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Cytometry

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https://www.readbyqxmd.com/read/28735481/single-cell-mass-cytometry-of-acute-myeloid-leukemia-and-leukemia-stem-progenitor-cells
#1
Zhihong Zeng, Marina Konopleva, Michael Andreeff
Mass cytometry time-of-flight (CyTOF) empowers us to understand acute myeloid leukemia (AML) biology at the single-cell level. This technology, combined with advanced data-analysis methods, enables identification and characterization of the rare AML stem/progenitor cells that play key roles in drug resistance and AML relapse. Here we provide a protocol for AML sample preparation for CyTOF and keynotes emphasizing critical steps in and troubleshooting strategies for this procedure.
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28735480/immunophenotyping-by-multiparameter-flow-cytometry
#2
Weina Chen, Hung S Luu
Multiparameter flow cytometry has become an indispensable tool for the diagnosis and classification of acute myeloid leukemia (AML). The basic method relies on the unique ability to detect immunophenotypic abnormalities on discrete subpopulations. The primary roles in the initial assessment of AML are to determine the immaturity of the leukemic population, define the lineage and the immunophenotypic aberrancies in blasts, and identify characteristic immunophenotypic features to predict important recurrent cytogenetic and genetic abnormalities and prognosis...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28735358/il-10-and-tgf-%C3%AE-unbalanced-levels-in-neutrophils-contribute-to-increase-inflammatory-cytokine-expression-in-childhood-obesity
#3
Nayara I Medeiros, Rafael T Mattos, Carlos A Menezes, Rafaelle C G Fares, André Talvani, Walderez O Dutra, Fabrício Rios-Santos, Rodrigo Correa-Oliveira, Juliana A S Gomes
PURPOSE: Obesity is a multifactorial disease, associated with metabolic disorders, chronic low-grade inflammation, and impaired immunity. This study aimed to evaluate the childhood obesity-associated effects on neutrophil activation and cytokine production. METHODS: We evaluated activation and recognition markers and cytokine production in neutrophils from the peripheral blood of children with obesity and normal weight using multicolor flow cytometry. RESULTS: We demonstrate a higher frequency of neutrophils in childhood obesity group (CO) compared to normal-weight group (NW)...
July 22, 2017: European Journal of Nutrition
https://www.readbyqxmd.com/read/28734845/leveraging-siglec-8-endocytic-mechanisms-to-kill-human-eosinophils-and-malignant-mast-cells
#4
Jeremy A O'Sullivan, Daniela J Carroll, Yun Cao, Adriano N Salicru, Bruce S Bochner
BACKGROUND: Sialic acid-binding immunoglobulin-like lectin (Siglec)-8 is a cell-surface protein expressed selectively on human eosinophils, mast cells, and basophils, making it an ideal target for the treatment of diseases involving these cell types. However, the effective delivery of therapeutic agents to these cells requires an understanding of the dynamics of Siglec-8 surface expression. OBJECTIVE: To determine whether Siglec-8 is endocytosed in human eosinophils and malignant mast cells, identify mechanisms underlying its endocytosis, and demonstrate whether a toxin can be targeted to Siglec-8-bearing cells to kill these cells...
July 19, 2017: Journal of Allergy and Clinical Immunology
https://www.readbyqxmd.com/read/28734583/kidney-transplantation-from-hla-incompatible-live-donors-efficiency-and-outcome-of-32-patients-after-desensitisation
#5
Constantino Fernández, María Calvo, Natacha Leite, Andrés López, Tamara Ferreiro, Roi Ribera, Rocío Seijo, Ángel Alonso
Desensitisation is a procedure undergone by the recipient of a kidney transplant from a donor who is cross-match positive. The aim of this study was to present the outcomes from our hospital of kidney transplant recipients from HLA-incompatible live donors after desensitisation. We studied 32 patients aged 46±14 years with a mean fluorescence intensity (MFI) versus class I HLA of 7979±4089 and 6825±4182 MFI versus class II and relative intensity scale (RIS) of 8.9±7.6. The complement-dependent cytotoxicity (CDC) cross-matching test was positive in 18 patients, flow cytometry was positive in 7 patients and donor-specific antibodies (DEA) were detected in 7...
