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Intracellular Flow Cytometry

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https://www.readbyqxmd.com/read/29145201/dietary-fructo-oligosaccharides-attenuate-early-activation-of-cd4-t-cells-which-produce-both-th1-and-th2-cytokines-in-the-intestinal-lymphoid-tissues-of-a-murine-food-allergy-model
#1
Masato Tsuda, Haruka Arakawa, Narumi Ishii, Chihiro Ubukata, Mana Michimori, Masanari Noda, Kyoko Takahashi, Shuichi Kaminogawa, Akira Hosono
BACKGROUND: Fructo-oligosaccharides (FOS) are prebiotic agents with immunomodulatory effects involving improvement of the intestinal microbiota and metabolome. In this study, we investigated the cellular mechanisms through which FOS modulate intestinal antigen-specific CD4+ T cell responses in food allergy, using OVA23-3 mice. METHODS: OVA23-3 mice were fed an experimental diet containing either ovalbumin (OVA) or OVA and FOS for 1 week. Body weight and mucosal mast cell protease 1 in the serum were measured as the indicator of intestinal inflammation...
November 17, 2017: International Archives of Allergy and Immunology
https://www.readbyqxmd.com/read/29143242/comparing-reactive-oxygen-species-and-dna-fragmentation-in-semen-samples-of-unexplained-infertile-and-healthy-fertile-men
#2
Zahra Zandieh, Akram Vatannejad, Mahmood Doosti, Sara Zabihzadeh, Mahnaz Haddadi, Leila Bajelan, Batool Rashidi, Saeid Amanpour
BACKGROUND: Male factor infertility has increased to more than 40% during the last decade. About 30% of these couples are diagnosed with unexplained infertility. In fact, reactive oxygen species (ROS), especially superoxide anion (O2(-)·) and hydrogen peroxide (H2O2), play a crucial role in regulation of physiological and pathological processes in spermatozoa. Moreover, since the diagnosis of unexplained infertility just through semen analysis is a matter of much controversy; we aimed to evaluate the levels of ROS and sperm DNA fragmentation in the semen samples of unexplained infertile and fertile control couples...
November 15, 2017: Irish Journal of Medical Science
https://www.readbyqxmd.com/read/29142995/screening-of-novel-rgd-peptides-to-modify-nanoparticles-for-targeted-cancer-therapy
#3
Liang Ge, Xinru You, Keqing Huang, Yang Kang, Yuejian Chen, Ying Zhu, Yuan Ren, Yuan Zhang, Jun Wu, Hai Qian
New targeted RGD peptides obtained by solid phase peptide synthesis (SPPS) were successfully screened by Molecular Operating Environment (MOE) and used for the building of the 6-O-carboxymethyl chitosan based carrier with an active target on the surface. CRGDYC-6-O-carboxymethyl chitosan based nanoparticles (NPs) loaded with doxorubicin hydrochloride (DOX) were successfully prepared by an ionic gelation method with the carrier synthesized before. Synthesis conditions and formulation parameters were optimized by determining the characteristics of nanoparticles including the particle size and drug encapsulation efficiency...
November 16, 2017: Biomaterials Science
https://www.readbyqxmd.com/read/29140786/selection-of-an-anticalin%C3%A2-against-the-membrane-form-of-hsp70-via-bacterial-surface-display-and-its-theranostic-application-in-tumour-models
#4
Lars Friedrich, Petra Kornberger, Claudia T Mendler, Gabriele Multhoff, Markus Schwaiger, Arne Skerra
We describe the selection of Anticalins against a common tumour surface antigen, human Hsp70, using functional display on live E. coli cells as fusion with a truncated EspP autotransporter. While found intracellularly in normal cells, Hsp70 is frequently exposed in a membrane-bound state on the surface of tumour cells and, even more pronounced, in metastases or after radiochemotherapy. Employing a recombinant Hsp70 fragment comprising residues 383-548 as target, Anticalins were selected from a naive bacterial library...
