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https://www.readbyqxmd.com/read/28720784/snap-tagged-chikungunya-virus-replicons-improve-visualisation-of-non-structural-protein-3-by-fluorescence-microscopy
#1
Roland Remenyi, Grace C Roberts, Carsten Zothner, Andres Merits, Mark Harris
Chikungunya virus (CHIKV), a mosquito-borne alphavirus, causes febrile disease, muscle and joint pain, which can become chronic in some individuals. The non-structural protein 3 (nsP3) plays essential roles during infection, but a complete understanding of its function is lacking. Here we used a microscopy-based approach to image CHIKV nsP3 inside human cells. The SNAP system consists of a self-labelling enzyme tag, which catalyses the covalent linking of exogenously supplemented synthetic ligands. Genetic insertion of this tag resulted in viable replicons and specific labelling while preserving the effect of nsP3 on stress granule responses and co-localisation with GTPase Activating Protein (SH3 domain) Binding Proteins (G3BPs)...
July 18, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28717648/response-detection-of-castrate-resistant-prostate-cancer-to-clinically-utilised-and-novel-treatments-by-monitoring-phospholipid-metabolism
#2
Tim A D Smith, Su M Phyu, Kholoud S Alzyoud, Chih-Chung Tseng
Androgen receptor (AR) activation is the primary driving factor in prostate cancer which is initially responsive to castration but then becomes resistant (castration-resistant prostate cancer (CRPC)). CRPC cells still retain the functioning AR which can be targeted by other therapies. A recent promising development is the use of inhibitors (Epi-1) of protein-protein interaction to inhibit AR-activated signalling. Translating novel therapies into the clinic requires sensitive early response indicators. Here potential response markers are explored...
2017: BioMed Research International
https://www.readbyqxmd.com/read/28710114/circadian-rhythm-disruption-impairs-tissue-homeostasis-and-exacerbates-chronic-inflammation-in-the-intestine
#3
René Pagel, Florian Bär, Torsten Schröder, Annika Sünderhauf, Axel Künstner, Saleh M Ibrahim, Stella E Autenrieth, Kathrin Kalies, Peter König, Anthony H Tsang, Dominik Bettenworth, Senad Divanovic, Hendrik Lehnert, Klaus Fellermann, Henrik Oster, Stefanie Derer, Christian Sina
Endogenous circadian clocks regulate 24 h rhythms of physiology and behavior. Circadian rhythm disruption (CRD) is suggested as a risk factor for inflammatory bowel disease. However, the underlying molecular mechanisms remain unknown. Intestinal biopsies from Per1/2 mutant and wild-type (WT) mice were investigated by electron microscopy, immunohistochemistry, and bromodeoxyuridine pulse-chase experiments. TNF-α was injected intraperitoneally, with or without necrostatin-1, into Per1/2 mice or rhythmic and externally desynchronized WT mice to study intestinal epithelial cell death...
July 14, 2017: FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology
https://www.readbyqxmd.com/read/28691764/cten-promotes-epithelial-mesenchymal-transition-through-post-transcriptional-stabilization-of-snail
#4
Hannah Thorpe, Abdulaziz Asiri, Maham Akhlaq, Mohammad Ilyas
Cten promotes cell migration however the knowledge of underlying signalling pathways is sparse. We have shown that Cten downregulates E-cadherin, a feature of epithelial to mesenchymal transition (EMT). This prompted us to investigate whether Cten further contributed to EMT processes to regulate cell motility. The regulation of Snail by Cten was investigated following overexpression, knockdown (by RNA-interference) or knockout of Cten in HCT116, Caco-2 and SW620 colorectal cancer (CRC) cell lines. Subsequently, the cycloheximide (CHX) pulse chase assay was used to investigate changes in Snail protein stability and the functional relevance of Cten-Snail signalling was investigated...
July 10, 2017: Molecular Carcinogenesis
https://www.readbyqxmd.com/read/28686945/trophic-transfer-of-134-cs-in-the-manila-clam-ruditapes-philippinarum
#5
Murat Belivermiş, Önder Kılıç, Narin Sezer, Gülşah Kalaycı, Marc Metian
Bioaccumulation of radiocaesium in many marine organisms occurs through complex trophic transfer mechanisms. The present study addresses the trophic transfer of (134)Cs in the widely distributed marine bivalve, the Manila clam Ruditapes philippinarum, by experimentally determining the assimilation efficiency (AE) and the specific role of food quality or diet on the AE in this marine invertebrate. Pulse-chase feeding experiments were carried out on this clam using the phytoplankton species Tetraselmis chuii, Phaeodactylum tricornutum and Isochrysis galbana...