July 19, 2017: Nefrología: Publicación Oficial de la Sociedad Española Nefrologia
https://www.readbyqxmd.com/read/28734132/high-dimensional-single-cell-mapping-of-cerium-distribution-in-the-lung-immune-microenvironment-of-an-active-smoker
#6
Adeeb H Rahman, Yonit Lavin, Soma Kobayashi, Andrew Leader, Miriam Merad
BACKGROUND: Mass cytometry leverages inductively coupled mass spectrometry to perform high dimensional single cell analyses using antibodies tagged with rare earth isotopes that are considered to be largely absent in biological samples. We have recently noted an unusual exception to this rule while analyzing tissue samples from patients undergoing surgical resection for early stage lung cancer, and here we present a detailed cytometric characterization of cerium in a clinical patient sample...
July 22, 2017: Cytometry. Part B, Clinical Cytometry
https://www.readbyqxmd.com/read/28733866/inhibitory-effect-and-mechanism-of-mesenchymal-stem-cells-on-melanoma-cells
#7
J Zhang, L Hou, D Zhao, M Pan, Z Wang, H Hu, J He
PURPOSE: To explore the inhibitory effect and mechanism of MSCs on melanoma proliferation. METHODS: The inhibitory effect of MSCs on melanoma A375 cells was detected by co-culture and conditioned medium (CM) experiments using MTT method. The cell cycle was analyzed by flow cytometry. Then, Western Blot experiment detected the expression of proteins related to NF-κB signaling in A375 cells. The expression of IL-1Ra in MSCs was proved by RT-PCR. The over-expression and silencing vector pcDNA3...
July 21, 2017: Clinical & Translational Oncology
https://www.readbyqxmd.com/read/28733655/lectin-mediated-reversible-immobilization-of-human-cells-into-a-glycosylated-macroporous-protein-hydrogel-as-a-cell-culture-matrix
#8
Nicholas Bodenberger, Dennis Kubiczek, Laura Trösch, Ali Gawanbacht, Susanne Wilhelm, Denis Tielker, Frank Rosenau
3D cell culture is a helpful approach to study cell-cell interaction in a native-like environment, but is often limited due the challenge of retrieving cells from the material. In this study, we present the use of recombinant lectin B, a sugar-binding protein with four binding cavities, to enable reversible cell integration into a macroporous protein hydrogel matrix. By functionalizing hydrogel precursors with saccharose, lectin B can both bind to sugar moieties on the cellular surface as well as to the modified hydrogel network...
July 21, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28733594/cd56-is-a-pathogen-recognition-receptor-on-human-natural-killer-cells
#9
Sabrina Ziegler, Esther Weiss, Anna-Lena Schmitt, Jan Schlegel, Anne Burgert, Ulrich Terpitz, Markus Sauer, Lorenzo Moretta, Simona Sivori, Ines Leonhardt, Oliver Kurzai, Hermann Einsele, Juergen Loeffler
Aspergillus (A.) fumigatus is an opportunistic fungal mold inducing invasive aspergillosis (IA) in immunocompromised patients. Although antifungal activity of human natural killer (NK) cells was shown in previous studies, the underlying cellular mechanisms and pathogen recognition receptors (PRRs) are still unknown. Using flow cytometry we were able to show that the fluorescence positivity of the surface receptor CD56 significantly decreased upon fungal contact. To visualize the interaction site of NK cells and A...
July 21, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28733476/mir-30e-targets-glipr-2-to-modulate-diabetic-nephropathy-in-vitro-and-in-vivo-experiments
#10
Dong Zhao, Jinhua Jia, Hong Shao
The objectives of this study are to investigate the effect of miR-30e targeting GLIPR-2 on the pathological mechanism of DN. The renal tissues of db/db and db/m mice at different age of weeks were stained with PAS. qRT-PCR was applied to detect the expression of miR-30e and GLIPR-2, not only in the renal tissues of mice but also in the renal tubular epithelial cells (RTECs). By luciferase reporter gene assays, we found the 3'-UTR of the GLIPR-2 mRNA as a direct target of miR-30e. The RTECs cultured in high glucose were divided into blank control, NC, miR-30e mimics, miR-30e inhibitors, miR-30e inhibitor + si-GLIPR-2 and si-GLIPR-2 groups...
August 2017: Journal of Molecular Endocrinology
https://www.readbyqxmd.com/read/28733206/omics-based-identification-of-the-combined-effects-of-idiosyncratic-drugs-and-inflammatory-cytokines-on-the-development-of-drug-induced-liver-injury
#11
J Jiang, K Mathijs, L Timmermans, S M Claessen, A Hecka, J Weusten, R Peters, J H van Delft, J C S Kleinjans, D G J Jennen, T M de Kok
The mechanisms of idiosyncratic drug-induced hepatotoxicity remain largely unclear. It has demonstrated that the drug idiosyncrasy is potentiated in the context of inflammation and intracellular ceramides may play a role in this process. To study the mechanisms, HepG2 cells were co-treated with high and low doses of three idiosyncratic (I) and three non-idiosyncratic (N) compounds, with (I+ and N+) or without (I- and N-) a cytokine mix. Microarray, lipidomics and flow cytometry were performed to investigate the genome-wide expression patterns, the intracellular ceramide levels and the induction of apoptosis...
