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Intracellular Flow Cytometry

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https://www.readbyqxmd.com/read/28736158/nanoliposomes-carrying-her2-neu-derived-peptide-ae36-with-cpg-odn-exhibit-therapeutic-and-prophylactic-activities-in-a-mice-tubo-model-of-breast-cancer
#1
Nastaran Barati, Amin Reza Nikpoor, Atefeh Razazan, Fatemeh Mosaffa, Ali Badiee, Atefeh Arab, Zahra Gholizadeh, Javad Behravan, Mahmoud Reza Jaafari
This study was designed to prepare and characterize nanoliposomal vaccine formulation encapsulating AE36 HER2/neu-derived peptide with or without CpG and evaluate the immunologic and therapeutic responses of that in BALB/c mice model of Her2 overexpressing breast cancer. AE36 was encapsulated in liposomes composed of DOTAP, DOPE and Cholesterol (DDC) or DD with. The formulations could induce both CD8+ and CD4+ responses and stimulate production of cytokines which was detected by Enzyme-linked immunospot assay (ELISpot) kits, cytotoxicity test and intracellular cytokine assay by flow cytometry...
July 20, 2017: Immunology Letters
https://www.readbyqxmd.com/read/28734845/leveraging-siglec-8-endocytic-mechanisms-to-kill-human-eosinophils-and-malignant-mast-cells
#2
Jeremy A O'Sullivan, Daniela J Carroll, Yun Cao, Adriano N Salicru, Bruce S Bochner
BACKGROUND: Sialic acid-binding immunoglobulin-like lectin (Siglec)-8 is a cell-surface protein expressed selectively on human eosinophils, mast cells, and basophils, making it an ideal target for the treatment of diseases involving these cell types. However, the effective delivery of therapeutic agents to these cells requires an understanding of the dynamics of Siglec-8 surface expression. OBJECTIVE: To determine whether Siglec-8 is endocytosed in human eosinophils and malignant mast cells, identify mechanisms underlying its endocytosis, and demonstrate whether a toxin can be targeted to Siglec-8-bearing cells to kill these cells...
July 19, 2017: Journal of Allergy and Clinical Immunology
https://www.readbyqxmd.com/read/28733206/omics-based-identification-of-the-combined-effects-of-idiosyncratic-drugs-and-inflammatory-cytokines-on-the-development-of-drug-induced-liver-injury
#3
J Jiang, K Mathijs, L Timmermans, S M Claessen, A Hecka, J Weusten, R Peters, J H van Delft, J C S Kleinjans, D G J Jennen, T M de Kok
The mechanisms of idiosyncratic drug-induced hepatotoxicity remain largely unclear. It has demonstrated that the drug idiosyncrasy is potentiated in the context of inflammation and intracellular ceramides may play a role in this process. To study the mechanisms, HepG2 cells were co-treated with high and low doses of three idiosyncratic (I) and three non-idiosyncratic (N) compounds, with (I+ and N+) or without (I- and N-) a cytokine mix. Microarray, lipidomics and flow cytometry were performed to investigate the genome-wide expression patterns, the intracellular ceramide levels and the induction of apoptosis...
July 18, 2017: Toxicology and Applied Pharmacology
https://www.readbyqxmd.com/read/28732504/ischaemic-stroke-and-the-recanalization-drug-tissue-plasminogen-activator-interfere-with-antibacterial-phagocyte-function
#4
Antje Vogelgesang, Claudia Lange, Lara Blümke, Georg Laage, Sarah Rümpel, Sönke Langner, Barbara M Bröker, Alexander Dressel, Johanna Ruhnau
BACKGROUND: Stroke induces immune alterations such as impaired oxidative burst and reduced release of neutrophil extracellular traps (NETs). We hypothesised that key enzymes of these defence mechanisms may be altered in ischaemic stroke. Therefore, we analysed the intra- and extracellular amounts of myeloperoxidase (MPO) and neutrophil elastase (NE) in patient sera and granulocytes and monocytes. Because the autonomous nervous system is thought to mediate stroke-induced immune alterations, we also studied the influence of stress hormones and acetylcholine on MPO and NE...
