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Intracellular Flow Cytometry

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https://www.readbyqxmd.com/read/29790576/memory-t-cell-subsets-in-healthy-gingiva-and-periodontitis-tissues
#1
Rangsini Mahanonda, Chantrakorn Champaiboon, Keskanya Subbalekha, Noppadol Sa-Ard-Iam, Arsarn Yongyuth, Benjarat Isaraphithakkul, Pimprapa Rerkyen, Orawan Charatkulangkun, Sathit Pichyangkul
BACKGROUND: In the gingival sulcus, effective and balanced innate and adaptive immune responses against subgingival plaque microbiome are crucial to maintain immune homeostasis. In this study, we investigated the memory T cell subsets in healthy gingiva and periodontitis tissues. METHODS: Anatomical localization of T cells (CD3+ , CD4+ , and CD8+ ) in healthy gingiva and periodontitis tissues were examined immunohistochemically. Subsets of memory T cells from isolated gingival cells were analyzed by flow cytometry using a cocktail of monoclonal antibodies (anti-CD69, anti-CD103, anti-CD45RA, anti-CCR7, anti-CD28, and anti-CD95)...
May 23, 2018: Journal of Periodontology
https://www.readbyqxmd.com/read/29787951/glutathione-triggered-dual-release-of-doxorubicin-and-camptothecin-for-highly-efficient-synergistic-anticancer-therapy
#2
Yiwen Li, Huailin Yang, Jiuxu Yao, Haiyang Yu, Xin Chen, Peng Zhang, Chunsheng Xiao
An amphiphilic biodegradable prodrug (PLG-g-mPEG/CPT) was synthesized by conjugating disulfide-containing camptothecin (CPT) to poly(L-glutamic acid)-graft-methoxy poly(ethylene glycol) (PLG-g-mPEG) through esterification reaction. The amphiphilic prodrugs could self-assemble into micellar nanoparticles and encapsulate doxorubicin (DOX) in aqueous solution at pH 7.4. The treatment of the nanoparticles with reducing glutathione (GSH) at cytosolic concentration (10 mM) significantly promoted the in vitro dual release of DOX and CPT from the micelles...
May 17, 2018: Colloids and Surfaces. B, Biointerfaces
https://www.readbyqxmd.com/read/29787559/peptidoglycan-suppresses-phagocytic-activities-and-apoptosis-of-macrophages-in-colonic-mucosa-tissues-of-crohn-s-disease-patients-and-in-vitro
#3
Yang Jing, Ying Ran, Jing Zhao, Zhe Zhou, Jun Zhang, Yiqi Qian, Zhiqi Yin, Mingfang Zhang, Zongshun Lv, Lu Zhou, Bangmao Wang
BACKGROUND Rac1 signaling plays a crucial role in controlling macrophage functions in CD. Peptidoglycan triggers several intracellular signaling pathways, including activation of Rac1, to regulate the function of macrophage. Suppressed Rac1 signaling in non-inflamed colonic mucosa of Crohn's disease patients has been shown to correlate with increased innate immunity. MATERIAL AND METHODS We examined the effect of peptidoglycan on Rac1 signaling in macrophages and mucosal tissue samples collected from 10 patients with active Crohn's disease and further investigated the effects of peptidoglycan on apoptosis and phagocytic activities of macrophages in vitro...
May 22, 2018: Medical Science Monitor: International Medical Journal of Experimental and Clinical Research
https://www.readbyqxmd.com/read/29786746/luteolin-induces-myelodysplastic-syndrome%C3%A2-derived-cell-apoptosis-via-the-p53%C3%A2-dependent-mitochondrial-signaling-pathway-mediated-by-reactive-oxygen-species
#4
Weimin Dong, Yan Lin, Yang Cao, Yue Liu, Xiaobao Xie, Weiying Gu
Luteolin, a common dietary flavonoid, induces the apoptosis of cells in several types of cancer. However, its role in myelodysplastic syndrome (MDS) and the potential underlying mechanisms remain to be elucidated. To evaluate the potential benefit and underlying mechanisms of luteolin in MDS cells, the viability of SKM‑1 cells and primary bone marrow (PBM) mononuclear cells from patients with intermediate‑ or high‑risk MDS were assessed using a Cell Counting Kit‑8 assay. The apoptotic features of cell morphology were assessed using Wright‑Giemsa staining, DNA fragmentation was analyzed by agarose gel electrophoresis, and the extent of apoptosis was quantified by flow cytometry (FCM)...
