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Intracellular Flow Cytometry

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https://www.readbyqxmd.com/read/28338196/study-on-the-proliferation-of-human-gastric-cancer-cell-ags-by-activation-of-egfr-in-h2o2
#1
Q Wang, W Shen, G-Q Tao, J Sun, L-P Shi
OBJECTIVE: This study is to investigate the effect of low concentration hydrogen peroxide (H2O2) on the proliferation of gastric cancer AGS cell line in vitro and the mechanism. MATERIALS AND METHODS: AGS cells were treated with different low concentrations of H2O2 (1, 0.1, 0.01, and 0.001 μm) for 48 hours. The effect of H2O2 concentration gradient on the activity of AGS cell activities was detected by methyl thiazolyl tetrazolium (MTT) method. The expression of the epidermal growth factor receptor (EGFR) and its downstream signaling pathway extracellular signal-regulated kinase (ERK) protein in H2O2 was detected by Western blot method; moreover, the effect of H2O2 on intracellular reactive oxygen species (ROS) in AGS cells was observed under the fluorescence microscope and quantitative analysis by flow cytometry...
March 2017: European Review for Medical and Pharmacological Sciences
https://www.readbyqxmd.com/read/28338173/valsartan-reduces-at1-aa-induced-apoptosis-through-suppression-oxidative-stress-mediated-er-stress-in-endothelial-progenitor-cells
#2
Z-C Wang, J Qi, L-M Liu, J Li, H-Y Xu, B Liang, B Li
OBJECTIVE: Valsartan has been reported to have the function of treating hypertension and improving the prognosis of patients. Many studies indicated that valsartan can also increase angiotensin II, andosterone and plasma renin activity (PRA). Autoantibodies against the angiotensin II type 1 receptor (AT1-AA) have been showed to increase reactive oxygen species (ROS) and calcium (Ca2+) and result in apoptosis in vascular smooth muscle cells. In this study, we attempted to explore the effect of valsartan on AT1-AA-induced apoptosis in endothelial progenitor cells...
March 2017: European Review for Medical and Pharmacological Sciences
https://www.readbyqxmd.com/read/28336310/regulatory-role-of-ccl5-rs2280789-and-cxcl10-rs56061981-gene-polymorphisms-on-intracellular-ccl5-and-cxcl10-expression-in-pulmonary-tuberculosis
#3
Brijendra Singh, Selvaraj Anbalagan, Paramasivam Selvaraj
Genetic variations in chemokine genes influence the chemoattractive properties of T cells which may be associated with outcome of infections. In present study, we have investigated the regulatory role played by In1.1T/C (rs2280789) polymorphism of CCL5 and -135G/A (rs56061981) polymorphism of CXCL10 gene on intracellular CCL5 and CXCL10 expression in T cells. Whole blood cell cultures were stimulated with culture filtrate antigen (CFA) and infected with live M. tuberculosis were used for intracellular CCL5 and CXCL10 expression using flow cytometry...
March 21, 2017: Human Immunology
https://www.readbyqxmd.com/read/28331079/characterization-and-vaccine-potential-of-membrane-vesicles-produced-by-francisella-noatunensis-sup-orientalis-in-an-adult-zebrafish-model
#4
Leidy Lagos, Julia I Tandberg, Urska Repnik, Preben Boysen, Erik Ropstad, Deepa Varkey, Ian T Paulsen, Hanne C Winther-Larsen
Vaccine development against extracellular bacteria has been important for the sustainability of the aquaculture industry. In contrast, infections with intracellular pathogens remain largely an unresolved problem. Francisella noatunensis subspecies orientalis (Fno) are Gram-negative, facultative intracellular bacteria that cause the disease francisellosis in fish. Francisellosis is commonly characterized as a chronic granulomatous disease with high morbidity and can result in high mortality depending on the host...
