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https://www.readbyqxmd.com/read/28655167/identification-of-transcript-regulatory-patterns-in-cell-differentiation
#1
Arief Gusnanto, John Paul Gosling, Christopher Pope
Motivation: Studying transcript regulatory patterns in cell differentiation is critical in understanding its complex nature of the formation and function of different cell types. This is done usually by measuring gene expression at different stages of the cell differentiation. However, if the gene expression data available are only from the mature cells, we have some challenges in identifying transcript regulatory patterns that govern the cell differentiation. Results: We propose to exploit the information of the lineage of cell differentiation in terms of correlation structure between cell types...
June 24, 2017: Bioinformatics
https://www.readbyqxmd.com/read/28654657/hek293t-cell-lines-defective-for-o-linked-glycosylation
#2
James M Termini, Zachary A Silver, Bryony Connor, Aristotelis Antonopoulos, Stuart M Haslam, Anne Dell, Ronald C Desrosiers
Here we describe derivatives of the HEK293T cell line that are defective in their ability to generate mucin-type O-linked glycosylation. Using CRISPR/Cas9 and a single-cell GFP-sorting procedure, the UDP-galactose-4-epimerase (GALE), galactokinase 1 (GALK1), and galactokinase 2 (GALK2) genes were knocked out individually and in combinations with greater than 90% of recovered clones having the desired mutations. Although HEK293T cells are tetraploid, we found this approach to be an efficient method to target and disrupt all 4 copies of the target gene...
2017: PloS One
https://www.readbyqxmd.com/read/28654217/dual-labelled-antibodies-for-flow-and-mass-cytometry-a-new-tool-for-cross-platform-comparison-and-enrichment-of-target-cells-for-mass-cytometry
#3
EDITORIAL
Sabine Baumgart, Axel R Schulz, Anette Peddinghaus, Silke Stanislawiak, Sarah Gillert, Heike Hirseland, Susanne Krauthäuser, Christian Dose, Henrik E Mei, Andreas Grützkau
Antibody conjugates applicable in both conventional flow and mass cytometry would offer interesting options for cross-platform comparison, as well as the enrichment of rare target cells by conventional flow cytometry (FC) sorting prior to deep phenotyping by mass cytometry (MC). Here, we introduce a simple method to generate dual fluorochrome/metal-labelled antibodies by consecutive orthogonal labelling. First we compared different fluorochrome-conjugated antibodies specific for CD4, such as FITC, Vio667, VioGreen or VioBlue for their compatibility with the conventional secondary MAXPAR(®) labelling protocol...
June 27, 2017: European Journal of Immunology
https://www.readbyqxmd.com/read/28654052/flow-sorting-and-exome-sequencing-of-the-reed-sternberg-cells-of-classical-hodgkin-lymphoma
#4
Jonathan B Reichel, Jason McCormick, Jonathan R Fromm, Olivier Elemento, Ethel Cesarman, Mikhail Roshal
The Hodgkin Reed-Sternberg cells of classical Hodgkin lymphoma are sparsely distributed within a background of inflammatory lymphocytes and typically comprise less than 1% of the tumor mass. Material derived from bulk tumor contains tumor content at a concentration insufficient for characterization. Therefore, fluorescence activated cell sorting using eight antibodies, as well as side- and forward-scatter, is described here as a method of rapidly separating and concentrating with high purity thousands of HRS cells from the tumor for subsequent study...
June 10, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28652308/b10-cells-alleviate-periodontal-bone-loss-in-experimental-periodontitis
#5
Yuhua Wang, Xiaoqian Yu, Jiang Lin, Yang Hu, Qian Zhao, Toshihisa Kawai, Martin A Taubman, Xiaozhe Han
B10 cells can regulate inflammatory responses in the innate immunity. Toll-like receptors (TLRs) play an important role in B cell-mediated immune responses in periodontal disease. This study was to determine the effects of TLR-activated B10 cells on periodontal bone loss in experimental periodontitis. Spleen B cells isolated from C57BL/6J mice were cultured with P. gingivalis LPS and cytosine-phospho-guanine oligodeoxynucleotides (CpG) for 48 hours. B10-enriched CD1d(hi)CD5(+) B cells were sorted by flow cytometry and were adoptively transferred to recipient mice through tail vein injection...
