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V O Gunko, T N Pogorelova, V A Linde
Mass-spectrometric profiling of the serum in women at weeks 16-17 of gestation was carried out in order to detect proteomic predictors of preterm delivery. Changes in the production of 25 proteins (down-regulation for 13 proteins and up-regulation for 12 proteins) were detected in the sera of women whose pregnancies eventuated in premature deliveries. Among them, proteins with various regulatory functions were distinguished: antioxidant enzymes, chaperons, cytoskeleton proteins, cell adhesion molecules, and proteins involved in angiogenesis, proteolysis, transcription, inflammation processes, binding and transportation of various ligands...
October 26, 2016: Bulletin of Experimental Biology and Medicine
Weilong Zhong, Bo Sun, Cheng Lu, Hengheng Yu, Changhua Wang, Lingfei He, Ju Gu, Shuang Chen, Yanrong Liu, Xiangyan Jing, Zhun Bi, Guang Yang, Honggang Zhou, Tao Sun, Cheng Yang
Small-molecule fluorescent probes have been widely used in target identification, but this method has many disadvantages. For example, the identified proteins are usually complex, and additional biochemical studies are needed to distinguish real targets from interference results. To address this problem, we propose a series of strategies for improving the efficiency of target identification. First, pretreatment with a lower concentration of hydrogen peroxide can shield against thiol interference. Second, the use of benzophenone as a photo-affinity group is not appropriate, and diazirines are preferred...
October 26, 2016: Scientific Reports
Petrina Koumarianou, Gonzalo Goméz-López, Pilar Santisteban
Organization of epithelial cells during follicular lumen formation is crucial for thyroid morphogenesis and function of the thyroid gland; however, the molecular mechanisms underlying this are poorly understood. To investigate this process, we established three-dimensional (3D) epithelial culture model systems using Fischer rat thyroid (FRT) cells or murine primary thyrocytes that developed polarized spherical structures with a central lumen mimicking thyroid follicles. Using microarray-based differential expression analysis of FRT cells grown under 2D or 3D conditions followed by RNA-mediated interference (RNAi) and morphogenetic analysis, we identified a key role for the thyroid transcription factor Pax8 and its target Cadherin-16 (Cdh16) in the generation of polarized follicle-like structures...
October 25, 2016: Journal of Cell Science
Chiara Tersigni, Fiorella Di Nicuolo, Giuseppe Maulucci, Alessandro Rolfo, Domenica Giuffrida, Manuela Veglia, Marco De Spirito, Giovanni Scambia, Tullia Todros, Nicoletta Di Simone
BACKGROUND: Pre-eclampsia (PE) is a major cause of maternal and perinatal morbidity and mortality worldwide. It is defined by new onset of hypertension and proteinuria after the 20th week of gestation and characterized by systemic exaggerated inflammatory response. D6 is a chemokines scavenger receptor that binds with high affinity CC chemokines, internalizes and targets the ligands for degradation. It is expressed in trophoblast-derived tissues and prevents excessive placenta leukocyte infiltration...
2016: PloS One
Cheng-Feng Chiang, Mike Flint, Jin-Mann S Lin, Christina F Spiropoulou
Andes virus (ANDV) is the major cause of hantavirus pulmonary syndrome (HPS) in South America. Despite a high fatality rate (up to 40%), no vaccines or antiviral therapies are approved to treat ANDV infection. To understand the role of endocytic pathways in ANDV infection, we used 3 complementary approaches to identify cellular factors required for ANDV entry into human lung microvascular endothelial cells. We screened an siRNA library targeting 140 genes involved in membrane trafficking, and identified 55 genes required for ANDV infection...
2016: PloS One
Wen-Lei Shi, Ting Zhang, Jiang-Rui Zhou, Yong-Hua Huang, Chun-Lei Jiang
Glucocorticoids (GCs) play a vital role in the regulation of blood pressure by their permissive effects in potentiating vasoactive responses to catecholamines through glucocorticoid receptors. GCs achieve this function by controlling vascular smooth muscle tone. Clinically, low to moderate doses of GCs are generally used in the treatment of septic shock in recent years. GCs are now known to have both genomic and non-genomic effects. While genomic effects of GCs were well studied, few non-genomic effects were reported, much less the non-genomic mechanisms...
October 22, 2016: Biomedicine & Pharmacotherapy, Biomédecine & Pharmacothérapie
Steven G Thomas, Natalie S Poulter, Danai Bem, Brenda Finney, Laura M Machesky, Stephen P Watson
A dynamic, properly organised actin cytoskeleton is critical for the production and haemostatic function of platelets. The Wiskott Aldrich Syndrome protein (WASp) and Actin-Related Proteins 2 & 3 Complex (Arp2/3 complex) are critical mediators of actin polymerisation and organisation in many cell types. In platelets and megakaryocytes, these proteins have been shown to be important for proper platelet production and function. The cortactin family of proteins (Cttn & HS1) are known to regulate WASp-Arp2/3-mediated actin polymerisation in other cell types and so here we address the role of these proteins in platelets using knockout mouse models...
