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Cytometry. Part B, Clinical Cytometry

Sanjay de Mel, Jenny Bei Li, Muhammad Bilal Abid, Tiffany P L Tang, Hui Ming Tay, Wen Chang Ting, Li Mei Poon, Tae Hoon Chung, Benjamin Mow, Allison Tso, Kiat Hoe Ong, Wee Joo Chng, Te Chih Liu
BACKGROUND: The WHO defines 3 categories of NK cell malignancies; extra nodal NK/T cell lymphoma (NKTCL), aggressive NK cell leukaemia, and the provisional entity chronic lymphoproliferative disorder of NK cells (CLPD-NK). Although the flow cytometric (FC) phenotype of CLPD-NK has been described, studies on FC phenotype of NKTCL are limited. To the best of our knowledge ours is the first study to compare the phenotype of NKTCL, CLPD-NK, reactive NK lymphocytosis (RNKL), and normal NK cells using eight colour (8C) FC...
April 21, 2017: Cytometry. Part B, Clinical Cytometry
Chris P Verschoor, Vikas Kohli, Cynthia Balion
BACKGROUND: Monitoring the frequency and phenotype of white blood cell subsets using flow cytometry (immunophenotyping) has proven to be an incredibly powerful tool in the assessment of health. Although improved technologies have aided in the practical implementation of immunophenotyping in clinical and epidemiological studies, the transportation of blood from the site of collection to a central laboratory for analysis within a reasonable timeframe may not be feasible. Hence, the purpose of the following study was to investigate the validity of cryopreserved whole blood as a simple to prepare and cost-effective biospecimen for multi-colour immunophenotyping in a large epidemiological study - namely, The Canadian Longitudinal Study on Aging (CLSA)...
April 5, 2017: Cytometry. Part B, Clinical Cytometry
Joan Puñet-Ortiz, José Vicente Hervás-García, Aina Teniente-Serra, Antonio Cano-Orgaz, Maria José Mansilla, Bibiana Quirant-Sánchez, Juan Navarro-Barriuso, Marco A Fernández-Sanmartín, Silvia Presas-Rodríguez, Cristina Ramo-Tello, Eva María Martínez-Cáceres
BACKGROUND: In natalizumab-treated relapsing-remitting MS (RRMS) patients, various extended interval dosing strategies are under evaluation to minimize severe treatment-associated side effects, mainly progressive multifocal leukoencephalopathy development. Up to now, it has not been presented any approach, even in form of assay design, to determine the optimal percentage of CD49d receptor occupancy (RO) associated with a favorable clinical, radiological, and immunological response. METHODS: A multiparametric quantitative flow cytometry method was settled to measure CD49d RO on peripheral blood lymphocytes...
April 5, 2017: Cytometry. Part B, Clinical Cytometry
Julien Guy, Orianne Wagner-Ballon, Olivier Pages, François Bailly, Jessica Borgeot, Marie-C Béné, Marc Maynadié
OBJECTIVES: Microscopic leukocyte differentials display many drawbacks. Several single 5 to 8-color tubes using multiparameter flow cytometry (MFC) are able to provide extended differentials with sequential gating-based analysis strategies. We investigated a new 5-color MFC method to perform an extended 8-part differential with a simplified gating strategy. METHODS: Whole blood was stained with a combination of antibodies including HLA-DR-FITC/CD19-PE/CD45-ECD/CD16-PC5+CD71-PC5/CD5-PC7...
March 21, 2017: Cytometry. Part B, Clinical Cytometry
Virginia Reyes-Núñez, Evelyn Galo-Hooker, Beatriz Pérez-Romano, Ricardo E Duque, Alejandro Ruiz-Arguelles, Javier Garcés-Eisele
BACKGROUND: The aim of this work was to simultaneously use multiplex ligation-dependent probe amplification (MLPA) assay and flow cytometric DNA ploidy analysis (FPA) to detect aneuploidy in patients with newly diagnosed acute leukemia. METHODS: MLPA assay and propidium iodide FPA were used to test samples from 53 consecutive patients with newly diagnosed acute leukemia referred to our laboratory for immunophenotyping. Results were compared by nonparametric statistics...
