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Nur-Vaizura Mohamad, Muhammad Afiq Amani Che Zulkepli, Krystine May Theseira, Norain Zulkifli, Nur Quraisha Shahrom, Nurul Amni Mohamad Ridzuan, Nor Aini Jamil, Ima-Nirwana Soelaiman, Kok-Yong Chin
Introduction: Orchidectomy is currently the preferred method to induce bone loss in preclinical male osteoporosis model. Gonadotropin-releasing hormone (GnRH) agonists used in prostate cancer treatment can induce testosterone deficiency but its effects on bone in preclinical male osteoporosis model are less studied. Objective: This study aimed to evaluate the skeletal effect of buserelin (a GnRH agonist) in male rats and compare it with orchidectomy. Methods: Forty-six three-month-old male Sprague-Dawley rats were divided into three experimental arms...
2018: International Journal of Medical Sciences
Molly K Hritzo, Jean-Paul Courneya, Amit Golding
While flow cytometry can reliably assess surface and intracellular marker expression within small cell populations, it does not provide any information on protein localization. Several key transcription factors (TF) downstream of lymphocyte surface receptors are regulated by nuclear versus cytoplasmic localization, and one such TF is Forkhead box O1 (FOXO1). FOXO1 integrates antigen-binding, co-receptor activation and metabolic signals in lymphocytes, leading to proliferation and differentiation. Importantly, the nuclear or cytoplasmic localization of FOXO1 is key for gene expression leading to different lymphocyte phenotypes...
March 2018: Journal of Immunological Methods
Henry Hui, Kathryn A Fuller, Hun Chuah, James Liang, Hasib Sidiqi, Dejan Radeski, Wendy N Erber
Chronic Lymphocytic Leukaemia (CLL), the most common leukaemia in the Western world, has a characteristic phenotype and prognosis largely defined by the presence of cytogenetic aberrations. The gold standard for detecting these cytogenetic abnormalities is interphase fluorescence in situ hybridisation (FISH) performed on cell smears or tissue sections on glass slides. Fluorescently labelled DNA probes bind to specific chromosomal regions and the signal detected by fluorescent microscopy. Generally only 200 cells are assessed and the limit of sensitivity is 3% positive cells...
February 1, 2018: Methods: a Companion to Methods in Enzymology
Laurent Muller, Patricia Simms, Chang-Sook Hong, Michael I Nishimura, Edwin K Jackson, Simon C Watkins, Theresa L Whiteside
Tumor-derived exosomes (TEX) are ubiquitously present in the tumor microenvironment and plasma of cancer patients. TEX carry a cargo of multiple stimulatory and inhibitory molecules and deliver them to recipient cells, serving as a communication network for the tumor. The mechanisms TEX use for delivering messages to recipient cells were evaluated using PKH26-labeled TEX produced by cultured human tumor cells, exosomes produced by dendritic cells-derived exosomes (DEX), or exosomes isolated from plasma of cancer patients (EXO)...
2017: Oncoimmunology
Michael S Misale, Linda Witek Janusek, Dina Tell, Herbert L Mathews
It is well-established that psychological distress reduces natural killer cell immune function and that this reduction can be due to the stress-induced release of glucocorticoids. Glucocorticoids are known to alter epigenetic marks associated with immune effector loci, and are also known to influence chromatin organization. The purpose of this investigation was to assess the effect of glucocorticoids on natural killer cell chromatin organization and to determine the relationship of chromatin organization to natural killer cell effector function, e...
January 2018: Brain, Behavior, and Immunity
Yasaman Alaghband, Janine L Kwapis, Alberto J López, André O White, Osasumwen V Aimiuwu, Amni Al-Kachak, Kasuni K Bodinayake, Nicole C Oparaugo, Richard Dang, Mariam Astarabadi, Dina P Matheos, Marcelo A Wood
Histone deacetylases (HDACs) are chromatin modifying enzymes that have been implicated as powerful negative regulators of memory processes. HDAC3has been shown to play a pivotal role in long-term memory for object location as well as the extinction of cocaine-associated memory, but it is unclear whether this function depends on the deacetylase domain of HDAC3. Here, we tested whether the deacetylase domain of HDAC3has a role in object location memory formation as well as the formation and extinction of cocaine-associated memories...
