realized by an additional flow of
helium gas under well-defined flow conditions, which deflects heavy particles
in the effluent without affecting the VUV and UV photons. Both components of
the effluent, the photons and the reactive species, can be used separately or
in combination for sample treatment. The results of treatment of a model plasma
polymer film and vegetative Bacillus subtilis and Escherichia coli cells are
shown and discussed. A simple model of the He gas flow and reaction kinetics of
oxygen atoms in the gas phase and at the surface is used to provide a better
understanding of the processes in the plasma effluent. The new jet
modification, called X-Jet for its appearance, will simplify the investigation
of interaction mechanisms of atmospheric pressure plasmas with biological
samples.Comment: 10 pages, 7 figures, submitted to Journal of Physics D: Applied
Physic
Three distinct molecular surfaces in ephrin-A5 are essential for a functional interaction with EphA3
Eph receptor tyrosine kinases (Ephs) function as molecular relays that interact with cell surface-bound ephrin ligands to direct the position of migrating cells. Structural studies revealed that, through two distinct contact surfaces on opposite sites of each protein, Eph and ephrin binding domains assemble into symmetric, circular heterotetramers. However, Eph signal initiation requires the assembly of higher order oligomers, suggesting additional points of contact. By screening a random library of EphA3 binding-compromised ephrin-A5 mutants, we have now determined ephrin-A5 residues that are essential for the assembly of high affinity EphA3 signaling complexes. In addition to the two interfaces predicted from the crystal structure of the homologous EphB2 center dot ephrin-B2 complex, we identified a cluster of 10 residues on the ephrin-A5 E alpha-helix, the E-F loop, the underlying H beta-strand, as well as the nearby B - C loop, which define a distinct third surface required for oligomerization and activation of EphA3 signaling. Together with a corresponding third surface region identified recently outside of the minimal ephrin binding domain of EphA3, our findings provide experimental evidence for the essential contribution of three distinct protein-interaction interfaces to assemble functional EphA3 signaling complexes
Protein dynamics and conformational selection in bidirectional signal transduction
Protein conformational dynamics simultaneously allow promiscuity and specificity in binding. The multiple conformations of the free EphA4 ligand-binding domain observed in two new EphA4 crystal structures provide a unique insight into the conformational dynamics of EphA4 and its signaling pathways. The heterogeneous ensemble and loop dynamics explain how the EphA4 receptor is able to bind multiple A- and B-ephrin ligands and small molecules via conformational selection, which helps to fine-tune cellular signal response in both receptor and ligand cells
Proapoptotic BH3-only proteins trigger membrane integration of prosurvival Bcl-w and neutralize its activity
Prosurvival Bcl-2–like proteins, like Bcl-w, are thought to function on organelles such as the mitochondrion and to be targeted to them by their hydrophobic COOH-terminal domain. We unexpectedly found, however, that the membrane association of Bcl-w was enhanced during apoptosis. In healthy cells, Bcl-w was loosely attached to the mitochondrial membrane, but it was converted into an integral membrane protein by cytotoxic signals that induce binding of BH3-only proteins, such as Bim, or by the addition of BH3 peptides to lysates. As the structure of Bcl-w has revealed that its COOH-terminal domain occupies the hydrophobic groove where BH3 ligands bind, displacement of that domain by a BH3 ligand would displace the hydrophobic COOH-terminal residues, allowing their insertion into the membrane. To determine whether BH3 ligation is sufficient to induce the enhanced membrane affinity, or to render Bcl-w proapoptotic, we mimicked their complex by tethering the Bim BH3 domain to the NH2 terminus of Bcl-w. The chimera indeed bound avidly to membranes, in a fashion requiring the COOH-terminal domain, but neither promoted nor inhibited apoptosis. These results suggest that ligation of a proapoptotic BH3-only protein alters the conformation of Bcl-w, enhances membrane association, and neutralizes its survival function
Effects of Benzopyrene-7,8-Diol-9,10-Epoxide (BPDE) In Vitro and of Maternal Smoking In Vivo on Micronuclei Frequencies in Fetal Cord Blood
Up to 20% of pregnant women smoke and there is indirect evidence that certain
tobacco-specific metabolites can cross the placental barrier and are genotoxic to
the fetus. The presence of micronuclei results from chromosome damage and
reflects the degree of underlying genetic instability. Fetal blood was obtained
from the cord blood of 143 newborns (102 from nonsmoking mothers and 41 from
mothers smoking >10 cigarettes/d during pregnancy). The micronucleus assay was
performed following the guidelines established by the Human MicroNucleus project
with modifications. To test the micronucleus assay, we evaluated the effect of a
range of benzopyrene-7,8-diol-9,10-epoxide concentrations (from 3.125 nM to 4
microM) on cord blood from nonsmoking mothers. This validation showed that the
number of micronuclei and apoptotic cells increased with
benzopyrene-7,8-diol-9,10-epoxide dose (p < 0.0001 and p = 0.001, respectively);
the minimal detectable effect was induced by 12.5 nM
benzopyrene-7,8-diol-9,10-epoxide. In our sample, the number of MN was
significantly higher in the 41 cord blood samples from mothers who smoked during
pregnancy [smokers: 4 (1; 10.5); nonsmokers: 3 (0; 8); p = 0.016]. Therefore, the
data reported herein support the hypothesis that tobacco compounds are able to
induce chromosomal losses and breaks that are detectable as an increased number
of micronuclei
Ephrin-A5 induces rounding, blebbing and deadhesion of EphA3-expressing 293T and melanoma cells by CrkII and Rho-mediated signalling
Eph receptor tyrosine kinases and ephrins regulate morphogenesis in the developing embryo where they effect adhesion and motility of interacting cells. Although scarcely expressed in adult tissues, Eph receptors and ephrins are overexpressed in a range of tumours. In malignant melanoma, increased Eph and ephrin expression levels correlate with metastatic progression. We have examined cellular and biochemical responses of EphA3-expressing melanoma cell lines and human epithelial kidney 293T cells to stimulation with polymeric ephrin-A5 in solution and with surfaces of defined ephrin-A5 densities. Within minutes, rapid reorganisation of the actin and myosin cytoskeleton occurs through activation of RhoA, leading to the retraction of cellular protrusions, membrane blebbing and detachment, but not apoptosis. These responses are inhibited by monomeric ephrin-A5, showing that receptor clustering is required for this EphA3 response. Furthermore, the adapter CrkII, which associates with tyrosine-phosphorylated EphA3 in vitro, is recruited in vivo to ephrin-A5-stimulated EphA3. Expression of an SH3-domain mutated CrkII ablates cell rounding, blebbing and detachment. Our results suggest that recruitment of CrkII and activation of Rho signalling are responsible for EphA3-mediated cell rounding, blebbing and de-adhesion, and that ephrin-A5-mediated receptor clustering and EphA3 tyrosine kinase activity are essential for this response
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