November 27, 2017: Biological Chemistry
https://www.readbyqxmd.com/read/29135969/hla-b27-correlates-with-the-intracellular-elimination-replication-and-trafficking-of-salmonella-enteritidis-collected-from-reactive-arthritis-patients
#5
Yi Zhao, Honghu Tang, Chunyu Tan, Hua Zhao, Yi Liu
BACKGROUND The aim of this study was to explore the correlation between HLA-B27 and the intracellular elimination, replication, and trafficking of Salmonella enteritidis (S. enteritidis) collected from patients with reactive arthritis. MATERIAL AND METHODS Confocal microscopy, flow cytometry, and sandwich enzyme-linked immunosorbent assay (ELISA) were employed in this study to evaluate the localization of proteins of interest, to assess the intracellular trafficking of S. enteritidis, and to measure the production of cytokines of interest...
November 14, 2017: Medical Science Monitor: International Medical Journal of Experimental and Clinical Research
https://www.readbyqxmd.com/read/29135959/intratumoral-delivery-of-doxorubicin-on-folate-conjugated-graphene-oxide-by-in-situ-forming-thermo-sensitive-hydrogel-for-breast-cancer-therapy
#6
Yi Teng Fong, Chih-Hao Chen, Jyh-Ping Chen
By taking advantage of the pH-sensitive drug release property of graphene oxide (GO) after intracellular uptake, we prepared folic acid (FA)-conjugated GO (GOFA) for targeted delivery of the chemotherapeutic drug doxorubicin (DOX). GOFA-DOX was further encapsulated in an injectable in-situ forming thermo-sensitive hyaluronic acid-chitosan-g-poly(N-isopropylacrylamide) (HACPN) hydrogel for intratumoral delivery of DOX. As the degradation time of HACPN could be extended up to 3 weeks, intratumoral delivery of GOFA-DOX/HACPN could provide controlled and targeted delivery of DOX through slow degradation HACPN and subsequent cellular uptake of released GOFA-DOX by tumor cells through interactions of GOFA with folate receptors on the tumor cell's surface...
November 14, 2017: Nanomaterials
https://www.readbyqxmd.com/read/29135655/neutrophil-activation-and-enhanced-release-of-granule-products-in-hiv-tb-immune-reconstitution-inflammatory-syndrome
#7
Justine K Nakiwala, Naomi F Walker, Collin R Diedrich, William Worodria, Graeme Meintjes, Robert J Wilkinson, Harriet Mayanja-Kizza, Robert Colebunders, Luc Kestens, Katalin A Wilkinson, David M Lowe
BACKGROUND: Tuberculosis Immune Reconstitution Inflammatory Syndrome (TB-IRIS) remains incompletely understood. Neutrophils are implicated in tuberculosis pathology but detailed investigations in TB-IRIS are lacking. We sought to further explore the biology of TB-IRIS and in particular the role of neutrophils. SETTING: Two observational, prospective cohort studies in HIV/TB co-infected patients starting antiretroviral therapy, one to analyze gene expression and subsequently one to explore neutrophil biology...
November 6, 2017: Journal of Acquired Immune Deficiency Syndromes: JAIDS
https://www.readbyqxmd.com/read/29135573/evidence-of-inflammasome-activation-and-formation-of-monocyte-derived-apoptosis-associated-speck-like-protein-containing-a-caspase-recruitment-domain-specks-in-hiv-1-positive-patients
#8
Fareed Ahmad, Neha Mishra, Gerrit Ahrenstorf, Bernardo S Franklin, Eicke Latz, Reinhold E Schmidt, Lukas Bossaller
OBJECTIVE: The formation of large intracellular protein aggregates of the inflammasome adaptor apoptosis-associated speck-like protein containing a caspase-recruitment domain (ASC) (PYCARD) is a hallmark of inflammasome activation. ASC speck-forming cells release the highly proinflammatory cytokine IL-1β in addition to ASC specks into the extracellular space during pyroptotic cell death. There ASC specks can propagate inflammation to other nonactivated cells or tissues. HIV-1 retroviral infection triggers inflammasome activation of abortively infected CD4 T cells in secondary lymphatic tissues...
