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https://www.readbyqxmd.com/read/28811610/antibiotic-induced-release-of-small-extracellular-vesicles-exosomes-with-surface-associated-dna
#1
Andrea Németh, Norbert Orgovan, Barbara W Sódar, Xabier Osteikoetxea, Krisztina Pálóczi, Katalin É Szabó-Taylor, Krisztina V Vukman, Ágnes Kittel, Lilla Turiák, Zoltán Wiener, Sára Tóth, László Drahos, Károly Vékey, Robert Horvath, Edit I Buzás
Recently, biological roles of extracellular vesicles (which include among others exosomes, microvesicles and apoptotic bodies) have attracted substantial attention in various fields of biomedicine. Here we investigated the impact of sustained exposure of cells to the fluoroquinolone antibiotic ciprofloxacin on the released extracellular vesicles. Ciprofloxacin is widely used in humans against bacterial infections as well as in cell cultures against Mycoplasma contamination. However, ciprofloxacin is an inducer of oxidative stress and mitochondrial dysfunction of mammalian cells...
August 15, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28808015/a-p53-dependent-response-limits-the-viability-of-mammalian-haploid-cells
#2
Teresa Olbrich, Cristina Mayor-Ruiz, Maria Vega-Sendino, Carmen Gomez, Sagrario Ortega, Sergio Ruiz, Oscar Fernandez-Capetillo
The recent development of haploid cell lines has facilitated forward genetic screenings in mammalian cells. These lines include near-haploid human cell lines isolated from a patient with chronic myelogenous leukemia (KBM7 and HAP1), as well as haploid embryonic stem cells derived from several organisms. In all cases, haploidy was shown to be an unstable state, so that cultures of mammalian haploid cells rapidly become enriched in diploids. Here we show that the observed diploidization is due to a proliferative disadvantage of haploid cells compared with diploid cells...
August 14, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28807817/assessment-of-glutathione-glutathione-disulphide-ratio-and-s-glutathionylated-proteins-in-human-blood-solid-tissues-and-cultured-cells
#3
REVIEW
Daniela Giustarini, Graziano Colombo, Maria Lisa Garavaglia, Emanuela Astori, Nicola Marcello Portinaro, Francesco Reggiani, Salvatore Badalamenti, Anna Maria Aloisi, Annalisa Santucci, Ranieri Rossi, Aldo Milzani, Isabella Dalle-Donne
Glutathione (GSH) is the major non-protein thiol in humans and other mammals, which is present in millimolar concentrations within cells, but at much lower concentrations in the blood plasma. GSH and GSH-related enzymes act both to prevent oxidative damage and to detoxify electrophiles. Under oxidative stress, two GSH molecules become linked by a disulphide bridge to form glutathione disulphide (GSSG). Therefore, assessment of the GSH/GSSG ratio may provide an estimation of cellular redox metabolism. Current evidence resulting from studies in human blood, solid tissues, and cultured cells suggests that GSH also plays a prominent role in protein redox regulation via S-glutathionylation, i...
August 11, 2017: Free Radical Biology & Medicine
https://www.readbyqxmd.com/read/28803732/reck-and-gpr124-are-essential-receptor-cofactors-for-wnt7a-wnt7b-specific-signaling-in-mammalian-cns-angiogenesis-and-blood-brain-barrier-regulation
#4
Chris Cho, Philip M Smallwood, Jeremy Nathans
Reck, a GPI-anchored membrane protein, and Gpr124, an orphan GPCR, have been implicated in Wnt7a/Wnt7b signaling in the CNS vasculature. We show here that vascular endothelial cell (EC)-specific reduction in Reck impairs CNS angiogenesis and that EC-specific postnatal loss of Reck, combined with loss of Norrin, impairs blood-brain barrier (BBB) maintenance. The most N-terminal domain of Reck binds to the leucine-rich repeat (LRR) and immunoglobulin (Ig) domains of Gpr124, and weakening this interaction by targeted mutagenesis reduces Reck/Gpr124 stimulation of Wnt7a signaling in cell culture and impairs CNS angiogenesis...
