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Flow cytometry—hematology

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https://www.readbyqxmd.com/read/28231690/downregulation-of-hur-inhibits-the-progression-of-esophageal-cancer-through-interleukin-18
#1
Xiaohui Xu, Cheng Song, Zhihua Chen, Chenxiao Yu, Yi Wang, Yiting Tang, Judong Luo
Purpose: To investigate the effect of HuR downregulation and the potential target genes of HuR on the progression of esophageal squamous cell carcinoma (ESCC). Materials and Methods: In this study, a proteomics assay was used to detect the expression of proteins after HuR downregulation, and a luciferase assay was used to detect the potential presence of a HuR binding site on the 3'-UTR of IL-18. In addition, colony formation assay, MTT, EdU incorporation assay, Western blot, flow cytometry, immunohistochemistry, transwell invasion assay and wound healing assay were used...
February 24, 2017: Cancer Research and Treatment: Official Journal of Korean Cancer Association
https://www.readbyqxmd.com/read/28231550/short-term-particulate-matter-exposure-induces-extracellular-vesicle-release-in-overweight-subjects
#2
Matteo Bonzini, Laura Pergoli, Laura Cantone, Mirjam Hoxha, Andrea Spinazzè, Luca Del Buono, Chiara Favero, Michele Carugno, Laura Angelici, Lucia Broggi, Andrea Cattaneo, Angela C Pesatori, Valentina Bollati
BACKGROUND: Extracellular vesicles (EVs) represent a plausible molecular mechanism linking particulate matter (PM) inhalation to its systemic effects. Microvesicles (MVs) are released from many cell types in response to various stimuli. Increased body mass index (BMI) could modify the response to PM exposure due to enhanced PM uptake and/or an underlying pro-oxidative state. We investigated the relationship between EV release and PM10/PM2.5 exposure in a cohort of 51 volunteers. Subjects were stratified based on their BMI to evaluate whether overweight BMI is a determinant of hypersusceptibility to PM effects...
February 20, 2017: Environmental Research
https://www.readbyqxmd.com/read/28231426/engineering-of-anti-cd133-tri-specific-molecule-capable-of-inducing-nk-expansion-and-driving-antibody-dependent-cell-mediated-cytotoxicity-adcc
#3
Jörg U Schmohl, Martin Felices, Felix Oh, Alexander J Lenvik, Aaron M Lebeau, Jayanth Panyam, Jeffrey S Miller, Daniel A Vallera
Purpose: The selective elimination of cancer stem cells (CSCs) in tumor patients is a crucial goal because CSCs cause drug refractory relapse. To improve the current conventional bispecific immune-engager platform, a 16133 BiKE, consisting of scFvs binding FcγRIII (CD16) on NK cells and CD133 on carcinoma cells, was first synthesized and a modified IL-15 crosslinker capable of stimulating NK effector cells was introduced. Materials and Methods: DNA shuffling and ligation techniques were used to assemble and synthesize the 1615133 trispecific NK cell engager (TriKE)...
February 20, 2017: Cancer Research and Treatment: Official Journal of Korean Cancer Association
https://www.readbyqxmd.com/read/28230780/a3r-phage-and-staphylococcus-aureus-lysate-do-not-induce-neutrophil-degranulation
#4
Jan Borysowski, Ryszard Międzybrodzki, Piotr Wierzbicki, Danuta Kłosowska, Grażyna Korczak-Kowalska, Beata Weber-Dąbrowska, Andrzej Górski
The objective of this study was to evaluate the effects of A3R phage and Staphylococcus aureus lysate obtained after phage infection on neutrophil degranulation. The exocytosis of primary and secondary granules from neutrophils was investigated in vitro in whole blood specimens by flow cytometry based on the expression of specific markers of exocytosis (CD63 for primary granules and CD66b for secondary granules). We found that both A3R and S. aureus lysate had no significant effect on the exocytosis of primary and secondary granules...
February 21, 2017: Viruses
https://www.readbyqxmd.com/read/28230645/clinical-scale-rapid-autologous-bk-virus-specific-t-cell-line-generation-from-kidney-transplant-recipients-with-active-viremia-for-adoptive-immunotherapy
#5
Caroline Lamarche, Julie Orio, Victoria Georges-Tobar, Thomas Pincez, Mathieu Goupil, Amina Dahmani, Cedric Carli C, Ann Brasey, Lambert Busque, Jean-Sébastien Delisle
BACKGROUND: Polyomavirus-associated nephropathy (PVAN) following BK virus reactivation in kidney transplant recipients (KTR) can compromise graft survival. Lowering immunosuppression is the only established approach to prevent or treat PVAN but nonspecifically increasing host immune competence also augments rejection risk. Ex vivo T cell stimulation/expansion offers the possibility to generate BK-specific T cell lines for adoptive immunotherapy. The objective of this study was to develop and characterize a clinical scale protocol to generate BK-specific T cell lines from viremic KTR...
