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A Zissler, B Ehrenfellner, E E Foditsch, F C Monticelli, S Pittner
An accurate estimation of the postmortem interval (PMI) is a central aspect in forensic routine. Recently, a novel approach based on the analysis of postmortem muscle protein degradation has been proposed. However, a number of questions remain to be answered until sensible application of this method to a broad variety of forensic cases is possible. To evaluate whether altered in vivo protein metabolism interferes with postmortem degradation patterns, we conducted a comparative study. We developed a standardized animal degradation model in rats, and collected additional muscle samples from animals recovering from muscle injury and from rats with developed disuse muscle atrophy after induced spinal cord injury...
March 2, 2018: International Journal of Legal Medicine
Poonam Sehgal, Xinyu Kong, Jun Wu, Raimon Sunyer, Xavier Trepat, Deborah Leckband
This study reports novel findings that link E-Cadherin-mediated force-transduction signaling to vinculin targeting to intercellular junctions via epidermal growth factor receptor (EGFR) and integrins. These results build on previous findings, which demonstrated that mechanically perturbed E-Cadherin receptors activate phosphoinositide-3-kinase and downstream integrins, in an EGFR-dependent manner. Results of this study show that this EGFR-mediated kinase cascade controls the force-dependent recruitment of vinculin to stressed E-Cadherin complexes--a key early signature of cadherin-based mechanotransduction...
February 27, 2018: Journal of Cell Science
Aleena Gladkikh, Anastasia Kovaleva, Anna Tvorogova, Ivan A Vorobjev
Cell-extracellular matrix (ECM) adhesion is an important property of virtually all cells in multicellular organisms. Cell-ECM adhesion studies, therefore, are very significant both for biology and medicine. Over the last three decades, biomedical studies resulted in a tremendous advance in our understanding of the molecular basis and functions of cell-ECM adhesion. Based on morphological and molecular criteria, several different types of model cell-ECM adhesion structures including focal adhesions, focal complexes, fibrillar adhesions, podosomes, and three-dimensional matrix adhesions have been described...
2018: Methods in Molecular Biology
Anna M Chizhik, Carina Wollnik, Daja Ruhlandt, Narain Karedla, Alexey I Chizhik, Lara Hauke, Dirk Hähnel, Ingo Gregor, Jörg Enderlein, Florian Rehfeldt
We report a novel method, dual color axial nanometric localization by Metal Induced Energy Transfer (dcMIET), and combine it with Förster Resonant Energy Transfer (FRET) for resolving structural details in cells on the molecular level. We demonstrate the capability of this method on cytoskeletal elements and adhesions in human mesenchymal stem cells (hMSCs). Our approach is based on Fluorescence-Lifetime-Imaging Microscopy (FLIM), and allows for precise determination of the 3D architecture of stress fibers anchoring at focal adhesions, thus yielding crucial information to understand cell-matrix mechanics...
February 14, 2018: Molecular Biology of the Cell
Elias H Barriga, Kristian Franze, Guillaume Charras, Roberto Mayor
Collective cell migration is essential for morphogenesis, tissue remodelling and cancer invasion. In vivo, groups of cells move in an orchestrated way through tissues. This movement involves mechanical as well as molecular interactions between cells and their environment. While the role of molecular signals in collective cell migration is comparatively well understood, how tissue mechanics influence collective cell migration in vivo remains unknown. Here we investigated the importance of mechanical cues in the collective migration of the Xenopus laevis neural crest cells, an embryonic cell population whose migratory behaviour has been likened to cancer invasion...
February 14, 2018: Nature
Martino V Franchi, Severin Ruoss, Paola Valdivieso, Kyle W Mitchell, Kenneth Smith, Philip J Atherton, Marco V Narici, Martin Flück
AIMS: We assessed focal adhesion kinase (FAK) response to concentric (CON) versus eccentric (ECC) resistance training (RT) at two vastus lateralis (VL) sites, and the relationships between FAK, muscle protein synthesis (MPS), and morphological remodeling. METHODS: Six young males trained both legs unilaterally 3 times/week for 8 weeks; one leg performed CON RT, the contralateral performed ECC RT. Muscle biopsies were collected after training from VL midbelly (MID) and distal (Distal) sites at 0, 4, 8 weeks...
February 13, 2018: Acta Physiologica
Dror S Chorev, Tova Volberg, Ariel Livne, Miriam Eisenstein, Bruno Martins, Zvi Kam, Brigitte M Jockusch, Ohad Medalia, Michal Sharon, Benny Geiger
Focal adhesions (FAs) are multi-protein complexes that connect the actin cytoskeleton to the extracellular matrix, via integrin receptors. The growth, stability and adhesive functionality of these structures are tightly regulated by mechanical stress, yet, despite the extensive characterization of the integrin adhesome, the detailed molecular mechanisms underlying FA mechanosensitivity are still unclear. Besides talin, another key candidate for regulating FA-associated mechanosensing, is vinculin, a prominent FA component, which possesses either closed ("auto-inhibited") or open ("active") conformation...
