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At least 109 records · Page 6

The regulation of sclerostin by cathepsin K in periodontal ligament cells

Highlights: • Sclerostin is degraded by cathepsin K in vitro. • Cathepsin K degradation of sclerostin is affected by hypoxia. Sclerostin is a clinically important protein with key functions in the musculoskeletal system playing a key role in bone formation and remodelling. Whilst a wide range of mechanisms have been identified which regulate sclerostin expression, little is known about the degradation of the protein. The aim of this study was to identify enzymes capable of degrading sclerostin in peridontal ligament (PDL) fibroblasts cells in vitro and to investigate the functionality of these enzymes. We have demonstrated that cathepsin K cleaves sclerostin in vitro in PDL. We have shown that cathepsin K and sclerostin are co-localised in PDL cells and, using cathepsin K knockdown experiments, we have shown that cathepsin K actively controls sclerostin levels in these cells, through a lysosomal mechanism that is affected by hypoxia. These results are the first description of the degradative control of sclerostin in musculoskeletally derived cells in vitro and suggest that degradation of the protein may well play an important role in the control of bone formation and remodelling.

60 APPLIED LIFE SCIENCES↗

miRNA-mediated apoptosis activation through TMEM 48 inhibition in A549 cell line

Highlights: • miR421 is able to inhibit TMEM48. • The suppression of TMEM48 triggers apoptosis. • TMEM48 plays vital roles in the stability of mitochondrial membrane. Lung has critic function in gas exchange, supplying oxygen to all cells. Rapid metastasis and the high rate of mortality characterises lung cancer. There are two types of this disease, small cell and non-small cell, which differs from each other according to histopathologic features. To date, many therapeutic approaches have been developed to destroy this deadly type of cancer, which one of them is mRNA targeted therapies through miRNA. miRNAs are 19–25 base paired molecules be able to suppress and destruct mRNA and found to be involved in development and progression of lung cancer. Transmembrane Protein 48 (TMEM48) is localised on nuclear pore complex and plays critic roles in nuclear traffic. Known that TMEM48 gene overexpressed in non-small lung cancer cells. Growing TMEM48 suppressed therapeutic studies indicated that decreased TMEM48 level might reveal a therapeutic effect for non-small cell lung cancers. TMEM48 studies based on the same strategy of gene-silencing, however, to our knowledge, any report has been published evaluates TMEM48's regulation by miRNAs. We aimed to clarify if miR-421 might be therapeutic player for non-small cancer cell lines (A549), hereby we suppressed TMEM48 by miR-421 and performed advanced molecular tests. Consequently, we recorded that while miR-421 is significantly suppressing TMEM48 expression; it increased apoptotic and tumor suppressor players CASPASE 3, PTEN and TP53 in A549 line, which is consistent with Annexin V – PI results: 30,6% of A549 observed to be apoptotic - 68,5% of A549 was in GO/G1. Our study indicated that miR-421 can suppress TMEM48 so that leads the cells to apoptosis. But it is not entirely clear how miR-421 triggers apoptosis and whether it interacts with the other cellular death pathways in A549.

60 APPLIED LIFE SCIENCES↗

Molecular basis for the lack of auto-inhibition of Plasmodium falciparum importin α

Highlights: • P. falciparum contains a single gene for importin α which lacks auto-inhibition. • Mutation of a single motif in P. falciparum importin α restores auto-inhibition. • Auto-inhibition is not crucial for viability of the parasite, unlike in yeast. • The parasite may employ unique strategies for cargo binding and release. • Lack of auto-inhibition may confer advantages to parasites for faster growth. Importin α is nuclear transport receptor that recognises nuclear localisation sequences (NLS). The protein has two domains: armadillo (ARM) repeats containing NLS-binding sites and the importin β-binding (IBB) domain. The IBB domain mimics an NLS and can bind to the ARM repeats, preventing NLS binding. This phenomenon, called auto-inhibition, is a key regulatory feature for binding and release of NLS-containing cargo by importin α and mutants that lack auto-inhibition show impaired viability in Saccharomyces cerevisiae. The genome of the human malaria parasite, Plasmodium falciparum, contains a single gene for importin α and here we show that the native protein expressed by this gene lacks auto-inhibition, suggesting that P. falciparum parasites possess unconventional mechanisms for regulation of cargo binding and release. Mutation of a single SKR motif (conserved in Plasmodium species) to KRR in P. falciparum importin α restores auto-inhibition. This is the first report of a single-celled eukaryote that has evolved with a single importin α isoform lacking auto-inhibition.