May 18, 2018: International Journal of Molecular Medicine
https://www.readbyqxmd.com/read/29783743/cell-propagation-of-cholera-toxin-cta-adp-ribosylating-factor-by-exosome-mediated-transfer
#5
Cristiana Zanetti, Angelo Gallina, Alessia Fabbri, Sofia Parisi, Angela Palermo, Katia Fecchi, Zaira Boussadia, Maria Carollo, Mario Falchi, Luca Pasquini, Maria Luisa Fiani, Massimo Sargiacomo
In this study, we report how the cholera toxin (CT) A subunit (CTA), the enzyme moiety responsible for signaling alteration in host cells, enters the exosomal pathway, secretes extracellularly, transmits itself to a cell population. The first evidence for long-term transmission of CT's toxic effect via extracellular vesicles was obtained in Chinese hamster ovary (CHO) cells. To follow the CT intracellular route towards exosome secretion, we used a novel strategy for generating metabolically-labeled fluorescent exosomes that can be counted by flow cytometry assay (FACS) and characterized...
May 19, 2018: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/29781747/sialylation-pattern-in-lung-epithelial-cell-line-and-siglecs-expression-in-monocytic-thp-1-cells-as-cellular-indicators-of-cigarette-smoke-induced-pathology-in-vitro
#6
Przemyslaw Wielgat, Emil Trofimiuk, Robert Czarnomysy, Adam Holownia, Jan J Braszko
PURPOSE: Cellular response to cigarette smoke (CS) involves activation of recognition receptors resulting in changes in immune status, oxidative stress and cell turnover. We investigated the effects of CS on sialic acid-binding immunoglobulin type lectins (Siglecs) expression and their sialylated ligands in human immune and non-immune cells. METHODS: Human monocytes (THP-1) and epithelial cells (A549) were cultured in CS-conditioned medium (CSM). Expression of Siglec-8 and Siglec-5/Siglec-14 was analysed in THP-1 cells using flow cytometry...
May 21, 2018: Experimental Lung Research
https://www.readbyqxmd.com/read/29781122/protective-effects-of-endothelial-progenitor-cell-derived-extracellular-mitochondria-in-brain-endothelium
#7
Kazuhide Hayakawa, Su Jing Chan, Emiri T Mandeville, Ji Hyun Park, Morgan Bruzzese, Joan Montaner, Ken Arai, Anna Rosell, Eng H Lo
Endothelial progenitor cells (EPCs) have been pursued as a potential cellular therapy for stroke and central nervous system injury. However, their underlying mechanisms remain to be fully defined. Recent experimental studies suggest that mitochondria may be released and transferred between cells. In this proof-of-concept study, we asked whether beneficial effects of EPCs may partly involve a mitochondrial phenomenon as well. First, EPC-derived conditioned medium was collected and divided into supernatant and particle fractions after centrifugation...
May 21, 2018: Stem Cells
https://www.readbyqxmd.com/read/29777675/brain-derived-microparticles-activate-microglia-macrophages-and-induce-neuroinflammation
#8
Hongtao Rong, Yueshan Fan, Mengchen Yang, Baoliang Zhang, Dongdong Sun, Zilong Zhao, Dong Wang, Weidong Fan, Jianhao Wang, Gang Gu, Fanjian Li, Xilei Liu, Chenxu Rao, Huijiao Chen, Yi Wang, Ye Tian, Jianning Zhang
Microparticles are cell fragments derived from damaged cells that are able to present an antigen from the parent cells to other cells to activate intracellular signaling pathways. Microparticles are closely related to the inflammatory response. Brain-derived microparticles (BDMPs) play an important role in brain injury. However, the inflammatory effect of BDMPs on microglia/macrophages remains unclear. The BDMPs were consumed by microglia/macrophages in vivo and in vitro. The BDMPs activated microglia/macrophages and changed their morphology in vitro...