November 2017: Neurobiology of Learning and Memory
S Denk, R P Taylor, R Wiegner, E M Cook, M A Lindorfer, K Pfeiffer, S Paschke, T Eiseler, M Weiss, E Barth, J D Lambris, M Kalbitz, T Martin, H Barth, D A C Messerer, F Gebhard, M S Huber-Lang
The complement and neutrophil defence systems, as major components of innate immunity, are activated during inflammation and infection. For neutrophil migration to the inflamed region, we hypothesized that the complement activation product C5a induces significant changes in cellular morphology before chemotaxis. Exposure of human neutrophils to C5a dose- and time-dependently resulted in a rapid C5a receptor-1 (C5aR1)-dependent shape change, indicated by enhanced flow cytometric forward-scatter area values. Similar changes were observed after incubation with zymosan-activated serum and in blood neutrophils during murine sepsis, but not in mice lacking the C5aR1...
September 2017: Scandinavian Journal of Immunology
Nana H Overgaard, Jazmina L Cruz, Jennifer A Bridge, Hendrik J Nel, Ian H Frazer, Nicole L La Gruta, Antje Blumenthal, Raymond J Steptoe, James W Wells
CD4(+)CD8(+) double-positive (DP), mature, peripheral T cells are readily detectable in a variety of species and tissues. Despite a common association with autoimmune and malignant skin disorders, however, little is understood about their role or function. Herein, we show that DP T cells are readily detectable in the blood, spleen, and peripheral lymph nodes of naïve C57BL/6 mice. DP T cells were also present in Jα18(-/-) and CD1d(-/-) mice, indicating that these cells are not NK-T cells. After skin administration of CASAC adjuvant, but not Quil A adjuvant, both total DP T cells and skin-infiltrating DP T cells increased in number...
September 2017: Journal of Leukocyte Biology
Mark Hildebrand, Aubrey Davis, Raffaela Abbriano, Haley R Pugsley, Jesse C Traller, Sarah R Smith, Roshan P Shrestha, Orna Cook, Eva L Sánchez-Alvarez, Kalpana Manandhar-Shrestha, Benjamin Alderete
The ability to image large numbers of cells at high resolution enhances flow cytometric analysis of cells and cell populations. In particular, the ability to image intracellular features adds a unique aspect to analyses, and can enable correlation between molecular phenomena resulting in alterations in cellular phenotype. Unicellular microalgae are amenable to high-throughput analysis to capture the diversity of cell types in natural samples, or diverse cellular responses in clonal populations, especially using imaging cytometry...
2016: Methods in Molecular Biology
David A Basiji
Over the last decade imaging flow cytometry (IFC) has become an established technique, as evidenced by its use in over 500 peer-reviewed scientific articles. Nevertheless, it is still an emerging technique with an installed base of less than 5 % that of conventional flow cytometers. In parallel with its adoption, the technology has evolved rapidly, increasing in speed, sensitivity, and real-time data analysis capacity by over an order of magnitude since its introduction. This chapter summarizes IFC's basic principles of operation and describes the current state of the art...
2016: Methods in Molecular Biology
Aldo Nascimento, Joanne Lannigan, David Kashatus
Mitochondria are highly dynamic organelles whose fusion and fission play an increasingly important role in a number of both normal and pathological cellular functions. Despite the increased interest in mitochondrial dynamics, robust, and quantitative methods to analyze mitochondrial fusion and fission activity in intact cells have not been developed. The current state-of-the art method to measure mitochondrial fusion activity is the polyethylene glycol (PEG) fusion assay in which cells expressing distinct mitochondrially-targeted fluorescent proteins (FPs) are fused together and mitochondrial fusion activity is determined by the rate at which color mixing occurs...
August 2016: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
I Tzoran, A Rebibo-Sabbah, B Brenner, A Aharon
INTRODUCTION: Acute myeloid leukemia (AML) is characterized by rapid growth of leukemic blast cells. Extracellular vesicles (EVs) are shed from normal and pathologic cells and express membrane proteins and antigens, reflecting their cellular origin. AIM: To explore whether bone marrow EVs of AML patients originate from blast cells and are capable of influencing hematopoietic stem cells (HSC) in a pseudo-natural microenvironment obtained by co-culture of HSC with mesenchymal stem cells (MSC)...
April 2016: Thrombosis Research
Feng Qian, Ruth R Montgomery
To demonstrate effects of aging visually requires a robust technique that can reproducibly detect small differences in efficiency or kinetics between groups. Investigators of aging will greatly appreciate the benefits of Amnis ImageStream technology (, which combines quantitative flow cytometry with simultaneous high-resolution digital imaging. Imagestream is quantitative, reproducible, feasible with limited samples, and it facilitates in-depth examination of cellular mechanisms between cohorts of samples...