October 25, 2016: Platelets
Sgb Furness, D L Hare, A Kourakis, A M Turnley, P J Wookey
We have discovered that the accumulation of an anti-calcitonin receptor (anti-CTR) antibody conjugated to a fluorophore (mAb2C4:AF568) provides a robust signal for cells undergoing apoptotic programmed cell death (PCD). PCD is an absolute requirement for normal development of metazoan organisms. PCD is a hallmark of common diseases such as cardiovascular disease and tissue rejection in graft versus host pathologies, and chemotherapeutics work by increasing PCD. This robust signal or high fluorescent events were verified by confocal microscopy and flow cytometry in several cell lines and a primary culture in which PCD had been induced...
2016: Cell Death Discovery
Midori Nakamura, Masanori Koide, Nobuyuki Udagawa
The objective of treating osteoporosis is to prevent fractures. Bisphosphonates inhibit bone resorption and increase bone density, thereby suppressing the occurrence of fractures. Bisphosphonates have a high affinity for hydroxyapatite. About 20-80%of bisphosphonates absorbed in vivo is adsorbed onto the bone surface. However, the absorption rates of orally ingested bisphosphonates are less than 1%. Bisphosphonates adhered to the bone surface are specifically incorporated into cells upon bone resorption by osteoclasts...
2016: Clinical Calcium
Brie Sorrenson, Emmanuelle Cognard, Kathryn L Lee, Waruni C Dissanayake, Yanyun Fu, Weiping Han, William E Hughes, Peter R Shepherd
The processes regulating glucose-stimulated insulin secretion (GSIS) and its modulation by incretins in pancreatic β-cells are only partly understood. Here we investigate the involvement of β-catenin in these processes. Reducing β-catenin levels using siRNA knockdown attenuated GSIS in a range of β-cell models and blocked the ability of GLP-1 agonists and the depolarizing agent KCl to potentiate this. This could be mimicked in both β-cell models and isolated islets by short-term exposure to the β-catenin inhibitory drug pyrvinium...
October 24, 2016: Journal of Biological Chemistry
Heledd M Davies, Konstantinos Thalassinos, Andrew R Osborne
Repetitive low-complexity sequences, mostly assumed to have no function, are common in proteins that are exported by the malaria parasite into its host erythrocyte. We identify a group of exported proteins containing short lysine-rich tandemly repeated sequences that are sufficient to localise to the erythrocyte periphery where key virulence-related modifications to the plasma membrane and the underlying cytoskeleton are known to occur. Efficiency of targeting is dependent on repeat number, indicating that novel targeting modules could evolve by expansion of short lysine-rich sequences...
October 24, 2016: Journal of Biological Chemistry
Jihye Chung, Shunsuke Aburaya, Wataru Aoki, Mitsuyoshi Ueda
In very early stages of cancer development, one or a few cells expressing cancer-associated genes appear among a much larger number of surrounding normal cells. To analyze the molecular changes induced by this co-existence, we artificially prepared transformed cells with complete loss of tumor suppressor gene, SCRIB, among normal human embryonic kidney (HEK293T) cells. A cell strain with SCRIB-knockout was successfully constructed by using Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-Cas9 nuclease system and co-cultured with normal cells...
October 21, 2016: Journal of Bioscience and Bioengineering
Rakesh Kumar
Professor Ferid Murad has been a remarkable colleague and a mentor. During our very first meeting, he not only shared unresolved puzzles in Nitric Oxide (NO) research but also listened to my questions pointing to protein nitration and nitrosylation. This was start of a new avenue in my laboratory involving protein nitration, inducible nitric oxide synthase and nitrite production in the context of signaling and gene expression in cancer cells. Dynamic changes in the cytoskeleton remodeling in response to the cell membrane generated signals are regulated by p21-activated kinase 1 (PAK1) which also feed into microtubules (MT) dynamic via phosphorylating Tubulin Cofactor B (CoB) on serine 65 and serine 128...
2016: Current Medicinal Chemistry
Huiying Fu, Renjie Wu, Yuanyuan Li, Lizong Zhang, Xiaofang Tang, Jue Tu, Weimin Zhou, Jianchao Wang, Qiyang Shou
Carthamus tinctorius L. is a traditional Chinese medicine that activates blood circulation and dissipates blood stasis, and has been extensively used as antitumor treatment in a clinical setting in single or in compound preparation form. However, empirical evidence and a better understanding of the possible mechanisms involved are still required. Here, we investigated the role of safflower yellow (SY), the active ingredient of C. tinctorius, in the pulmonary metastasis of breast cancer, and the underlying mechanism of action...