May 16, 2018: Brain Research
https://www.readbyqxmd.com/read/29777275/overexpression-of-abcb4-contributes-to-acquired-doxorubicin-resistance-in-breast-cancer-cells-in-vitro
#9
Jia-Feng Huang, Chun-Jie Wen, Guo-Zhi Zhao, Yi Dai, Ying Li, Lan-Xiang Wu, Hong-Hao Zhou
PURPOSE: Doxorubicin is one of the most active agents in the first-line therapy for metastatic breast cancer, but its utility is partially limited by the frequent emergence of doxorubicin resistance. In this study, we aimed to investigate the role of ATP-binding cassette sub-family B, member 4 (ABCB4) in acquired doxorubicin resistance in breast cancer cells, as well as its potential mechanism. METHODS: In doxorubicin-sensitive and -resistant breast cancer cell lines MCF-7 and MDA-MB-231, the expression levels of ABCB4 were detected using real-time quantitative PCR and Western blot analysis, the DNA methylation and histone acetylation status of ABCB4 gene were investigated by bisulfite-sequencing PCR (BSP) and chromatin immunoprecipitation (ChIP) assays, and the doxorubicin sensitivity and intracellular doxorubicin accumulation were observed using cell cytotoxicity assay and flow cytometry...
May 18, 2018: Cancer Chemotherapy and Pharmacology
https://www.readbyqxmd.com/read/29774078/targeting-intracellular-mmps-efficiently-inhibits-tumor-metastasis-and-angiogenesis
#10
Yaqi Lv, Xiangmei Zhao, Lidan Zhu, Sijia Li, Qingqing Xiao, Wei He, Lifang Yin
Treatment for metastatic cancer is a great challenge throughout the world. Commonly, directed inhibition of extracellular matrix metalloproteinases (MMPs) secreted by cancer cells can reduce metastasis. Here, a novel nanoplatform (HPMC NPs) assembled from hyaluronic acid (HA)-paclitaxel (PTX) prodrug and marimastat (MATT)/β-casein (CN) complexes was established to cure a 4T1 metastatic cancer model via targeting CD44 and intracellular, rather than extracellular, MMPs. Methods: HPMC NPs were prepared by assembling the complexes and prodrug under ultrasonic treatment, which the interaction between them was evaluated by förster resonance energy transfer, circular dichroism and fluorescence spectra...
2018: Theranostics
https://www.readbyqxmd.com/read/29774032/protein-o-glcnac-modification-increases-in-white-blood-cells-after-a-single-bout-of-physical-exercise
#11
Tamás Nagy, Emese Kátai, Viktória Fisi, Tamás Tibor Takács, Antal Stréda, István Wittmann, Attila Miseta
Background: Protein O-linked N -acetylglucosamine (O-GlcNAc) is a dynamic posttranslational modification influencing the function of many intracellular proteins. Recently it was revealed that O-GlcNAc regulation is modified under various stress states, including ischemia and oxidative stress. Aside from a few contradictory studies based on animal models, the effect of exercise on O-GlcNAc is unexplored. Purpose: To evaluate O-GlcNAc levels in white blood cells (WBC) of human volunteers following physical exercise...
