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Clinical flow cytometry

Gabriele Picco, Consalvo Petti, Francesco Sassi, Katia Grillone, Giorgia Migliardi, Teresa Rossi, Claudio Isella, Federica Di Nicolantonio, Ivana Sarotto, Anna Sapino, Alberto Bardelli, Livio Trusolino, Andrea Bertotti, Enzo Medico
BACKGROUND: The NEDD8 conjugation pathway modulates the ubiquitination and activity of a wide range of intracellular proteins, and its blockade by pevonedistat is emerging as a promising therapeutic approach in various cancer settings. However, systematic characterization of pevonedistat efficacy in specific tumor types and definition of response predictors are still missing. METHODS: We investigated in vitro sensitivity to pevonedistat in 122 colorectal cancer (CRC) cell lines by an ATP-based proliferation assay and evaluated apoptosis and DNA content by flow cytometry...
February 2017: Journal of the National Cancer Institute
Sergio Montserrat-de la Paz, M Carmen Naranjo, Sergio Lopez, Rocio Abia, Francisco J Garcia Muriana, Beatriz Bermudez
Niacin is a broad-spectrum lipid-regulating drug used for clinical therapy of chronic high-grade inflammatory diseases. However, the mechanisms by which either niacin or the byproducts of its catabolism ameliorate these inflammatory diseases are not clear yet. Human circulating monocytes and mature macrophages were used to analyze the effects of niacin and its metabolites (NAM, NUA and 2-Pyr) on oxidative stress, plasticity and inflammatory response by using biochemical, flow cytometry, quantitative real-time PCR and Western blot technologies...
October 1, 2016: Journal of Nutritional Biochemistry
J Fotschki, A M Szyc, J M Laparra, L H Markiewicz, B Wróblewska
The aim of this study was to examine immune adaptive changes, the expression of innate biomarkers and variations in intestinal microbiota composition after horse-milk administration in BALB/c mice, which were sensitized intraperitoneally using cow β-lactoglobulin and α-casein with aluminum adjuvant. We measured serum antibody IgE levels and the expression of MCP-1, IL-4, and TNF-α in duodenal samples. Changes in immune cell populations in peripheral blood were quantified using flow cytometry, and intestinal microbiota composition was assessed using real-time PCR...
October 19, 2016: Journal of Dairy Science
Deying Liu, Jinxiang Liu, Jinghua Wang, Congcong Liu, Sirui Yang, Yanfang Jiang
BACKGROUND: Circulating follicular helper T (Tfh) cells are a heterogeneous population of CD4(+) helper T cells that promotes pathogenic immune responses in autoimmune diseases. In this study, we examined the status of different subpopulations of Tfh cells in peripheral circulation and their associations with various clinical characteristics of IgA vasculitis (IgAV). METHODS: According to the phenotypic expressions of different molecules, focus was given on six subpopulations of Tfh cells: CD4(+)CXCR5(+), CD4(+)CXCR5(+)ICOS(+), CD4(+)CXCR5(+)ICOS(+)PD-1(+), CD4(+)CXCR5(+)ICOS(high)PD-1(high), CD4(+)CXCR5(+)ICOS(-)PD-1(+), and CXCR5(+)CD45RA(-)IL-21(+)...
October 21, 2016: BMC Immunology
A L Litvinenko, A E Moskalensky, N A Karmadonova, V M Nekrasov, D I Strokotov, A I Konokhova, M A Yurkin, E A Pokushalov, A V Chernyshev, V P Maltsev
Whereas commercially available hematological analyzers measure volume of individual platelets, angle-resolved light-scattering provides unique ability to additionally measure their shape index. We utilized the scanning flow cytometer to measure light-scattering profiles (LSPs) of individual platelets taken from 16 healthy donors and the solution of the inverse light-scattering problem to retrieve the volume and shape index of each platelet. In normal conditions, the platelet shape index distribution (PSID) demonstrates three peaks, which relate to resting, partially activated, and fully activated platelets...