October 25, 2016: American Journal of Chinese Medicine
Christine Gross, Veit Wiesmann, Sebastian Millen, Martina Kalmer, Thomas Wittenberg, Jan Gettemans, Andrea K Thoma-Kress
The delta-retrovirus Human T-cell leukemia virus type 1 (HTLV-1) preferentially infects CD4+ T-cells via cell-to-cell transmission. Viruses are transmitted by polarized budding and by transfer of viral biofilms at the virological synapse (VS). Formation of the VS requires the viral Tax protein and polarization of the host cytoskeleton, however, molecular mechanisms of HTLV-1 cell-to-cell transmission remain incompletely understood. Recently, we could show Tax-dependent upregulation of the actin-bundling protein Fascin (FSCN-1) in HTLV-1-infected T-cells...
October 2016: PLoS Pathogens
Lulu Jiang, Charles C T Hindmarch, Mark Rogers, Colin Campbell, Christy Waterfall, Jane Coghill, Peter W Mathieson, Gavin I Welsh
Glucocorticoids are steroids that reduce inflammation and are used as immunosuppressive drugs for many diseases. They are also the mainstay for the treatment of minimal change nephropathy (MCN), which is characterised by an absence of inflammation. Their mechanisms of action remain elusive. Evidence suggests that immunomodulatory drugs can directly act on glomerular epithelial cells or 'podocytes', the cell type which is the main target of injury in MCN. To understand the nature of glucocorticoid effects on non-immune cell functions, we generated RNA sequencing data from human podocyte cell lines and identified the genes that are significantly regulated in dexamethasone-treated podocytes compared to vehicle-treated cells...
October 24, 2016: Scientific Reports
Timothy K Lee, Kevin Meng, Handuo Shi, Kerwyn Casey Huang
The peptidoglycan cell wall is an integral organelle critical for bacterial cell shape and stability. Proper cell wall construction requires the interaction of synthesis enzymes and the cytoskeleton, but it is unclear how the activities of individual proteins are coordinated to preserve the morphology and integrity of the cell wall during growth. To elucidate this coordination, we used single-molecule imaging to follow the behaviours of the two major peptidoglycan synthases in live, elongating Escherichia coli cells and after perturbation...
October 24, 2016: Nature Communications
Niya Xiong, Shun Li, Kai Tang, Hongxia Bai, Yueting Peng, Hong Yang, Chunhui Wu, Yiyao Liu
Tumor cells translocating to distant sites are subjected to hemodynamic shear forces during their passage in the blood vessels. Low shear stress (LSS) plays a critical role in the regulation of various aspects of tumor cells functions, including motility and adhesion. Beyond its structural role, caveolin-1 (Cav-1), the important component of caveolae, represents a modulator of several cancer-associated functions as tumor progression and metastasis. However, the role of Cav-1 in regulating tumor cells response to shear stress remains poorly explored...
October 20, 2016: Biochimica et Biophysica Acta
Eliza Pelrine, Sara Diana Pasik, Leyla Bayat, Debora Goldschmiedt, Elizabeth P Bauer
Selective serotonin reuptake inhibitors (SSRIs) are widely prescribed to treat anxiety and depression, yet they paradoxically increase anxiety during initial treatment. Acute administration of these drugs prior to learning can also enhance Pavlovian cued fear conditioning. This potentiation has been previously reported to depend upon the bed nucleus of the stria terminalis (BNST). Here, using temporary inactivation, we confirmed that the BNST is not necessary for the acquisition of cued or contextual fear memory...
October 20, 2016: Neurobiology of Learning and Memory
Monika S Brill, Tatjana Kleele, Laura Ruschkies, Mengzhe Wang, Natalia A Marahori, Miriam S Reuter, Torben J Hausrat, Emily Weigand, Matthew Fisher, Andrea Ahles, Stefan Engelhardt, Derron L Bishop, Matthias Kneussel, Thomas Misgeld
Developmental axon remodeling is characterized by the selective removal of branches from axon arbors. The mechanisms that underlie such branch loss are largely unknown. Additionally, how neuronal resources are specifically assigned to the branches of remodeling arbors is not understood. Here we show that axon branch loss at the developing mouse neuromuscular junction is mediated by branch-specific microtubule severing, which results in local disassembly of the microtubule cytoskeleton and loss of axonal transport in branches that will subsequently dismantle...
October 18, 2016: Neuron
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