March 18, 2017: Cytometry. Part B, Clinical Cytometry
L E Scott, L Kestens, K Pattanapanyasat, K Sukapirom, W S Stevens
BACKGROUND: Method comparison tools are used to determine the accuracy, precision, agreement and clinical relevance of a new or improved technology versus a reference technology. Guidelines for the most appropriate method comparison tools as well as their acceptable limits are lacking and not standardised for CD4 counting technologies. METHODS: Different method comparison tools were applied to a previously published CD4 data set (n= 150 data pairs) evaluating five different CD4 counting technologies (TruCOUNT, Dual Platform, FACSCount, Easy CD4, CyFlow) on a single specimen...
March 10, 2017: Cytometry. Part B, Clinical Cytometry
Katayoon Jafari, Anne Tierens, Amr Rajab, Rumina Musani, André Schuh, Anna Porwit
BACKGROUND: The enormous potential of complex data files generated by 10-color flow cytometry (FC) is hindered by the requirement for exhaustive manual gating and the complexity of multidimensional data visualization. We propose a model using radar plots (RPs), to improve FC data visualization by capturing multidimensionality and integration of FC findings. METHOD: We analysed 12 normal/reactive bone marrow (N/R BM) samples and 12 BM samples from patients with myelodysplasia (MDS) with 10-color FC...
March 3, 2017: Cytometry. Part B, Clinical Cytometry
Olga Mizrahi, Eliran Ish Shalom, Michal Baniyash, Yair Klieger
BACKGROUND: Quantitative flow cytometry (QFCM) can be an important element within the developing toolbox for clinical diagnostics which relies on precise and rapid tests that provide a conclusive answer for physicians. The FC technology combines all of these features. Until recently, this imperative discipline was based on qualitative assessments of cell populations. However, due to the enormous advancement in FC technology, which allows the quantification of a number of antigens on cell surface and within the cells by units of median fluorescence intensity (MFI), this method becomes meaningful and fits the clinical needs...
February 11, 2017: Cytometry. Part B, Clinical Cytometry
Marc Sorigue, Clara Maluquer, Jordi Junca
BACKGROUND: Trisomy 12 chronic lymphocytic leukemia (CLL) is phenotypically different from the rest of CLL cytogenetic subgroups. However, it is unknown whether this is also the case for trisomy 12 CLL-phenotype monoclonal B-cell lymphocytosis (MBL). METHODS: We analyzed the expression of several markers in a series of 89 cytogenetically characterized MBL (including 17 trisomy 12 cases). Additionally, we compared the expression of these markers between trisomy 12 MBL, trisomy 12 CLL and a series of cases with trisomy 12 but fulfilling only 3 of the 5 Moreau CLL diagnostic criteria...
February 10, 2017: Cytometry. Part B, Clinical Cytometry
Michael N Dworzak, Barbara Buldini, Giuseppe Gaipa, Richard Ratei, Ondrej Hrusak, Drorit Luria, Eti Rosenthal, Jean-Pierre Bourquin, Mary Sartor, Angela Schumich, Leonid Karawajew, Ester Mejstrikova, Oscar Maglia, Georg Mann, Wolf-Dieter Ludwig, Andrea Biondi, Martin Schrappe, Giuseppe Basso
Immunophenotyping by flow cytometry (FCM) is a worldwide mainstay in leukemia diagnostics. For concordant multicentric application, however, a gap exists between available classification systems, technologic standardization, and clinical needs. The AIEOP-BFM consortium induced an extensive standardization and validation effort between its nine national reference laboratories collaborating in immunophenotyping of pediatric acute lymphoblastic leukemia (ALL). We elaborated common guidelines which take advantage of the possibilities of multi-color FCM: marker panel requirements, immunological blast gating, in-sample controls, tri-partite antigen expression rating (negative vs...