March 22, 2017: Clinical and Vaccine Immunology: CVI
https://www.readbyqxmd.com/read/28329114/pcsk9-monoclonal-antibodies-reverse-the-pro-inflammatory-profile-of-monocytes-in-familial-hypercholesterolaemia
#5
Sophie J Bernelot Moens, Annette E Neele, Jeffrey Kroon, Fleur M van der Valk, Jan Van den Bossche, Marten A Hoeksema, Renate M Hoogeveen, Johan G Schnitzler, Marie T Baccara-Dinet, Garen Manvelian, Menno P J de Winther, Erik S G Stroes
Aims: Migration of monocytes into the arterial wall contributes to arterial inflammation and atherosclerosis progression. Since elevated low-density lipoprotein cholesterol (LDL-C) levels have been associated with activation of plasma monocytes, intensive LDL-C lowering may reverse these pro-inflammatory changes. Using proprotein convertase subtilisin/kexin type 9 (PCSK9) monoclonal antibodies (mAbs) which selectively reduce LDL-C, we studied the impact of LDL-C lowering on monocyte phenotype and function in patients with familial hypercholesterolaemia (FH) not using statins due to statin-associated muscle symptoms...
February 18, 2017: European Heart Journal
https://www.readbyqxmd.com/read/28326454/quercetin-and-ascorbic-acid-suppress-fructose-induced-nlrp3-inflammasome-activation-by-blocking-intracellular-shuttling-of-txnip-in-human-macrophage-cell-lines
#6
Jung-Yoon Choe, Seong-Kyu Kim
The aim of this study was to identify the role of thioredoxin-interacting protein (TXNIP) and its interaction with antioxidants in the activation of the fructose-induced NOD-like receptor protein 3 (NLRP3) inflammasome in human macrophages. The study was performed with U937 and THP-1 macrophage cell lines. Total reactive oxygen species (ROS) were measured by flow cytometry. Interleukin-1β (IL-1β), IL-18, NLRP3, TXNIP, and caspase-1 protein expression was detected using western blotting. Quantitative real-time polymerase chain reaction was used to detect IL-1β, IL-18, and caspase-1 gene expression...
March 22, 2017: Inflammation
https://www.readbyqxmd.com/read/28326227/uptake-of-the-proteins-htra1-and-htra2-by-cells-mediated-by-calcium-phosphate-nanoparticles
#7
Olga Rotan, Katharina N Severin, Simon Pöpsel, Alexander Peetsch, Melisa Merdanovic, Michael Ehrmann, Matthias Epple
The efficient intracellular delivery of (bio)molecules into living cells remains a challenge in biomedicine. Many biomolecules and synthetic drugs are not able to cross the cell membrane, which is a problem if an intracellular mode of action is desired, for example, with a nuclear receptor. Calcium phosphate nanoparticles can serve as carriers for small and large biomolecules as well as for synthetic compounds. The nanoparticles were prepared and colloidally stabilized with either polyethyleneimine (PEI; cationic nanoparticles) or carboxymethyl cellulose (CMC; anionic nanoparticles) and loaded with defined amounts of the fluorescently labelled proteins HTRA1, HTRA2, and BSA...
2017: Beilstein Journal of Nanotechnology
https://www.readbyqxmd.com/read/28322741/evidence-of-k-homeostasis-disruption-in-cellular-dysfunction-triggered-by-7-ketocholesterol-24s-hydroxycholesterol-and-tetracosanoic-acid-c24-0-in-158n-murine-oligodendrocytes
#8
Maryem Bezine, Meryam Debbabi, Thomas Nury, Rym Ben-Khalifa, Mohammad Samadi, Mustapha Cherkaoui-Malki, Anne Vejux, Jérôme de Sèze, Thibault Moreau, Mohamed El-Ayeb, Gérard Lizard
Imbalance in the homeostasis of K(+) ions has been reported to contribute to the pathogenesis of neurodegenerative diseases. 7-ketocholesterol (7KC), 24S-hydroxycholesterol (24S-OHC), and tetracosanoic acid (C24:0), often found at increased levels in patients with Alzheimer's disease, Multiple Sclerosis and X-ALD, are able to trigger numerous nerve cell dysfunctions. We therefore studied the impact of 7KC, 24S-OHC, and C24:0 on 158N murine oligodendrocytes, and determined their impact on K(+) homeostasis. The effects of 7KC, 24S-OHC and C24:0 on lipid membrane organization and membrane potential were examined with merocyanine 540 (MC540) and bis-(1,3-diethylthiobarbituric acid) trimethine oxonol (DiSBAC2(3)), respectively...