November 10, 2017: AIDS
https://www.readbyqxmd.com/read/29135121/hepcidin-and-iron-metabolism-in-the-pathogenesis-of-prostate-cancer
#9
Feng Wang, An Liu, Ruyu Bai, Binbin Zhang, Yongsheng Jin, Wei Guo, Yi Li, Jixue Gao, Longqiang Liu
PURPOSE: To investigate the relationship between Hepcidin and iron metabolism, and cell proliferation, migration, and apoptosis in prostate cancer cells. METHODS: PC3 prostate cancer cells were cultured in vitro and divided into the control group, Hepcidin overexpression group, and Hepcidin low expression group. Prostate specific antigen (PSA) and soluble transferrin receptor (sTfR) levels were measured by ELISA. The levels of the Hepcidin receptor membrane transporter, Ferroportin, were determined by Western blot...
September 2017: Journal of B.U.ON.: Official Journal of the Balkan Union of Oncology
https://www.readbyqxmd.com/read/29134553/biological-characterization-of-a-novel-hybrid-copolymer-carrier-system-based-on-glycogen
#10
Markéta Jirátová, Aneta Pospíšilová, Maria Rabyk, Martin Pařízek, Jan Kovář, Andrea Gálisová, Martin Hrubý, Daniel Jirák
The effective drug delivery systems for cancer treatment are currently on high demand. In this paper, biological behavior of the novel hybrid copolymers based on polysaccharide glycogen were characterized. The copolymers were modified by fluorescent dyes for flow cytometry, confocal microscopy, and in vivo fluorescence imaging. Moreover, the effect of oxazoline grafts on degradation rate was examined. Intracellular localization, cytotoxicity, and internalization route of the modified copolymers were examined on HepG2 cell line...
November 13, 2017: Drug Delivery and Translational Research
https://www.readbyqxmd.com/read/29132302/immunomodulatory-role-for-membrane-vesicles-released-by-thp-1-macrophages-and-respiratory-pathogens-during-macrophage-infection
#11
Charlotte Volgers, Birke J Benedikter, Gert E Grauls, Paul H M Savelkoul, Frank R M Stassen
BACKGROUND: During infection, inflammation is partially driven by the release of mediators which facilitate intercellular communication. Amongst these mediators are small membrane vesicles (MVs) that can be released by both host cells and Gram-negative and -positive bacteria. Bacterial membrane vesicles are known to exert immuno-modulatory and -stimulatory actions. Moreover, it has been proposed that host cell-derived vesicles, released during infection, also have immunostimulatory properties...
November 13, 2017: BMC Microbiology
https://www.readbyqxmd.com/read/29131464/interferon-regulatory-factor-4-5-signaling-impacts-on-microglial-activation-after-ischemic-stroke-in-mice
#12
Abdullah Al Mamun, Anjali Chauhan, Haifu Yu, Yan Xu, Romana Sharmeen, Fudong Liu
Microglial activation is a key element in initiating and perpetuating inflammatory responses to stroke. Interferon regulatory factor 5 (IRF5) and IRF4 signaling have been found critical in mediating macrophage pro-inflammatory (M1) and anti-inflammatory (M2) phenotypes respectively in peripheral inflammation. We hypothesize that the IRF5/4 regulatory axis also mediates microglial activation after stroke. C57BL6 mice of 8-12 weeks were subject to a 90-minute middle cerebral artery occlusion and the brains evaluated at 24h, 3d, 10d and 30d after reperfusion...
