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Mass cytometry

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https://www.readbyqxmd.com/read/29148254/connecting-color-with-assembly-in-the-fluorescent-b-phycoerythrin-protein-complex
#1
Aneika C Leney, Aline Tschanz, Albert J R Heck
Phycoerythrin is the major light-harvesting pigment protein in red algae and is nowadays widely used as a fluorescent probe in biotechnological applications such as flow cytometry and immunofluorescence microscopy. In addition, it has had substantial economic impact due to its potential as a natural food colorant. However, knowledge on the precise molecular composition of phycoerythrin is limited. Here, we use a combination of high resolution native mass spectrometry (MS) and fluorescence spectroscopy to characterize the assembly properties of the B-phycoerythrin protein complex from Porphyridium cruentum...
November 17, 2017: FEBS Journal
https://www.readbyqxmd.com/read/29137329/development-of-a-radiolabeled-caninized-anti-egfr-antibody-for-comparative-oncology-trials
#2
Judit Fazekas-Singer, Neydher Berroterán-Infante, Christina Rami-Mark, Monika Dumanic, Miroslawa Matz, Michael Willmann, Fritz Andreae, Josef Singer, Wolfgang Wadsak, Markus Mitterhauser, Erika Jensen-Jarolim
Due to large homology of human and canine EGFR, dogs suffering from spontaneous EGFR+ cancer can be considered as ideal translational models. Thereby, novel immunotherapeutic compounds can be developed for both human and veterinary patients. This study describes the radiolabeling of a canine anti-EGFR IgG antibody (can225IgG) with potential diagnostic and therapeutic value in comparative clinical settings. Can225IgG was functionalized with DTPA for subsequent chelation with the radionuclide (99m)Tc. Successful coupling of 10 DTPA molecules per antibody on average was proven by significant mass increase in MALDI-TOF spectroscopy, gel electrophoresis and immunoblots...
October 10, 2017: Oncotarget
https://www.readbyqxmd.com/read/29135861/in-vitro-and-in-vivo-antimelanoma-effect-of-ethyl-ester-cyclohexyl-analog-of-ethylenediamine-dipropanoic-acid
#3
Andjelka M Isakovic, Sasa M Petricevic, Slavica M Ristic, Dusan M Popadic, Tamara K Kravic-Stevovic, Nevena S Zogovic, Jelena M Poljarevic, Tatjana V Zivanovic Radnic, Tibor J Sabo, Aleksandra J Isakovic, Ivanka D Markovic, Vladimir S Trajkovic, Sonja T Misirlic-Dencic
Melanoma, an aggressive skin tumor with high metastatic potential, is associated with high mortality and increasing morbidity. Multiple available chemotherapeutic and immunotherapeutic modalities failed to improve survival in advanced disease, and the search for new agents is ongoing. The aim of this study was to investigate antimelanoma effects of O,O-diethyl-(S,S)-ethylenediamine-N,N'di-2-(3-cyclohexyl) propanoate dihydrochloride (EE), a previously synthesized and characterized organic compound. Mouse melanoma B16 cell viability was assessed using acid phosphatase, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, sulforhodamine B, and lactate dehydrogenase assays...
November 13, 2017: Melanoma Research
https://www.readbyqxmd.com/read/29133210/high-speed-flow-cytometry-allows-the-detection-of-circulating-endothelial-cells-in-hemangioblastoma-patients
#4
Sara De Biasi, Lara Gibellini, Alberto Feletti, Giacomo Pavesi, Elena Bianchini, Domenico Lo Tartaro, Simone Pecorini, Anna De Gaetano, Rosalberta Pullano, Milena Nasi, Marcello Pinti, Andrea Cossarizza
Circulating endothelial cells (CECs) detach from the intima monolayer after endothelial damages. Their circulating endothelial progenitors (CEPs) represent less than 0.01% of nucleated blood cells. Increased levels of CECs and CEPs have been detected in patients with several types of cancer, suggesting that they could be a useful blood-based marker for detecting a tumor, or for monitoring its clinical course. However, their routine monitoring is time consuming and technically challenging. Here, we present a flow cytometry method for quantifying such cells in a cohort of patients with hemangioblastoma (HB)...
November 10, 2017: Methods: a Companion to Methods in Enzymology
https://www.readbyqxmd.com/read/29128078/applications-of-mass-cytometry-in-clinical-medicine-the-promise-and-perils-of-clinical-cytof
#5
REVIEW
Gregory K Behbehani
Mass cytometry is a novel technology similar to flow cytometry in which antibodies are tagged with heavy metal molecules rather than fluorophores and then detected with time-of-flight mass spectrometry. This enables measurement of up to 50 simultaneous parameters with no autofluorescent background and little or no spillover or required compensation. Mass cytometry has tremendous potential for the analysis of highly complex clinical samples for the diagnosis and monitoring of malignant and autoimmune disorders...