2018: Frontiers in Immunology
https://www.readbyqxmd.com/read/29774023/ferritin-h-deficiency-in-myeloid-compartments-dysregulates-host-energy-metabolism-and-increases-susceptibility-to-mycobacterium-tuberculosis-infection
#12
Vineel P Reddy, Krishna C Chinta, Vikram Saini, Joel N Glasgow, Travis D Hull, Amie Traylor, Fernanda Rey-Stolle, Miguel P Soares, Rajhmun Madansein, Md Aejazur Rahman, Coral Barbas, Kievershen Nargan, Threnesan Naidoo, Pratistadevi K Ramdial, James F George, Anupam Agarwal, Adrie J C Steyn
Iron is an essential factor for the growth and virulence of Mycobacterium tuberculosis ( Mtb) . However, little is known about the mechanisms by which the host controls iron availability during infection. Since ferritin heavy chain (FtH) is a major intracellular source of reserve iron in the host, we hypothesized that the lack of FtH would cause dysregulated iron homeostasis to exacerbate TB disease. Therefore, we used knockout mice lacking FtH in myeloid-derived cell populations to study Mtb disease progression...
2018: Frontiers in Immunology
https://www.readbyqxmd.com/read/29773195/measuring-zinc-in-biological-nanovesicles-by-multiple-analytical-approaches
#13
Francesco Piacenza, Antje Biesemeier, Marco Farina, Francesco Piva, Xin Jin, Eleonora Pavoni, Lorenzo Nisi, Maurizio Cardelli, Laura Costarelli, Robertina Giacconi, Andrea Basso, Elisa Pierpaoli, Mauro Provinciali, James C M Hwang, Antonio Morini, Andrea di Donato, Marco Malavolta
Exosomes are nanovesicles known to mediate intercellular communication. Although it is established that zinc ions can act as intracellular signaling factors, the measurement of zinc in circulating nanovesicles has not yet been attempted. Providing evidence of the existence of this zinc fraction and methods for its measurement might be important to advance our knowledge of zinc status and its relevance in diseases. Exosomes from 0.5 ml of either fresh or frozen human plasma were isolated by differential centrifugation...
July 2018: Journal of Trace Elements in Medicine and Biology
https://www.readbyqxmd.com/read/29771573/increased-circulating-proinflammatory-t-lymphocytes-in-children-with-different-forms-of-anterior-uveitis-results-from-a-pilot-study
#14
Karoline Walscheid, Lisa Neekamp, Arnd Heiligenhaus, Toni Weinhage, Carsten Heinz, Dirk Foell
PURPOSE: To characterize peripheral blood T cells in juvenile idiopathic arthritis-associated uveitis (JIAU). METHODS: Blood samples were taken from children with JIAU (n = 18), JIA without ocular involvement (n = 11), idiopathic anterior uveitis (IAU, n = 12), and healthy controls (n = 11). Cells were stained for T cell surface markers, and intracellular cytokine staining was performed after cell stimulation and analyzed by flow cytometry. RESULTS: The Th1/Th2 ratio was increased in JIAU patients...
May 17, 2018: Ocular Immunology and Inflammation
https://www.readbyqxmd.com/read/29771149/preparation-and-evaluation-of-tumour-microenvironment-response-multistage-nanoparticles-for-epirubicin-delivery-and-deep-tumour-penetration
#15
Jialing Dai, Shangcong Han, Fang Ju, Mei Han, Lisa Xu, Ruoyu Zhang, Yong Sun
Poor tumour penetration became a major challenge for the use of nanoparticles in anticancer therapy. To further enhance the tumour penetration efficiency, we developed a tumour-microenvironment-responsive multistage drug delivery system which was formed layer by layer via electrostatic interaction with cationic drug-loaded nanoparticles, hyaluronidase (HAase) and iRGD-modified gelatin (G-iRGD). The drug-loaded nanoparticles were formed by self-assembling mPEG-PDPA-PG and encapsulation with epirubicin (EPI)...
May 17, 2018: Artificial Cells, Nanomedicine, and Biotechnology
https://www.readbyqxmd.com/read/29768981/a-scalable-pipeline-for-high-throughput-flow-cytometry
#16
Aaron C Wilson, Ioannis K Moutsatsos, Gary Yu, Javier J Pineda, Yan Feng, Douglas S Auld
Flow cytometry (FC) provides high-content data for a variety of applications, including phenotypic analysis of cell surface and intracellular markers, characterization of cell supernatant or lysates, and gene expression analysis. Historically, sample preparation, acquisition, and analysis have presented as a bottleneck for running such types of assays at scale. This article will outline the solutions that have been implemented at Novartis which have allowed high-throughput FC to be successfully conducted and analyzed for a variety of cell-based assays...
