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Intracellular Flow Cytometry

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https://www.readbyqxmd.com/read/28525970/the-immunoregulatory-role-of-alpha-enolase-in-dendritic-cell-function-during-chlamydia-infection
#1
Khamia Ryans, Yusuf Omosun, Danielle N McKeithen, Tankya Simoneaux, Camilla C Mills, Nathan Bowen, Francis O Eko, Carolyn M Black, Joseph U Igietseme, Qing He
BACKGROUND: We have previously reported that interleukin-10 (IL-10) deficient dendritic cells (DCs) are potent antigen presenting cells that induced elevated protective immunity against Chlamydia. To further investigate the molecular and biochemical mechanism underlying the superior immunostimulatory property of IL-10 deficient DCs we performed proteomic analysis on protein profiles from Chlamydia-pulsed wild-type (WT) and IL-10(-/-) DCs to identify differentially expressed proteins with immunomodulatory properties...
May 19, 2017: BMC Immunology
https://www.readbyqxmd.com/read/28521061/extracellular-camp-activates-molecular-signalling-pathways-associated-with-sperm-capacitation-in-bovines
#2
Carlos Agustín I Alonso, Claudia E Osycka-Salut, Luciana Castellano, Andreína Cesari, Nicolás Di Siervi, Adrián Mutto, Anders Johannisson, Jane M Morrell, Carlos Davio, Silvina Perez-Martinez
STUDY QUESTION: Is extracellular cAMP involved in the regulation of signalling pathways in bovine sperm capacitation? SUMMARY ANSWER: Extracellular cAMP-induced sperm capacitation through the activation of different signalling pathways that involve phospholipase C (PLC), PKC/ERK1-2 signalling and an increase in sperm Ca2+ levels, as well as soluble AC and cAMP/protein kinase A (PKA) signalling. WHAT IS KNOWN ALREADY: In order to fertilize the oocyte, ejaculated spermatozoa must undergo a series of changes in the female reproductive tract, known as capacitation...
May 18, 2017: Molecular Human Reproduction
https://www.readbyqxmd.com/read/28518171/fish-flow-a-protocol-for-the-concurrent-detection-of-mrna-and-protein-in-single-cells-using-fluorescence-in-situ-hybridization-and-flow-cytometry
#3
Riccardo Arrigucci, Yuri Bushkin, Felix Radford, Karim Lakehal, Pooja Vir, Richard Pine, December Martin, Jeffrey Sugarman, Yanlin Zhao, George S Yap, Alfred A Lardizabal, Sanjay Tyagi, Maria Laura Gennaro
We describe a flow-cytometry-based protocol for intracellular mRNA measurements in nonadherent mammalian cells using fluorescence in situ hybridization (FISH) probes. The method, which we call FISH-Flow, allows for high-throughput multiparametric measurements of gene expression, a task that was not feasible with earlier, microscopy-based approaches. The FISH-Flow protocol involves cell fixation, permeabilization and hybridization with a set of fluorescently labeled oligonucleotide probes. In this protocol, surface and intracellular protein markers can also be stained with fluorescently labeled antibodies for simultaneous protein and mRNA measurement...
June 2017: Nature Protocols
https://www.readbyqxmd.com/read/28516182/intracellular-accumulation-and-immunological-responses-of-lipid-modified-magnetic-iron-nanoparticles-in-mouse-antigen-processing-cells
#4
Chenmeng Qiao, Jun Yang, Lei Chen, Jie Weng, Xin Zhang
Understanding the effects of magnetic iron nanoparticles (MINPs) on the immune response is vitally important for biomedical applications such as cancer therapy, disease diagnosis and novel cancer imaging. In this study, lipid modified MINPs were designed and prepared by introducing the neutral lipid DSPE-PEG or the zwitterionic lipid DSPE-PCB into hydrophobic MINPs through hydrophobic interaction (L-MINPs and ZL-MINPs, respectively). The effect of L-MINPs and ZL-MINPs on the intracellular accumulation and immune responses of three kinds of antigen processing cells was examined...