August 7, 2017: Neuron
https://www.readbyqxmd.com/read/28802127/investigations-on-the-molecular-mode-of-action-of-the-novel-immunostimulator-zelnate-activation-of-the-cgas-sting-pathway-in-mammalian-cells
#5
Thomas Ilg
Bovine respiratory disease (BRD) is usually prevented or treated with vaccines and/or antibiotics. The use of antibiotics is, however, of concern due to the potential promotion of microbial resistance and the occurrence of residues. Recently an alternative aid in the treatment of BRD, the cationic lipid/bacterial plasmid DNA liposome-based immunomodulator ZelNate, has entered the veterinary market. In the present study, we provide data on the molecular mode of action of ZelNate. Despite the presence of numerous non-methylated CpG motifs in its plasmid DNA, ZelNate proved to be inactive on human and mouse toll-like receptor 9 (TLR9) in cell culture, in both recombinant and natural cellular receptor settings...
August 9, 2017: Molecular Immunology
https://www.readbyqxmd.com/read/28800637/biologically-active-recombinant-human-erythropoietin-expressed-in-hairy-root-cultures-and-regenerated-plantlets-of-nicotiana-tabacum-l
#6
Poornima Devi Gurusamy, Holger Schäfer, Siva Ramamoorthy, Michael Wink
Hairy root culture is a potential alternative to conventional mammalian cell culture to produce recombinant proteins due to its ease in protein recovery, low costs and absence of potentially human pathogenic contaminants. The current study focussed to develop a new platform of a hairy root culture system from Nicotiana tabacum for the production of recombinant human EPO (rhEPO), which is regularly produced in mammalian cells. The human EPO construct was amplified with C-terminal hexahistidine tag from a cDNA of Caco-2 cells...
2017: PloS One
https://www.readbyqxmd.com/read/28798413/structural-basis-for-ccd1-auto-inhibition-in-the-wnt-pathway-through-homomerization-of-the-dix-domain
#7
Shin-Ichi Terawaki, Shohei Fujita, Takuya Katsutani, Kensuke Shiomi, Kazuko Keino-Masu, Masayuki Masu, Kaori Wakamatsu, Naoki Shibata, Yoshiki Higuchi
Wnt signaling plays an important role in governing cell fate decisions. Coiled-coil-DIX1 (Ccd1), Dishevelled (Dvl), and Axin are signaling proteins that regulate the canonical pathway by controlling the stability of a key signal transducer β-catenin. These proteins contain the DIX domain with a ubiquitin-like fold, which mediates their interaction in the β-catenin destruction complex through dynamic head-to-tail polymerization. Despite high sequence similarities, mammalian Ccd1 shows weaker stimulation of β-catenin transcriptional activity compared with zebrafish (z) Ccd1 in cultured cells...
August 10, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28796235/development-of-fertile-mouse-oocytes-from-mitotic-germ-cells-in-vitro
#8
Kanako Morohaku, Yuji Hirao, Yayoi Obata
Mammalian fetal ovaries contain numerous primordial germ cells (PGCs), although few mature oocytes are obtained from females, owing to apoptosis and follicle atresia. The regulatory mechanisms underlying oogenesis/folliculogenesis remain unknown. Development of methods for obtaining mature oocytes from PGCs in fetal ovaries in vitro could contribute to clarifying these mechanisms. The failure of follicle assembly has been found to be the most challenging aspect in conventional culture conditions. Recently, we established novel culture conditions that enable successful follicle assembly, sustaining interactions between the oocyte and somatic cells, and, in turn, promoting oocyte growth and maturation...
September 2017: Nature Protocols
https://www.readbyqxmd.com/read/28796234/electron-microscopy-using-the-genetically-encoded-apex2-tag-in-cultured-mammalian-cells
#9
Jeffrey D Martell, Thomas J Deerinck, Stephanie S Lam, Mark H Ellisman, Alice Y Ting
Electron microscopy (EM) is the premiere technique for high-resolution imaging of cellular ultrastructure. Unambiguous identification of specific proteins or cellular compartments in electron micrographs, however, remains challenging because of difficulties in delivering electron-dense contrast agents to specific subcellular targets within intact cells. We recently reported enhanced ascorbate peroxidase 2 (APEX2) as a broadly applicable genetic tag that generates EM contrast on a specific protein or subcellular compartment of interest...
September 2017: Nature Protocols
https://www.readbyqxmd.com/read/28795516/the-potential-of-liquid-marbles-for-biomedical-applications-a-critical-review
#10
REVIEW
Nuno M Oliveira, Rui L Reis, João F Mano
Liquid marbles (LM) are freestanding droplets covered by micro/nanoparticles with hydrophobic/hydrophilic properties, which can be manipulated as a soft solid. The phenomenon that generates these soft structures is regarded as a different method to generate a superhydrophobic behavior in the liquid/solid interface without modifying the surface. Several applications for the LM have been reported in very different fields, however the developments for biomedical applications are very recent. At first, the LM properties are reviewed, namely shell structure, LM shape, evaporation, floatability and robustness...