May 1, 2018: SLAS Discovery
https://www.readbyqxmd.com/read/29768491/plasmodium-falciparum-dipeptidyl-aminopeptidase-3-activity-is-important-for-efficient-erythrocyte-invasion-by-the-malaria-parasite
#17
Christine Lehmann, Michele Ser Ying Tan, Laura E de Vries, Ilaria Russo, Mateo Isidrio Sanchez, Daniel E Goldberg, Edgar Deu
Parasite egress from infected erythrocytes and invasion of new red blood cells are essential processes for the exponential asexual replication of the malaria parasite. These two tightly coordinated events take place in less than a minute and are in part regulated and mediated by proteases. Dipeptidyl aminopeptidases (DPAPs) are papain-fold cysteine proteases that cleave dipeptides from the N-terminus of protein substrates. DPAP3 was previously suggested to play an essential role in parasite egress. However, little is known about its enzymatic activity, intracellular localization, or biological function...
May 16, 2018: PLoS Pathogens
https://www.readbyqxmd.com/read/29768261/n-cadherin-attenuates-high-glucose-induced-nucleus-pulposus-cell-senescence-through-regulation-of-the-ros-nf-%C3%AE%C2%BAb-pathway
#18
Gang Hou, Huiqing Zhao, Haijun Teng, Pei Li, Wenbin Xu, Junbin Zhang, Lulu Lv, Zhiliang Guo, Li Wei, Hui Yao, Yichun Xu
BACKGROUND/AIMS: Diabetes mellitus (DM) is a potential etiology of disc degeneration. N-cadherin (N-CDH) helps maintain the cell viability, cell phenotype and matrix biosynthesis of nucleus pulposus (NP) cells. Here, we mainly aimed to investigate whether N-CDH can attenuate high glucose-induced NP cell senescence and its potential mechanism. METHODS: Rat NP cells were cultured in a base culture medium and base culture medium with a 0.2 M glucose concentration. Recombinant lentiviral vectors were used to enhance N-CDH expression in NP cells...
May 11, 2018: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/29766327/characterization-of-carfilzomib-resistant-non-small-cell-lung-cancer-cell-lines
#19
Neale T Hanke, Elliot Imler, Marilyn T Marron, Bruce E Seligmann, Linda L Garland, Amanda F Baker
PURPOSE: We previously showed that carfilzomib (CFZ) has potent anti-proliferative and cytotoxic activity in a broad range of lung cancer cell lines. Here we investigate possible mechanisms of CFZ acquired resistance in lung cancer cell lines. METHODS: CFZ-resistant non-small cell lung cancer (NSCLC) cell lines were developed by exposing A549 and H520 cells to stepwise increasing concentrations of CFZ. Resistance to CFZ and cross-resistance to bortezomib and other chemotherapy drugs was measured using the MTT assay...
May 15, 2018: Journal of Cancer Research and Clinical Oncology
https://www.readbyqxmd.com/read/29764507/aldolase-a-promotes-proliferation-and-g-1-s-transition-via-the-egfr-mapk-pathway-in-non-small-cell-lung-cancer
#20
Hailu Fu, Huijun Gao, Xiaoyu Qi, Lei Zhao, Donghua Wu, Yuxin Bai, Huimin Li, Xuan Liu, Jun Hu, Shujuan Shao
BACKGROUND: Our previous study demonstrated that aldolase A (ALDOA) is overexpressed in clinical human lung squamous cell carcinoma and that ALDOA promotes epithelial-mesenchymal transition and tumorigenesis. The present study aimed to explore the function of ALDOA in the modulation of non-small cell lung cancer (NSCLC) proliferation and cell cycle progression and the potential mechanism. METHODS: ALDOA was knocked down by short hairpin RNA in H520 and H1299 cells...
May 8, 2018: Cancer communications
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