June 26, 2017: Infection and Immunity
https://www.readbyqxmd.com/read/28652265/a-massively-parallel-fluorescence-assay-to-characterize-the-effects-of-synonymous-mutations-on-tp53-expression
#6
Geetha Bhagavatula, Matthew S Rich, David L Young, Maximillian Marin, Stanley Fields
Although synonymous mutations can affect gene expression, they have generally not been considered in genomic studies that focus on mutations that increase the risk of cancer. However, mounting evidence implicates some synonymous mutations as driver mutations in cancer. Here a massively parallel assay, based on cell sorting of a reporter containing a segment of p53 fused to green fluorescent protein (GFP), was used to measure the effects of nearly all synonymous mutations in exon 6 of TP53. In this reporter context, several mutations within the exon caused strong expression changes including mutations that may cause potential gain or loss of function...
June 26, 2017: Molecular Cancer Research: MCR
https://www.readbyqxmd.com/read/28651892/construction-of-a-recombinant-full-length-membrane-associated-igg-library
#7
Zehua Sun, Shiqiang Lu, Zheng Yang, Jingjing Li, Mei-Yun Zhang
HIV/AIDS has become a global pandemic. Development of an effective HIV-1 vaccine eliciting broadly neutralizing monoclonal antibodies (bnmAbs) remains a big challenge. Before an effective vaccine comes out, passive treatment for prevention and protection of HIV-1 infection may alleviate the burden caused by the pandemic. Numerous bnmAbs have been isolated against different epitopes in HIV-1 envelope glycoprotein via phage/yeast display, EBV-immortalization, single cell sorting and micro neutralization. Recombinant antibody library with extended diversity and enlarged size of units are applicable by phage/yeast display and mammalian cell display for monoclonal antibody isolation...
June 23, 2017: Virus Research
https://www.readbyqxmd.com/read/28650531/cover-image-volume-114-number-8-august-2017
#8
Xuan Droz, Niamh Harraghy, Etienne Lançon, Valérie Le Fourn, David Calabrese, Thierry Colombet, Pascal Liechti, Amar Rida, Pierre-Alain Girod, Nicolas Mermod
Cover Legend The cover image, by Xuan Droz et al., is based on the Article Automated microfluidic sorting of mammalian cells labeled with magnetic microparticles for those that efficiently express and secrete a protein of interest, DOI: 10.1002/bit.26270.
August 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28650530/microfluidic-based-process-to-sort-cells-efficiently-secreting-a-protein
#9
(no author information available yet)
No abstract text is available yet for this article.
August 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28645775/altered-populations-of-natural-killer-cells-cytotoxic-t-lymphocytes-and-regulatory-t-cells-in-major-depressive-disorder-association-with-sleep-disturbance
#10
Hideo Suzuki, Jonathan Savitz, T Kent Teague, Siva K Gandhapudi, Chibing Tan, Masaya Misaki, Brett A McKinney, Michael R Irwin, Wayne C Drevets, Jerzy Bodurka
A subset of individuals with major depressive disorder (MDD) have impaired adaptive immunity characterized by a greater vulnerability to viral infection and a deficient response to vaccination along with a decrease in the number and/or activity of T cells and natural killer cells (NKC). Nevertheless, it remains unclear which specific subsets of lymphocytes are altered in MDD, a shortcoming we address here by utilizing an advanced fluorescence-activated cell sorting (FACS) method that allows for the differentiation of important functionally-distinct lymphocyte sub-populations...
June 20, 2017: Brain, Behavior, and Immunity
https://www.readbyqxmd.com/read/28642819/adoptive-cell-therapy-using-pd-1-myeloma-reactive-t-cells-eliminates-established-myeloma-in-mice
#11
Weiqing Jing, Jill A Gershan, Grace C Blitzer, Katie Palen, James Weber, Laura McOlash, Matthew Riese, Bryon D Johnson
BACKGROUND: Adoptive cellular therapy (ACT) with cancer antigen-reactive T cells following lymphodepletive pre-conditioning has emerged as a potentially curative therapy for patients with advanced cancers. However, identification and enrichment of appropriate T cell subsets for cancer eradication remains a major challenge for hematologic cancers. METHODS: PD-1(+) and PD-1(-) T cell subsets from myeloma-bearing mice were sorted and analyzed for myeloma reactivity in vitro...