March 17, 2017: Chemistry and Physics of Lipids
https://www.readbyqxmd.com/read/28320438/regulatory-t-cell-frequency-but-not-plasma-il-33-levels-represents-potential-immunological-biomarker-to-predict-clinical-response-to-intravenous-immunoglobulin-therapy
#9
Mohan S Maddur, Emmanuel Stephen-Victor, Mrinmoy Das, Praveen Prakhar, Varun K Sharma, Vikas Singh, Magalie Rabin, Jamma Trinath, Kithiganahalli N Balaji, Francis Bolgert, Jean-Michel Vallat, Laurent Magy, Srini V Kaveri, Jagadeesh Bayry
BACKGROUND: Intravenous immunoglobulin (IVIG) is a polyspecific pooled immunoglobulin G preparation and one of the commonly used therapeutics for autoimmune diseases including those of neurological origin. A recent report in murine model proposed that IVIG expands regulatory T (Treg) cells via induction of interleukin 33 (IL-33). However, translational insight on these observations is lacking. METHODS: Ten newly diagnosed Guillain-Barré syndrome (GBS) patients were treated with IVIG at the rate of 0...
March 20, 2017: Journal of Neuroinflammation
https://www.readbyqxmd.com/read/28318884/udp-p2y6-receptor-signaling-regulates-ige-dependent-degranulation-in-human-basophils
#10
Manabu Nakano, Koichi Ito, Takeo Yuno, Nobuyuki Soma, Syun Aburakawa, Kosuke Kasai, Toshiya Nakamura, Hideki Takami
BACKGROUND: P2Y purinergic receptors (P2YR) are G protein-coupled receptors that are stimulated by extracellular nucleotides. They mediate cellular effects by regulating cAMP production, protein kinase C activation, inositol trisphosphate generation, and Ca(2+) release from intracellular stores. The P2Y6 receptor of this family is selectively stimulated by UDP, and selectively inhibited by MRS2578. In the present study, we examined the effect of UDP/P2Y6 receptor signaling on IgE-dependent degranulation in human basophils...
March 16, 2017: Allergology International: Official Journal of the Japanese Society of Allergology
https://www.readbyqxmd.com/read/28317806/mycobacterium-lepraemurium-uses-tlr-6-and-mr-but-not-lipid-rafts-or-dc-sign-to-gain-access-into-mouse-macrophages
#11
Mayra Silva-Miranda, Patricia Arce-Paredes, Oscar Rojas-Espinosa
OBJECTIVE/BACKGROUND: Mycobacterium lepraemurium (MLM), the etiologic agent of murine leprosy, is an intracellular parasite of macrophages; the mechanism used by this bacterium to enter macrophages is not known. The fate of the MLM phagosome inside macrophages is also unknown. This study was conducted to investigate how MLM enters macrophages and to define the maturation process of MLM phagosome inside macrophages. MATERIALS AND METHODS: Peritoneal macrophages were incubated in the presence of mannan-bovine serum albumin (BSA), and antibodies to known macrophage receptors, including, anti-FcγRIII/RII (anti-CD16/32), anti-CD35 (anti-CR1), anti-TLR2, anti-TLR4, anti-TLR6, anti-CD14, and anti-dendritic cell-specific intercellular adhesion molecule-3-grabbing nonintegrin (DC-SIGN)...
January 2017: International Journal of Mycobacteriology
https://www.readbyqxmd.com/read/28317209/the-effects-of-cryopreservation-on-the-expression-of-canine-regulatory-t-cell-markers
#12
Noemi Tarpataki, Marcin Wawrzyniak, Cezmi A Akdis, Beate Rückert, Marina L Meli, Nina M Fischer, Claude Favrot, Ana Rostaher
BACKGROUND: Regulatory T (Treg) cells have been described as key regulators in various immunological processes and are of growing interest in veterinary allergy. Cryopreservation of immune cells is performed routinely in human basic science research and in clinical studies. As such, it allows batch testing of collected samples at a single time point, resulting in a significant reduction in sample variability. Data which describe the effects of cryopreservation on Treg cell frequency and functionality in the canine species are important to inform future research...