December 2017: Clinics in Laboratory Medicine
https://www.readbyqxmd.com/read/29125132/chitosan-inhibits-platelet-mediated-clot-retraction-increases-platelet-derived-growth-factor-release-and-increases-residence-time-and-bioactivity-of-platelet-rich-plasma-in-vivo
#6
Gabrielle Deprés-Tremblay, Anik Chevrier, Nicolas Tran-Khanh, Monica Nelea, Michael D Buschmann
Platelet-rich plasma (PRP) has been used to treat different orthopedic conditions, however, the clinical benefits of using PRP remain uncertain. Chitosan (CS)-PRP implants have been shown to improve meniscus, rotator cuff and cartilage repair in pre-clinical models. The purpose of this current study was to investigate in vitro and in vivo mechanisms of action of CS-PRP implants. Freeze-dried formulations containing 1% (w/v) CS (80% degree of deacetylation and number average molar mass 38 kDa), 1% (w/v) trehalose as a lyoprotectant and 42...
November 10, 2017: Biomedical Materials
https://www.readbyqxmd.com/read/29124436/postnatal-deletion-of-%C3%AE-catenin-in-osterix-expressing-cells-is-necessary-for-bone-growth-and-intermittent-pth-induced-bone-gain
#7
Caixia Yu, Miao Xuan, Mingzhu Zhang, Qianqian Yao, Keqin Zhang, Xiuzhen Zhang, Jun Guo, Lige Song
wnt/β-catenin signaling has been shown to influence bone homeostasis and is important for parathyroid hormone (PTH)-induced bone gain. To further understand the role of β-catenin in the early stages of osteoblastic lineage cells for postnatal bone homeostasis and the anabolic actions of PTH on bone, we examined mice with postnatal disruption of β-catenin in osterix-expressing cells (β-catenin KO mice) by mating floxed β-catenin mice with transgenic mice expressing cre under the control of the osterix promoter suppressible by doxycycline...
November 9, 2017: Journal of Bone and Mineral Metabolism
https://www.readbyqxmd.com/read/29118058/induced-pluripotent-stem-cell-ipsc-derived-extracellular-vesicles-are-safer-and-more-effective-for-cardiac-repair-than-ipscs
#8
Marta Adamiak, Guangming Cheng, Sylwia Bobis-Wozowicz, Lin Zhao, Sylwia Kedracka-Krok, Anweshan Samanta, Elzbieta Karnas, Yu-Ting Xuan, Bozena Skupien-Rabian, Xing Chen, Urszula Jankowska, Magdy Girgis, Malgorzata Sekula, Arash Davani, Slawomir Lasota, Robert J Vincent, Michal Sarna, Kathy L Newell, Ou-Li Wang, Nathaniel Dudley, Zbigniew Madeja, Buddhadeb Dawn, Ewa K Zuba-Surma
Rationale: Extracellular vesicles (EVs) are tiny membrane-enclosed droplets released by cells through membrane budding or exocytosis. The myocardial reparative abilities of EVs derived from induced pluripotent stem cells (iPSCs) have not been directly compared with the source iPSCs. Objective: To examine whether iPSC-derived EVs can influence the biological functions of cardiac cells in vitro; and compare the safety and efficacy of iPSC-EVs and iPSCs for cardiac repair in vivo. Methods and Results: Murine iPSCs were generated and EVs isolated from culture supernatants by sequential centrifugation...
November 8, 2017: Circulation Research
https://www.readbyqxmd.com/read/29110464/joule-heating-induced-dispersion-in-open-microfluidic-electrophoretic-cytometry
#9
Julea Vlassakis, Amy E Herr
While protein electrophoresis conducted in capillaries and microchannels offers high-resolution separations, such formats can be cumbersome to parallelize for single-cell analysis. One approach for realizing large numbers of concurrent separations is open microfluidics (i.e., no microchannels). In an open microfluidic device adapted for single-cell electrophoresis, we perform 100s to 1000s of simultaneous separations of endogenous proteins. The microscope slide-sized device contains cells isolated in microwells located in a ∼40 μm polyacrylamide gel...
November 15, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/29107632/keratin-biomaterials-augment-anti-inflammatory-macrophage-phenotype-in-vitro
#10
Michele Waters, Pamela VandeVord, Mark Van Dyke
Tissue regeneration following injury is mediated by macrophage recruitment and differentiation in response to environmental signals. In general, macrophages adopt either a classically (M[IFN-γ, LPS]) or alternatively activated (M[IL-4, IL-13] and M[IL-10]) phenotype. Recent studies have highlighted the importance of alternatively activated macrophages in tissue remodeling and repair as well as the contribution of an imbalance of classically and alternatively activated macrophages to tissue degeneration and disease progression...