May 18, 2017: Biomaterials Science
https://www.readbyqxmd.com/read/28515228/nk1-1-cells-promote-sustained-tissue-injury-and-inflammation-after-trauma-with-hemorrhagic-shock
#5
Shuhua Chen, Rosemary A Hoffman, Melanie Scott, Joanna Manson, Patricia Loughran, Mostafa Ramadan, Anthony J Demetris, Timothy R Billiar
Various cell populations expressing NK1.1 contribute to innate host defense and systemic inflammatory responses, but their role in hemorrhagic shock and trauma remains uncertain. NK1.1(+) cells were depleted by i.p. administration of anti-NK1.1 (or isotype control) on two consecutive days, followed by hemorrhagic shock with resuscitation and peripheral tissue trauma (HS/T). The plasma levels of IL-6, MCP-1, alanine transaminase (ALT), and aspartate aminotransferase (AST) were measured at 6 and 24 h. Histology in liver and gut were examined at 6 and 24 h...
May 17, 2017: Journal of Leukocyte Biology
https://www.readbyqxmd.com/read/28510277/analysis-of-cytotoxic-effects-of-chlorhexidine-gluconate-as-antiseptic-agent-on-human-blood-lymphocytes
#6
Ahmad Salimi, Bahare Alami, Jalal Pourahmad
The aim of this study was to assess the cytotoxicity of chlorhexidine gluconate (CHG) on human blood lymphocytes as a useful ex vivo model for accelerated human toxicity studies. Using biochemical and flow cytometry assessments, we demonstrated that addition of CHG at 1 μM concentration to human blood lymphocytes induced cytotoxicity following 6 h. The CHG-induced cytotoxicity on human blood lymphocytes was associated with intracellular reactive oxygen species generation, mitochondrial membrane potential collapse, lysosomal membrane injury, lipid peroxidation, and depletion of glutathione...
May 16, 2017: Journal of Biochemical and Molecular Toxicology
https://www.readbyqxmd.com/read/28504144/low-molecular-weight-xanthan-gum-suppresses-oxidative-stress-induced-apoptosis-in-rabbit-chondrocytes
#7
Qixin Chen, Xintian Shao, Peixue Ling, Fei Liu, Huarong Shao, Aibin Ma, Jixu Wu, Wei Zhang, Fuyan Liu, Guanying Han, Fengshan Wang
We have previously reported the application of low molecular weight XG(LM-XG), with molecular weights ranging from 1×10(6)Da to 1.5×10(6)Da for treating osteoarthritis. In this study, we investigated the anti-apoptotic activity of LM-XG under oxidative stress conditions, activated by hydrogen peroxide (H2O2)-treated chondrocytes in vitro. Chondrocytes were pretreated with various doses of LM-XG (0, 10, 100, 500, or 1000μg/mL) or 1000μg/mL sodium hyaluronate for 12h, and then exposed to 0.5mmol/L H2O2 for another 12h...
August 1, 2017: Carbohydrate Polymers
https://www.readbyqxmd.com/read/28503167/the-impact-of-space-flight-on-survival-and-interaction-of-cupriavidus-metallidurans-ch34-with-basalt-a-volcanic-moon-analog-rock
#8
Bo Byloos, Ilse Coninx, Olivier Van Hoey, Charles Cockell, Natasha Nicholson, Vyacheslav Ilyin, Rob Van Houdt, Nico Boon, Natalie Leys
Microbe-mineral interactions have become of interest for space exploration as microorganisms could be used to biomine from extra-terrestrial material and extract elements useful as micronutrients in life support systems. This research aimed to identify the impact of space flight on the long-term survival of Cupriavidus metallidurans CH34 in mineral water and the interaction with basalt, a lunar-type rock in preparation for the ESA spaceflight experiment, BIOROCK. Therefore, C. metallidurans CH34 cells were suspended in mineral water supplemented with or without crushed basalt and send for 3 months on board the Russian FOTON-M4 capsule...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28500977/endocytosis-and-intracellular-traffic-of-cholesterol-pdmaema-liposome-complexes-in-human-epithelial-like-cells
#9
F Szymanowski, A A Hugo, P Alves, P N Simões, A Gómez-Zavaglia, Pablo F Pérez
Liposomes are generally used as delivery systems, as they are capable of encapsulating a wide variety of molecules (i.e. plasmids, recombinant proteins, therapeutic drugs). However, liposomal drug delivery have to fulfill different requirements, such as the effective internalization by the target cells and avoidance of the degradative activity of the intracellular compartments. The use of polymer lipid complexes (PLCs), by including different polymers in the liposome formulation, could improve internalization and intracellular release of drugs...