July 21, 2017: Journal of Neuroinflammation
https://www.readbyqxmd.com/read/28732161/an-elegant-ph-responsive-nanovehicle-for-drug-delivery-based-on-triazine-dendrimer-modified-magnetic-nanoparticles
#5
Amir Landarani-Isfahani, Majid Moghadam, Shima Mohammadi, Maryam Royvaran, Naimeh Moshtael-Arani, Saghar Rezaei, Shahram Tangestaninejad, Valiollah Mirkhani, Iraj Mohammadpoor-Baltork
Owing to properties of magnetic nanoparticles and elegant three-dimensional macromolecule architectural features, dendrimeric structures have been investigated as nanoscale drug delivery systems. In this paper, a novel magnetic nanocarrier, generation two (G2) triazine dendrimer modified Fe3O4@SiO2 magnetic nanoparticles (MNP-G2), was designed, fabricated and characterized by Fourier transform infrared (FT-IR), thermal gravimetric analysis (TGA), vibrating sample magnetometer (VSM), field emission scanning electron microscopy (FE-SEM), transmission electron microscopy (TEM) and dynamic light scattering (DLS)...
July 21, 2017: Langmuir: the ACS Journal of Surfaces and Colloids
https://www.readbyqxmd.com/read/28730808/determination-of-the-relationship-between-expression-and-functional-activity-of-multidrug-resistance-associated-protein-1-using-scanning-electrochemical-microscopy
#6
David Polcari, Javier Alejandro Hernandez-Castro, Kebin Li, Matthias Geissler, Janine Mauzeroll
Cancer cells can develop multidrug resistance (MDR) after prolonged exposure to chemotherapeutic drugs, which is a severe impediment to successful treatment. MDR is typically associated with transmembrane proteins mediating efflux of administered drugs, thereby keeping their intracellular concentration below the threshold required to kill cells. Although expression assays based on flow cytometry and immunostaining have shown that multidrug resistance-associated protein 1 (MRP1) is prevalent in many cancer types, the functional activity of this efflux pump is more difficult to elucidate, especially at the single-cell level...
July 21, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28730337/memantine-can-reduce-ethanol-induced-caspase-3-activity-and-apoptosis-in-h4-cells-by-decreasing-intracellular-calcium
#7
Xiaolong Wang, Jiajun Chen, Hongbo Wang, Hao Yu, Changliang Wang, Jiabin You, Pengfei Wang, Chunmei Feng, Guohui Xu, Xu Wu, Rui Zhao, Guohua Zhang
Caspase-3 activation and apoptosis are associated with various neurodegenerative disorders. Calcium activation is an important factor in promoting apoptosis. We, therefore, assessed the role of intracellular calcium in ethanol-induced activation of caspase-3 in H4 human neuroglioma cells and the protective effect of the NMDA receptor antagonist, memantine, on ethanol-induced apoptosis in H4 cells. H4 cells were treated with 100 mM EtOH (in culture medium) for 2 days. For interaction studies, cells were treated with memantine (4 μM), EDTA (1 mM), or BAPTA-AM (10 μM) before treatment with EtOH...
July 20, 2017: Journal of Molecular Neuroscience: MN
https://www.readbyqxmd.com/read/28729093/pim-kinases-1-2-and-3-in-intracellular-lif-signalling-proliferation-and-apoptosis-in-trophoblastic-cells
#8
Stella Mary Photini, Wittaya Chaiwangyen, Maja Weber, Boodor Al-Kawlani, Rodolfo R Favaro, Udo Jeschke, Ekkehard Schleussner, Diana M Morales-Prieto, Udo R Markert
Proviral insertion in murine (PIM) lymphoma proteins are mainly regulated by the Janus Kinase/Signal Transducer Activator of Transcription (JAK/STAT) signaling pathway, which can be activated by members of the Interleukin-6 (IL-6) family, including Leukemia Inhibitory Factor (LIF). Aim of the study was to compare PIM1, PIM2 and PIM3 expression and potential cellular functions in human first and third trimester trophoblast cells, the immortalized first trimester extravillous trophoblast cell line HTR8/SVneo and the choriocarcinoma cell line JEG-3...