October 28, 2017: Acta Biomaterialia
https://www.readbyqxmd.com/read/29097918/gd-3-asparagine-anionic-linear-globular-dendrimer-second-generation-g2-complexes-novel-nanobiohybrid-theranostics
#11
Nasim Hashempour Alamdari, Mahmood Alaei-Beirami, Seyed Ataollah Sadat Shandiz, Hadi Hejazinia, Rahimeh Rasouli, Mostafa Saffari, Seyed Esmaeil Sadat Ebrahimi, Artin Assadi, Mehdi Shafiee Ardestani
Designing a unique theranostic biocompatible, biodegradable, and cost-effective agent which is easy to be synthesized as a biohybrid material was the aim of this study. In this matter, asparagine attached to anionic linear globular dendrimer G2 (as a biocompatible, biodegradable, and cost-effective agent which is negatively charged nanosized and water soluble polymer that outweighs other traditionally used dendrimers) and finally contrast agent (Gd(3+)) was loaded (which made complexes) in synthesized asparagine-dendrimer...
2017: Contrast Media & Molecular Imaging
https://www.readbyqxmd.com/read/29092906/application-and-utility-of-mass-cytometry-in-vaccine-development
#12
REVIEW
Patrick M Reeves, Ann E Sluder, Susan Raju Paul, Anja Scholzen, Satoshi Kashiwagi, Mark C Poznansky
Mass cytometry enables highly multiplexed profiling of cellular immune responses in limited-volume samples, advancing prospects of a new era of systems immunology. The capabilities of mass cytometry offer expanded potential for deciphering immune responses to infectious diseases and to vaccines. Several studies have used mass cytometry to profile protective immune responses, both postinfection and postvaccination, although no vaccine-development program has yet systematically employed the technology from the outset to inform both candidate design and clinical evaluation...
November 1, 2017: FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology
https://www.readbyqxmd.com/read/29078283/humanized-mouse-model-supports-development-function-and-tissue-residency-of-human-natural-killer-cells
#13
Dietmar Herndler-Brandstetter, Liang Shan, Yi Yao, Carmen Stecher, Valerie Plajer, Melanie Lietzenmayer, Till Strowig, Marcel R de Zoete, Noah W Palm, Jie Chen, Catherine A Blish, Davor Frleta, Cagan Gurer, Lynn E Macdonald, Andrew J Murphy, George D Yancopoulos, Ruth R Montgomery, Richard A Flavell
Immunodeficient mice reconstituted with a human immune system represent a promising tool for translational research as they may allow modeling and therapy of human diseases in vivo. However, insufficient development and function of human natural killer (NK) cells and T cell subsets limit the applicability of humanized mice for studying cancer biology and therapy. Here, we describe a human interleukin 15 (IL15) and human signal regulatory protein alpha (SIRPA) knock-in mouse on a Rag2(-/-)Il2rg(-/-) background (SRG-15)...
October 25, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/29073617/preimplantation-factor-pif-promotes-hla-g-e-f-c-expression-in-jeg-3-choriocarcinoma-cells-and-endogenous-progesterone-activity
#14
Miya Soukaina Hakam, Jose Maria Miranda-Sayago, Soren Hayrabedyan, Krassimira Todorova, Patrick Simon Spencer, Asma Jabeen, Eytan R Barnea, Nelson Fernandez
BACKGROUND/AIMS: Pregnancy success requires mandatory maternal tolerance of the semi/ allogeneic embryo involving embryo-derived signals. Expression levels of PreImplantation Factor (PIF), a novel peptide secreted by viable embryos, correlate with embryo development, and its early detection in circulation correlates with a favourable pregnancy outcome. PIF enhances endometrial receptivity to promote embryo implantation. Via the p53 pathway, it increases trophoblast invasion, improving cell survival / immune privilege...
October 27, 2017: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/29071674/mass-cytometry-assays-for-antigen-specific-t-cells-using-cytof
#15
Dongxia Lin, Holden T Maecker
T Cells specific for a single antigen tend to be rare, even after expansion of memory cells. They are commonly detected by in vitro stimulation with peptides or protein, followed by staining for intracellular cytokines. In this protocol, we use CyTOF(®) mass cytometry to collect single-cell data on a large number of cytokines/chemokines, as well as cell-surface proteins that characterize T cells and other immune cells. We also include a method for magnetic bead enrichment of antigen-stimulated T cells, based on their expression of CD154 and CD69...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29070130/-establishment-of-a-myelodysplastic-syndrome-transformed-to-leukemia-cell-line-stably-expressing-green-fluorescent-protein
#16
Lin Zhuang, Yan Ma, Qian Wang, Xiao-Ping Xu, Jing Zhang, Chen Zhu, Jing-Wen Gu, Wen-Jiao Qin
OBJECTIVE: To establish a myelodysplastic syndrome transformed to leukemia cell line stably expressing green fluorescent protein (GFP), and to evaluate its biological characteristics and applications. METHODS: SKM-1 cells were transfected by lentiviral particles with vector of GFP. The GFP positive single cell clone was isolated by limiting dilution and continued being cultured. The cells were injected into mice subcutaneously and were screened in vivo. Then SKM-1/GFP cells were obtained after tumour plaque was separated and cultivated...