July 4, 2017: Journal of Environmental Radioactivity
https://www.readbyqxmd.com/read/28684414/kinetic-processivity-of-the-two-step-oxidations-of-progesterone-and-pregnenolone-to-androgens-by-human-cytochrome-p450-17a1
#6
Eric Gonzalez, F Peter Guengerich
Cytochrome P450 (P450, CYP) 17A1 plays a critical role in steroid metabolism, catalyzing both the 17α-hydroxylation of pregnenolone and progesterone and the subsequent 17α,20-lyase reactions to form dehydroepiandrosterone (DHEA) and androstenedione (Andro), respectively, critical for generating glucocorticoids and androgens. Human P450 17A1 reaction rates examined are enhanced by the accessory protein cytochrome b5 (b5), but the exact role of b5 in P450 17A1-catalyzed reactions is unclear as are several details of these reactions...
July 6, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28650345/preexisting-endothelial-cells-mediate-cardiac-neovascularization-after-injury
#7
Lingjuan He, Xiuzhen Huang, Onur Kanisicak, Yi Li, Yue Wang, Yan Li, Wenjuan Pu, Qiaozhen Liu, Hui Zhang, Xueying Tian, Huan Zhao, Xiuxiu Liu, Shaohua Zhang, Yu Nie, Shengshou Hu, Xiang Miao, Qing-Dong Wang, Fengchao Wang, Ting Chen, Qingbo Xu, Kathy O Lui, Jeffery D Molkentin, Bin Zhou
The mechanisms that promote the generation of new coronary vasculature during cardiac homeostasis and after injury remain a fundamental and clinically important area of study in the cardiovascular field. Recently, it was reported that mesenchymal-to-endothelial transition (MEndoT) contributes to substantial numbers of coronary endothelial cells after myocardial infarction. Therefore, the MEndoT has been proposed as a paradigm mediating neovascularization and is considered a promising therapeutic target in cardiac regeneration...
June 26, 2017: Journal of Clinical Investigation
https://www.readbyqxmd.com/read/28645366/optimization-of-native-and-formaldehyde-ipond-techniques-for-use-in-suspension-cells
#8
Nathaniel E Wiest, Alan E Tomkinson
The isolation of proteins on nascent DNA (iPOND) technique developed by the Cortez laboratory allows a previously unparalleled ability to examine proteins associated with replicating and newly synthesized DNA in mammalian cells. Both the original, formaldehyde-based iPOND technique and a more recent derivative, accelerated native iPOND (aniPOND), have mostly been performed in adherent cell lines. Here, we describe modifications to both protocols for use with suspension cell lines. These include cell culture, pulse, and chase conditions that optimize sample recovery in both protocols using suspension cells and several key improvements to the published aniPOND technique that reduce sample loss, increase signal to noise, and maximize sample recovery...
2017: Methods in Enzymology
https://www.readbyqxmd.com/read/28630424/characterization-of-slow-cycling-corneal-limbal-epithelial-cells-identifies-putative-stem-cell-markers
#9
R Sartaj, C Zhang, P Wan, Z Pasha, V Guaiquil, A Liu, J Liu, Y Luo, E Fuchs, M I Rosenblatt
In order to identify reliable markers of corneal epithelial stem cells, we employed an inducible transgenic "pulse-chase" murine model (K5Tta × TRE-H2BGFP) to localize, purify, and characterize slow cycling cells in the cornea. The retention of GFP labeling in slowly dividing cells allowed for localization of these cells to the corneal limbus and their subsequent purification by FACS. Transcriptome analysis from slow cycling cells identified differentially expressed genes when comparing to GFP(-) faster-dividing cells...
June 19, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28616130/quantitative-investigation-of-human-cell-surface-n-glycoprotein-dynamics
#10
Haopeng Xiao, Ronghu Wu
Surface glycoproteins regulate nearly every extracellular event and they are dynamic for cells to adapt to the ever-changing extracellular environment. These glycoproteins contain a wealth of information on cellular development and disease states, and have significant biomedical implications. Systematic investigation of surface glycoproteins will result in a better understanding of surface protein functions, cellular activities and the molecular mechanisms of disease. However, it is extraordinarily challenging to specifically and globally analyze surface glycoproteins...