May 1, 2017: Colloids and Surfaces. B, Biointerfaces
https://www.readbyqxmd.com/read/28499237/the-protective-effect-of-propofol-against-tnf-%C3%AE-induced-apoptosis-was-mediated-via-inhibiting-inos-no-production-and-maintaining-intracellular-ca-2-homeostasis-in-mouse-hippocampal-ht22-cells
#10
Zheng Xu, Yan Lu, Jiaqiang Wang, Xiaowei Ding, Jiawei Chen, Changhong Miao
AIM: Inflammation cytokine tumor necrosis factor-α (TNF-α) induces apoptosis in neuronal cells. We hypothesized that propofol may attenuate TNF-α-induced apoptosis in mouse hippocampal HT22 cells and aimed to explore the underlying mechanisms. METHODS: Mouse hippocampal HT22 cells were pretreated with propofol, and then stimulated with TNF-α. Cell viability was measured by cell counting kit 8 (CCK8). Cell apoptosis was examined by flow cytometry analysis. The effect of propofol on TNF-α-modulated nitric oxide production was measured by a nitrate reductase assay kit, intracellular calcium release and mitochondrial membrane potential (MMP) depolarization were measured by flow cytometry analysis, and the expression of inducible nitric oxide synthase (iNOS), C/EBP homologous protein (CHOP), B-cell lymphoma 2 (Bcl2) family and caspases were detected by Western blot...
May 9, 2017: Biomedicine & Pharmacotherapy, Biomédecine & Pharmacothérapie
https://www.readbyqxmd.com/read/28499202/novel-dual-ves-phospholipid-self-assembled-liposomes-with-an-extremely-high-drug-loading-efficiency
#11
Yongpeng Hou, Chen Yao, Longbing Ling, Yawei Du, Ruiyu He, Muhammad Ismail, Ying Zhang, Zhenglin Fu, Xinsong Li
Vitamin E succinate (VES), a unique selective anti-cancer drug, has attracted much attention for its ability to induce apoptosis in various cancer cells. Importantly, it has been reported that VES is largely non-toxic to normal cells. However, poor aqueous solubility and bioavailability extensively restricted its clinical utility. In this report, dual VES phospholipid conjugate (di-VES-GPC) prodrug based liposomes were prepared in order to develop an efficient delivery system for VES. Di-VES-GPC was first synthesized by conjugating VES with l-α-glycerophosphorylcholine (GPC) using N,N'-dicyclohexylcarbodiimide (DCC) as a coupling agent...
May 2, 2017: Colloids and Surfaces. B, Biointerfaces
https://www.readbyqxmd.com/read/28498840/a-third-generation-vaccine-for-human-visceral-leishmaniasis-and-post-kala-azar-dermal-leishmaniasis-first-in-human-trial-of-chad63-kh
#12
Mohamed Osman, Anoop Mistry, Ada Keding, Rhian Gabe, Elizabeth Cook, Sarah Forrester, Rebecca Wiggins, Stefania Di Marco, Stefano Colloca, Loredana Siani, Riccardo Cortese, Deborah F Smith, Toni Aebischer, Paul M Kaye, Charles J Lacey
BACKGROUND: Visceral leishmaniasis (VL or kala azar) is the most serious form of human leishmaniasis, responsible for over 20,000 deaths annually, and post kala azar dermal leishmaniasis (PKDL) is a stigmatizing skin condition that often occurs in patients after successful treatment for VL. Lack of effective or appropriately targeted cell mediated immunity, including CD8+ T cell responses, underlies the progression of VL and progression to PKDL, and can limit the therapeutic efficacy of anti-leishmanial drugs...