February 23, 2017: Transplantation
https://www.readbyqxmd.com/read/28230642/strategies-to-obtain-diverse-and-specific-human-monoclonal-antibodies-from-transgenic-animals
#6
Marianne Brüggemann, Michael J Osborn, Biao Ma, Roland Buelow
Techniques to obtain large quantities of antigen-specific monoclonal antibodies, mAbs, were first established in the 1970s when Georges Köhler and César Milstein immortalized antibody-producing mouse B-lymphocytes by fusion with myeloma cells (http://www.whatisbiotechnology.org/exhibitions/milstein). Combined with the expression of human antibodies in transgenic animals, this technique allowed upon immunization the generation of highly specific fully human mAbs for therapeutic applications. Apart from being extremely beneficial, mAbs are a huge success commercially...
February 23, 2017: Transplantation
https://www.readbyqxmd.com/read/28230290/a3-adenosine-receptor-agonist-inhibited-survival-of-breast-cancer-stem-cells-via-gli-1-and-erk1-2-pathway
#7
Seyyed Mehdi Jafari, Mojtaba Panjehpour, Mahmoud Aghaei, Hamid Reza Joshaghani, Seyed Ehsan Enderami
Numerous studies have demonstrated the role of A3 adenosine receptor (A3AR) and signaling pathways in the multiple aspects of the tumor. However, there is a little study about the function of A3AR in the biological processes of cancer stem cells (CSCs). CSCs have a critical role in the maintenance and survival of breast cancer. The aim of current study was to investigate the effect of A3AR agonist on breast cancer stem cells (BCSCs). XTT assay showed antiprolierative effect of A3AR agonist (Cl-IB-MECA) on BCSCs...
February 23, 2017: Journal of Cellular Biochemistry
https://www.readbyqxmd.com/read/28230011/changes-of-cd4-t-cell-subsets-after-radiofrequency-ablation-in-lung-cancer-and-its-significance
#8
Wang Shaobin, Xiong Yu, Lin Jiatian, Chen Zaizhong, Dai Luping, Chen Junhui
AIMS: To analyze the changes of CD4+ T-cell subsets following radiofrequency ablation (RFA) in lung cancer and the impact of RFA on the antitumor immunity. SUBJECTS AND METHODS: Flow cytometry was employed to detect CD4+ T-cell subsets in the peripheral blood from 45 healthy controls and 45 lung cancer patients before and after RFA. The correlation between CD4+ T-cell subsets and the clinical characteristics was discussed by comparing the results between different groups...
December 2016: Journal of Cancer Research and Therapeutics
https://www.readbyqxmd.com/read/28229962/overexpression-of-hsa-mir-939-follows-by-ngfr-down-regulation-and-apoptosis-reduction
#9
Fahimeh Hosseini Aghdaei, Bahram M Soltani, Sadat Dokanehiifard, Seyed Javad Mowla, Masoud Soleimani
Neurotrophin receptors play a crucial role in neuronal survival, differentiation and regeneration. Nerve growth factor receptor (NGFR) or P75(NTR) is a neurotrophin receptor that is involved in many pathological conditions including cancers. Genetic factors that are involved in regulation of neurotrophin receptors are under intense investigation. MiRNAs are novel regulators of signalling pathways that are candidates for regulation of neurotrophin receptors. Computational programs predicted that NGFR gene is a bona fide target for hsa-miR- 939...