February 9, 2018: Scientific Reports
Katarzyna I Jankowska, Edward K Williamson, Nathan H Roy, Daniel Blumenthal, Vidhi Chandra, Tobias Baumgart, Janis K Burkhardt
Full T cell activation requires coordination of signals from multiple receptor-ligand pairs that interact in parallel at a specialized cell-cell contact site termed the immunological synapse (IS). Signaling at the IS is intimately associated with actin dynamics; T cell receptor (TCR) engagement induces centripetal flow of the T cell actin network, which in turn enhances the function of ligand-bound integrins by promoting conformational change. Here, we have investigated the effects of integrin engagement on actin flow, and on associated signaling events downstream of the TCR...
2018: Frontiers in Immunology
Qin Wang, Julie Delcorde, Tracy Tang, Gregory P Downey, Christopher A McCulloch
IL-1 signaling is adhesion-restricted in many cell types, but the mechanism that drives it is not defined. We screened for proteins recruited to nascent adhesions in IL-1-treated human fibroblasts with tandem mass tag-mass spectrometry. We used fibronectin bead preparations to enrich 10 actin-associated proteins. There was a 1.2 times log 2-fold enrichment of actin capping protein (ACP) at 30 min after IL-1 stimulation. Knockdown (KD) of ACP by siRNA reduced IL-1-induced ERK activation(by 56%, matrix metalloproteinase-3 (MMP-3) expression by 48%, and MMP-9 expression by 62% (in all reductions, P < 0...
January 22, 2018: FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology
Eric S Orwoll, Jack Wiedrick, Jon Jacobs, Erin S Baker, Paul Piehowski, Vladislav Petyuk, Yuqian Gao, Tujin Shi, Richard D Smith, Douglas C Bauer, Steven R Cummings, Carrie M Nielson, Jodi Lapidus
The biological perturbations associated with incident mortality are not well elucidated, and there are limited biomarkers for the prediction of mortality. We used a novel high-throughput proteomics approach to identify serum peptides and proteins associated with 5-year mortality in community-dwelling men age ≥65 years who participated in a longitudinal observational study of musculoskeletal aging (Osteoporotic Fractures in Men: MrOS). In a discovery phase, serum specimens collected at baseline in 2473 men were analyzed using liquid chromatography-ion mobility-mass spectrometry, and incident mortality in the subsequent 5 years was ascertained by tri-annual questionnaire...
February 5, 2018: Aging Cell
Baiyun Liu, Jason I Kilpatrick, Bartlomiej Lukasz, Suzanne P Jarvis, Fiona McDonnell, Deborah M Wallace, Abbot F Clark, Colm J O'Brien
Purpose: Alteration in the extracellular matrix (ECM) of the optic nerve head (ONH) causes lamina cribrosa (LC) fibrosis and affects the mechanical integrity of the ONH. Increased ECM tissue stiffness drives myofibroblast activation leading to tissue fibrosis throughout the body. Here using primary human LC cells, we investigate the effect of substrate stiffness on profibrotic changes, which might be a key molecular mechanism driving ECM remodeling of the LC in primary open-angle glaucoma (POAG) glaucoma...
February 1, 2018: Investigative Ophthalmology & Visual Science
Rahuman S Malik-Sheriff, Sarah Imtiaz, Hernán E Grecco, Eli Zamir
Focal adhesions anchor contractile actin fibers with the extracellular matrix, sense the generated tension and respond to it by changing their morphology and composition. Here we ask how this mechanosensing is enabled at the protein-network level, given the modular assembly and multitasking of focal adhesions. To address this, we applied a sensitive 4-color live cell imaging approach, enabling monitoring patterns of molecular changes in single focal adhesions. Co-imaging zyxin, FAK, vinculin and paxillin revealed heterogeneities in their responses to Rho-associated kinase (ROCK)-mediated perturbations of actomyosin contractility...
February 1, 2018: Scientific Reports
M Ojansivu, L Hyväri, M Kellomäki, L Hupa, S Vanhatupa, S Miettinen
Bioactive glasses (BaGs) are widely utilised in bone tissue engineering (TE) but the molecular response of cells to BaGs is poorly understood. To elucidate the mechanisms of cell attachment to BaGs and BaG-induced early osteogenic differentiation, we cultured human adipose stem cells (hASCs) on discs of two silica-based BaGs S53P4 (23.0 Na2O - 20.0 CaO - 4.0 P2O5 - 53.0 SiO2 (wt-%)) and 1-06 (5.9 Na2O - 12.0 K2O - 5.3 MgO - 22.6 CaO - 4.0 P2O5 - 0.2 B2O3 - 50.0 SiO2) in the absence of osteogenic supplements...