January 1, 2017: Chemical Science
https://www.readbyqxmd.com/read/28615413/breaking-the-color-barrier-a-multi-selective-antibody-reporter-offers-innovative-strategies-of-fluorescence-detection
#11
Eugenio Gallo, Jonathan W Jarvik
A novel bi-partite fluorescence platform exploits the high affinity and selectivity of antibody scaffolds to capture and activate small molecule fluorogens. In this report we investigated the property of multi-selectivity activation from a single antibody against diverse cyanine-family fluorogens. Our fluorescence screen identified three cell impermeant fluorogens; each with unique emission spectra (blue, green, and red) and nanomolar affinities. Most importantly, as a protein fusion-tag to G-protein-coupled receptors the antibody biosensor retained full activity - displaying bright fluorogen signals with minimal background on live cells...
June 14, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/28578400/lipid-droplet-associated-hydrolase-promotes-lipid-droplet-fusion-and-enhances-atgl-degradation-and-triglyceride-accumulation
#12
Young-Hwa Goo, Se-Hee Son, Antoni Paul
Lipid droplet (LD)-associated hydrolase (LDAH) is a newly identified LD protein abundantly expressed in tissues that predominantly store triacylglycerol (TAG). However, how LDAH regulates TAG metabolism remains unknown. We found that upon oleic acid loading LDAH translocalizes from the ER to newly formed LDs, and induces LD coalescence in a tubulin-dependent manner. LDAH overexpression and downregulation in HEK293 cells increase and decrease, respectively, TAG levels. Pulse and chase experiments show that LDAH enhances TAG biogenesis, but also decreases TAG turnover and fatty acid release from cells...
June 2, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28530236/psilac-mass-spectrometry-reveals-zfp91-as-imid-dependent-substrate-of-the-crl4-crbn-ubiquitin-ligase
#13
Jian An, Charles M Ponthier, Ragna Sack, Jan Seebacher, Michael B Stadler, Katherine A Donovan, Eric S Fischer
Thalidomide and its derivatives lenalidomide and pomalidomide (IMiDs) are effective treatments of haematologic malignancies. It was shown that IMiDs impart gain-of-function properties to the CUL4-RBX1-DDB1-CRBN (CRL4(CRBN)) ubiquitin ligase that enable binding, ubiquitination and degradation of key therapeutic targets such as IKZF1, IKZF3 and CSNK1A1. While these substrates have been implicated as efficacy targets in multiple myeloma (MM) and 5q deletion associated myelodysplastic syndrome (del(5q)-MDS), other targets likely exist...
May 22, 2017: Nature Communications
https://www.readbyqxmd.com/read/28526082/proof-of-concept-non-invasive-estimation-of-peripheral-venous-oxygen-saturation
#14
Musabbir Khan, Chris G Pretty, Alexander C Amies, Joel Balmer, Houda E Banna, Geoffrey M Shaw, J Geoffrey Chase
BACKGROUND: Pulse oximeters continuously monitor arterial oxygen saturation. Continuous monitoring of venous oxygen saturation (SvO2) would enable real-time assessment of tissue oxygen extraction (O2E) and perfusion changes leading to improved diagnosis of clinical conditions, such as sepsis. METHODS: This study presents the proof of concept of a novel pulse oximeter method that utilises the compliance difference between arteries and veins to induce artificial respiration-like modulations to the peripheral vasculature...
May 19, 2017: Biomedical Engineering Online
https://www.readbyqxmd.com/read/28515092/human-cd62l-dim-neutrophils-identified-as-a-separate-subset-by-proteome-profiling-and-in-vivo-pulse-chase-labelling
#15
Tamar Tak, Patrick Wijten, Marjolein Heeres, Peter Pickkers, Arjen Scholten, Albert J R Heck, Nienke Vrisekoop, Luke P Leenen, Jos Eacute A M Borghans, Kiki Tesselaar, Leo Koenderman
During acute inflammation three neutrophil subsets are found in the blood: neutrophils with a conventional segmented nucleus, neutrophils with a banded nucleus and T-cell-suppressing CD62L(dim) neutrophils with a high number of nuclear lobes. In this study (clinicaltrials.gov NCT01766414) we compared the in vivo kinetics and proteomes of banded, mature and hypersegmented neutrophils to determine whether these cell types represent truly different neutrophil subsets or reflect changes induced by LPS activation...