July 18, 2017: Toxicology and Applied Pharmacology
https://www.readbyqxmd.com/read/28733192/pharmacological-evaluation-of-phytochemicals-from-south-indian-black-turmeric-curcuma-caesia-roxb-to-target-cancer-apoptosis
#12
K S Mukunthan, R S Satyan, T N Patel
Curcuma caesia Roxb. (Black turmeric), a perennial herb of the family Zingiberaceae is indigenous to India. C. caesia is used as a spice, food preservative and coloring agent commonly in the Indian subcontinent. Functional parametric pharmacological evaluations like drug ability and toxicity profile of this endangered species is poorly documented. In our present study, among all the extracts of dried C. caesia rhizome viz- hexane, ethyl acetate, methanol and water tested for free radical scavenging capacity by total antioxidant activity (TAO) method, Hexane Rhizome Extract (HRE) wzs found to possess remarkable activity (1200mg ascorbic acid equivalent/100g)...
July 18, 2017: Journal of Ethnopharmacology
https://www.readbyqxmd.com/read/28732393/microrna-30a-suppresses-tumor-progression-by-blocking-ras-raf-mek-erk-signaling-pathway-in-hepatocellular-carcinoma
#13
Kun Zhou, Xiaoyu Luo, Yu Wang, Dachun Cao, Gang Sun
Emerging reports suggest microRNAs (miRNAs) play a vital role in the progression of malignant tumors. MiR-30a is downregulated in a variety of cancers and acts as a tumor suppressing gene. However, the molecular mechanisms of miRNA-30a in hepatocellular carcinoma (HCC) are still unclear. Hereby, in this study, we detected that miR-30a expression was significantly down-regulated in both HCC tissues compared with adjacent non-cancerous liver tissues, and we also observed that miR-30a expression was lower in HCC cell lines than that of normal controls...
July 17, 2017: Biomedicine & Pharmacotherapy, Biomédecine & Pharmacothérapie
https://www.readbyqxmd.com/read/28732355/checkpoint-kinase-1-inhibition-sensitises-transformed-cells-to-dihydroorotate-dehydrogenase-inhibition
#14
Stéphanie Arnould, Geneviève Rodier, Gisèle Matar, Charles Vincent, Nelly Pirot, Yoann Delorme, Charlène Berthet, Yoan Buscail, Jean Yohan Noël, Simon Lachambre, Marta Jarlier, Florence Bernex, Hélène Delpech, Pierre Olivier Vidalain, Yves L Janin, Charles Theillet, Claude Sardet
Reduction in nucleotide pools through the inhibition of mitochondrial enzyme dihydroorotate dehydrogenase (DHODH) has been demonstrated to effectively reduce cancer cell proliferation and tumour growth. The current study sought to investigate whether this antiproliferative effect could be enhanced by combining Chk1 kinase inhibition. The pharmacological activity of DHODH inhibitor teriflunomide was more selective towards transformed mouse embryonic fibroblasts than their primary or immortalised counterparts, and this effect was amplified when cells were subsequently exposed to PF477736 Chk1 inhibitor...
July 12, 2017: Oncotarget
https://www.readbyqxmd.com/read/28732161/an-elegant-ph-responsive-nanovehicle-for-drug-delivery-based-on-triazine-dendrimer-modified-magnetic-nanoparticles
#15
Amir Landarani-Isfahani, Majid Moghadam, Shima Mohammadi, Maryam Royvaran, Naimeh Moshtael-Arani, Saghar Rezaei, Shahram Tangestaninejad, Valiollah Mirkhani, Iraj Mohammadpoor-Baltork
Owing to properties of magnetic nanoparticles and elegant three-dimensional macromolecule architectural features, dendrimeric structures have been investigated as nanoscale drug delivery systems. In this paper, a novel magnetic nanocarrier, generation two (G2) triazine dendrimer modified Fe3O4@SiO2 magnetic nanoparticles (MNP-G2), was designed, fabricated and characterized by Fourier transform infrared (FT-IR), thermal gravimetric analysis (TGA), vibrating sample magnetometer (VSM), field emission scanning electron microscopy (FE-SEM), transmission electron microscopy (TEM) and dynamic light scattering (DLS)...
