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Journal articles on the topic 'Cytofluorimetry'

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Published: 4 June 2021
Last updated: 14 March 2023

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1

Kit, O. I., N. K. Guskova, O. N. Selyutina, V. V. Dmitrieva, I. A. Novikova, I. S. Torpujyan, and V. R. Zakharchenko. "FEATURES OF A DIFFERENTIAL DIAGNOSIS OF PEDIATRIC ACUTE MONOCYTIC LEUKEMIA (AML-M5a) ON THE EXAMPLE OF A CLINICAL CASE." South Russian Journal of Cancer 1, no. 1 (March 7, 2020): 76–83. http://dx.doi.org/10.37748/2687-0533-2020-1-1-7.

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The aim of this work was to assess the significance of investigating clinical and laboratory parameters for diagnosing acute monocytic leukemia in children on the basis of a clinical case. The article demonstrates specific features of differentiating AML M5a from other forms of acute myeloid leukemia. According to the results of hematological, morphological and cytofluorimetric studies of blood and bone marrow samples, the diagnosis of acute myeloid leukemia was established. The morphological and phenotypic characteristics of blast cells hampered the diagnosis of an AML form. However, a comprehensive analysis of the expressed CD antigens allowed acute monocytic leukemia to be identified, which diagnosis was subsequently confirmed by a cytochemical study. Thus, the clinical diagnosis was established over a short period of time. This was of importance given the rising severity of the patient’s condition requiring immediate treatment, the initial hyperleukocytosis and the development of life-threatening complications associated with leukostasis in the lungs and the central nervous system. In the presented case, the clinical manifestations of the underlying disease and the results of flow cytofluorimetry were determining factors in initiating timely specific therapy.
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2

Mbida, A. D., B. Pozzetto, O. Sabido, Y. Akono, F. Grattard, M. Habib, and O. G. Gaudin. "Competition binding studies with biotinylated echovirus 11 in cytofluorimetry analysis." Journal of Virological Methods 35, no. 2 (November 1991): 169–76. http://dx.doi.org/10.1016/0166-0934(91)90132-j.

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3

Sabido, Odile, D. Mbida André, Bruno Pozetto, Odette Gaudin, and François Berthoux. "COMPETITION BINDING STUDIES WITH BIOTINYLATED ECHOVIRUS 11 IN CYTOFLUORIMETRY ANALYSIS." Biology of the Cell 73, no. 2-3 (January 1991): 47a. http://dx.doi.org/10.1016/0248-4900(91)90252-i.

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4

Jacob, Marie-Christine, Mireille Favre, and Jean-Claude Bensa. "EVALUATION OF CD4 CD45RA LYMPHOCYTES IN MALIGNANT LYMPHOMA BY CYTOFLUORIMETRY." Biology of the Cell 73, no. 2-3 (January 1991): 50a. http://dx.doi.org/10.1016/0248-4900(91)90263-m.

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5

Dimitropoulos, K., J. M. Rolland, and R. C. Nairn. "Flow cytofluorimetry of fluorescein fluorescence polarization to assay lymphocyte activation." Biochemical and Biophysical Research Communications 136, no. 3 (May 1986): 1021–29. http://dx.doi.org/10.1016/0006-291x(86)90435-3.

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6

Winstanley, F. P. "Detection of TNF alpha receptors on ovine leucocytes by flow cytofluorimetry." Research in Veterinary Science 52, no. 1 (January 1992): 129–31. http://dx.doi.org/10.1016/0034-5288(92)90074-c.

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7

Ambrosio, Maria Rosaria, Giusy Mosca, Teresa Migliaccio, Domenico Liguoro, Gisella Nele, Fabrizio Schonauer, Francesco D’Andrea, et al. "Glucose Enhances Pro-Tumorigenic Functions of Mammary Adipose-Derived Mesenchymal Stromal/Stem Cells on Breast Cancer Cell Lines." Cancers 14, no. 21 (November 3, 2022): 5421. http://dx.doi.org/10.3390/cancers14215421.

