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https://www.readbyqxmd.com/read/29236307/interleukin-4-induces-a-cd44high-cd49bhigh-pc3-subpopulation-with-tumor-initiating-characteristics
#1
Holger H H Erb, Fabian Guggenberger, Frédéric R Santer, Zoran Culig
Pro- and anti-inflammatory cytokines may influence proliferation, migration, invasion and other cellular events of prostate cancer (PCa) cells. The hyaluronan receptor CD44, which is regulated by Interleukin (IL)-4, is a prostate basal cell marker. CD44high /CD49bhigh expressing cells have been demonstrated to have tumor-initiating characteristics. Here, we aimed to analyze the effects of long-term IL-4 treatment on CD44/CD49b expression, migration, proliferation and clonogenic potential of basal-like PCa cells...
December 13, 2017: Journal of Cellular Biochemistry
https://www.readbyqxmd.com/read/29235974/processing-fixed-and-stored-adipose-derived-stem-cells-for-quantitative-protein-array-assays
#2
Jessica S Sadick, Eric M Darling
Accurately characterizing cellular subpopulations is essential for elucidating the mechanisms underlying normal and pathological biology. Isolation of specific cell types can be accomplished by labeling unique cell-associated proteins with fluorescent antibodies. Cell fixation is commonly used to prepare these samples and allow for long-term storage, but this poses challenges for subsequent protein analysis. We previously established the FITSAR (formaldehyde-fixed intracellular target-sorted antigen retrieval) method, in which protein can be isolated and characterized from fixed, enriched cell subpopulations...
December 1, 2017: BioTechniques
https://www.readbyqxmd.com/read/29235543/purification-of-replicating-pancreatic-%C3%AE-cells-for-gene-expression-studies
#3
Reyes Carballar, Maria de Lluc Canyelles, Claudia Fernández, Yasmina Martí, Sarah Bonnin, Esther Castaño, Eduard Montanya, Noèlia Téllez
β-cell proliferation is a rare event in adult pancreatic islets. To study the replication-related β-cell biology we designed a replicating β-cells sorting system for gene expression experiments. Replicating β-cells were identified by EdU incorporation and purified by flow cytometry. For β-cell separation islet cells were sorted by size, granularity and Newport Green fluorescence emission that was combined with emitted fluorescence for EdU-labelled replicating cells sorting. The purity of the resulting sorted populations was evaluated by insulin staining and EdU for β-cell identification and for replicating cells, respectively...
December 13, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29235444/contact-guidance-for-cardiac-tissue-engineering-using-3d-bioprinted-gelatin-patterned-hydrogel
#4
Ajay Tijore, Scott Alexander Irvine, Udi Sarig, Priyadarshini Mhaisalkar, Vrushali Baisane, Subbu S Venkatraman
Here, we have developed a 3D bioprinted microchanneled gelatin hydrogel that promotes human mesenchymal stem cell (hMSC) myocardial commitment and supports native cardiomyocytes (CMs) contractile functionality. Firstly, we studied the effect of bioprinted microchanneled hydrogel on the alignment, elongation, and differentiation of hMSC. Notably, the cells displayed well defined F-actin anisotropy and elongated morphology on the microchanneled hydrogel, hence showing the effects of topographical control over cell behavior...
December 13, 2017: Biofabrication
https://www.readbyqxmd.com/read/29234079/a-genetic-screen-to-discover-sumoylated-proteins-in-living-mammalian-cells
#5
Maki Komiya, Akihiro Ito, Mizuki Endo, Daisuke Hiruma, Mitsuru Hattori, Hisato Saitoh, Minoru Yoshida, Takeaki Ozawa
Post-translational modification by the Small Ubiquitin-related Modifier (SUMO) is indispensable for diverse biological mechanisms. Although various attempts have been made to discover novel SUMO substrate proteins to unveil the roles of SUMOylation, the reversibility of SUMOylation, and the differences in the SUMOylation level still makes it difficult to explore infrequently-SUMOylated proteins in mammalian cells. Here, we developed a method to screen for mammalian SUMOylated proteins using the reconstitution of split fluorescent protein fragments in living mammalian cells...