May 12, 2017: PLoS Neglected Tropical Diseases
https://www.readbyqxmd.com/read/28498411/mitochondrial-activity-and-oxidative-stress-functions-are-influenced-by-the-activation-of-ahr-induced-cyp1a1-overexpression-in-cardiomyocytes
#13
Bing Zhou, Xi Wang, Feng Li, Yingting Wang, Lei Yang, Xiaolong Zhen, Wuhong Tan
There is an endemic cardiomyopathy currently occurring in China, termed, Keshan disease (KD). The authors previously compared mitochondrial‑associated gene expression profiles of peripheral blood mononuclear cells (PBMCs) derived from KD patients and normal controls, using mitochondria‑focused cDNA microarray technology. The results detected an upregulation of the enzyme‑associated CYP1A1 gene, (ratios ≥2.0). The aryl hydrocarbon receptor (AhR) regulates the expression of numerous cytochrome P450 (CYP) genes including members of the CYP1 family; CYP1A1 and CYP1A2...
May 12, 2017: Molecular Medicine Reports
https://www.readbyqxmd.com/read/28494925/comparison-of-humoral-and-t-cell-mediated-immune-responses-to-a-single-dose-of-bovela-%C3%A2-live-double-deleted-bvdv-vaccine-or-to-a-field-bvdv-strain
#14
Ratree Platt, Lyle Kesl, Christian Guidarini, Chong Wang, James A Roth
The objective of this study was to determine and compare the humoral and cellular immune responses of calves exposed to a single dose of Bovela(®) bovine viral diarrhea virus (BVDV) live double deleted vaccine or a field strain virus (FSV) of BVDV type 2 (strain 890). Thirty seronegative, colostrum-deprived 5 month-old Holstein steer calves that tested negative for persistent BVDV by ear notch immunohistochemistry and seronegative to BVDV types 1 and 2 were used. Calves were screened by multi-parameter flow cytometry (MP-FCM) 1 week before vaccination to ensure that they were negative for T cell responses to the BVDV types 1 and 2 viruses in the Bovela(®) vaccine...
May 2017: Veterinary Immunology and Immunopathology
https://www.readbyqxmd.com/read/28494534/ferroptosis-a-novel-anti-tumor-action-for-cisplatin
#15
Jipeng Guo, Bingfei Xu, Qi Han, Hongxia Zhou, Yun Xia, Chongwen Gong, Xiaofang Dai, Zhenyu Li, Gang Wu
Purpose: Ferroptosis is a new mode of regulated cell death, which is completely distinct from other cell death modes based on morphological, biochemical, and genetic criteria. This study evaluated the therapeutic role of ferroptosis in classic chemotherapy drugs, including the underlying mechanism. Materials and Methods: Cell viability was detected by using the Methylthiazoltetrazlium dye uptake method. RNAi was used to knockout iron-responsive element binding protein 2, and PCR, western blot was used to evaluate the efficiency...
May 10, 2017: Cancer Research and Treatment: Official Journal of Korean Cancer Association
https://www.readbyqxmd.com/read/28492340/alterations-of-zinc-homeostasis-in-response-to-cryptococcus-neoformans-in-a-murine-macrophage-cell-line
#16
Francine Melise Dos Santos, Alícia Corbellini Piffer, Rafael de Oliveira Schneider, Nicole Sartori Ribeiro, Ane Wichine Acosta Garcia, Augusto Schrank, Lívia Kmetzsch, Marilene Henning Vainstein, Charley Christian Staats
AIM: To evaluate alterations of zinc homeostasis in macrophages exposed to Cryptococcus neoformans. Materials & methods: Using a fluorescent zinc probe-based flow cytometry and atomic absorption spectrometry, zinc levels were evaluated in J774.A1 cell lines exposed to C. neoformans H99 cells. The transcription profile of macrophage zinc related homeostasis genes - metallothioneins and zinc transporters (ZnTs) of the SLC30 and SLC39 (Zrt-Irt-protein) families - was analyzed by quantitative PCR...