60 APPLIED LIFE SCIENCES↗

Fracturing process of micro-concrete under uniaxial and triaxial compression: Insights from in-situ X-ray mechanical tests

This paper presents an experimental study of concrete at meso-scale (aggregates, macro-pores and mortar matrix) in order to get a better understanding of the local failure mechanisms known to drive the macroscopic mechanical behaviour of the material. The main originality comes from conducting in-situ X-ray mechanical tests on micro-concrete samples of realistic composition (including cement, sand, aggregates and water), under uniaxial compression and, for the first time, under triaxial compression at 5 MPa, 10 MPa and 15 MPa confining pressures. A timeseries analysis of the set of 3D images coming from each test allows for the measurement of the 3D kinematic fields (displacement and strain fields) throughout the experiments. The different failure patterns observed for each loading path are discussed, along with a quantification of the 3D fracturing processes at the scale of the largest heterogeneities (aggregates and macro-pores). With an increasing level of confinement, the transition from brittle to ductile response is observed, as well as an increase of the strength of the material. The pronounced impact of the meso-scale heterogeneities of concrete on their local failure mechanisms is highlighted. It is shown that strain localisation mainly originates between aggregates and mortar matrix, with the shape and location of the largest aggregates and macro-pores essentially driving the propagation of the cracking network.

36 MATERIALS SCIENCE↗

Resilience framework and metrics for energy master planning of communities

Changes in the nature, intensity, and frequency of climate-related extreme events have imposed a higher risk of failure on energy systems, especially those at the community level. Furthermore, the evolving energy demand patterns and transition towards renewable and localised energy supply can affect energy system resilience. How can an energy system be planned and reconfigured to address these challenges without compromising the system's resilience against chronic stresses and extreme events? Unlike energy system reliability, resilience is neither a common nor an explicit consideration in energy master planning at the community level. In addition, there is no universally agreed-upon method or metrics for measuring or estimating resilience and defining mitigation strategies. This paper introduces a multi-layered energy resilience framework and set of metrics for energy master planning of communities, including the new generation of district energy systems. The potential system disturbances and their short and long-term impacts on various components of the energy system are discussed for commonly expected and extreme events. Three layers of energy resilience are discussed: engineering-designed resilience, operational resilience, and community-societal resilience. A starting set of energy resilience metrics to support engineering design and energy master planning for communities is identified. Implications for future research and practice are noted.

24 POWER TRANSMISSION AND DISTRIBUTION↗

Towards data-driven constitutive modelling for granular materials via micromechanics-informed deep learning

The analytical description of path-dependent elastic-plastic responses of a granular system is highly complicated because of continuously evolving microstructures and strain localisation within the system undergoing deformation. This study offers an alternative to the current analytical paradigm by developing micromechanics-informed machine-learning based constitutive modelling approaches for granular materials. A set of critical variables associated with the constitutive behaviour of granular materials are identified through an incremental stress-strain relationship analysis. Depending on the strategy to exploit the priori micromechanical knowledge, three different training strategies are explored. The first model uses only the measurable external variables to make stress predictions; the second model utilises a directed graph to link all the external strain sequences and internal microstructural evolution variables into a single prediction model comprised of a series of sub-mappings, and the third model explicitly integrates the physically important non-temporal properties with external strain paths into training through an enhanced Gated Recurrent Unit (GRU). These three models show satisfactory agreement with unseen test specimens based on multi-directional loading cases. The basic features and potential applications of each model are explained. Lastly, the key factors for constitutive training and limitations of the current work are also discussed in detail.