October 2017: Zhongguo Shi Yan Xue Ye Xue za Zhi
https://www.readbyqxmd.com/read/29064419/ni-ii-complexes-with-schiff-base-ligands-preparation-characterization-dna-protein-interaction-and-cytotoxicity-studies
#17
Hui Yu, Wei Zhang, Qing Yu, Fu-Ping Huang, He-Dong Bian, Hong Liang
In this study, two Ni(II) complexes, namely [Ni(HL1)₂(OAc)₂] (1) and [Ni(L2)₂] (2) (where HL1 and HL2 are (E)-1-((1-(2-hydroxyethyl)-1H-pyrazol-5-ylimino)methyl)-naphthalen-2-ol) and (E)-ethyl-5-((2-hydroxynaphthalen-1-yl)methyleneamino)-1-methyl-1H-pyrazole-4-carboxylate, respectively), were synthesized and characterized by X-ray crystallography, Electrospray Ionization Mass Spectrometry (ESI-MS), elemental analysis, and IR. Their uptake in biological macromolecules and cancer cells were preliminarily investigated through electronic absorption (UV-Vis), circular dichroism (CD) and fluorescence quenching measurements...
October 24, 2017: Molecules: a Journal of Synthetic Chemistry and Natural Product Chemistry
https://www.readbyqxmd.com/read/29061960/single-cell-mass-cytometry-and-transcriptome-profiling-reveal-the-impact-of-graphene-on-human-immune-cells
#18
Marco Orecchioni, Davide Bedognetti, Leon Newman, Claudia Fuoco, Filomena Spada, Wouter Hendrickx, Francesco M Marincola, Francesco Sgarrella, Artur Filipe Rodrigues, Cécilia Ménard-Moyon, Gianni Cesareni, Kostas Kostarelos, Alberto Bianco, Lucia G Delogu
Understanding the biomolecular interactions between graphene and human immune cells is a prerequisite for its utilization as a diagnostic or therapeutic tool. To characterize the complex interactions between graphene and immune cells, we propose an integrative analytical pipeline encompassing the evaluation of molecular and cellular parameters. Herein, we use single-cell mass cytometry to dissect the effects of graphene oxide (GO) and GO functionalized with amino groups (GONH2) on 15 immune cell populations, interrogating 30 markers at the single-cell level...
October 24, 2017: Nature Communications
https://www.readbyqxmd.com/read/29054027/sapofectosid-ensuring-non-toxic-and-effective-dna-and-rna-delivery
#19
Simko Sama, Gerold Jerz, Peter Schmieder, Eric Woith, Matthias F Melzig, Alexander Weng
Different methods are being deployed for non-viral DNA/RNA delivery. However non-viral formulations for DNA/RNA-delivery are often accompanied by severe toxicity and thus low efficiency. Particular costly cell culture media are required as well. Here we introduce sapofection as a valuable enhancing method for non-viral DNA/RNA delivery. Sapofection is based on the application of DNA/RNA nanoplexes and sapofectosid, a plant derived natural transfection reagent. Sapofectosid was produced from plant raw material by chromatographic methods and characterized by tandem mass spectrometry and intensive one and two dimensional NMR-spectroscopy...
October 17, 2017: International Journal of Pharmaceutics
https://www.readbyqxmd.com/read/29041935/a-long-non-coding-rna-hottip-expression-is-associated-with-disease-progression-and-predicts-outcome-in-small-cell-lung-cancer-patients
#20
Yanqin Sun, Yuanyuan Zhou, Yifeng Bai, Qiongyao Wang, Jiarong Bao, Yingshan Luo, Ying Guo, Linlang Guo
BACKGROUND: Despite progress in treatment of small cell lung cancer (SCLC), the biology of the tumor still remains poorly understood. Recently, we globally investigated the contributions of lncRNA in SCLC with a special focus on sponge regulatory network. Here we report lncRNA HOTTIP, which is specifically amplified in SCLC, is associated with SCLC proliferation and poor prognosis of patients. METHODS: RT-qPCR was used to investigate the expression of HOTTIP in SCLC tissues and cell lines...
October 17, 2017: Molecular Cancer
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