February 10, 2017: Cytometry. Part B, Clinical Cytometry
Ji Yuan, Vasantha L Gali, Deborah A Perry, Kai Fu, Hina Qureishi, Catalina Amador-Ortiz, Timothy Greiner, Samuel J Pirruccello
BACKGROUND: Normal thymocyte precursors in secondary lymphoid organs have previously been described. It is important to recognize normal thymocyte precursors by flow cytometry to differentiate them from T-cell lymphoblastic leukemia. METHODS: A 3-year-old boy status 2 years post-allogenic cardiac transplant underwent adenoidectomy to exclude post-transplant lymphoproliferative disorder. Microscopic, immunohistochemical, and flow cytometry analyses of the adenoid were performed...
February 6, 2017: Cytometry. Part B, Clinical Cytometry
Andreas Lundqvist, Veronika Kremer
No abstract text is available yet for this article.
March 2017: Cytometry. Part B, Clinical Cytometry
Genny Del Zotto, Emanuela Marcenaro, Paola Vacca, Simona Sivori, Daniela Pende, Mariella Della Chiesa, Francesca Moretta, Tiziano Ingegnere, Maria Cristina Mingari, Alessandro Moretta, Lorenzo Moretta
Natural killer (NK) cells, the most important effectors of the innate lymphoid cells (ILCs), play a fundamental role in tumor immune-surveillance, defense against viruses and, in general, in innate immune responses. NK cell activation is mediated by several activating receptors and co-receptors able to recognize ligands on virus-infected or tumor cells. To prevent healthy cells from auto-aggression, NK cells are provided with strong inhibitory receptors (KIRs and NKG2A) which recognize HLA class I molecules on target cells and, sensing their level of expression, allow killing of targets underexpressing HLA-class I...
March 2017: Cytometry. Part B, Clinical Cytometry
Daniel N Tran, Sandy A B C Smith, David A Brown, Andrew J C Parker, Joanne E Joseph, Nicola Armstrong, William A Sewell
BACKGROUND: There is an emerging role for flow cytometry (FC) in the assessment of small populations of plasma cells (PC). However, FC's utility has been questioned due to consistent underestimation of the percentage of PC compared to microscopy. METHODS: A retrospective study was performed on bone marrow samples analysed by 8-colour FC. Plasma cell populations were classified as polyclonal or monoclonal based on FC analysis. FC findings were compared with microscopy of aspirates, histology and immunohistochemistry of trephine biopsies, and immunofixation (IFX) of serum and/or urine...
March 2017: Cytometry. Part B, Clinical Cytometry
C Wyatt Shields, Korine A Ohiri, Luisa M Szott, Gabriel P López
Advances in microfluidic cell sorting have revolutionized the ways in which cell-containing fluids are processed, now providing performances comparable to, or exceeding, traditional systems, but in a vastly miniaturized format. These technologies exploit a wide variety of physical phenomena to manipulate cells and fluid flow, such as magnetic traps, sound waves and flow-altering micropatterns, and they can evaluate single cells by immobilizing them onto surfaces for chemotherapeutic assessment, encapsulate cells into picoliter droplets for toxicity screenings and examine the interactions between pairs of cells in response to new, experimental drugs...