36 MATERIALS SCIENCE↗

Characterization of high-quality kerfless epitaxial silicon for solar cells: Defect sources and impact on minority-carrier lifetime

We investigate the types and origins of structural defects in thin (<100 μm) kerfless epitaxial single crystal silicon grown on top of reorganized porous silicon layers. Although the structural defect density is low (has average defect density < 10 4 cm -2 ), localized areas with a defect density > 10 5 cm -2 are observed. Cross-sectional and systematic plan-view defect etching and microscopy reveals that the majority of stacking faults and dislocations originate at the interface between the porous silicon layer and the epitaxial wafer. Localised dislocation clusters are observed in regions of collapsed/deformed porous silicon and at decorated stacking faults. In localized regions of high extended defect density, increased minority-carrier recombination activity is observed. Evidence for impurity segregation to the extended defects (internal gettering), which is known to exacerbate carrier recombination is demonstrated. In conclusion, the impact of the defects on material performance and substrate re-use is also discussed.

14 SOLAR ENERGY↗

Structural, optical, and ferromagnetic characterization of Sm-doped LaOCl nanocrystalline synthesized by solvothermal route: Significant effect of hydrogen post treatment

Pure and Sm-doped lanthanum oxychloride (LaOCl) nanomaterials were synthesized by solvothermal route followed by a subsequent heat treatment process. The objective of the present work is to study and develop conditions required to create stable room-temperature ferromagnetic (RT-FM) properties in LaOCl. To achieve that aim, magnetic samarium Sm{sup 3+} ions were used as dopant sources for stable FM properties. Systematic structural, optical, and magnetic properties of undoped and Sm-doped LaOCl samples were investigated as function of post-annealing conditions (temperature and atmosphere). The optical absorption properties were studied by diffuse reflection spectroscopy (DRS). The magnetic measurements reveal that Sm-doped LaOCl nanopowders have partial RT-FM properties due to the doped ions. The variations of magnetic properties with pre-annealing temperature were investigated. Furthermore, the electronic medium of host LaOCl crystalline lattice, which carries the spin-spin (S.S) exchange interaction between localised dopant Sm{sup 3+}(4f{sup 5}) spins, was developed by annealing in hydrogen gas (hydrogenation). It was established that annealing in hydrogen atmosphere boosts the RT-FM properties so that the saturation magnetisation could be increased by more than 100%. Physical explanations and discussions were given in this paper. Thus, it was proved that the magnetic properties could be tailored to diamagnetic LaOCl compound by Sm-doping and post treatment under H{sub 2} atmosphere. Therefore, LaOCl nanocrystals could be used as a potential candidate for optical phosphor applications with magnetic properties. - Graphical abstract: M-H dependence of Sm-doped LaOCl powders. Study the effect of hydrogenation. - Highlights: • Synthesis of Sm-doped LaOCl nanoparticles. • DM LaOCl transforms to FM with dilute concentration of Sm doping. • Annealing under H{sub 2} atmosphere induces drastic boost in the FM properties. • Saturation magnetization attained 29 memu/g with little doping and hydrogenation.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Synthesis and magnetic structure of the layered manganese oxide selenide Sr{sub 2}MnO{sub 2}Ag{sub 1.5}Se{sub 2}

The synthesis of a high-purity sample of the layered oxide selenide Sr{sub 2}MnO{sub 2}Ag{sub 1.5}Se{sub 2} is reported. At ambient temperature it crystallises in the space group I4/mmm with two formula units in the unit cell and lattice parameters a=4.08771(1) Å, c=19.13087(8) Å. The compound displays mixed-valent manganese in a formal oxidation state close to +2.5 and powder neutron diffraction measurements reveal that below the Néel temperature of 63(1) K this results in an antiferromagnetic structure which may be described as A-type, modelled in the magnetic space group P{sub I}4/mnc (128.410 in the Belov, Neronova and Smirnova (BNS) scheme) in which localised Mn moments of 3.99(2) μ{sub B} are arranged in ferromagnetic layers which are coupled antiferromagnetically. In contrast to the isostructural compound Sr{sub 2}MnO{sub 2}Cu{sub 1.5}S{sub 2}, Sr{sub 2}MnO{sub 2}Ag{sub 1.5}Se{sub 2} does not display long range ordering of coinage metal ions and vacancies, nor may significant amounts of the coinage metal readily be deintercalated using soft chemical methods. - Graphical abstract: Sr{sub 2}MnO{sub 2}Ag{sub 1.5}Se{sub 2} containing mixed valent Mn ions undergoes magnetic ordering with ferromagnetic coupling within MnO{sub 2} sheets and antiferromagnetic coupling between MnO{sub 2} sheets. - Highlights: • High purity sample of Sr{sub 2}MnO{sub 2}Ag{sub 1.5}Se{sub 2} obtained. • Magnetic structure determined. • Compared with related mixed-valent manganite oxide chalcogenides.