2017: Journal for Immunotherapy of Cancer
https://www.readbyqxmd.com/read/28641640/-application-of-cd138-immunomagnetic-sorting-myeloma-cells-combined-with-fluorescence-in-situ-hybridization-for-detecting-cytogenetic-abnormalities-of-multiple-myeloma
#12
Lu Gao, Qing Liu, Yan Shi, Hui Dang, Qi He, Zheng Wang, Lin Feng, Ye Li, Xiao-Yan Wang, Na Li, Wen-Jie Song, Yan-Lin Wang, Shu Kong, Jin Lu, Xiao-Jun Huang, Yue-Yun Lai
OBJECTIVE: To investigate the efficiency of direct fluorescence in situ hybridization (D-FISH) versus FISH on CD138 immunomagnetic sorting myeloma cells (MACS-FISH) to detect the cytogenetic abnormalities of multiple myeloma. METHODS: Thirty-one patients with multiple myeloma (MM) were detected by D-FISH and MACS-FISH, using 5 probes, including 1q21, D13S319, RB1, IgH, P53. The IgH rearrangement positive patients were further examined by 3 IgH rearrangement subtype FISH probes including IgH/FGFR3, IgH/MAF and IgH/CCND1...
June 2017: Zhongguo Shi Yan Xue Ye Xue za Zhi
https://www.readbyqxmd.com/read/28641613/-prostaglandin-e2-receptor-4-agonist-promotes-human-cd34-cell-proliferation-in-vitro-by-activating-wnt-%C3%AE-catenin-signaling-pathway
#13
Ya-Qun Wang, Fang-Jie Liu, Hui-Zhen Chen, He-Hua Wang, Wai-Yi Zou, Chang Su, Juan Li, Duo-Rong Xu
OBJECTIVE: To investigate the potential signaling pathway that regulates the proliferation of human CD34(+) cells stimulated by prostaglandin E2 receptor 4 agonist (EP4A) in vitro. METHODS: Twenty samples of peripheral blood containing stem cells were collected from the G-CSF mobilized healthy donors in our department of hematology. Human CD34(+) cells were isolated by magnetic activated cell sorting (MACS) microbeads kit. The Cell Counting Kit-8 (CCK8) assay was used to determine the optimal concentration and time of EP4A to promote human CD34(+) cell proliferation in vitro...
June 2017: Zhongguo Shi Yan Xue Ye Xue za Zhi
https://www.readbyqxmd.com/read/28640924/correction-accuracy-of-the-fluorescence-activated-cell-sorting-assay-for-the-aquaporin-4-antibody-aqp4-ab-comparison-with-the-commercial-aqp4-ab-assay-kit
#14
Jiwon Yang, Sung Min Kim, Yoo-Jin Kim, So Young Cheon, Boram Kim, Kyeong Cheon Jung, Kyung Seok Park
[This corrects the article DOI: 10.1371/journal.pone.0162900.].
2017: PloS One
https://www.readbyqxmd.com/read/28640894/characterization-and-analysis-of-long-non-coding-rna-lncrna-in-in-vitro-and-ex-vivo-derived-cardiac-progenitor-cells
#15
Baron Arnone, Jake Y Chen, Gangjian Qin
Recent advancements in cell-based therapies for the treatment of cardiovascular disease (CVD) show continuing promise for the use of transplanted stem and cardiac progenitor cells (CPCs) to promote cardiac restitution. However, a detailed understanding of the molecular mechanisms that control the development of these cells remains incomplete and is critical for optimizing their use in such therapy. Long non-coding (lnc) RNA has recently emerged as a crucial class of regulatory molecules involved in directing a variety of critical biological processes including development, homeostasis and disease...
2017: PloS One
https://www.readbyqxmd.com/read/28640401/two-stage-approach-for-risk-estimation-of-fetal-trisomy-21-and-other-aneuploidies-using-computational-intelligence-systems
#16
A C Neocleous, A Syngelaki, K H Nicolaides, C N Schizas
OBJECTIVE: To estimate the risk for fetal trisomy 21 (T21) and other chromosomal abnormalities at 11-13 week's gestation using computational intelligence classification methods. METHODS: As a first step, we train the artificial neural networks with 72054 euploid pregnancies, 295 cases of T21 and 305 of other chromosomal abnormalities (OCA). Then, we sort the cases into two categories of "no-risk" and "risk". The cases of "no-risk" are no further examined, while the cases with "risk" are forwarded in Stage 2 for further examination where we classify them in three types of risk, namely "no-risk", "moderate-risk" and "high-risk"...