January 30, 2018: European Cells & Materials
Peng Chen, Toshihiro Aso, Ryuichiro Sasaki, Yusuke Tsutsumi, Maki Ashida, Hisashi Doi, Takao Hanawa
To clarify the effects of micron/submicron hybrid topography on cell morphology and functionalization, we investigated the adhesion and differentiation of human mesenchymal stem cells (hMSCs) to titanium (Ti) surfaces with three different topographies: micron, submicron, and hybrid grooves created using a femtosecond laser. hMSCs cultured on Ti specimens showed high alignment on micron and hybrid surfaces after 6 h of incubation, whereas cells attached to submicron and hybrid surfaces were elongated. An examination of vinculin-positive adhesion plaques indicated that micron grooves affected cellular alignment by modifying the initial cell polarization, whereas submicron grooves affected cellular extension...
March 2017: Journal of Biomedical Nanotechnology
Koichiro Maki, Sung-Woong Han, Yoshinori Hirano, Shigenobu Yonemura, Toshio Hakoshima, Taiji Adachi
Adherens junctions (AJs) adaptively change their intensities in response to intercellular tension; therefore, they integrate tension generated by individual cells to drive multicellular dynamics, such as morphogenetic change in embryos. Under intercellular tension, α-catenin, which is a component protein of AJs, acts as a mechano-chemical transducer to recruit vinculin to promote actin remodeling. Although in vivo and in vitro studies have suggested that α-catenin-mediated mechanotransduction is a dynamic molecular process, which involves a conformational change of α-catenin under tension to expose a cryptic vinculin binding site, there are no suitable experimental methods to directly explore the process...
January 25, 2018: Scientific Reports
Prahalathan Pichavaram, Wen Yin, Kirk W Evanson, Jonathan H Jaggar, Salvatore Mancarella
Aims: Stromal interaction molecule 1 (STIM1) has emerged as an important player in the regulation of growth and proliferation of smooth muscle cells. Therefore, we hypothesized that STIM1 plays a crucial role in the maintenance of vascular integrity. The objective of this study was to evaluate whether reduced expression of STIM1 could modify the structure and function of the vasculature, leading to changes in blood pressure (BP). Methods and results: Smooth muscle-specific STIM1 knockout (sm-STIM1 KO) in mice resulted in arteries with ∼80% reduced STIM1 protein expression as compared to control mice...
January 19, 2018: Cardiovascular Research
Jian Lu, Miaomiao Tang, Yi Liu, Jin Wang, Zhanao Wu
Chromium (Cr) is a highly toxic, common heavy metal used in industrial production. There are two types of Cr in nature: hexavalent chromium (Cr(VI)) and chromium trichloride (Cr(III)). Cr(III) is involved in the metabolism of sugars and lipids, whereas Cr(VI) is absorbed through the respiratory tract and skin and generates free radicals that result in secondary toxicity. Cr(VI) leads to cancer in the occupational population and is therefore recognized as a human carcinogen by the International Agency for Research on Cancer...
January 16, 2018: Biological Trace Element Research
Marta Pokrywczynska, Arkadiusz Jundzill, Karolina Warda, Lukasz Buchholz, Marta Rasmus, Jan Adamowicz, Magdalena Bodnar, Andrzej Marszalek, Anna Helmin-Basa, Jacek Michalkiewicz, Maciej Gagat, Alina Grzanka, Malgorzata Frontczak-Baniewicz, Agata Magdalena Gastecka, Tomasz Kloskowski, Maciej Nowacki, Camillo Ricordi, Tomasz Drewa
A variety of tissue engineering techniques utilizing different cells and biomaterials are currently being explored to construct urinary bladder walls de novo, but so far no approach is clearly superior. The aim of this study was to determine whether mesenchymal stem cells (MSCs) isolated from different sources, (bone marrow [BM-MSCs] and adipose tissue [ADSCs]), differ in their potential to regenerate smooth muscles in tissue-engineered urinary bladders and to determine an optimal number of MSCs for urinary bladder smooth muscle regeneration...
November 2017: Cell Transplantation
Sara Proietti, Angela Catizone, Maria Grazia Masiello, Simona Dinicola, Gianmarco Fabrizi, Mirko Minini, Giulia Ricci, Roberto Verna, Russel J Reiter, Alessandra Cucina, Mariano Bizzarri
Through activation of the ERK pathway nicotine, in both normal MCF-10A and low malignant breast cancer cells (MCF7), promotes increased motility and invasiveness. Melatonin antagonizes both these effects by inhibiting almost completely ERK phosphorylation. As melatonin has no effect on not-stimulated cells, it is likely that melatonin can counteract ERK-activation only downstream of nicotine-induced activation. This finding suggests that melatonin hampers ERK phosphorylation presumably by targeting a still unknown intermediate factor that connects nicotine stimulation to ERK phosphorylation...
January 16, 2018: Journal of Pineal Research
Joshua H Cook, Norikiyo Ueno, Melissa B Lodoen
The motility of blood monocytes is orchestrated by the activity of cell surface integrins, which translate extracellular signals into cytoskeletal changes to mediate adhesion and migration. Toxoplasma gondii is an intracellular parasite that infects migratory cells and enhances their motility, but the mechanisms underlying T. gondii-induced hypermotility are incompletely understood. We have investigated the molecular basis for the hypermotility of primary human peripheral blood monocytes and THP-1 cells infected with T...
January 2, 2018: Journal of Biological Chemistry
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