November 13, 2017: European Journal of Neuroscience
https://www.readbyqxmd.com/read/29129818/inter-and-intra-subject-variability-of-nitric-oxide-levels-in-leukocyte-subpopulations
#13
Sheena Maharaj, Kim Lu, Shlomit Aizik, Frank Zaldivar, Fadia Haddad, Hye-Won Shin, Szu-Yun Leu, Eliezer Nussbaum, Inderpal Randhawa, Dan Cooper
Assessment of nitric oxide (NO) dynamics in immune cells, commonly measured using NO surrogates such as inducible nitric oxide synthase (iNOS) rather than NO itself, has been effective in understanding pathophysiology across a wide range of diseases. Although the intracellular measurement of NO is now feasible, many technical issues remain unresolved. The principle aim of our study was to determine the effect of storage time of whole blood on nitric oxide (NO) level expression in leukocytes. This is important because immune cells remain chemically dynamic even after they are removed from the circulation, and the impact of storage time must be known to optimally quantify the effect of a disease or condition on NO dynamics in circulating leukocytes...
November 9, 2017: Nitric Oxide: Biology and Chemistry
https://www.readbyqxmd.com/read/29127442/increased-frequencies-of-circulating-cxcl10-cxcl8-and-ccl4-producing-monocytes-and-siglec-3-expressing-myeloid-dendritic-cells-in-systemic-sclerosis-patients
#14
Tiago Carvalheiro, Sara Horta, Joel A G van Roon, Mariana Santiago, Maria J Salvador, Hélder Trindade, Timothy R D J Radstake, José A P da Silva, Artur Paiva
OBJECTIVE: To investigate the ex vivo pro-inflammatory properties of classical and non-classical monocytes as well as myeloid dendritic cells (mDCs) in systemic sclerosis (SSc) patients. METHODS: Spontaneous production of CXCL10, CCL4, CXCL8 and IL-6 was intracellularly evaluated in classical, non-classical monocytes and Siglec-3-expressing mDCs from peripheral blood of SSc patients and healthy controls (HC) through flow cytometry. In addition, production of these cytokines was determined upon toll-like receptor (TLR) 4 plus Interferon-γ (IFN-γ) stimulation...
November 10, 2017: Inflammation Research: Official Journal of the European Histamine Research Society ... [et Al.]
https://www.readbyqxmd.com/read/29123248/engineering-cell-robots-for-parallel-and-highly-sensitive-screening-of-biomolecules-under-in-vivo-conditions
#15
Lifu Song, An-Ping Zeng
Cells are capable of rapid replication and performing tasks adaptively and ultra-sensitively and can be considered as cheap "biological-robots". Here we propose to engineer cells for screening biomolecules in parallel and with high sensitivity. Specifically, we place the biomolecule variants (library) on the bacterial phage M13. We then design cells to screen the library based on cell-phage interactions mediated by a specific intracellular signal change caused by the biomolecule of interest. For proof of concept, we used intracellular lysine concentration in E...
November 9, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29122079/sinonasal-t-cell-expression-of-cytotoxic-mediators-granzyme-b-and-perforin-is-reduced-in-patients-with-chronic-rhinosinusitis
#16
Sarah E Smith, Rodney J Schlosser, James R Yawn, Jose L Mattos, Zachary M Soler, Jennifer K Mulligan
BACKGROUND: CD8+ T cells and natural killer (NK) cells are cytotoxic cells that use granzyme B (GrB) and perforin. Defective cytotoxic function is known to play a role in dysregulated immune response as seen in chronic sinusitis, also referred to as chronic rhinosinusitis (CRS). However, to our knowledge, in the United States, neither GrB or perforin expression has been reported in patients with CRS. OBJECTIVE: The aim of this study was to investigate sinonasal cytotoxic cells, their mediators, and cell-specific distribution of these mediators in patients with CRS with nasal polyp (CRSwNP) and in patients with CRS without nasal polyp (CRSsNP)...