June 22, 2017: Ultrasound in Obstetrics & Gynecology
https://www.readbyqxmd.com/read/28640302/influence-of-substituents-on-dna-and-protein-binding-of-cyclometalated-ir-iii-complexes-and-anticancer-activity
#17
Sujay Mukhopadhyay, Roop Shikha Singh, Rajendra Prasad Paitandi, Gunjan Sharma, Biplob Koch, Daya Shankar Pandey
Synthesis of terpyridyl based ligands 3-([2,2':6',2''-terpyridin]-4'-yl)-7-methoxy-2-(methylthio)-quinolone, (L1); 3-([2,2':6',2''-terpyridin]-4'-yl)-6-methoxyquinolin-2(1H)-one, (L2); 3-([2,2'-:6',2''-terpyridin]-4'-yl)-6-methylquinolin-2(1H)-one (L3) and cyclometalated iridium(iii) complexes [[Ir(ppy)2L1](+)PF6(-) (1), [Ir(ppy)2L2](+)PF6(-) (2), [Ir(ppy)2L3](+)PF6(-) (3) (2-phenylpyridine = Hppy)] involving these ligands has been described. The ligands L1-L3 and complexes 1-3 have been thoroughly characterized by elemental analyses, spectral studies (IR, (1)H, (13)C NMR, UV/vis and fluorescence) ESI-MS, and the structure of 3 has been unambiguously authenticated by single crystal X-ray analyses...
June 22, 2017: Dalton Transactions: An International Journal of Inorganic Chemistry
https://www.readbyqxmd.com/read/28637924/vp4-and-vp7-specific-antibodies-mediate-heterotypic-immunity-to-rotavirus-in-humans
#18
Nitya Nair, Ningguo Feng, Lisa K Blum, Mrinmoy Sanyal, Siyuan Ding, Baoming Jiang, Adrish Sen, John M Morton, Xiao-Song He, William H Robinson, Harry B Greenberg
Human rotaviruses (RVs) are the leading cause of severe diarrhea in young children worldwide. The molecular mechanisms underlying the rapid induction of heterotypic protective immunity to RV, which provides the basis for the efficacy of licensed monovalent RV vaccines, have remained unknown for more than 30 years. We used RV-specific single cell-sorted intestinal B cells from human adults, barcode-based deep sequencing of antibody repertoires, monoclonal antibody expression, and serologic and functional characterization to demonstrate that infection-induced heterotypic immunoglobulins (Igs) primarily directed to VP5*, the stalk region of the RV attachment protein, VP4, are able to mediate heterotypic protective immunity...
June 21, 2017: Science Translational Medicine
https://www.readbyqxmd.com/read/28637895/cd4-cd8%C3%AE-double-positive-t-cells-in-skin-draining-lymph-nodes-respond-to-inflammatory-signals-from-the-skin
#19
Nana H Overgaard, Jazmina L Cruz, Jennifer A Bridge, Hendrik J Nel, Ian H Frazer, Nicole L La Gruta, Antje Blumenthal, Raymond J Steptoe, James W Wells
CD4(+)CD8(+) double-positive (DP), mature, peripheral T cells are readily detectable in a variety of species and tissues. Despite a common association with autoimmune and malignant skin disorders, however, little is understood about their role or function. Herein, we show that DP T cells are readily detectable in the blood, spleen, and peripheral lymph nodes of naïve C57BL/6 mice. DP T cells were also present in Jα18(-/-) and CD1d(-/-) mice, indicating that these cells are not NK-T cells. After skin administration of CASAC adjuvant, but not Quil A adjuvant, both total DP T cells and skin-infiltrating DP T cells increased in number...
June 21, 2017: Journal of Leukocyte Biology
https://www.readbyqxmd.com/read/28637874/direct-demonstration-of-a-neonatal-fc-receptor-fcrn-driven-endosomal-sorting-pathway-for-cellular-recycling-of-albumin
#20
Esben G W Schmidt, Michael L Hvam, Filipa Antunes, Jason Cameron, Dorthe Viuff, Birgitte Andersen, Nanna N Kristensen, Kenneth A Howard
Albumin is the most abundant plasma protein involved in the transport of many compounds, such as fatty acids, bilirubin, and heme. The endothelial cellular neonatal Fc receptor (FcRn) has been suggested to play a central role in maintaining the high albumin plasma levels through a cellular recycling pathway. However, direct mapping of this process is still lacking. This work presents the use of wild type and engineered recombinant albumins with either decreased or increased FcRn affinity in combination with a low or high FcRn-expressing endothelium cell line to clearly define the FcRn involvement, intracellular pathway and kinetics of albumin trafficking by flow cytometry, quantitative confocal microscopy, and an albumin-recycling assay...
June 21, 2017: Journal of Biological Chemistry
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