March 2017: Journal of Biosciences
https://www.readbyqxmd.com/read/28229418/measurement-of-autophagy-by-flow-cytometry
#10
Silvia Zappavigna, Angela Lombardi, Gabriella Misso, Anna Grimaldi, Michele Caraglia
Autophagy activation is characterized by the accumulation of double-membrane autophagic vesicles (autophagosomes) in the cytoplasm. The mere presence of autophagosomes in the cytoplasm does not necessarily indicate an increased level of autophagy, since the blockade of any step downstream of autophagosome formation increases the number of autophagosomes. Therefore, quantitative methods for the detection of cytoplasmic protein turnover should be employed in addition to autophagosome monitoring, to verify increased levels of autophagy...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28229417/a-simplified-and-systematic-method-to-isolate-culture-and-characterize-multiple-types-of-human-dental-stem-cells-from-a-single-tooth
#11
Mohammed Bakkar, Younan Liu, Dongdong Fang, Camille Stegen, Xinyun Su, Murali Ramamoorthi, Li-Chieh Lin, Takako Kawasaki, Nicholas Makhoul, Huan Pham, Yoshinori Sumita, Simon D Tran
This chapter describes a simplified method that allows the systematic isolation of multiple types of dental stem cells such as dental pulp stem cells (DPSC), periodontal ligament stem cells (PDLSC), and stem cells of the apical papilla (SCAP) from a single tooth. Of specific interest is the modified laboratory approach to harvest/retrieve the dental pulp tissue by minimizing trauma to DPSC by continuous irrigation, reduction of frictional heat from the bur rotation, and reduction of the bur contact time with the dentin...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28229416/human-induced-pluripotent-stem-cell-derived-mesenchymal-stem-cells-as-an-individual-specific-and-renewable-source-of-adult-stem-cells
#12
Glen Lester Sequiera, Sekaran Saravanan, Sanjiv Dhingra
This chapter deals with the employment of human-induced pluripotent stem cells (hiPSCs) as a candidate to differentiate into mesenchymal stem cells (MSCs). This would enable to help establish a regular source of human MSCs with the aim of avoiding the problems associated with procuring the MSCs either from different healthy individuals or patients, limited extraction potentials, batch-to-batch variations or from diverse sources such as bone marrow or adipose tissue. The procedures described herein allow for a guided and ensured approach for the regular maintenance of hiPSCs and their subsequent differentiation into MSCs using the prescribed medium...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28229408/isolation-and-in-vitro-characterization-of-epidermal-stem-cells
#13
Kasper S Moestrup, Marianne S Andersen, Kim B Jensen
Colony-forming assays represent prospective methods, where cells isolated from enzymatically dissociated tissues or from tissue cultures are assessed for their proliferative capacity in vitro. Complex tissues such as the epithelial component of the skin (the epidermis) are characterized by a substantial cellular heterogeneity. Analysis of bulk populations of cells by colony-forming assays can consequently be convoluted by a number of factors that are not controlled for in population wide studies. It is therefore advantageous to refine in vitro growth assays by sub-fractionation of cells using flow cytometry...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28229168/cd14-cd16-nonclassical-monocytes-are-associated-with-endothelial-dysfunction-in-patients-with-coronary-artery-disease
#14
Karol Urbanski, Dominik Ludew, Grzegorz Filip, Magdalena Filip, Agnieszka Sagan, Piotr Szczepaniak, Grzegorz Grudzien, Jerzy Sadowski, Barbara Jasiewicz-Honkisz, Tomasz Sliwa, Boguslaw Kapelak, Eilidh McGinnigle, Tomasz Mikolajczyk, Tomasz J Guzik
Endothelial dysfunction and inflammation are key mechanisms of vascular disease. We hypothesised that heterogeneity of monocyte subpopulations may be related to the development of vascular dysfunction in coronary artery disease (CAD). Therefore, we examined the relationships between monocyte subsets (CD14(++)CD16(-) "classical - Mon1", CD14(++)CD16(+) "intermediate - Mon2" and CD14(+)CD16(++) "nonclassical - Mon3"), endothelial function and risk factor profiles in 130 patients with CAD undergoing coronary artery bypass grafting...
February 23, 2017: Thrombosis and Haemostasis
https://www.readbyqxmd.com/read/28229135/the-colonic-metabolites-dihydrocaffeic-acid-and-dihydroferulic-acid-are-more-effective-inhibitors-of-in-vitro-platelet-activation-than-their-phenolic-precursors
#15
Gema Baeza, Eva-Maria Bachmair, Sharon Wood, Raquel Mateos, Laura Bravo, Baukje de Roos
Cardiovascular disease (CVD) is the major cause of morbidity and mortality worldwide. The consumption of a healthy diet rich in polyphenols has been inversely associated with the development of CVD. This study evaluated the effects of green coffee bean extract (GCBE) and yerba mate phenolic extract (YMPE), the main phenolic and methylxanthine constituents (5-caffeoylquinic acid, 3,5-dicaffeoylquinic acid, caffeine, and theobromine), and their main metabolites (caffeic acid, ferulic acid, dihydrocaffeic acid (DHCA) and dihydroferulic acid (DHFA)) on platelet activation in vitro...
