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cytometri of flow

Razavi Hashemi M, Vahabpour R, Ranji N, Sanati Mh, Mapar M, Sadat Sm
BACKGROUND: Acute T Cell Leukemia (ALL) is an aggressive and prevalent human malignancy. Chemotherapy is the most frequent therapeutic strategy; however, cytotoxicity and recurrence of cancer are the main concerns. The discovery of microRNA (miRNA) drew the attention of scientists who were working on targeted cancer therapy to use the potential of gene regulation by using this small RNAs. METHODS: miRanda, TargetScan, miRDB databases, and miRWalk software were applied to find probable miRNAs targeting 3'UTR of JAK1, STAT3, SOCS6, AKT1, APAF, BID, and Caspase9 mRNA...
October 1, 2018: Clinical Laboratory
Neha Sharma, Lisamarie Moore, Shravanthi Chidambaram, Nicholas W Colangelo, Sonia M de Toledo, Edouard I Azzam
Neural stem cells are attracting enormous attention in regenerative medicine due to their ability to self-renew and differentiate into the cell lineages that constitute the central nervous system. However, little is known about the mechanism underlying the regulation of their redox environment, which is essential for homeostatic cellular functions. The redox-modulated c-Jun N-terminal kinases (JNK) are a molecular switch in stress signal transduction and are involved in numerous brain functions. Using a selective but broad-spectrum inhibitor of JNK 1/2/3, we investigated the role of JNK in regulating the levels of reactive oxygen species in mitochondria, mitochondrial membrane potential, viability, proliferation and lineage alterations in human H9-derived neural stem/progenitor cells (NSPs)...
October 18, 2018: Developmental Neuroscience
Jalil Mehrzad, Saman Hosseinkhani, Amir Mohammad Malvandi
OBJECTIVE: Knowledge regarding interactions of AFB1 with the human nervous system and how a naturally occurring level of AFB1 could potentially induce neuroimmune dysregulation is very limited. To assess the cellular effects of AFB1 on the human brain, we used the human microglia cell line CHME5 as a model to pinpoint its potential in vivo translation. METHODS: We used the CHME5 cell line culture system, multiplex qPCR, (chemi)bioluminescence, Luminex ELISA, and flow cytometry assays to evaluate the toxic effects of a naturally occurring level of AFB1 on human microglia...
October 18, 2018: Neuroimmunomodulation
Yuanyuan Lu, Xinglou Liu, Yuan Huang, Yi Liao, Ting Xi, Yanan Zhang, Sainan Shu, Feng Fang
OBJECTIVE: The aim of this study was to determine which factors lead to the susceptibility to mouse cytomegalovirus (MCMV) in the spleens of BALB/c mice. METHODS: BALB/c and C57BL/6 mice were randomly divided into a control group and an infection group and sacrificed on day 0, 1, 3, 7, 14, and 28 postinfection. The cytotoxicity of NK cells was determined by evaluating lactate dehydrogenase contents. Flow cytometry was used to analyze activated NK cells, IFNγ+ NK cells, and total NK cells in the spleen...
October 18, 2018: Intervirology
Enguang Lv, Jiawang Ding, Wei Qin
Peptide-based sandwich assays are promising tools in molecular detection, but may be restricted by the availability of "pairs" of affinity peptides. Herein, a new potentiometric sandwich assay for bacteria involving peptide pairs derived from an antimicrobial peptide (AMP) ligand is demonstrated. As a model, the original AMP with well-defined structure for Listeria monocytogenes (LM) can be split into two fragments to serve as the peptide pairs for the sandwich assay. The recognition and binding of the short peptide pairs to the target can be verified by circular dichroism, flow cytometry, fluorometry, and optical microscopy...
October 18, 2018: Analytical Chemistry
Julia Fernández-Pérez, Mark Ahearne
PURPOSE: To identify biochemical cues that could promote a keratocyte-like phenotype in human corneal stromal cells that had become fibroblastic when expanded in serum-supplemented media while also examining the effect on cell proliferation and migration. METHODS: Proliferation was assessed by PrestoBlueTM , morphology was monitored by phase contrast microscopy, phenotype was analysed by real-time polymerase chain reaction (qPCR), immunochemistry and flow cytometry, and migration was studied with a scratch assay...
