345 research outputs found

    Corrosion induced cracking modelled by a coupled transport-structural approach

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    Transport of corrosion products into pores and cracks in concrete must be considered when predicting corrosion induced cracking in reinforced concrete structures, since this transport significantly delays the onset of cracking and spalling by reducing the amount of radial displacement imposed on the concrete at the steel/concrete interface. We aim to model this process by means of a coupled transport-structural approach, whereby the transport of corrosion products is determined by a pressure gradient generated by the confined volumetric expansion due to the transformation of steel into corrosion products. This pressure driven transport was studied by using both an axisymmetric thick-walled cylinder model and a network approach. The network approach was then applied to corrosion induced cracking experiments reported in the literature

    McKenzie, Ellen

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    This interview features Ellen McKenzie, an African-American lesbian woman living in Portland, Maine. Having lived in Portland for almost her entire life, Ellen can provide insight on growing up in one of the only black families in her community, the intersections between race and sexuality, co-parenting children from a spouse’s previous marriage and generally navigating the world and her career as a queer woman of color. Throughout this interview, we hear a lot about her childhood and her family’s history as civil rights activists in Maine, her relationship with her spouse and and co-parenting their children with both her spouse, and the children’s father and stepmother. This insight into LGBTQ blended families is insightful and interesting. We also hear about the loss of one of their sons to suicide. Lastly, she provides insight into her career as a social worker, and how to navigate the workplace as a woman of color and how she handles racism and discrimination. Citation Please cite as: Querying the Past: LGBTQ Maine Oral History Project Collection, Lesbian, Gay, Bisexual, Transgender, and Queer+ Collection, Jean Byers Sampson Center for Diversity in Maine, University of Southern Maine Libraries. For more information about the Querying the Past: Maine LGBTQ Oral History Project, please contact Dr. Wendy Chapkis.https://digitalcommons.usm.maine.edu/querying_ohproject/1033/thumbnail.jp

    On a 2D hydro-mechanical lattice approach for modelling hydraulic fracture

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    A 2D lattice approach to describe hydraulic fracturing is presented. The interaction of fluid pressure and mechanical response is described by Biot's theory. The lattice model is applied to the analysis of a thick-walled cylinder, for which an analytical solution for the elastic response is derived. The numerical results obtained with the lattice model agree well with the analytical solution. Furthermore, the coupled lattice approach is applied to the fracture analysis of the thick-walled cylinder. It is shown that the proposed lattice approach provides results that are independent of the mesh size. Moreover, a strong geometrical size effect on nominal strength is observed which lies between analytically derived lower and upper bounds. This size effect decreases with increasing Biot's coefficient

    Differing calcification processes in cultured vascular smooth muscle cells and osteoblasts

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    © 2019 Published by Elsevier Inc.Arterial medial calcification (AMC) is the deposition of calcium phosphate mineral, often as hydroxyapatite, inthe medial layer of the arteries. AMC shares some similarities to skeletal mineralisation and has been associatedwith the transdifferentiation of vascular smooth muscle cells (VSMCs) towards an osteoblast-like phenotype. Thisstudy used primary mouse VSMCs and calvarial osteoblasts to directly compare the established and widely usedin vitromodels of AMC and bone formation. Significant differences were identified between osteoblasts andcalcifying VSMCs. First, osteoblasts formed large mineralised bone nodules that were associated with widespreaddeposition of an extracellular collagenous matrix. In contrast, VSMCs formed small discrete regions of calcifi-cation that were not associated with collagen deposition and did not resemble bone. Second, calcifying VSMCsdisplayed a progressive reduction in cell viability over time (≤7-fold), with a 50% increase in apoptosis,whereas osteoblast and control VSMCs viability remained unchanged. Third, osteoblasts expressed high levels ofalkaline phosphatase (TNAP) activity and TNAP inhibition reduced bone formation by to 90%. TNAP activity incalcifying VSMCs was∼100-fold lower than that of bone-forming osteoblasts and cultures treated withβ-gly-cerophosphate, a TNAP substrate, did not calcify. Furthermore, TNAP inhibition had no effect on VSMC calci-fication. Although, VSMC calcification was associated with increased mRNA expression of osteoblast-relatedgenes (e.g. Runx2, osterix, osteocalcin, osteopontin), the relative expression of these genes was up to 40-foldlower in calcifying VSMCs versus bone-forming osteoblasts. In summary, calcifying VSMCsin vitrodisplay somelimited osteoblast-like characteristics but also differ in several key respects: 1) their inability to form collagen-containing bone; 2) their lack of reliance on TNAP to promote mineral deposition; and, 3) the deleterious effectof calcification on their viability.Peer reviewedFinal Published versio