December 12, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29234006/enhancing-size-based-size-separation-through-vertical-focus-microfluidics-using-secondary-flow-in-a-ridged-microchannel
#6
Bushra Tasadduq, Wilbur Lam, Alexander Alexeev, A Fatih Sarioglu, Todd Sulchek
High throughput size based separation and sorting of bioparticles and cells is critical to a variety of biomedical processing steps for medical diagnostics and pharmaceutical purification. Improving microfluidic size-based particle/cell sorting is a challenge to better address the need for generating more homogeneous subpopulations for study and use. We propose a novel advance to microfluidic sorting devices that uses three-dimensional focusing of the sample to optimally position particles to amplify the size-dependent differences in trajectories caused by differential secondary flows...
December 12, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29233871/interplay-of-autophagy-receptor-tyrosine-kinase-signalling-and-endocytic-trafficking
#7
REVIEW
Jane Fraser, Ainara G Cabodevilla, Joanne Simpson, Noor Gammoh
Vesicular trafficking events play key roles in the compartmentalization and proper sorting of cellular components. These events have crucial roles in sensing external signals, regulating protein activities and stimulating cell growth or death decisions. Although mutations in vesicle trafficking players are not direct drivers of cellular transformation, their activities are important in facilitating oncogenic pathways. One such pathway is the sensing of external stimuli and signalling through receptor tyrosine kinases (RTKs)...
December 12, 2017: Essays in Biochemistry
https://www.readbyqxmd.com/read/29233551/generation-of-adrenal-chromaffin-like-cells-from-human-pluripotent-stem%C3%A2-cells
#8
Kwaku Dad Abu-Bonsrah, Dongcheng Zhang, Andrew R Bjorksten, Mirella Dottori, Donald F Newgreen
Adrenomedullary chromaffin cells are catecholamine (CA)-producing cells originating from trunk neural crest (NC) via sympathoadrenal progenitors (SAPs). We generated NC and SAPs from human embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) in vitro via BMP2/FGF2 exposure, ascertained by qPCR and immunoexpression of SOX10, ASCL1, TFAP2α, and PHOX2B, and by fluorescence-activated cell sorting selection for p75NTR and GD2, and confirmed their trunk-like HOX gene expression. We showed that continuing BMP4 and curtailing FGF2 in vitro, augmented with corticosteroid mimetic, induced these cells to upregulate the chromaffin cell-specific marker PNMT and other CA synthesis and storage markers, and we demonstrated noradrenaline and adrenaline by Faglu and high-performance liquid chromatography...
November 27, 2017: Stem Cell Reports
https://www.readbyqxmd.com/read/29232382/charting-the-transcriptional-landscape-of-cells-of-renin-lineage-following-podocyte-depletion
#9
Aaron D McClelland, Julia Lichtnekert, Diana G Eng, Jeffrey W Pippin, Kenneth W Gross, Sina A Gharib, Stuart J Shankland
Renin producing cells of the juxtaglomerulus, herein called cells of renin lineage (CoRL), have garnered recent interest for their propensity to act as a progenitor source for various kidney cell types including podocytes. Despite recent advances, the process of transdifferentiation of CoRL to podocytes is poorly understood. In this study, we employed a transgenic reporter mouse line which permanently labels CoRL with ZsGreen fluorescent protein, allowing for isolation by fluorescence-activated cell sorting...
2017: PloS One
https://www.readbyqxmd.com/read/29232025/hospital-volume-and-outcomes-of-robot-assisted-partial-nephrectomy
#10
Leilei Xia, Jose E Pulido, Raju R Chelluri, Marshall C Strother, Benjamin L Taylor, Jay D Raman, Thomas J Guzzo
OBJECTIVE: To evaluate the impact of hospital volume on outcomes of robot-assisted partial nephrectomy (RAPN). MATERIALS AND METHODS: Patients with renal cell carcinoma who underwent RAPN between 2010 and 2013 were identified in the National Cancer Database. Hospital yearly RAPN volume was categorized into five groups by most closely sorting patients into five groups of equal size (quintiles): very low, low, medium, high, and very high. Outcomes included 30-day mortality, 90-day mortality, open conversion, prolonged length of stay (PLOS, >3 days), 30-day readmission, and positive surgical margin (PSM) rates...