November 1, 2017: American Journal of Rhinology & Allergy
https://www.readbyqxmd.com/read/29121457/drugs-implicated-in-systemic-autoimmunity-modulate-neutrophil-extracellular-trap-formation
#17
Jorge A Irizarry-Caro, Carmelo Carmona-Rivera, Daniella M Schwartz, Sami S Khaznadar, Mariana J Kaplan, Peter C Grayson
OBJECTIVE: Aberrant neutrophil extracellular trap (NET) formation has been implicated as a mechanism to induce auto-reactivity in at-risk individuals. The study objective was to assess whether medications implicated in cases of drug-induced autoimmunity (hydralazine and procainamide) and medications less commonly associated with drug-induced autoimmunity (minocycline and clozapine) induce NET formation and/or prevent NET degradation. METHODS: Human neutrophils were incubated with the drugs of interest and resultant NET formation was quantified by fluorescent microscopy...
November 9, 2017: Arthritis & Rheumatology
https://www.readbyqxmd.com/read/29118757/decreased-immunity-to-varicella-zoster-virus-in-giant-cell-arteritis
#18
Christien Rondaan, Kornelis S M van der Geest, Elisabeth Eelsing, Annemieke M H Boots, Nicolaas A Bos, Johanna Westra, Elisabeth Brouwer
Introduction: Herpes zoster, which can have a major impact on quality of life, results from reactivation of a latent varicella zoster virus (VZV) infection. We hypothesized that giant cell arteritis (GCA) patients are at increased risk of herpes zoster because of treatment with high-dose glucocorticoids and advanced age. Aim of the study, therefore, was to determine cell-mediated and humoral immunity to VZV in patients with GCA, patients with closely related disease polymyalgia rheumatica (PMR; treated with lower doses of glucocorticoids) and healthy controls (HCs)...
2017: Frontiers in Immunology
https://www.readbyqxmd.com/read/29115423/apoptosis-in-human-liver-carcinoma-caused-by-gold-nanoparticles-in-combination-with-carvedilol-is-mediated-via-modulation-of-mapk-akt-mtor-pathway-and-egfr-faad-proteins
#19
Raimundo F De Araújo, Jonas B Pessoa, Luis J Cruz, Alan B Chan, Emílio De Castro Miguel, Rômulo S Cavalcante, Gerly Anne C Brito, Heloiza Fernada O Silva, Luiz H S Gasparotto, Paulo M M Guedes, Aurigena A Araújo
In cancers, apoptosis signaling pathways and cell survival and growth pathways responsible for resistance to conventional treatments, such as Pi3K/Akt/mTOR and mitogen-activated protein kinase (MAPK) become dysregulated. Recently, alternative treatments to promote tumor cell death have become important. The present study reports on the antitumor and cytoprotective action of gold nanoparticles (GNPs) and carvedilol in combination and in isolated application. Apoptosis was analyzed by FITC/propidium iodide staining flow cytometry; caspase-3, caspase-8, Bcl-2 and MAPK/ERK activity by immunofluorescence microscopy; gene expression of proteins related to cell death as Akt, mTOR, EGFR, MDR1, survivin, FADD and Apaf, by the real-time PCR; and western blot analysis for MAPK/ERK, Akt and mTOR...
October 27, 2017: International Journal of Oncology
https://www.readbyqxmd.com/read/29113727/effect-of-single-layer-centrifugation-porcicoll-70-80-and-90-or-supplementation-with-reduced-glutathione-seminal-plasma-and-bovine-serum-albumin-on-frozen-thawed-boar-sperm
#20
I Crespo-Félez, A Castañeda-Sampedro, D I Sánchez, E Fernández-Alegre, M Álvarez-Rodríguez, J C Domínguez, J M Morrell, F Martínez-Pastor
Selecting the optimal sperm population is essential for success with reproductive techniques. Porcicoll (formerly Androcoll-P) is a colloid formulation for selection of high-quality boar spermatozoa by single layer centrifugation (SLC). To date, most studies have been carried out with fresh semen and large volumes. We carried out 2 experiments to test the use of Porcicoll for thawed boar semen in small volumes. In Experiment 1, cryopreserved semen doses were thawed, split in 200-μL aliquots and layered on 1mL of Porcicoll 70%, 80% or 90%, or buffer without colloid...
November 4, 2017: Animal Reproduction Science
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