October 21, 2016: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
Kristian Jensen, Rikke Krusenstjerna-Hafstrøm, Jesper Lohse, Kenneth H Petersen, Helene Derand
In clinical routine pathology today, detection of protein in intact formalin-fixed, paraffin-embedded tissue is limited to immunohistochemistry, which is semi-quantitative. This study presents a new and reliable quantitative immunohistochemistry method, qIHC, based on a novel amplification system that enables quantification of protein directly in formalin-fixed, paraffin-embedded tissue by counting of dots. The qIHC technology can be combined with standard immunohistochemistry, and assessed using standard bright-field microscopy or image analysis...
October 21, 2016: Modern Pathology: An Official Journal of the United States and Canadian Academy of Pathology, Inc
Eeva-Liisa Paattiniemi, Santra Karumaa, Anna-Mari Viita, Pauliina Kärpänoja, Marjukka Mäkelä, Jaana Isojärvi, Hannu Sarkkinen
Urine culture samples comprise a large proportion of the workload in clinical microbiology laboratories, and most of the urine samples show no growth or insignificant growth. A flow cytometry-based analyzer (Sysmex Corporation, Japan) has been used to screen out negative urine samples prior to culture in the Päijät-Häme district. We applied decision analytic modelling to analyze, from a laboratory perspective, the economic feasibility of the screening method as compared to culture only (conventional method) for diagnosis of urinary tract infection...
October 21, 2016: Infectious Diseases
Mikkel Staberg, Signe Regner Michaelsen, Rikke Darling Rasmussen, Mette Villingshøj, Hans Skovgaard Poulsen, Petra Hamerlik
PURPOSE: Glioblastoma (GBM) ranks among the deadliest solid cancers worldwide and its prognosis has remained dismal, despite the use of aggressive chemo-irradiation treatment regimens. Limited drug delivery into the brain parenchyma and frequent resistance to currently available therapies are problems that call for a prompt development of novel therapeutic strategies. While only displaying modest efficacies as mono-therapy in pre-clinical settings, histone deacetylase inhibitors (HDACi) have shown promising sensitizing effects to a number of cytotoxic agents...
October 20, 2016: Cellular Oncology (Dordrecht)
Alicia Martinez-Varea, Roberto Romero, Yi Xu, Derek Miller, Ahmed I Ahmed, Piya Chaemsaithong, Noppadol Chaiyasit, Lami Yeo, Majid Shaman, Kia Lannaman, Benjamin Cher, Sonia S Hassan, Nardhy Gomez-Lopez
OBJECTIVES: 1) To characterize the cellular composition of the amniotic fluid of patients diagnosed with clinical chorioamnionitis at term, as a function of the presence or absence of microorganisms determined by cultivation techniques, and 2) to characterize the cytokine production by white blood cells present in the amniotic fluid using flow cytometry-based techniques. MATERIALS AND METHODS: Amniotic fluid samples from 20 women who had the diagnosis of clinical chorioamnionitis at term were analyzed using cultivation techniques (for aerobic and anaerobic bacteria as well as genital Mycoplasmas)...
October 20, 2016: Journal of Perinatal Medicine
Seyed Hadi Mousavi, Azar Hosseini, Elham Bakhtiari, Hassan Rakhshandeh
OBJECTIVE: Doxorubicin (DOX) is an effective anticancer drug but its clinical application is limited because it induces apoptosis in cardiomyocytes and leads to permanent degenerative cardiomyopathy and heart failure possibly due to oxidative stress. Recent studies showed that Capparis spinosa (C. spinose) exhibits potent antioxidant activity. So, in this study, we explored the protective effect of hydro-alcoholic extract of C. spinosa against DOX-induced cytotoxicity in H9c2 cells. MATERIALS AND METHODS: Cell viability was quantified by MTT assay...