36 MATERIALS SCIENCE↗

Enabling high resolution strain mapping in zirconium alloys

Highlights: • Optimisation of speckle patterns suitable for high resolution digital image correlation in a zirconium alloy • Evidence of distinct strain patterning at the macro and micro scale • Analysis of strain heterogeneity in a neighbouring low and high strain band • Paved the way for future studies on strain localization in zirconium alloys subjected to hydriding and irradiation damage - Abstract: High Resolution Digital Image Correlation (HRDIC) has been established recently as a novel displacement mapping technique during mechanical loading experiments to quantify strain localisation down to the level of individual slip traces. This is achieved by the creation of a nano-scale gold pattern naturally formed during remodelling of a thin gold layer that was sputtered onto the region of interest. To date, the gold remodelling is carried out in a water vapourisation environment, which excludes the technique to be applied to materials that readily form noticeable oxide layers in such environments. The current paper describes a recently developed gold remodelling technique using a styrene-argon environment at substantially lower temperatures than the water-vapour based technique. The material used in the present work is a zirconium alloy where we first demonstrate the problem of oxide formation during remodelling in water vapour and the benefit of the modified remodelling procedure. The error associated with the spatial drift was assessed for different interrogation window sizes followed by detailed analysis of the subsequent strain maps produced using the styrene remodelled patterns after tensile deformation to nominal applied strains of ~3.5% and ~7.0%. The level of detail captured demonstrate the suitability of styrene-argon-based remodelling for materials like Zr alloys with the strain maps showing clear strain patterning on both a transgranular and single grain scale with the possibility of quantifying strain across a single slip trace.

36 MATERIALS SCIENCE↗

Acoustic emission monitoring of crack propagation in additively manufactured and conventional titanium components

We report that additive manufacturing (AM) is a novel and innovative production technology that can produce complex and lightweight engineering products. In AM components, as in all engineering materials, fatigue is considered as one of the principle causes of unexpected failure. In order to detect, localise and characterise cracks in various material components and metals, acoustic emission (AE) is used as a non-destructive monitoring technique. One of the main advantages of AE is that it can be also used for dynamic damage characterisation and specifically for crack propagation monitoring. In this research, we use AE to monitor the fatigue crack growth behaviour of Ti6Al4V components under four-point bending. The samples were produced by means of AM as well as conventional material. Notched and unnotched specimens were investigated with respect to the crack severity and crack detection using AE. The main AE signal parameters –such as cumulative events, hits, duration, average frequency and rise time– were evaluated and indicate sensitivity to damage propagation in order to lead to a warning against the final fracture occurrence. Finally, this is the first time that AE is applied in AM components under fatigue.

36 MATERIALS SCIENCE↗

Crystallographic ordering of Al and Sn in α-Ti

Increasing attention is being paid to α (2) Ti 3 (Al,Sn) precipitation from the α phase of titanium alloys owing to its effect on slip band formation, localisation and the implications for fatigue performance in jet engine titanium. However, the early stages of α 2 precipitation have historically been difficult to observe by electron microscopy, neutron diffraction or atom probe analysis. Here, small angle X-ray scattering is used to reexamine the phase boundary in binary Ti-Al and Ti-Sn alloys with around 500 ppmw O. It is found that the phase boundaries in the literature are approximately correct, at 6.2 wt.% Al and 16.9 wt.% Sn, and that this favours the use of Al as a solid solution strengthener over Sn for ambient temperature applications. However, once O content and phase partitioning in α + β alloys are taken into account, this implies that Al eq limits for future alloy design of critical rotating parts should be lowered substantially.