December 12, 2017: BJU International
https://www.readbyqxmd.com/read/29231738/affinity-purification-and-single-molecule-analysis-of-integral-membrane-proteins-from-crude-cell-membrane-preparations
#11
Anders Oskar Lundgren, Björn Johansson Fast, Stephan Block, Björn Agnarsson, Erik Reimhult, Anders Gunnarsson, Fredrik Höök
The function of integral membrane proteins is critically dependent on their naturally surrounding lipid membrane. Detergent-solubilized and purified membrane proteins are therefore often reconstituted into cell-membrane mimics and analysed for their function with single-molecule microscopy. Expansion of this approach towards a broad range of pharmaceutically interesting drug targets and biomarkers however remains hampered by the fact that these proteins have low expression levels, and that detergent solubilisation and reconstitution often cause protein conformational changes and loss of membrane-specific co-factors, which may impair protein function...
December 12, 2017: Nano Letters
https://www.readbyqxmd.com/read/29228587/chronic-myeloid-leukemia-progenitor-cells-require-autophagy-when-leaving-hypoxia-induced-quiescence
#12
Angela Ianniciello, Pierre-Yves Dumas, Claire Drullion, Amélie Guitart, Arnaud Villacreces, Yan Peytour, Jean Chevaleyre, Philippe Brunet de la Grange, Isabelle Vigon, Vanessa Desplat, Muriel Priault, Persio Dello Sbarba, Zoran Ivanovic, François-Xavier Mahon, Jean-Max Pasquet
Albeit tyrosine kinase inhibitors anti-Abl used in Chronic Myeloid Leukemia (CML) block the deregulated activity of the Bcr-Abl tyrosine kinase and induce remission in 90% of patients, they do not eradicate immature hematopoietic compartments of leukemic stem cells. To elucidate if autophagy is important for stem cell survival and/or proliferation, we used culture in low oxygen concentration (0.1% O2 for 7 days) followed back by non-restricted O2 supply (normoxic culture) to mimic stem cell proliferation and commitment...
November 14, 2017: Oncotarget
https://www.readbyqxmd.com/read/29228265/complete-motif-analysis-of-sequence-requirements-for-translation-initiation-at-non-aug-start-codons
#13
Alexander J Diaz de Arce, William L Noderer, Clifford L Wang
The initiation of mRNA translation from start codons other than AUG was previously believed to be rare and of relatively low impact. More recently, evidence has suggested that as much as half of all translation initiation utilizes non-AUG start codons, codons that deviate from AUG by a single base. Furthermore, non-AUG start codons have been shown to be involved in regulation of expression and disease etiology. Yet the ability to gauge expression based on the sequence of a translation initiation site (start codon and its flanking bases) has been limited...
December 8, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/29228002/comparative-cell-cycle-transcriptomics-reveals-synchronization-of-developmental-transcription-factor-networks-in-cancer-cells
#14
Johan Boström, Zuzana Sramkova, Alena Salašová, Helena Johard, Diana Mahdessian, Radek Fedr, Carolyn Marks, Jiřina Medalová, Karel Souček, Emma Lundberg, Sten Linnarsson, Vítězslav Bryja, Petra Sekyrova, Mikael Altun, Michael Andäng
The cell cycle coordinates core functions such as replication and cell division. However, cell-cycle-regulated transcription in the control of non-core functions, such as cell identity maintenance through specific transcription factors (TFs) and signalling pathways remains unclear. Here, we provide a resource consisting of mapped transcriptomes in unsynchronized HeLa and U2OS cancer cells sorted for cell cycle phase by Fucci reporter expression. We developed a novel algorithm for data analysis that enables efficient visualization and data comparisons and identified cell cycle synchronization of Notch signalling and TFs associated with development...
2017: PloS One
https://www.readbyqxmd.com/read/29226566/probing-the-importance-of-clonality-single-cell-subcloning-of-clonally-derived-cho-cell-lines-yields-widely-diverse-clones-differing-in-growth-productivity-and-product-quality
#15
Peggy Ko, Shahram Misaghi, Zhilan Hu, Dejin Zhan, Joni Tsukuda, Mandy Yim, Mark Sanford, David Shaw, Masaru Shiratori, Brad Snedecor, Michael Laird, Amy Shen
In the past few decades, a large variety of therapeutic antibodies and proteins have been expressed in Chinese hamster ovary (CHO) cells. This mammalian expression system is robust, scalable, relatively inexpensive, and importantly allows for post-translational modifications that are important for some therapeutic proteins. Historically, CHO cell lines were derived from colonies of cells grown in semi-solid or liquid plates using either serum-containing or serum-free media. Current advancements in cell sorting and imaging technologies have allowed for isolating and imaging single cell progenitors at the seeding step, significantly increasing the probability of isolating clonally-derived cell lines...