September 2016: Avicenna Journal of Phytomedicine
Monika Ryba-Stanisławowska, Paulina Werner, Maria Skrzypkowska, Agnieszka Brandt, Jolanta Myśliwska
IL-33 is an IL-1 cytokine family member, with ability to induce both Th1 and Th2 immune responses. It binds to ST2 receptor, whose deficiency is associated with enhanced inflammatory response. The most recent studies have shown the immunoregulatory effect of IL-33 on Tregs in animal models. As type 1 diabetes is an autoimmune, inflammatory disease, where Treg defects have been described, we aimed to analyze the in vitro influence of recombinant IL-33 on quantitative properties of regulatory CD4(+)CD25(high)FOXP3(+) T cells...
2016: Mediators of Inflammation
Geraint P Williams, Peter Nightingale, Sue Southworth, Alastair K O Denniston, Paul J Tomlins, Stephen Turner, John Hamburger, Simon J Bowman, S John Curnow, Saaeha Rauz
Purpose: Ocular mucous membrane pemphigoid (OcMMP) is a rare autoimmune disorder resulting in progressive conjunctival fibrosis and ocular surface failure leading to sight loss in up to 50%. This study was designed to optimize an ocular surface sampling technique for identification of novel biomarkers associated with disease activity and/or progressive fibrosis. Methods: Fifty-seven patients with OcMMP underwent detailed examination of conjunctival inflammation and fibrosis using fornix depth measurement...
October 1, 2016: Investigative Ophthalmology & Visual Science
Adam J Mitchell, Warren D Gray, Max Schroeder, Hong Yi, Jeannette V Taylor, Rebecca S Dillard, Zunlong Ke, Elizabeth R Wright, David Stephens, John D Roback, Charles D Searles
BACKGROUND: Red blood cell (RBC) transfusions are a common, life-saving therapy for many patients, but they have also been associated with poor clinical outcomes. We identified unusual, pleomorphic structures in human RBC transfusion units by negative-stain electron microscopy that appeared identical to those previously reported to be bacteria in healthy human blood samples. The presence of viable, replicating bacteria in stored blood could explain poor outcomes in transfusion recipients and have major implications for transfusion medicine...
2016: PloS One
H Zöllner, R Jouni, S Panzer, A Khadour, L Janzen, J Wesche, M Ten Berg, S Schellong, A Heinken, A Greinacher, T Bakchoul
BACKGROUND: Protamine (PRT) is used to stabilize insulin in Neutral Protamine Hagedorn (NPH) insulin, a commonly used therapeutic agent for diabetes mellitus. Immunization against PRT/heparin-complexes is common in diabetic patients. OBJECTIVES: To investigate the impact of NPH-insulin on the interaction between anti-PRT/heparin antibodies and platelets. METHODS: The interaction between NPH-insulin and anti-PRT/heparin antibodies was tested using in-house enzyme immunoassays...
October 19, 2016: Journal of Thrombosis and Haemostasis: JTH
Katarzyna Sznurkowska, Anton Żawrocki, Jacek Sznurkowski, Maciej Zieliński, Piotr Landowski, Katarzyna Plata-Nazar, Ewa Iżycka-Świeszewska, Piotr Trzonkowski, Agnieszka Szlagatys-Sidorkiewicz, Barbara Kamińska
BACKGROUND/AIMS: To determine the proportion of T-regulatory cells (CD4(+)CD25(high)FOXP3(+) cells) in peripheral blood and the number of FOXP3(+) cells in intestinal mucosa of children with inflammatory bowel disease (IBD), and to verify whether these parameters correlate with the activity of the disease. MATERIAL AND METHODS: 24 patients newly diagnosed for IBD were included in the study: ulcerative colitis (UC; n = 13) and Crohn's disease (CD; n = 11). Seventeen healthy controls (HC) and 16 patients with irritable bowel syndrome (IBS) served as a control group for peripheral and intestinal Tregs assessment, respectively...