July 17, 2017: Experimental Cell Research
https://www.readbyqxmd.com/read/28726600/age-and-sex-associated-differences-in-phenotypic-and-functional-characteristics-of-peripheral-blood-lymphocytes-in-chimpanzees-pan-troglodytes
#9
Pramod Nehete, Elizabeth Magden, Bharti Nehete, Lawrence Williams, Christian Abee, K Sastry
Chimpanzees are the closest phylogenetic relatives to humans, sharing more than 98% genetic sequence identity. Thesegenetic similarities prompted the belief that chimpanzees can serve as an ideal model for human disease conditions andvaccine development. However, in light of the recent NIH decision to phase out biomedical research in chimpanzees and retire NIH-supported chimpanzees, data from the present study will continue to provide value for the care of aged and sick chimpanzees located in zoos, sanctuaries, and primate centers...
July 19, 2017: Journal of the American Association for Laboratory Animal Science: JAALAS
https://www.readbyqxmd.com/read/28720095/fibrocyte-measurement-in-peripheral-blood-correlates-with-number-of-cultured-mature-fibrocytes-in-vitro-and-is-a-potential-biomarker-for-interstitial-lung-disease-in-rheumatoid-arthritis
#10
Søren Andreas Just, Hanne Lindegaard, Eva Kildall Hejbøl, Jesper Rømhild Davidsen, Niels Bjerring, Søren Werner Karlskov Hansen, Henrik Daa Schrøder, Inger Marie Jensen Hansen, Torben Barington, Christian Nielsen
BACKGROUND: Interstitial lung disease (ILD) can be a severe extra-articular disease manifestation in Rheumatoid Arthritis (RA). A potential role of fibrocytes in RA associated ILD (RA-ILD) has not previously been described. We present a modified faster method for measuring circulating fibrocytes, without intracellular staining. The results are compared to the traditional culture method, where the number of monocytes that differentiate into mature fibrocytes in vitro are counted. The results are following compared to disease activity in patients with severe asthma, ILD, RA (without diagnosed ILD) and RA with verified ILD (RA-ILD)...
July 18, 2017: Respiratory Research
https://www.readbyqxmd.com/read/28717895/correlating-uptake-and-activity-of-proline-rich-antimicrobial-peptides-in-escherichia-coli
#11
Luzia Holfeld, Ralf Hoffmann, Daniel Knappe
Increasing death tolls accounted for by antimicrobial drug resistance demand novel antibiotic lead compounds. Among different promising candidate classes, proline-rich antimicrobial peptides (PrAMPs) are very favorable due to their intracellular mechanism, i.e., binding to the 70S ribosome and DnaK, after active uptake relying on bacterial transporters like SbmA and MdtM. Studies on peptide internalization as the first step of their complex mode of action rely typically on fluorophore or radioactive labeling and quantification using microscopy, flow cytometry, or radioactivity...
July 17, 2017: Analytical and Bioanalytical Chemistry
https://www.readbyqxmd.com/read/28715391/isolation-of-primary-murine-retinal-ganglion-cells-rgcs-by-flow-cytometry
#12
Sumana R Chintalapudi, Need N Patel, Zachary K Goldsmith, Levon Djenderedjian, Xiang Di Wang, Tony N Marion, Monica M Jablonski, Vanessa M Morales-Tirado
Neurodegenerative diseases often have a devastating impact on those affected. Retinal ganglion cell (RGC) loss is implicated in an array of diseases, including diabetic retinopathy and glaucoma, in addition to normal aging. Despite their importance, RGCs have been extremely difficult to study until now due in part to the fact that they comprise only a small percentage of the wide variety of cells in the retina. In addition, current isolation methods use intracellular markers to identify RGCs, which produce non-viable cells...