July 21, 2017: Langmuir: the ACS Journal of Surfaces and Colloids
https://www.readbyqxmd.com/read/28732145/evaluation-of-primary-mediastinal-large-b-cell-lymphoma-by-flow-cytometry
#16
Sindhu Cherian, Jonathan R Fromm
BACKGROUND: Primary mediastinal large B cell lymphoma (PMLBCL) is a B cell non-Hodgkin lymphoma (B-NHL) that shows morphologic, immunophenotypic, and genetic similarities to classical Hodgkin lymphoma (CHL). We evaluated the neoplastic and reactive infiltrate of PMLBCL by flow cytometry (FC). METHODS: 24 cases of PMLBCL were retrospectively characterized using FC combinations for B-NHL, T-NHL, and CHL. RESULTS: The CHL assay identified the neoplastic population (NP) in all cases, while the B-NHL assay identified the NP in 18 of 24 cases...
July 21, 2017: Cytometry. Part B, Clinical Cytometry
https://www.readbyqxmd.com/read/28732140/fibroblast-growth-factor-2-fgf2-is-present-in-human-spermatozoa-and-is-related-with-sperm-motility-the-use-of-recombinant-fgf2-to-improve-motile-sperm-recovery
#17
D J Garbarino Azúa, L Saucedo, S Giordana, M L Magri, M G Buffone, F Neuspiller, M H Vazquez-Levin, C I Marín-Briggiler
Fibroblast growth factors (FGFs) and their receptors (FGFRs) regulate several functions of somatic cells. In a previous work, we reported FGFR expression in human spermatozoa and their involvement in motility. This study aimed to evaluate the presence and localization of fibroblast growth factor 2 (FGF2) in human spermatozoa, to determine the relationship of FGF2 levels with conventional semen parameters and to assess the effect of recombinant FGF2 (rFGF2) on sperm recovery in a selection procedure. Western immunoblotting analysis using an antibody against FGF2 revealed an 18-kDa band in sperm protein extracts...
July 21, 2017: Andrology
https://www.readbyqxmd.com/read/28731485/th17-regulatory-t-cell-imbalance-in-sepsis-patients-with-multiple-organ-dysfunction-syndrome-attenuated-by-high-volume-hemofiltration
#18
Jiguang Guo, Wu Tao, Dan Tang, Jianbin Zhang
BACKGROUND: We assessed the Th17 (T-helper cell)/Treg (Regulatory T cell) imbalance in sepsis patients with multiple organ dysfunction syndrome (MODS) and the clinical benefits of continuous high-volume hemofiltration (HVHF). METHODS: 48 sepsis patients, including 22 patients with MODS (MODS group, n = 22) and 26 without (non-MODS group, n = 26), and 20 healthy volunteer controls were enrolled. The patients in MODS group were randomly divided into the continuous blood purification (CBP) group (n = 11) receiving conventional comprehensive treatment plus high-volume hemofiltration and the non-CBP group (n = 11) receiving conventional comprehensive treatment only...
July 14, 2017: International Journal of Artificial Organs
https://www.readbyqxmd.com/read/28731458/flow-cytometry-based-monocyte-subset-analysis-accurately-distinguishes-chronic-myelomonocytic-leukemia-from-myeloproliferative-neoplasms-with-associated-monocytosis
#19
M M Patnaik, M M Timm, R Vallapureddy, T L Lasho, R P Ketterling, N Gangat, M Shi, A Tefferi, E Solary, K K Reichard, D Jevremovic
No abstract text is available yet for this article.
July 21, 2017: Blood Cancer Journal
https://www.readbyqxmd.com/read/28731226/curcumin-converts-foxp3-regulatory-t-cells-to-t-helper-1-cells-in-patients-with-lung-cancer
#20
JianYong Zou, ChunHua Su, HongHe Luo, YiYan Lei, Bo Zeng, HaoShuai Zhu, ZhenGuang Chen
The regulatory T cells (Treg) play an important role in the tumor tolerance. The methods to regulate the Treg population in cancer-bearing hosts are limited currently. The effect of curcumin on inhibiting cancer has been recognized, but the mechanism remains elusive. This study tests a hypothesis that administration of curcumin down regulates Tregs in lung cancer (LC) patients. In this study, a group of LC patients was treated with curcumin. The peripheral Tregs and T helper (Th) 1 cells were analyzed by flow cytometry...
July 21, 2017: Journal of Cellular Biochemistry
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