    Prenatal Androgen Effects as a Proximate Mechanism Underpinning Variation in Social Behavior Among Female Nonhuman Primates

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    While the role of ecological factors in shaping primate social systems has been a central focus for decades, less attention has been given to phylogenetic relationships and the potential role of underlying proximate mechanisms. This study aimed to investigate the relationship between one such proximate mechanism, prenatal androgen effects (PAEs), and aspects of social behavior in female nonhuman primates using the 2D:4D ratio as a proxy for PAEs and phylogenetically controlled methods. In general, female 2D:4D ratios were highest in monogamous species (low inferred PAEs) and lowest in polygynandrous and polygynous species (high inferred PAEs). 2D:4D ratios also varied with the form of polygyny/polygynandry, potentially with regard to the need for competitive over cooperative behaviors and the intensity of female reproductive competition. Species characterized by female dominance had lower 2D:4D ratios than species characterized by male dominance or codominance. There were no significant relationships between 2D:4D ratio and either degree of frugivory or group size. Relationships between 2D:4D ratios and the directional consistency index and 2D:4D ratios and rates of female–female agonism were also nonsignificant although sample sizes for both of these variables were small. Female social relationships are a manifestation of complex competitive and cooperative behaviors and the results suggest thatPAEs may act as a proximate mechanism underlying the expression of certain aspects of behavior in female primates in ways that are adaptive to their social system

    Direct activation of NADPH oxidase 2 by 2-deoxyribose-1-phosphate triggers nuclear factor kappa B-dependent angiogenesis.

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    AbstractAims: Deoxyribose-1-phosphate (dRP) is a proangiogenic paracrine stimulus released by cancer cells, platelets, and macrophages and acting on endothelial cells. The objective of this study was to clarify how dRP stimulates angiogenic responses in human endothelial cells.Results: Live cell imaging, electron paramagnetic resonance, pull-down of dRP-interacting proteins, followed by immunoblotting, gene silencing of different NADPH oxidases (NOXs), and their regulatory cosubunits by small interfering RNA (siRNA) transfection, and experiments with inhibitors of the sugar transporter glucose transporter 1 (GLUT1) were utilized to demonstrate that dRP acts intracellularly by directly activating the endothelial NOX2 complex, but not NOX4. Increased reactive oxygen species generation in response to NOX2 activity leads to redox-dependent activation of the transcription factor nuclear factor kappa B (NF-κB), which, in turn, induces vascular endothelial growth factor receptor 2 (VEGFR2) upregulation. Using endothelial tube formation assays, gene silencing by siRNA, and antibody-based receptor inhibition, we demonstrate that the activation of NF-κB and VEGFR2 is necessary for the angiogenic responses elicited by dRP. The upregulation of VEGFR2 and NOX2-dependent stimulation of angiogenesis by dRP were confirmed in excisional wound and Matrigel plug vascularization assays in vivo using NOX2−/− mice.Innovation: For the first time, we demonstrate that dRP acts intracellularly and stimulates superoxide anion generation by direct binding and activation of the NOX2 enzymatic complex.Conclusions: This study describes a novel molecular mechanism underlying the proangiogenic activity of dRP, which involves the sequential activation of NOX2 and NF-κB and upregulation of VEGFR2. Antioxid. Redox Signal. 28, 110–130