May 11, 2017: Future Microbiology
https://www.readbyqxmd.com/read/28490863/development-and-evaluation-of-a-new-modular-nanotransporter-for-drug-delivery-into-nuclei-of-pathological-cells-expressing-folate-receptors
#17
Tatiana A Slastnikova, Andrey A Rosenkranz, Yuri V Khramtsov, Tatiana S Karyagina, Sergey A Ovechko, Alexander S Sobolev
PURPOSE: Modular nanotransporters (MNTs) are artificial multifunctional systems designed to facilitate receptor-specific transport from the cell surface into the cell nucleus through inclusion of polypeptide domains for accomplishing receptor binding and internalization, as well as sequential endosomal escape and nuclear translocation. The objective of this study was to develop a new MNT targeted at folate receptors (FRs) for precise delivery of therapeutic cargo to the nuclei of FR-positive cells and to evaluate its potential, particularly for delivery of therapeutic agents (eg, the Auger electron emitter (111)In) into the nuclei of target cancer cells...
2017: Drug Design, Development and Therapy
https://www.readbyqxmd.com/read/28489280/synthesis-and-evaluation-of-cytotoxic-activities-of-artemisinin-derivatives
#18
Qian Sun, Jin Wang, Yao Li, Jingjing Zhuang, Qian Zhang, Xiao Sun, Dequn Sun
Artemisinin is naturally occurring antimalarials which has shown potent anticancer activity. In the present work, a new kind of artemisinin derivatives with piperazino group were synthesized. The cytotoxic activities of derivatives 5a-5d were evaluated by MTT assay against ten cell lines. The result showed that 5a-5d were more effective in inhibiting cancer cells growth than artemisinin. 5d was the most active against HepG2 and PLC-PRF-5 cells, and presented no cytotoxicity on L-02 cells. Hoechst33342 staining and flow cytometry experiment revealed that 5d could induce HepG2 and PLC-PRF-5 cells apoptosis...
May 10, 2017: Chemical Biology & Drug Design
https://www.readbyqxmd.com/read/28488596/the-impact-of-microfluidic-mixing-of-triblock-micelleplexes-on-in-vitro-formula-see-text-in-vivo-gene-silencing-and-intracellular-trafficking
#19
Daniel P Feldmann, Yuran Xie, Steven K Jones, Dongyue Yu, Anna Moszczynska, Olivia M Merkel
The triblock copolymer polyethylenimine-polycaprolactone-polyethylene glycol (PEI-PCL-PEG) has been shown to spontaneously assemble into nano-sized particulate carriers capable of complexing with nucleic acids for gene delivery. The objective of this study was to investigate micelleplex characteristics, their in vitro and in vivo fate following microfluidic preparation of siRNA nanoparticles compared to the routinely used batch reactor mixing technique. Herein, PEI-PCL-PEG nanoparticles were prepared with batch reactor or microfluidic mixing techniques and characterized by various biochemical assays and in cell culture...
June 2, 2017: Nanotechnology
https://www.readbyqxmd.com/read/28488248/response-to-treatment-with-tnf%C3%AE-inhibitors-in-rheumatoid-arthritis-is-associated-with-high-levels-of-gm-csf-and-gm-csf-t-lymphocytes
#20
Jonas Bystrom, Felix I Clanchy, Taher E Taher, Mohammed M Al-Bogami, Hawzheen A Muhammad, Saba Alzabin, Pamela Mangat, Ali S Jawad, Richard O Williams, Rizgar A Mageed
Biologic TNFα inhibitors are a mainstay treatment option for patients with rheumatoid arthritis (RA) refractory to other treatment options. However, many patients either do not respond or relapse after initially responding to these agents. This study was carried out to identify biomarkers that can distinguish responder from non-responder patients before the initiation of treatment. The level of cytokines in plasma and those produced by ex vivo T cells, B cells and monocytes in 97 RA patients treated with biologic TNFα inhibitors was measured before treatment and after 1 and 3 months of treatment by multiplex analyses...
May 9, 2017: Clinical Reviews in Allergy & Immunology
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