July 5, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28713935/curcumin-pretreatment-protects-against-pm2-5%C3%A2-induced-oxidized-low%C3%A2-density-lipoprotein%C3%A2-mediated-oxidative-stress-and-inflammation-in-human-microvascular-endothelial-cells
#13
Jun Shi, Huiping Deng, Min Zhang
A previous study demonstrated that particulate matter (≤2.5 µm in diameter; PM2.5) may promote atherosclerosis. However, the underlying mechanisms of PM2.5 in human microvascular endothelial cells (HMEC‑1) remain to be elucidated. It has been reported that inflammation and oxidative stress can be reduced by curcumin, and in the present study, the aim was to investigate the protective effects of curcumin on PM2.5‑induced oxidative stress and inflammatory response in HMEC‑1. HMEC‑1 were stimulated with curcumin and PM2...
September 2017: Molecular Medicine Reports
https://www.readbyqxmd.com/read/28710313/ex-vivo-pd-l1-pd-1-pathway-blockade-reverses-dysfunction-of-circulating-cea-specific-t-cells-in-pancreatic-cancer-patients
#14
Yuan Chen, Shao-An Xue, Goran Hamid Mohammad, Stephen P Pereira, Emma Morris, Shahriar Behboudi
Carcinoembryonic antigen (CEA) is a candidate target for cellular immunotherapy of pancreatic cancer (PC). In this study, we have characterised the antigen-specific function of autologous cytotoxic T lymphocytes (CTL) specific for the HLA-A2 restricted peptide, pCEA691-699, isolated from the peripheral T cell repertoire of PC patients and sought to determine if ex vivo PD-L1 & TIM3 blockade could enhance CTL function. CD8(+) T cell lines were generated from peripheral blood mononuclear cells (PBMCs) of 18 HLA-A2(+) patients with PC and from 15 healthy controls...
July 14, 2017: Clinical Cancer Research: An Official Journal of the American Association for Cancer Research
https://www.readbyqxmd.com/read/28709892/liuwei-dihuang-soft-capsules-attenuates-endothelial-cell-apoptosis-to-prevent-atherosclerosis-through-gpr30-mediated-regulation-in-ovariectomized-apoe-deficient-mice
#15
Yi Jing, Danfeng Cai, Qi Chen, Qingping Xiong, Tianhui Hu, Yuan Yao, Chao Lin, Xin Sun, Ying Lu, Xueyun Kong, Xiang Wu, Yu Li, Huimin Bian
ETHNOPHARMACOLOGICAL RELEVANCE: Liuwei Dihuang (LWDH), a classical traditional Chinese medicine prescription, has been widely used to prevent and to treat various diseases with symptoms of 'Kidney-Yin' deficiency syndrome for over 1000 years in China. It is commonly used to treat functional decline associated with senile disease and menopausal syndrome, especially memory decline, insomnia, diabetes and osteoporosis. Modern experimental pharmacological studies indicated that the mechanism of LWDH treatment of menopausal syndrome may be associated with enhanced estrogenic effects...
July 12, 2017: Journal of Ethnopharmacology
https://www.readbyqxmd.com/read/28706956/ethyl-caffeate-ameliorates-collagen-induced-arthritis-by-suppressing-th1-immune-response
#16
Shikui Xu, Aixue Zuo, Zengjun Guo, Chunping Wan
The present study was designed to assess the antiarthritic potential of ECF in collagen-induced arthritis (CIA) and explore its underlying mechanism. Methods. In vitro, lymphocyte proliferation assay was measured by CCK-8 kit. In vivo, the therapeutic potential of ECF on CIA was investigated; surface marker, Treg cell, and intracellular cytokines (IL-17A and IFN-γ) were detected by flow cytometry. Th1 cell differentiation assay was performed, and mRNA expression in interferon-γ-related signaling was examined by q-PCR analysis...