    Inhibition of vascular smooth muscle cell calcification by ATP analogues

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    Arterial medial calcification (AMC) has been associated with phenotypic changes in vascular smooth muscle cells (VSMCs) that reportedly makes them more osteoblast-like. Previous work has shown that ATP/UTP can inhibit AMC directly via P2 receptors and indirectly by NPP1-mediated hydrolysis to produce the mineralisation inhibitor, pyrophosphate (PPi). This study investigated the role of P2X receptors in the inhibitory effects of extracellular nucleotides on VSMC calcification. We found that Bz-ATP, α,β-meATP and β,γ-meATP, inhibited calcification by up to 100%. Culture in a high phosphate medium (2mM) was associated with increased VSMC death and apoptosis; treatment with Bz-ATP, α,β-meATP and β,γ-meATP reduced apoptosis to levels seen in non-calcifying cells. Calcification was also associated with alterations in the protein levels of VSMC (e.g. SM22α, SMA) and osteoblast-associated (e.g. Runx2, osteopontin) markers; Bz-ATP, α,β-meATP and β,γ-meATP attenuated these changes in protein expression. Long-term culture with Bz-ATP, α,β-meATP and β,γ-meATP resulted in lower extracellular ATP levels and an increased rate of ATP breakdown. P2X receptor antagonists failed to prevent the inhibitory effects of these analogues suggesting that they act via P2X receptor-independent mechanisms. In agreement, the breakdown products of α,β-meATP and β,γ-meATP (α,β-meADP and methylene diphosphonate, respectively) also dose dependently inhibited VSMC calcification. Furthermore, the actions of Bz-ATP, α,β-meATP and β,γ-meATP were unchanged in VSMCs isolated from NPP1 knockout mice, suggesting that the functional effects of these compounds do not involve NPP1-mediated generation of PPi. Together, these results indicate that the inhibitory effects of ATP analogues on VSMC calcification and apoptosis in vitro may be mediated, at least in part, by mechanisms that are independent of purinergic signalling and PPi

    Modulation of endoglin expression in islets of langerhans by VEGF reveals a novel regulator of islet endothelial cell function

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    Background: endoglin/CD105 is an auxiliary receptor for transforming growth factor-? with established roles in vascular remodelling. It has recently been shown that heterozygous endoglin deficiency in mice decreases insulin secretion in an animal model of obesity, highlighting a potential role for endoglin in the regulation of islet function. We have previously identified two different populations of endoglin expressing cells in human and mouse islets which are: (i) endothelial cells (ECs) and (ii) islet mesenchymal stromal cells. The contribution of islet EC endoglin expression to islet development and sensitivity to VEGF is unknown and is the focus of this study.Results: in vitro culture of mouse islets with VEGF164 for 48 h increased endoglin mRNA levels above untreated controls but VEGF did not modulate VEGFR2, CD31 or CD34 mRNA expression or islet viability. Removal of EC-endoglin expression in vivo reduced islet EC area but had no apparent effect on islet size or architecture.Conclusion: EC-specific endoglin expression in islets is sensitive to VEGF and plays partial roles in driving islet vascular development, however such regulation appears to be distinct to mechanisms required to modulate islet viability and siz

    A thin layer angiogenesis assay: a modified basement matrix assay for assessment of endothelial cell differentiation

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    BACKGROUND: Basement matrices such as Matrigel™ and Geltrex™ are used in a variety of cell culture assays of anchorage-dependent differentiation including endothelial cell tube formation assays. The volumes of matrix recommended for these assays (approximately 150 μl/cm(2)) are costly, limit working distances for microscopy, and require cell detachment for subsequent molecular analysis. Here we describe the development and validation of a thin-layer angiogenesis (TLA) assay for assessing the angiogenic potential of endothelial cells that overcomes these limitations. RESULTS: Geltrex™ basement matrix at 5 μl/cm(2) in 24-well (10 μl) or 96-well (2 μl) plates supports endothelial cell differentiation into tube-like structures in a comparable manner to the standard larger volumes of matrix. Since working distances are reduced, high-resolution single cell microscopy, including DIC and confocal imaging, can be used readily. Using MitoTracker dye we now demonstrate, for the first time, live mitochondrial dynamics and visualise the 3-dimensional network of mitochondria present in differentiated endothelial cells. Using a standard commercial total RNA extraction kit (Qiagen) we also show direct RNA extraction and RT-qPCR from differentiated endothelial cells without the need to initially detach cells from their supporting matrix. CONCLUSIONS: We present here a new thin-layer assay (TLA) for measuring the anchorage-dependent differentiation of endothelial cells into tube-like structures which retains all the characteristics of the traditional approach but with the added benefit of a greatly lowered cost and better compatibility with other techniques, including RT-qPCR and high-resolution microscopy. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12860-014-0041-5) contains supplementary material, which is available to authorized users
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