2015: Methods in Molecular Biology
Christopher N Parris, Sheba Adam Zahir, Hussein Al-Ali, Emma C Bourton, Christina Plowman, Piers N Plowman
Accurate and rapid methods for the detection of DNA damage foci in eukaryotic cells are central to DNA repair studies, which identify differences in DNA repair capacity in cell lines. Such assays have been important in delineating mechanisms of DNA repair in human cells. Previously we were the first to demonstrate the use of imaging flow cytometry for the detection of γ-H2AX foci in cells exposed to ionizing radiation causing the induction of DNA strand breaks. In this report we extend these studies and show an enhancement of foci quantitation and image resolution using next generation imaging flow cytometry with the Amnis Imagestream(X) Mark II...
August 2015: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
Vanessa Lagal, Márcia Dinis, Dominique Cannella, Daniel Bargieri, Virginie Gonzalez, Nicole Andenmatten, Markus Meissner, Isabelle Tardieux
The apical membrane antigen 1 (AMA1) protein was believed to be essential for the perpetuation of two Apicomplexa parasite genera, Plasmodium and Toxoplasma, until we genetically engineered viable parasites lacking AMA1. The reduction in invasiveness of the Toxoplasma gondii RH-AMA1 knockout (RH-AMA1(KO)) tachyzoite population, in vitro, raised key questions about the outcome associated with these tachyzoites once inoculated in the peritoneal cavity of mice. In this study, we used AMNIS technology to simultaneously quantify and image the parasitic process driven by AMA1(KO) tachyzoites...
June 2015: Infection and Immunity
Sarah E Headland, Hefin R Jones, Adelina S V D'Sa, Mauro Perretti, Lucy V Norling
Interest in extracellular vesicle biology has exploded in the past decade, since these microstructures seem endowed with multiple roles, from blood coagulation to inter-cellular communication in pathophysiology. In order for microparticle research to evolve as a preclinical and clinical tool, accurate quantification of microparticle levels is a fundamental requirement, but their size and the complexity of sample fluids present major technical challenges. Flow cytometry is commonly used, but suffers from low sensitivity and accuracy...
June 10, 2014: Scientific Reports
Giuseppina Bologna, Paola Lanuti, Primiano D'Ambrosio, Lucia Tonucci, Laura Pierdomenico, Carlo D'Emilio, Nicola Celli, Marco Marchisio, Nicola d'Alessandro, Eugenio Santavenere, Mario Bressan, Sebastiano Miscia
Breast cancer represents the second cause of death in the European female population. The lack of specific therapies together with its high invasive potential are the major problems associated to such a tumor. In the last three decades platinum-based drugs have been considered essential constituents of many therapeutic strategies, even though with side effects and frequent generation of drug resistance. These drugs have been the guide for the research, in last years, of novel platinum and ruthenium based compounds, able to overcome these limitations...
June 2014: Biometals: An International Journal on the Role of Metal Ions in Biology, Biochemistry, and Medicine
Carrie A Franzen, Patricia E Simms, Adam F Van Huis, Kimberly E Foreman, Paul C Kuo, Gopal N Gupta
Bladder tumors represent a special therapeutic challenge as they have a high recurrence rate requiring repeated interventions and may progress to invasive or metastatic disease. Exosomes carry proteins implicated in bladder cancer progression and have been implicated in bladder cancer cell survival. Here, we characterized exosome uptake and internalization by human bladder cancer cells using Amnis ImageStreamX, an image cytometer. Exosomes were isolated by ultracentrifugation from bladder cancer culture conditioned supernatant, labeled with PKH-26, and analyzed on the ImageStreamX with an internal standard added to determine concentration...
2014: BioMed Research International
Amny N Mendenhall, Susan Frauenholtz
No abstract text is available yet for this article.
October 2013: Social Work
Iris Marangon, Nicole Boggetto, Cécilia Ménard-Moyon, Nathalie Luciani, Claire Wilhelm, Alberto Bianco, Florence Gazeau
Carbon-based nanomaterials, like carbon nanotubes (CNTs), belong to this type of nanoparticles which are very difficult to discriminate from carbon-rich cell structures and de facto there is still no quantitative method to assess their distribution at cell and tissue levels. What we propose here is an innovative method allowing the detection and quantification of CNTs in cells using a multispectral imaging flow cytometer (ImageStream, Amnis). This newly developed device integrates both a high-throughput of cells and high resolution imaging, providing thus images for each cell directly in flow and therefore statistically relevant image analysis...
2013: Journal of Visualized Experiments: JoVE
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