October 19, 2016: Immunological Investigations
Monique C Minnema, Eva Kimby, Shirley D'Sa, Luc-Matthieu Fornecker, Stéphanie Poulain, Tom J Snijders, Efstathios Kastritis, Stéphane Kremer, Aikaterini Fitsiori, Laurence Simon, Frédéric Davi, Michael Lunn, Jorge J Castillo, Christopher J Patterson, Magali Le Garff-Tavernier, Myrto Costopoulos, Véronique Leblond, Marie-José Kersten, Meletios A Dimopoulos, Steven P Treon
Bing Neel syndrome is a rare disease manifestation of Waldenstrom macroglobulinemia that results from infiltration of the central nervous system by malignant lymphoplasmacytic cells. In this guideline, we describe the clinical symptoms, as well as the appropriate laboratory and radiological studies that can aid in the diagnosis. The presentation of Bing Neel syndrome may be very diverse, and include headaches, cognitive deficits, paresis, and psychiatric symptoms. The syndrome can present in patients with known Waldenstrom macroglobulinemia, even in the absence of systemic progression, but also in previously undiagnosed patients...
October 6, 2016: Haematologica
Troels Holz Borch, Lotte Engell-Noerregaard, Trine Zeeberg Iversen, Eva Ellebaek, Özcan Met, Morten Hansen, Mads Hald Andersen, Per Thor Straten, Inge Marie Svane
INTRODUCTION: Vaccination with dendritic cells (DCs) has generally not fulfilled its promise in cancer immunotherapy due to ineffective translation of immune responses into clinical responses. A proposed reason for this is intrinsic immune regulatory mechanisms, such as regulatory T cells (Tregs). A metronomic regimen of cyclophosphamide (mCy) has been shown to selectively deplete Tregs. To test this in a clinical setting, we conducted a phase I trial to evaluate the feasibility and safety of vaccination with DCs transfected with mRNA in combination with mCy in patients with metastatic malignant melanoma (MM)...
2016: Oncoimmunology
Radosław Januchowski, Karolina Sterzyńska, Katarzyna Zaorska, Patrycja Sosińska, Andrzej Klejewski, Maciej Brązert, Michał Nowicki, Maciej Zabel
BACKGROUND: Multiple drug resistance (MDR) of cancer cells is the main reason of intrinsic or acquired insensitivity to chemotherapy in many cancers. In this study we used ovarian cancer model of acquired drug resistance to study development of MDR. We have developed eight drug resistant cell lines from A2780 ovarian cancer cell line: two cell lines resistant to each drug commonly used in ovarian cancer chemotherapy: cisplatin (CIS), paclitaxel (PAC), doxorubicin (DOX) and topotecan (TOP)...
October 18, 2016: Journal of Ovarian Research
Gerrit Ahrenstorf, Hui-Zhi Low, Katja Kniesch, David Ordonez, Dirk Meyer-Olson, Fareed Ahmad, Claudia Kücherer, Barbara Gunsenheimer-Bartmeyer, Matthias Stoll, Torsten Matthias, Reinhold E Schmidt, Torsten Witte
OBJECTIVE: To analyze the influence of LILRA3 and the genetic LILRA3 deletion on transmission and clinical course of HIV infection. DESIGN: Case and control study. METHODS: LILRA3 genotypes were determined by polymerase chain reaction. HIV patients were categorized into short-term progressors, normal progressors and long-term non-progressors according to the clinical course. Functional studies were performed using real-time PCR, intracellular flow cytometry and ELISA...
October 14, 2016: AIDS
Hisatake Matsumoto, Kazuma Yamakawa, Hiroshi Ogura, Taichin Koh, Naoya Matsumoto, Takeshi Shimazu
Activated immune cells such as monocytes are key factors in systemic inflammatory response syndrome (SIRS) following trauma and sepsis. Activated monocytes induce almost all tissue factor (TF) expression contributing to inflammation and coagulation. TF and CD13 double-positive microparticles (TF/CD13MPs) are predominantly released from these activated monocytes. This study aimed to evaluate TF/CD13MPs and assess their usefulness as a biomarker of pathogenesis in early SIRS following trauma and sepsis. This prospective study comprising 24 trauma patients, 25 severe sepsis patients and 23 healthy controls was conducted from November 2012 to February 2015...
October 6, 2016: Shock
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