October 18, 2018: Current Eye Research
Lina Marcela Barrera, León Darío Ortiz, Hugo Grisales, Mauricio Rojas, Mauricio Camargo
Introduction: The quantification of chromosomal instability is an important parameter to assess genotoxicity and radiosensitivity. Most conventional techniques require cell cultures or laborious microscopic analyses of chromosomes or nuclei. However, a flow cytometry that selects the reticulocytes has been developed as an alternative for in vivo studies, which expedites the analytical procedures and increases up to 20 times the number of target cells to be analyzed. Objectives: To standardize the flow cytometry parameters for selecting and quantifying the micronucleated reticulocytesCD71+ (MN-RET) from freshly drawn peripheral blood and to quantify the frequency of this abnormal cell subpopulation as a measure of cytogenetic instability in populations of healthy volunteers (n =25), and patients (n=25), recently diagnosed with high-grade gliomas before the onset of treatment...
September 1, 2018: Biomédica: Revista del Instituto Nacional de Salud
Nhora Silva, Natalia Calvache, Elizabeth Arrieta, Juan Carlos Bravo, Camilo Salazar, Joaquín Rosales, Francisco Jaramillo, Juan Guillermo Restrepo
The small lymphocytic lymphoma is a mature B cell neoplasm with a broad spectrum of clinical presentations. Opportunistic infections that are not related to the treatment, even in advanced stages, have a low incidence rate. There are few case reports in the medical literature of patients who have not received immunosuppressive therapy and present with small lymphocytic lymphoma associated with disseminated histoplasmosis at diagnosis. A female 82-year-old patient was admitted due to an intermittent dry cough, asthenia, and adynamia that had persisted for one month...
September 1, 2018: Biomédica: Revista del Instituto Nacional de Salud
Amy Galvin, Meredith Weglarz, Kat Folz-Donahue, Maris Handley, Misa Baum, Michael Mazzola, Hannah Litwa, David T Scadden, Lev Silberstein
Maintenance of hematopoietic stem cell (HSC) quiescence is critical for self-renewal and differentiation into mature lineages. Therefore, the ability to reliably detect abnormal HSC cycling is essential for experiments that seek to investigate abnormalities of HSC function. The ability to reproducibly evaluate cell cycle status in a rare cell subset requires careful optimization of multiple parameters during cell preparation and sample processing. Here, we describe a method where data acquisition parameters and fluorochrome combination for long-term HSC staining have been specifically designed for concurrent use with DAPI and Ki-67 antibodies...
October 18, 2018: Current Protocols in Cytometry
Janine van Duijn, Eva Kritikou, Naomi Benne, Thomas van der Heijden, Gijs H van Puijvelde, Mara J Kröner, Frank H Schaftenaar, Amanda C Foks, Anouk Wezel, Harm Smeets, Hideo Yagita, Ilze Bot, Wim Jiskoot, Johan Kuiper, Bram Slütter
Aims: T lymphocytes play an important role in atherosclerosis development, but the role of the CD8+ T-cell remains debated, especially in the clinically relevant advanced stages of atherosclerosis development. Here, we set out to determine the role of CD8+ T-cells in advanced atherosclerosis. Methods and Results: Human endarterectomy samples analyzed by flow cytometry showed a negative correlation between the percentage of CD8+ T-cells and macrophages, suggesting a possible protective role for these cells in lesion development...
October 18, 2018: Cardiovascular Research
Chen-Jan Chiu, Chih-Wen Chi, Hui-Ru Hsieh, Yu-Chuen Huang, Huey-June Wu, Yu-Jen Chen
The aim of this study was to examine the effect of the level-1 Yo-Yo intermittent recovery test (YYIRT1) on polarization of macrophages in young football players.Fourteen male football players (19.9 ± 1.4 years old) were enrolled in this study. YYIRT1 was performed with 20-meter shuttle runs at increasing speeds and 10-second active recovery in a 5-meter distance between runs till exhaustion. Fasting blood samples were collected before and immediately after YYIRT1. Analysis for macrophage polarization by flow cytometry, reactive oxygen species (ROS) by flow cytometry, biochemical parameters by chemical reactions, and serum cytokines by ELISA were performed...