March 2017: Cytometry. Part B, Clinical Cytometry
Tomasz J Slebioda, Agnieszka Bojarska-Junak, Marta Cyman, Piotr Landowski, Barbara Kaminska, Krzysztof Celinski, Zbigniew Kmiec
BACKGROUND: Interaction between TL1A and death receptor 3 (DR3) is associated with the pathogenesis of inflammatory bowel disease (IBD), although their role in the development of this disease remains not fully explained. Some studies showed elevated expression of TL1A and DR3 in inflamed intestinal tissue but currently there are no reports concerning expression of DR3 on peripheral blood mononuclear cells (PBMCs) of IBD patients which was the subject of our study. METHODS: We performed flow cytometry analysis of DR3 expression on CD4(+), CD8(+), CD11c(+), CD14(+) or CD20(+) PBMCs of adults and children with IBD and healthy volunteers with respect to C-reactive protein (CRP) levels in blood...
March 2017: Cytometry. Part B, Clinical Cytometry
Miriam Ciáurriz, Lorea Beloki, Eva Bandrés, Cristina Mansilla, Amaya Zabalza, Estela Pérez-Valderrama, Mercedes Lachén, Berta Ibáñez, Eduardo Olavarría, Natalia Ramírez
BACKGROUND: Multimer technology is widely used to screen antigen-specific immune recovery after allogeneic hematopoietic stem cell transplantation (allo-HSCT) as it enables identification, enumeration, phenotypic characterization and isolation of virus-specific T-cells. Novel approaches of multimerization might improve on classical tetramer staining; however, their use as standard monitoring technique to quantify antigen-specific cells has not been validated yet. We have compared two of these available multimeric complexes: pentamer and streptamer to select the best strategy for the incorporation into clinical monitoring practice...
March 2017: Cytometry. Part B, Clinical Cytometry
Katharina Lisenko, Stefan Schönland, Ute Hegenbart, Katrin Wallenwein, Ute Braun, Elias K Mai, Jens Hillengass, Hartmut Goldschmidt, Anna Jauch, Anthony D Ho, Marc Raab, Michael Hundemer
The discovery of new targets for tailored therapy is a major improvement in oncology, and tools for the rapid and reliable detection of these targets are essential. Clinical trials demonstrated the benefit of recently developed antibodies against antigens on malignant B-cells. The aim of this study was to assess patients with plasma cell (PC) disorders for expression of antigens on malignant PCs that have exhibited promise in targeted cancer therapy. We retrospectively analyzed the expression of CD20, CD22, CD27, CD30, CD38, CD52, CD81, CD138, and SLAMF7 on PCs by flow cytometry in 103 patients with PC disorders...
March 2017: Cytometry. Part B, Clinical Cytometry
Melanie Bremm, Claudia Cappel, Stephanie Erben, Andrea Jarisch, Michael Schumm, Andreas Arendt, Halvard Bonig, Thomas Klingebiel, Ulrike Koehl, Peter Bader, Sabine Huenecke
BACKGROUND: The depletion of TCRαβ(+) T cells and CD19(+) B cells is a graft purification method for haploidentical stem cell transplantation (HSCT) retaining stem cells, NK cells and TCRγδ(+) T cells. To avoid treatment-related occurrence of severe GvHD a precise quantification of residual TCRαβ(+) T cells in the graft is of essential importance. METHODS: Nine stem cell grafts were purified immunomagnetically on a CliniMACS device and flow cytometric quality control (QC) was performed before and after TCRαβ/CD19-depletion...
March 2017: Cytometry. Part B, Clinical Cytometry
Yael Shahal-Zimra, Zohar Rotem, Judith Chezar, Nino Oniashvili, Avi Leader, Pia Raanani, Esther Rabizadeh
BACKGROUND: We present a pre B-ALL patient with the rare clinical manifestation of extramedullary disease, and a normal hemogram. This patient's blasts expressed bright CD45 and high side scatter (SSc) placing the cells in the monocyte gate. METHODS: Samples from peripheral blood and bone marrow (BM) aspirate from a 50-year-old female patient were immunophenotyped by multiparametric flow cytometry. RESULTS: Flow cytometry studies of the BM aspirate showed a large monocyte gate with 90-95% of the cells expressing an abnormal B cell phenotype...
March 2017: Cytometry. Part B, Clinical Cytometry
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