March 19, 2017: Veterinary Dermatology
https://www.readbyqxmd.com/read/28314956/evolution-of-mhc-based-technologies-used-for-detection-of-antigen-responsive-t-cells
#13
REVIEW
Amalie Kai Bentzen, Sine Reker Hadrup
T cell-mediated recognition of peptide-major histocompatibility complex (pMHC) class I and II molecules is crucial for the control of intracellular pathogens and cancer, as well as for stimulation and maintenance of efficient cytotoxic responses. Such interactions may also play a role in the development of autoimmune diseases. Novel insights into this mechanism are crucial to understanding disease development and establishing new treatment strategies. MHC multimers have been used for detection of antigen-responsive T cells since the first report by Altman et al...
March 17, 2017: Cancer Immunology, Immunotherapy: CII
https://www.readbyqxmd.com/read/28306698/oral-application-of-magnesium-l-threonate-attenuates-vincristine-induced-allodynia-and-hyperalgesia-by-normalization-of-tumor-necrosis-factor-%C3%AE-nuclear-factor-%C3%AE%C2%BAb-signaling
#14
Ting Xu, Dai Li, Xin Zhou, Han-Dong Ouyang, Li-Jun Zhou, Hang Zhou, Hong-Mei Zhang, Xu-Hong Wei, Guosong Liu, Xian-Guo Liu
BACKGROUND: Antineoplastic agents, including vincristine, often induce neuropathic pain and magnesium deficiency clinically, but the causal link between them has not been determined. No drug is available for treating this form of neuropathic pain. METHODS: Injection of vincristine (0.1 mg · kg · day, intraperitoneally, for 10 days) was used to induce nociceptive sensitization, which was accessed with von Frey hairs and the plantar tester in adult male Sprague-Dawley rats...
March 17, 2017: Anesthesiology
https://www.readbyqxmd.com/read/28304296/pcsk9-promotes-oxldl-induced-pc12-cell-apoptosis-through-the-bcl-2-bax-caspase-9-3-signaling-pathway
#15
Lu-Shan Liu, Xue-Qin Bai, Ya Gao, Qi Wu, Zhong Ren, Qing Li, Li-Hong Pan, Ni-Ya He, Juan Peng, Zhi-Han Tang
BACKGROUND: Hyperlipidemia is a risk factor for neurodegenerative diseases. Proprotein convertase subtilisin / Kexin type 9 (PCSK9) degrades hepatic low-density lipoprotein receptor (LDLR) to regulate lipid metabolism. It is unclear if PCSK9 plays a role in neurodegenerative diseases. OBJECTIVE: This study was designed to determine whether PCSK9 is crucial between hyperlipidemia and Alzheimer's disease. The interrelationship between PCSK9 and neuronal apoptosis was explored in PC12 cells in response to treatment with oxidized low-density lipoprotein (oxLDL)...
March 10, 2017: Journal of Alzheimer's Disease: JAD
https://www.readbyqxmd.com/read/28303575/isoform-localization-of-dectin-1-regulates-the-signaling-quality-of-anti-fungal-immunity
#16
Mike Fischer, Jörg P Müller, Bärbel Spies-Weisshart, Christine Gräfe, Oliver Kurzai, Kerstin Hünniger, Andreas Hochhaus, Sebastian Scholl, Ulf Schnetzke
Dectin-1 is recognized as a major receptor for fungal ß-glucans and contributes to anti-fungal immunity. Human monocyte populations express Dectin-1 isoforms A and B, which differ by the presence of a stalk region and its N-linked glycosylation site. Here, we analyzed the expression of both isoforms in human monocyte-derived cells. The cellular localization on cell lines stably expressing either Dectin-1 isoform A or B was studied by flow cytometry and confocal laser scanning microscopy. Intracellular protein signaling and cytokine production was analyzed by immunoblotting and cytometric bead arrays, respectively...