May 17, 2017: Blood
https://www.readbyqxmd.com/read/28474479/quantification-of-epithelial-cell-proliferation-cell-dynamics-and-cell-kinetics-in-vivo
#16
REVIEW
Robert A Goodlad
The measurement of cell proliferation in vivo is usually carried out by the examination of static measures. These comprise the mitotic index or labeling indices using incorporation of DNA synthesis markers such as bromodeoxyuridine or tritiated thymidine, or intrinsic markers, such as Ki67 and proliferative cell nuclear antigen (PCNA). But static measures only provide a 'snapshot' of cell proliferation. Rate measures, including double labeling methods and the metaphase arrest method, can actually measure cell production rates but they are far less utilized at present...
May 5, 2017: Wiley Interdisciplinary Reviews. Developmental Biology
https://www.readbyqxmd.com/read/28464260/cellular-migration-ability-is-modulated-by-extracellular-purines-in-ovarian-carcinoma-skov-3-cells
#17
A S Martínez-Ramírez, M Díaz-Muñoz, A M Battastini, A Campos-Contreras, A Olvera, L Bergamin, T Glaser, C E Jacintho Moritz, H Ulrich, F G Vázquez-Cuevas
Extracellular nucleotides and nucleosides have emerged as important elements regulating tissue homeostasis. Acting through specific receptors, have the ability to control gene expression patterns to direct cellular fate. We observed that SKOV-3 cells express the ectonucleotidases: ectonucleotide pyrophosphatase 1 (ENPP1), ecto-5'-nucleotidase (NT5E), and liver alkaline phosphatase (ALPL). Strikingly, in pulse and chase experiments supplemented with ATP, SKOV-3 cells exhibited low catabolic efficiency in the conversion of ADP into AMP, but they were efficient in converting AMP into adenosine...
May 2, 2017: Journal of Cellular Biochemistry
https://www.readbyqxmd.com/read/28440336/ultrahigh-energy-density-harvested-from-domain-engineered-relaxor-ferroelectric-single-crystals-under-high-strain-rate-loading
#18
Sergey I Shkuratov, Jason Baird, Vladimir G Antipov, Evgueni F Talantsev, Jay B Chase, Wesley Hackenberger, Jun Luo, Hwan R Jo, Christopher S Lynch
Relaxor ferroelectric single crystals have triggered revolution in electromechanical systems due to their superior piezoelectric properties. Here the results are reported on experimental studies of energy harvested from (1-y-x)Pb(In1/2Nb1/2)O3-(y)Pb(Mg1/3Nb2/3)O3-(x)PbTiO3 (PIN-PMN-PT) crystals under high strain rate loading. Precise control of ferroelectric properties through composition, size and crystallographic orientation of domains made it possible to identify single crystals that release up to three times more electric charge density than that produced by PbZr0...
April 25, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28438216/improved-pressure-contour-analysis-for-estimating-cardiac-stroke-volume-using-pulse-wave-velocity-measurement
#19
Shun Kamoi, Christopher Pretty, Joel Balmer, Shaun Davidson, Antoine Pironet, Thomas Desaive, Geoffrey M Shaw, J Geoffrey Chase
BACKGROUND: Pressure contour analysis is commonly used to estimate cardiac performance for patients suffering from cardiovascular dysfunction in the intensive care unit. However, the existing techniques for continuous estimation of stroke volume (SV) from pressure measurement can be unreliable during hemodynamic instability, which is inevitable for patients requiring significant treatment. For this reason, pressure contour methods must be improved to capture changes in vascular properties and thus provide accurate conversion from pressure to flow...
April 24, 2017: Biomedical Engineering Online
https://www.readbyqxmd.com/read/28425971/%C3%AF-%C3%A2-cells-feature-de-ubiquitination-of-slc-transporters-and-increased-levels-and-fluxes-of-amino-acids
#20
André Bordinassi Medina, Marcin Banaszczak, Yang Ni, Ina Aretz, David Meierhofer
Solute carrier (SLC) transporters are a diverse group of membrane transporter proteins that regulate the cellular flux and distribution of endogenous and xenobiotic compounds. Post-translational modifications (PTMs), such as ubiquitination, have recently emerged as one of the major regulatory mechanisms in protein function and localization. Previously, we showed that SLC amino acid transporters were on average 6-fold de-ubiquitinated and increased amino acid levels were detected in ρ⁰ cells (lacking mitochondrial DNA, mtDNA) compared to parental cells...
April 20, 2017: International Journal of Molecular Sciences
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