October 2018: Medicine (Baltimore)
Astrid Muyldermans, Lisa Florin, Helena Devos, Barbara Cauwelier, Jan Emmerechts
OBJECTIVES: The diagnosis of myelodysplastic syndrome (MDS) is not always straightforward in the absence of objective markers such as ringed sideroblasts, an excess of blasts or clonal cytogenetic abnormalities. Moreover, the lack of specificity of morphological dysplasia makes the differentiation between MDS and other causes of peripheral cytopenia difficult. The WHO 2016 classification of MDS recognizes multiparameter flow cytometry (MFC) as an adjuvant tool for MDS diagnosis. An easily applicable MFC protocol based on CD34 and CD45 is proposed by Ogata et al...
October 18, 2018: Hematology (Amsterdam, Netherlands)
Fatemeh Ghahremani, Amirhosein Kefayat, Daryoush Shahbazi-Gahrouei, Hasan Motaghi, Masoud A Mehrgardi, Shaghayegh Haghjooy-Javanmard
AIM: Herein, the AS1411 aptamer-targeted ultrasmall gold nanoclusters (GNCs) were assessed at different aspects as a radiosensitizer. MATERIALS & METHODS: AS1411 aptamer-conjugated gold nanoclusters (Apt-GNCs) efficacy was evaluated at cancer cells targeting, radiosensitizing effect, tumor targeting, and biocompatibility in breast tumor-bearing mice. RESULTS: Flow cytometry and fluorescence microscopy exhibited more cellular uptake for Apt-GNCs in comparison with GNCs...
October 18, 2018: Nanomedicine
Yanle Ye, Feng Zhang, Qingxia Chen, Zhiyang Huang, Meijun Li
This investigation was purposed to extrapolate whether and how lncRNA MALAT1, miR-194-5p and ACVR2B altered development of clear cell kidney carcinoma (KIRC). We totally gathered 318 pairs of KIRC tissues and adjacent normal tissues, and also purchased human KIRC cell lines and normal human proximal tubular epithelial cell line. Besides, si-MALAT1, pcDNA-MALAT1, miR-194-5p mimic, miR-194-5p inhibitor and negative control (NC) were, respectively, transfected into KIRC cells. The viability, proliferation and apoptosis of the cells were determined with CCK-8 assay, colony formation assay and flow cytometry...
October 18, 2018: Molecular Carcinogenesis
Dimitrios C Ziogas, Meletios A Dimopoulos, Efstathios Kastritis
Multiple myeloma (MM) is characterized by notable inter-patient and intra-clonal heterogeneity that is gradually decoded over the last decade. Despite the deeper and better understanding of its biology and the development of novel therapeutic strategies that have prolonged overall survival, MM still retains a poor prognosis in patient subgroups with certain high-risk features. Areas covered: This paper summarizes currently identified features that stratified patients in high-risk myeloma with impaired prognosis and discuss available therapeutic options that may partially overcome the impact of these adverse factors in patients' outcome...
October 18, 2018: Expert Review of Hematology
Marina Garcia-Prat, Daniel Álvarez-Sierra, Aina Aguiló-Cucurull, Sandra Salgado-Perandrés, Sara Briongos-Sebastian, Clara Franco-Jarava, Andrea Martin-Nalda, Roger Colobran, Isabel Montserrat, Manuel Hernández-González, Ricardo Pujol-Borrell, Pere Soler-Palacin, Mónica Martínez-Gallo
BACKGROUND: For the accurate diagnosis of immunodeficiencies is crucial to compare patients' immunology laboratory values with age-sex matched controls, yet there is a paucity of normal values for most populations. OBJECTIVES: To define appropriate reference values of extended lymphocyte subpopulations and T-cell receptor excision circle (TRECs) levels in healthy pediatric donors between 1 month and 18 years of age. METHODS: Extended immunophenotyping values were obtained by analysis of multiparameter flow cytometry panels for the following subpopulations: CD4+ and CD8+ Naive, Effector, Effector Memory and Central Memory, T helper subpopulations and their degrees of activation, T Regulatory cells, Recent Thymic Emigrants (RTE), B Lymphocyte subpopulations (Transitional, Naive, Preswitch-Memory, Switch-Memory, Plasmablasts, CD21low, and Exhausted), and subpopulations for Monocytes, NK cells and Dendritic Cells...