August 10, 2017: Advanced Healthcare Materials
https://www.readbyqxmd.com/read/28795515/multiplexed-optogenetic-stimulation-of-neurons-with-spectrum-selective-upconversion-nanoparticles
#11
Xudong Lin, Ying Wang, Xian Chen, Runhuai Yang, Zixun Wang, Jingyu Feng, Haitao Wang, King W C Lai, Jufang He, Feng Wang, Peng Shi
Optical modulation of nervous system becomes increasingly popular as the wide adoption of optogenetics. For these applications, upconversion materials hold great promise as novel photonic elements. This study describes an upconversion based strategy for combinatorial neural stimulation both in vitro and in vivo by using spectrum-selective upconversion nanoparticles (UCNPs). NaYF4 based UCNPs are used to absorb near-infrared (NIR) energy and to emit visible light for stimulating neurons expressing different channelrhodopsin (ChR) proteins...
August 10, 2017: Advanced Healthcare Materials
https://www.readbyqxmd.com/read/28794402/dec1-negatively-regulates-the-expression-of-cxcl10-and-ccl5-induced-by-poly-ic-in-normal-human-mesangial-cells
#12
Qiang Liu, Tadaatsu Imaizumi, Keishu Murakami, Hiroshi Tanaka, Yunyan Wu, Tadashi Yoshizawa, Satoko Morohashi, Hiroko Seino, Hiroshi Kijima
The functions of differentiated embryonic chondrocyte gene (DEC) 1, a basic helix-loop-helix (bHLH) transcription factor, have been reported to be associated with the regulation of mammalian circadian rhythms, differentiation of chondrocytes and skeletal muscles, apoptosis, hypoxia-induced reactions and epithelial mesenchymal transition. Our previous report showed that another bHLH transcription factor DEC2 constitutes a negative feedback loop in Toll-like receptor 3 (TLR3)/interferon (IFN)-β-mediated inflammatory responses in human mesangial cells...
2017: Biomedical Research
https://www.readbyqxmd.com/read/28782927/going-deeper-biomolecular-tools-for-acoustic-and-magnetic-imaging-and-control-of-cellular-function
#13
Dan I Piraner, Arash Farhadi, Hunter C Davis, Di Wu, David Maresca, Jerzy O Szablowski, Mikhail G Shapiro
Most cellular phenomena of interest to mammalian biology occur within the context of living tissues and organisms. However, today's most advanced tools for observing and manipulating cellular function, based on fluorescent or light-controlled proteins, work best in cultured cells, transparent model species, or small, surgically accessed anatomical regions. Their reach into deep tissues and larger animals is limited by photon scattering. To overcome this limitation, we must design biochemical tools that interface with more penetrant forms of energy...
August 7, 2017: Biochemistry
https://www.readbyqxmd.com/read/28776750/generation-of-a-non-transmissive-borna-disease-virus-vector-lacking-both-matrix-and-glycoprotein-genes
#14
Kan Fujino, Yusuke Yamamoto, Takuji Daito, Akiko Makino, Tomoyuki Honda, Keizo Tomonaga
Borna disease virus (BoDV), a prototype of mammalian bornavirus, is a non-segmented, negative strand RNA virus that often causes severe neurological disorders in infected animals, including horses and sheep. Unique among animal RNA viruses, BoDV transcribes and replicates non-cytopathically in the cell nucleus, leading to the establishment of a long-lasting persistent infection. This striking feature of BoDV indicates its potential as an RNA virus vector system. In a previous study, we demonstrated that recombinant BoDV (rBoDV) lacking an envelope glycoprotein (G) gene develops a persistent infection in the transduced cells without loss of the viral genome...
August 4, 2017: Microbiology and Immunology
https://www.readbyqxmd.com/read/28770472/vacuole-inducing-compounds-that-disrupt-endolysosomal-trafficking-stimulate-production-of-exosomes-by-glioblastoma-cells
#15
Zehui Li, Nneka E Mbah, William A Maltese
Exosomes are produced from mammalian cells when multivesicular endosomes fuse with the plasma membrane, releasing their intralumenal vesicles. In this study we assessed the effects of MOPIPP, a novel indole-based chalcone, and vacuolin-1, a distinct triazine-based compound, on exosome production in cultured glioblastoma and 293T cells. Both compounds promote vacuolization of late endosome compartments and interfere with trafficking of late endosomes to lysosomes, without significant cytotoxicity. The results show that vacuolated cells treated with these compounds release exosomes with morphologies similar to untreated controls...