December 11, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/29226516/sost-an-lngfr-target-inhibits-the-osteogenic-differentiation-of-rat-ectomesenchymal-stem-cells
#16
Gang Li, Junyu Liu, Manzhu Zhao, Yingying Wang, Kun Yang, Chang Liu, Yong Xiao, Xiujie Wen, Luchuan Liu
OBJECTIVES: The aim of this study was to investigate whether sclerostin (SOST) regulates the osteogenic differentiation of rat ectomesenchymal stem cells (EMSCs) and whether SOST and low-affinity nerve growth factor receptor (LNGFR) regulate the osteogenic differentiation of EMSCs. MATERIALS AND METHODS: EMSCs were isolated from embryonic facial processes from an embryonic 12.5-day (E12.5d) pregnant Sprague-Dawley rat. LNGFR+ EMSCs and LNGFR- EMSCs were obtained by fluorescence-activated cell sorting and were subsequently induced to undergo osteogenic differentiation in vitro...
December 10, 2017: Cell Proliferation
https://www.readbyqxmd.com/read/29226183/in-vitro-homeostatic-proliferation-of-human-cd8-t-cells
#17
Hossam A Abdelsamed, Caitlin C Zebley, Ben Youngblood
Long-lived T-cell-mediated immunity requires persistence of memory T cells in an antigen-free environment while also maintaining a heightened capacity to recall effector functions. Such antigen-independent homeostatic proliferation is mediated in part by the common gamma-chain cytokines IL-7 and IL-15. To further explore the mechanisms governing maintenance of effector functions in long-lived memory T cells during antigen-independent proliferation, human naïve and memory CD8 T cells can be sorted from peripheral blood mononuclear cells (PBMCs), labeled with the proliferation-tracking dye carboxyfluorescein succinimidyl ester (CFSE), and then purified based on their levels of cell division...
November 20, 2017: Bio-protocol
https://www.readbyqxmd.com/read/29225785/golgi-structure-formation-function-and-post-translational-modifications-in-mammalian-cells
#18
REVIEW
Shijiao Huang, Yanzhuang Wang
The Golgi apparatus is a central membrane organelle for trafficking and post-translational modifications of proteins and lipids in cells. In mammalian cells, it is organized in the form of stacks of tightly aligned flattened cisternae, and dozens of stacks are often linked laterally into a ribbon-like structure located in the perinuclear region of the cell. Proper Golgi functionality requires an intact architecture, yet Golgi structure is dynamically regulated during the cell cycle and under disease conditions...
2017: F1000Research
https://www.readbyqxmd.com/read/29224352/functional-characterization-of-human-peptide-histidine-transporter-1-in-stably-transfected-mdck-cells
#19
Feifeng Song, Yongjun Hu, Yuqing Wang, David E Smith, Huidi Jiang
The proton-coupled oligopeptide transporter PHT1 (SLC15A4), which facilitates cross-membrane transport of histidine and small peptides from inside the endosomes or lysosomes to cytosol, plays an important role in intracellular peptide homeostasis and innate immune response. However, it remains a challenge to elucidate functional properties of the PHT1 transporter because of its subcellular localization. The purpose of this study was to re-sort hPHT1 protein from the subcellular to outer cell membrane of MDCK cells stably transfected with human PHT1 mutants, and to characterize its functional activity in these cells...
December 11, 2017: Molecular Pharmaceutics
https://www.readbyqxmd.com/read/29224141/tagmentation-based-library-preparation-for-low-dna-input-whole-genome-bisulfite-sequencing
#20
Dieter Weichenhan, Qi Wang, Andrew Adey, Stephan Wolf, Jay Shendure, Roland Eils, Christoph Plass
Aberrations of the DNA methylome contribute to onset and progression of diseases. Whole genome bisulfite sequencing (WGBS) is the only analytical method covering the complete methylome. Alternative methods requiring less DNA than WGBS analyze only a minor portion of the methylome and do not cover important regulatory features like enhancers and noncoding RNAs. In tagmentation-based WGBS (TWGBS), several DNA and time-consuming steps of the conventional WGBS library preparation are circumvented by the use of a hyperactive transposase, which simultaneously fragments DNA and appends sequencing adapters...
2018: Methods in Molecular Biology
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