October 17, 2018: Cytometry. Part B, Clinical Cytometry
Ru Feng, Vijaya Raj Bhatt, Kai Fu, Samuel Pirruccello, Ji Yuan
OBJECTIVES: The purpose of this study was to determine whether immunophenotypic profiles detected by flow cytometry are useful in differentiating chronic myelomonocytic leukemia (CMML) from reactive monocytosis, and between CMML subtypes. METHODS: Eight-color flow cytometry was used to immunophenotype blasts, monocytes, and granulocytes in the bone marrow of 34 patients with CMML and 12 patients with reactive monocytosis. RESULTS: Bone marrow myeloblast, promonocyte, and monocyte counts by flow cytometry were significantly higher in the CMML group than in the reactive monocytosis group...
October 17, 2018: Cytometry. Part B, Clinical Cytometry
Pei Zhang, Jicong Du, Hainan Zhao, Ying Cheng, Suhe Dong, Yanyong Yang, Bailong Li, Fu Gao, Xuejun Sun, Jianming Cai, Cong Liu
BACKGROUND: Ionizing radiation often causes severe injuries to radiosensitive tissues, especially haematopoietic system. Novel radioprotective drugs with low toxicity and high effectiveness are required. Prolyl hydroxylases domain (PHD) inhibitors have been reported to protect against radiation-induced gastrointestinal toxicity. In this study, we demonstrated the protective effects of a PHD inhibitor, roxadustat (FG-4592), against radiation-induced haematopoietic injuries in vitro and in vivo...
October 18, 2018: Journal of Cellular and Molecular Medicine
Nikolaos Gardikas, Myrofora Vikentiou, Evgenia Konsta, Christos K Kontos, Sotirios G Papageorgiou, Aris Spathis, Efthimia Bazani, Anthi Bouchla, Violetta Kapsimali, Katherina Psarra, Periklis Foukas, George Dimitriadis, Vasiliki Pappa
BACKGROUND: Myelodysplastic syndromes (MDS) are clonal hematopoietic stem cell disorders with unknown aetiology. Multiparameter flow cytometry (MFC) is being evaluated for the diagnosis and prognosis of MDS. METHODS: In the present study, five-color MFC was performed on bone marrow aspirates of 50 untreated patients, newly diagnosed with MDS and 27 age matched control samples. Patients were classified according to World Health Organization 2016, International Prognostic Scoring System (IPSS), and Revised IPSS (IPSS-R)...
October 17, 2018: Cytometry. Part B, Clinical Cytometry
Catarina Simões, Isabel Silva, Anabela Carvalho, Sandra Silva, Susana Santos, Gilberto Marques, André Ribeiro, Adriana Roque, José Carda, A B Sarmento-Ribeiro, Maria do Rosário Domingues, Letícia Ribeiro, Artur Paiva
BACKGROUND: Vδ1+ T cells, a subset of γδ T cells, are responsible for innate-like immune responses. Recently, an anti-tumor function mediated by MHC-unrestricted recognition of lipid and stress molecules, has also been described in these cells. This study aimed to quantify and phenotypically characterize circulating Vδ1+ T cells in B cell Chronic Lymphocytic Leukemia (CLL) and Monoclonal B cell lymphocytosis (MBL). METHODS: This study enrolled 58 individuals distributed in five groups: Binet B and C CLL (n = 9), Binet A CLL (n = 26), High count-MBL (n = 10), Low count-MBL (n = 5), and a control group (n = 8)...
October 17, 2018: Cytometry. Part B, Clinical Cytometry
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