Browsing by Subject "Flow cytometry"

Now showing 1 - 4 of 4
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    Cadmium- and lead-induced apoptosis in mallard erythrocytes (Anas platyrhynchos)
    (Elsevier, 2009-01) Romero García, Diego; Hernández-García, A.; Tagliati, C. A.; Martínez-López, E.; García Fernández, Antonio Juan; Ciencias Sociosanitarias
    Cadmium, lead and cadmium–lead (1:10) induced apoptosis were studied using mallard blood cells. The allowable range in concentrations were: 0.01–0.5, 0.1–5.0, and 0.01:0.10–0.50:5.00 mM, for cadmium, lead and cadmium–lead, respectively. The lowest EC50 achieved was for cadmium (0.2270.04 mM). Two doses from each treatment group were chosen to study apoptosis and the presence of metals in cells. The percentage of apoptotic cells increased as the concentration of metals increased. The percentage of cells with intracellular metals was high for both exposure levels and the quantity of intracellular metal was greater for exposure to high concentrations. Morphological alterations for all types of exposure were related to the diverse range of effects that these metals have on membranes. We suggest that the decrease in the number of erythrocytes observed in specimens suffering from lead and cadmium poisoning is related to the induction of apoptosis.
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    Cytotoxicity and alterations at transcriptional level caused by metals on fish erythrocytes in vitro
    (Springer, 2016-03-15) Morcillo, Patricia; Romero, Diego; Meseguer, José; Cuesta Peñafiel, Alberto; Esteban Abad, María de los Ángeles; Ciencias Sociosanitarias
    The in vitro use of fish erythrocytes to test the toxicity of aquatic pollutants could be a valuable alternative to fish bioassays but has received little attention. In this study, erythrocytes from marine gilthead sea bream (Sparus aurata L.) and European sea bass (Dicentrarchus labrax L.) specimens were exposed for 24 h to Cd, Hg, Pb and As and the resulting cytotoxicity was evaluated. Exposure to metals produced a dose-dependent reduction in the viability, and mercury showed the highest toxicity followed by MeHg, Cd, As and Pb. Moreover, fish erythrocytes incubated with each one of the metals exhibited alteration in gene expression profile of metallothionein, superoxide dismutase, catalase, peroxiredoxin, glutathione reductase, heat shock proteins 70 and 90, Bcl2-associated X protein and calpain1 indicating cellular protection, stress and apoptosis death as well as oxidative stress. This study points to the benefits for evaluating the toxicological mechanisms of marine pollution using fish erythrocytes in vitro.
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    Immunophenotype profile by flow cytometry reveals different subtypes of extracellular vesicles in porcine seminal plasma
    (2024-01-23) Barranco, Isabel; Alvarez-Barrientos, Alberto; Parra, Ana; Martinez-Diaz, Pablo; Lucas, Xiomara; Roca, Jordi; Medicina y Cirugía Animal
    Background: Porcine seminal plasma (SP) is endowed with a heterogeneous population of extracellular vesicles (sEVs). This study evaluated the immunophenotypic profile by high-sensitivity flow cytometry of eight sEV subpopulations isolated according to their size (small [S-sEVs] and large [L-sEVs]) from four different SP sources, namely three ejaculate fractions (the first 10 mL of the sperm rich fraction [SRF-P1], the remaining SRF [SRF-P2], and the post-SRF [PSRF]) and entire ejaculate (EE). Methods: Seminal EVs were isolated using a size exclusion chromatography-based protocol from six SP pools (five ejaculates/pool) of each SP source and characterized using complementary approaches including total protein (BCA™assay), particle size distribution (dynamic light scattering), morphology (transmission electron microscopy), and purity (albumin by Western blot). Expression of CD9, CD63, CD81, CD44 and HSP90β was analyzed in all sEV subpopulations by high-sensitivity flow cytometry according to MIFlowCyt-EV guidelines, including an accurate calibration, controls, and discrimination by CFSE-labelling. Results: Each sEV subpopulation exhibited a specific immunophenotypic profile. The percentage of sEVs positive for CD9, CD63, CD81 and HSP90β differed between S- and L-sEVs (P < 0.0001). Specifically, the percentage of sEVs positive for CD9 and CD63 was higher and that for CD81 was lower in S- than L-sEVs in the four SP sources. However, the percentage of HSP90β-positive sEVs was lower in S-sEVs than L-sEVs in the SRF-P1 and EE samples. The percentage of sEVs positive for CD9, CD63, and CD44 also differed among the four SP sources (P < 0.0001), being highest in PSRF samples. Notably, virtually all sEV subpopulations expressed CD44 (range: 88.04-98.50%). Conclusions: This study demonstrated the utility of high-sensitivity flow cytometry for sEV immunophenotyping, allowing the identification of distinct sEV subpopulations that may have different cellular origin, cargo, functions, and target cells.
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    Manual de software flowing para microaula de citometría de flujo
    (2017-01-24) Rubio Pedraza, Gonzalo; Bioquímica y Biología Molecular "B" e Inmunología; Facultad de Química