February 23, 2017: Food & Function
https://www.readbyqxmd.com/read/28229025/effect-of-interleukin-il-35-on-il-17-expression-and-production-by-human-cd4-%C3%A2-t-cells
#16
Kosuke Okada, Takeki Fujimura, Takeshi Kikuchi, Makoto Aino, Yosuke Kamiya, Ario Izawa, Yuki Iwamura, Hisashi Goto, Iichiro Okabe, Eriko Miyake, Yoshiaki Hasegawa, Makio Mogi, Akio Mitani
BACKGROUND: Interleukin (IL)-17 produced by mainly T helper 17 (Th17) cells may play an important destructive role in chronic periodontitis (CP). Thus, anti-inflammatory cytokines, such as IL-35, might have a beneficial effect in periodontitis by inhibiting differentiation of Th17 cells. Th17 differentiation is regulated by the retinoic acid receptor-related orphan receptor (ROR) α (encoded by RORA) and RORγt (encoded by RORC). However, the role of IL-35 in periodontitis is not clear and the effect of IL-35 on the function of Th17 cells is still incompletely understood...
2017: PeerJ
https://www.readbyqxmd.com/read/28228758/a-simple-and-rapid-method-for-quality-control-of-major-histocompatibility-complex-peptide-monomers-by-flow-cytometry
#17
P Anoop Chandran, Sonja Heidu, Henning Zelba, Barbara Schmid-Horch, Hans-Georg Rammensee, Steve Pascolo, Cécile Gouttefangeas
Major histocompatibility complex (MHC) multimers are essential tools in T cell immunomonitoring, which are employed both in basic and clinical research, as well as for assessing clinical samples during therapy. The generation of MHC monomers loaded with synthetic peptides is an elaborate and time-consuming process. It would be beneficial to assess the quality of these monomers prior to downstream applications. In this technical note, we describe a novel flow cytometry-based, cell-free, quick, and robust assay to check the quality of MHC monomers directly after refolding or after long-term storage...
2017: Frontiers in Immunology
https://www.readbyqxmd.com/read/28228489/age-dependent-shift-in-macrophage-polarisation-causes-inflammation-mediated-degeneration-of-enteric-nervous-system
#18
Laren Becker, Linh Nguyen, Jaspreet Gill, Subhash Kulkarni, Pankaj Jay Pasricha, Aida Habtezion
OBJECTIVE: The enteric nervous system (ENS) undergoes neuronal loss and degenerative changes with age. The cause of this neurodegeneration is poorly understood. Muscularis macrophages residing in close proximity to enteric ganglia maintain neuromuscular function via direct crosstalk with enteric neurons and have been implicated in the pathogenesis of GI motility disorders like gastroparesis and postoperative ileus. The aim of this study was to assess whether ageing causes alterations in macrophage phenotype that contributes to age-related degeneration of the ENS...
February 21, 2017: Gut
https://www.readbyqxmd.com/read/28228322/anti-tumour-research-of-recombinant-bcg-using-bzlf1-and-hgm-csf-fusion-genes
#19
Qing-Jie Xue, Yun-Qing Li, Chun-Qing Yang, Ting Chen, Xiu-Zhen Li, Baohua Cheng, Chun-Mei Wang
The random primer Oligo(dT)15 was used in RT-PCR to obtain cDNA from the human granulocyte macrophage colony stimulating factor (hGM-CSF) gene and the Epstein-Barr virus (EBV) gene BZLF1. Then, the sequence splicing overlap extension method was used to obtain a GCBF fusion gene containing a linker sequence that encoded the polypeptide (Gly4Ser)3. The GCBF fusion gene was inserted into pMV261, which was then transformed into competent E. coli DH5 alpha cells, and positive cells were selected based on kanamycin resistance on LB plates...
February 19, 2017: Vaccine
https://www.readbyqxmd.com/read/28228218/-thioridazine-sensitizes-apoptotic-effect-of-trail-in-human-lung-cancer-pc9-cells-through-er-stress-mediated-up-regulation-of-dr5
#20
Juan Li, Yi Wang, Liu Liu, Yuan Yuan, Yangyi Bao
BACKGROUND: Tumor necrosis factor-related apoptosis-inducting ligand (TRAIL) can induce apoptosis of tumor cells, however, various of tumor cells may survive because of resistance to TRAIL-mediated apoptosis. This study is to observe the proliferation inhibition effect of TRAIL sensitized by thioridazine on PC9 cells through endoplasmic reticulum (ER) stress mediated up-regulation of death receptor 5 (DR5) and investigate its mechanism. METHODS: PC9 cells were treated with different concentrations of thioridazine and TRAIL alone or in combination...
February 20, 2017: Zhongguo Fei Ai za Zhi, Chinese Journal of Lung Cancer
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