36 MATERIALS SCIENCE↗

HCV RNA traffic and association with NS5A in living cells

The spatiotemporal dynamics of Hepatitis C Virus (HCV) RNA localisation are poorly understood. To address this we engineered HCV genomes harbouring MS2 bacteriophage RNA stem-loops within the 3′-untranslated region to allow tracking of HCV RNA via specific interaction with a MS2-Coat-mCherry fusion protein. Despite the impact of these insertions on viral fitness, live imaging revealed that replication of tagged-HCV genomes induced specific redistribution of the mCherry-tagged-MS2-Coat protein to motile and static foci. Further analysis showed that HCV RNA was associated with NS5A in both static and motile structures while a subset of motile NS5A structures was devoid of HCV RNA. Further investigation of viral RNA traffic with respect to lipid droplets (LDs) revealed HCV RNA-positive structures in close association with LDs. These studies provide new insights into the dynamics of HCV RNA traffic with NS5A and LDs and provide a platform for future investigations of HCV replication and assembly. - Highlights: • HCV can tolerate can bacteriophage MS2 stem-loop insertions within the 3′ UTR. • MS2 stem-loop containing HCV genomes allow for real-time imaging of HCV RNA. • HCV RNA is both static and motile and associates with NS5A and lipid droplets.

60 APPLIED LIFE SCIENCES↗

Overexpression of microRNA-1288 in oesophageal squamous cell carcinoma

Purpose: This study aims to examine the expression profiles miR-1288 in oesophageal squamous cell carcinoma (ESCC). The cellular implications and target interactions of ESCC cells following miR-1288 overexpression was also examined. Methods: In total, 120 oesophageal tissues (90 primary ESCCs and 30 non-neoplastic tissues) were recruited for miR-1288 expression analysis using qRT-PCR. An exogenous miR-1288 mimic and its inhibitor were used to explore the in-vitro effects of miR-1288 on ESCC cells by performing cell proliferation, colony formation, cell invasion and migration assays. Localisation and modulatory changes of various miR-1288 regulated proteins such as FOXO1, p53, TAB3, BCL2 and kRAS was examined using immunofluorescence and western blot. Results: Overexpression of miR-1288 was more often noted in ESCC tissues when compared to non-neoplastic oesophageal tissues. High expression was often noted in high grade carcinomas and with metastases. Patients with high levels of miR-1288 expression showed a slightly better survival compared to patients with low miR-1288 levels. Furthermore, overexpression of miR-1288 showed increased cell proliferation and colony formation, improved cell migration and enhanced cell invasion properties in ESCC cells. In addition, miR-1288 overexpression in ESCC cells showed repression of cytoplasmic tumour suppressor FOXO1 protein expression. Inversely, inhibition of miR-1288 expression exhibited remarkable upregulation of FOXO1 protein, while expressions of other tested proteins remain unchanged. Conclusions: Up regulation of miR-1288 expression in ESCC tissues and miR-1288 induced oncogenic features of ESCC cells in-vitro indicates the oncogenic roles of miR-1288 in ESCCs. Overexpression of miR-1288 play a key role in the pathogenesis of ESCCs and its modulation may have potential therapeutic value in patients with ESCC. - Highlights: • miR-1288 was more often noted in neoplastic than non-neoplastic tissue. • miR-1288 overexpression increased proliferative/invasive activities of ESCC. • miR-1288 overexpression showed repression of FOXO1 protein expression. • miR-1288 functions as an oncogenic miRNA in ESCCs.

60 APPLIED LIFE SCIENCES↗

Multi-tasking Sulf1/Sulf2 enzymes do not only facilitate extracellular cell signalling but also participate in cell cycle related nuclear events