2017: Journal of Immunology Research
https://www.readbyqxmd.com/read/28704808/enhanced-reactive-oxygen-species-production-acidic-cytosolic-ph-and-upregulated-na-h-exchanger-nhe-in-dicer-deficient-cd4-t-cells
#17
Yogesh Singh, Yuetao Zhou, Shaqiu Zhang, Khalid N M Abdelazeem, Bernat Elvira, Madhuri S Salker, Florian Lang
BACKGROUND: MicroRNAs (miRNAs) negatively regulate gene expression at a post-transcriptional level. Dicer, a cytoplasmic RNase III enzyme, is required for the maturation of miRNAs from precursor miRNAs. Dicer, therefore, is a critical enzyme involved in the biogenesis and processing of miRNAs. Several biological processes are controlled by miRNAs, including the regulation of T cell development and function. T cells generate reactive oxygen species (ROS) with parallel H+ extrusion accomplished by the Na+/H+-exchanger 1 (NHE1)...
July 14, 2017: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/28702480/dataset-on-transcriptional-profiles-and-the-developmental-characteristics-of-pdgfr%C3%AE-expressing-lung-fibroblasts
#18
Mehari Endale, Shawn Ahlfeld, Erik Bao, Xiaoting Chen, Jenna Green, Zach Bess, Matthew Weirauch, Yan Xu, Anne Karina Perl
The following data are derived from key stages of acinar lung development and define the developmental role of lung interstitial fibroblasts expressing platelet-derived growth factor alpha (PDGFRα). This dataset is related to the research article entitled "Temporal, spatial, and phenotypical changes of PDGFRα expressing fibroblasts during late lung development" (Endale et al., 2017) [1]. At E16.5 (canalicular), E18.5 (saccular), P7 (early alveolar) and P28 (late alveolar), PDGFRα(GFP) mice, in conjunction with immunohistochemical markers, were utilized to define the spatiotemporal relationship of PDGFRα(+) fibroblasts to endothelial, stromal and epithelial cells in both the proximal and distal acinar lung...
August 2017: Data in Brief
https://www.readbyqxmd.com/read/28698276/a-novel-single-cell-fish-flow-assay-identifies-effector-memory-cd4-t-cells-as-a-major-niche-for-hiv-1-transcription-in-hiv-infected-patients
#19
Judith Grau-Expósito, Carla Serra-Peinado, Lucia Miguel, Jordi Navarro, Adrià Curran, Joaquin Burgos, Imma Ocaña, Esteban Ribera, Ariadna Torrella, Bibiana Planas, Rosa Badía, Josep Castellví, Vicenç Falcó, Manuel Crespo, Maria J Buzon
Cells that actively transcribe HIV-1 have been defined as the "active viral reservoir" in HIV-infected individuals. However, important technical limitations have precluded the characterization of this specific viral reservoir during both treated and untreated HIV-1 infections. Here, we used a novel single-cell RNA fluorescence in situ hybridization-flow cytometry (FISH-flow) assay that requires only 15 million unfractionated peripheral blood mononuclear cells (PBMCs) to characterize the specific cell subpopulations that transcribe HIV RNA in different subsets of CD4(+) T cells...
July 11, 2017: MBio
https://www.readbyqxmd.com/read/28697396/novel-mutations-in-the-exon-5-intron-2-and-3-utr-regions-of-il-12b-gene-were-observed-in-clinically-proven-tuberculosis-patients-of-south-india
#20
Pallipamu Prakash Babu, Pasupuleti Santhosh Kumar, Alladi Mohan, Bhattaram Siddhartha Kumar, Potukuchi Venkata Gurunadha Krishna Sarma
Interleukin-12 (IL-12) is formed by the interaction of IL-12p35 and IL-12p40 expressed independently from IL-12A and IL-12B genes. This interleukin plays prominent role in the T-helper type-1 (Th1) response against intracellular pathogens. Variations in IL-12B gene causes disruption of various activities one of them is suppression of Th1 response and is one of the characteristic features observed in patients with active tuberculosis. Hence, in the present study IL-12B gene status was evaluated in 50 new sputum smear-positive pulmonary tuberculosis patients (NSP-PTB) as identified by Ziehl-Nielsen (ZN) staining and 50 apparently healthy control subjects (HCS) who were sputum smear-negative...
July 8, 2017: Cytokine
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