March 16, 2017: European Journal of Immunology
https://www.readbyqxmd.com/read/28303331/implication-of-vegf-and-aquaporin-4-mediating-m%C3%A3-ller-cell-swelling-to-diabetic-retinal-edema
#17
Teruyo Kida, Hidehiro Oku, Taeko Horie, Masanori Fukumoto, Yoshitaka Okuda, Seita Morishita, Tsunehiko Ikeda
PURPOSE: Aquaporin 4 (AQP4), a water channel protein, is known to be expressed in retinal Müller cells. The purpose of this study was to determine the effects of VEGF and AQP4 channels on the volumetric changes in Müller cells. METHODS: Retinas from diabetic rats and a cultured Müller cell line, TR-MUL5, were used in this study. Intravitreal injections of VEGF or PBS were performed on either streptozotocin (STZ)-induced diabetic or normoglycemic rats. Retinal sections were immunostained for anti-glial fibrillary acidic protein (GFAP), anti-AQP4, and anti-VEGF...
March 16, 2017: Graefe's Archive for Clinical and Experimental Ophthalmology
https://www.readbyqxmd.com/read/28302038/current-application-of-quantum-dots-qd-in-cancer-therapy-a-review
#18
Lavanya Thilak Babu, Priyankar Paira
Semiconductor quantum dots proved themselves as an efficient fluorescent probes in cancer detection and treatment. Their size, high stability, non-photobleaching and water solubility made them a unique fluorophore in place of conventional organic dyes. Newly emerged theranoistic drug delivery system using quantum dots helped us in better understanding of the drug delivery mechanism inside the cells. Surface modified Quantum dots and their applications became wide in bioimaging, immunohistochemistry, tracking intracellular drug and intracellular molecules target...
March 15, 2017: Mini Reviews in Medicinal Chemistry
https://www.readbyqxmd.com/read/28300604/altered-activation-of-peripheral-cd8-t-cells-in-pediatric-crohn-s-disease
#19
Csaba Orbán, Dolóresz Szabó, Anna Bajnok, Barna Vásárhelyi, Tivadar Tulassay, András Arató, Gábor Veres, Gergely Toldi
AIM: Although Crohn's disease (CD) is an extensively investigated autoimmune condition, knowledge on early phase activation of lymphocytes, especially CD8+ Tc cells is scarce. Our aim was to investigate the calcium influx characteristics of CD8+ cells upon activation as well as the expression and function of Kv1.3 and IKCa1 lymphocyte potassium channels. METHODS: We took peripheral blood from 12 healthy controls, 23 CD children on conventional therapy and 6 severe CD children before and after infliximab therapy...
March 11, 2017: Immunology Letters
https://www.readbyqxmd.com/read/28298211/fty720-inhibits-mesothelioma-growth-in-vitro-and-in-a-syngeneic-mouse-model
#20
Agata Szymiczek, Sandra Pastorino, David Larson, Mika Tanji, Laura Pellegrini, Jiaming Xue, Shuangjing Li, Carlotta Giorgi, Paolo Pinton, Yasutaka Takinishi, Harvey I Pass, Hideki Furuya, Giovanni Gaudino, Andrea Napolitano, Michele Carbone, Haining Yang
BACKGROUND: Malignant mesothelioma (MM) is a very aggressive type of cancer, with a dismal prognosis and inherent resistance to chemotherapeutics. Development and evaluation of new therapeutic approaches is highly needed. Immunosuppressant FTY720, approved for multiple sclerosis treatment, has recently raised attention for its anti-tumor activity in a variety of cancers. However, its therapeutic potential in MM has not been evaluated yet. METHODS: Cell viability and anchorage-independent growth were evaluated in a panel of MM cell lines and human mesothelial cells (HM) upon FTY720 treatment to assess in vitro anti-tumor efficacy...
March 15, 2017: Journal of Translational Medicine
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