Highlights: • Dynamic SULF1/2 mobilisation to nucleus during cell cycle. • SULF1/2 co-ordinate extracellular cell signalling with nuclear role in cell cycle. • SULF1/2 overexpression increases cell proliferation while inhibition reduces growth. This study demonstrates highly dynamic spatial and temporal pattern of SULF1/SULF2 expression in a number of neuronal cell types growing in normal culture medium that included their transient nuclear mobilisation. Their nuclear translocation became particularly apparent during cell proliferation as both SULF1/SULF2 demonstrated not only cell membrane associated expression, their known site of function but also transient nuclear mobilisation during nuclear cell division. Nuclear localisation was apparent not only by immunocytochemical staining but also confirmed by immunoblotting staining of isolated nuclear fractions of C6, U87 and N2A cells. Immunocytochemical analysis demonstrated rapid nuclear exit of both SULF1/SULF2 following cell division that was slightly delayed but not blocked in a fraction of the polyploid cells observed in C6 cells. The overexpression of both Sulf1 and Sulf2 genes in C6 and U87 cells markedly promoted in vitro growth of these cells accompanied by nuclear mobilisation while inhibition of both these genes inhibited cell proliferation with little or no nuclear SULF1/SULF2 mobilisation. SULF1/SULF2 activity in these cells thus demonstrated a clear co-ordination of extracellular cell signalling with nuclear events related to cell proliferation.

60 APPLIED LIFE SCIENCES↗

Talin is a substrate for SUMOylation in migrating cancer cells

Highlights: • Talin is SUMOylated in cancer cells. • SUMOylation may regulate focal adhesion disassembly and cell migration. • SUMOylation may also regulate cleavage of talin. • Other focal adhesion proteins have been identified as potential SUMO targets including filamin-A and vinculin. Focal adhesions (FAs) play an important role in cancer cell migration and metastasis by linking the actin cytoskeleton to the extracellular matrix, allowing the cell to generate traction. SUMOylation is a post-translational modification of proteins on lysine residues that can affect protein localisation, turnover and protein-protein interactions. In this study, we demonstrate that talin, a key component of FAs, can be post-translationally modified by SUMOylation in MDA-MB-231 breast cancer cells and U2OS osteosarcoma cells. Furthermore we demonstrate that SUMOylation regulates the dynamic activities of FAs including their number, size and turnover rate. Inhibiting SUMOylation significantly reduced the speed of cell migration. The identification of talin as a SUMO target provides insight into the mechanisms regulating focal adhesion formation and turnover and potentially identifies a novel mechanism underlying cell migration.

60 APPLIED LIFE SCIENCES↗

IL-1β promotes transendothelial migration of PBMCs by upregulation of the FN/α5β1 signalling pathway in immortalised human brain microvascular endothelial cells

Highlights: • IL-1β-induced inflammation of THBMECs increases cell-matrix adhesion. • Active β{sub 1} integrins are redistributed under inflammatory conditions. • IL-1β activates the FN/α{sub 5}β{sub 1} integrin/FAK/Src-signalling pathway. • Blocking the α{sub 5} integrin subunit reduces IL-1β-induced transendothelial migration. Neuroinflammation is often associated with pathological changes in the function of the blood-brain barrier (BBB) caused by disassembly of tight and adherens junctions that under physiological conditions are important for the maintenance of the BBB integrity. Consequently, in inflammation the BBB becomes dysfunctional, facilitating leukocyte traversal of the barrier and accumulation of immune cells within the brain. The extracellular matrix (ECM) also contributes to BBB integrity but the significance of the main ECM receptors, the β{sub 1} integrins also expressed on endothelial cells, is less well understood. To evaluate whether β{sub 1} integrin function is affected during inflammation and impacts barrier function, we used a transformed human brain microvascular endothelial cell (THBMEC)-based Interleukin 1β (IL-1β)-induced inflammatory in vitro BBB model. We demonstrate that IL-1β increases cell-matrix adhesion and induces a redistribution of active β{sub 1} integrins to the basal surface. In particular, binding of α{sub 5}β{sub 1} integrin to its ligand fibronectin is enhanced and α{sub 5}β{sub 1} integrin-dependent signalling is upregulated. Additionally, localisation of the tight junction protein claudin-5 is altered. Blockade of the α{sub 5}β{sub 1} integrin reduces the IL-1β-induced transendothelial migration of peripheral blood mononuclear cells (PBMCs). These data imply that IL-1β-induced inflammation not only destabilizes tight junctions but also increases α{sub 5}β{sub 1} integrin-dependent cell-matrix adhesion to fibronectin.

60 APPLIED LIFE SCIENCES↗