August 2, 2017: Molecular and Cellular Biochemistry
https://www.readbyqxmd.com/read/28769797/over-expression-of-a-codon-optimized-yeast-cytosolic-pyruvate-carboxylase-pyc2-in-cho-cells-for-an-augmented-lactate-metabolism
#16
Sanjeev K Gupta, Ankit Sharma, Hiralal Kushwaha, Pratyoosh Shukla
Monoclonal antibodies are the most demanding biotherapeutic drugs now a days used for the cure of various critical illnesses. Chinese hamster ovary (CHO) cells are one of the main hosts used for the large scale production of these antibodies. However, the cell line and production processes are the key factors to determine the cost and affordability of these antibodies. The metabolic waste lactic acid and ammonium are accumulated during a cell culture process and adversely affects productivity as well as product quality...
2017: Frontiers in Pharmacology
https://www.readbyqxmd.com/read/28768720/angiotensin-ii-regulates-phosphorylation-of-actin-associated-proteins-in-human-podocytes
#17
Laura K Schenk, Annika Möller-Kerutt, Rafael Klosowski, Dirk Wolters, Elisabeth Schaffner-Reckinger, Thomas Weide, Hermann Pavenstädt, Beate Vollenbröker
Within the kidney, angiotensin II (AngII) targets different cell types in the vasculature, tubuli, and glomeruli. An important part of the renal filtration barrier is composed of podocytes with their actin-rich foot processes. In this study, we used stable isotope labeling with amino acids in cell culture coupled to mass spectrometry to characterize relative changes in the phosphoproteome of human podocytes in response to short-term treatment with AngII. In 4 replicates, we identified a total of 17,956 peptides that were traceable to 2081 distinct proteins...
August 2, 2017: FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology
https://www.readbyqxmd.com/read/28768195/attenuation-of-synaptic-potentials-in-dendritic-spines
#18
Taekyung Kwon, Masayuki Sakamoto, Darcy S Peterka, Rafael Yuste
Dendritic spines receive the majority of excitatory inputs in many mammalian neurons, but their biophysical properties and exact role in dendritic integration are still unclear. Here, we study spine electrical properties in cultured hippocampal neurons using an improved genetically encoded voltage indicator (ArcLight) and two-photon glutamate uncaging. We find that back-propagating action potentials (bAPs) fully invade dendritic spines. However, uncaging excitatory post-synaptic potentials (uEPSPs) generated by glutamate photorelease, ranging from 4 to 27 mV in amplitude, are attenuated by up to 4-fold as they propagate to the parent dendrites...
August 1, 2017: Cell Reports
https://www.readbyqxmd.com/read/28767220/engineering-intracellularly-retained-gaussia-luciferase-reporters-for-improved-biosensing-and-molecular-imaging-applications
#19
Shuchi Gaur, Aarohi Bhargava-Shah, Sharon Hori, Rayhaneh Afjei, Thillai V Sekar, Sanjiv S Gambhir, Tarik F Massoud, Ramasamy Paulmurugan
Gaussia luciferase (GLUC) is a bioluminescent reporter protein of increasing importance. As a secretory protein, it has increased sensitivity in vitro and in vivo (∼20 000-fold, and ∼1000-fold, respectively) over its competitor, secreted alkaline phosphatase. Unfortunately, this same advantageous secretory nature of GLUC limits its usefulness for many other possible intracellular applications, e.g., imaging signaling pathways in intact cells, in vivo imaging, and in developing molecular imaging biosensors to study protein-protein interactions and protein folding...
August 10, 2017: ACS Chemical Biology
https://www.readbyqxmd.com/read/28767065/do-multiwell-plate-high-throughput-assays-measure-loss-of-cell-viability-following-exposure-to-genotoxic-agents
#20
Razmik Mirzayans, Bonnie Andrais, David Murray
Cell-based assays in multiwell plates are widely used for radiosensitivity and chemosensitivity assessment with different mammalian cell types. Despite their relative ease of performance, such assays lack specificity as they do not distinguish between the cytostatic (reversible/sustained growth arrest) and cytotoxic (loss of viability) effects of genotoxic agents. We recently reported studies with solid tumor-derived cell lines demonstrating that radiosensitivity as measured by multiwell plate colorimetric (e...
August 2, 2017: International Journal of Molecular Sciences
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