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Adiposity and diabetes affect breast cancer (BC) progression. We addressed whether glucose may affect the interaction between mammary adipose tissue-derived mesenchymal stromal/stem cells (MAT-MSCs) and BC cells. Two-dimensional co-cultures and spheroids were established in 25 mM or 5.5 mM glucose (High Glucose-HG or Low Glucose-LG) by using MAT-MSCs and MCF7 or MDA-MB231 BC cells. Gene expression was measured by qPCR, while protein levels were measured by cytofluorimetry and ELISA. CD44high/CD24low BC stem-like sub-population was quantified by cytofluorimetry. An in vivo zebrafish model was assessed by injecting spheroid-derived labeled cells. MAT-MSCs co-cultured with BC cells showed an inflammatory/senescent phenotype with increased abundance of IL-6, IL-8, VEGF and p16INK4a, accompanied by altered levels of CDKN2A and LMNB1. BC cells reduced multipotency and increased fibrotic features modulating OCT4, SOX2, NANOG, αSMA and FAP in MAT-MSCs. Of note, these co-culture-mediated changes in MAT-MSCs were partially reverted in LG. Only in HG, MAT-MSCs increased CD44high/CD24low MCF7 sub-population and promoted their ability to form mammospheres. Injection in zebrafish embryos of HG spheroid-derived MCF7 and MAT-MSCs was followed by a significant cellular migration and caudal dissemination. Thus, MAT-MSCs enhance the aggressiveness of BC cells in a HG environment.
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8

Mustafin, I. G., V. Kh Fazylov, and A. O. Baryshnikov. "Phenotype of peripheral blood lymphocytes in angina." Kazan medical journal 81, no. 2 (February 2, 2022): 103–7. http://dx.doi.org/10.17816/kazmj96271.

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The phenotype of peripheric blood lymphocytes is studied in 87 patients with angina by cytofluorimetry. It is shown that the course of the disease is accompanied by significant changes of the phenotype of lymphocytes: the increase of expression of activation markers, the increase of the content of CD16+cells. However, the changes of the phenotype of lymphocytes in groups of patients with lacunar and ulceromembranous anginas are ambiguous in the acute period of the disease as well as late in the observation.
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9

Dini, L., A. Lentini, G. D. Diez, M. Rocha, L. Falasca, L. Serafino, and F. Vidal-Vanaclocha. "Phagocytosis of apoptotic bodies by liver endothelial cells." Journal of Cell Science 108, no. 3 (March 1, 1995): 967–73. http://dx.doi.org/10.1242/jcs.108.3.967.

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Using electron microscopy and cytofluorimetry we studied the role of carbohydrate-specific recognition systems in the interaction of apoptotic bodies with normal and interleukin 1-activated sinusoidal endothelial cells. Microfluorimetric observation of liver tissue sections revealed octadecylrhodamine B-labelled apoptotic body binding to the sinusoidal wall of mouse liver, when they were injected intraportally. Plate-scanning cytofluorimetry demonstrated that about 20–25% of Acridine Orange-labelled apoptotic bodies could adhere specifically to cultured endothelial cells after 15 minutes of incubation. Adhesion increased to 30% when the cells were incubated for 60 minutes. Using a mixture of galactose/N-acetylglucosamine/mannose as competition solution apoptotic body adhesion was significantly reduced especially after longer times of incubation, when the percentage of inhibition reached 50%. Following 4 hours exposure of liver endothelial cells to 1 ng/ml human recombinant interleukin-1 beta adhesion markedly increased after 60 minutes of incubation, whereas the co-incubation of interleukin-1 beta with the inhibitors brings down the adhesion to basal values obtained in controls. Electron microscopic observation of the adhesion process showed that the number of endothelial cells binding apoptotic bodies gradually increased from low to high values with time. After 60 minutes of incubation, the majority of apoptotic bodies were seen inside phagosomes and only a few remained at the cell surface. Liver endothelial cells bound and endocytosed apoptotic bodies through carbohydrate-specific receptors. Moreover, this scavenger action was interleukin-1 enhanced, thus suggesting its possible activation during inflammatory and immune processes.
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10

Pistillo, M. P., P. L. Tazzari, O. Mazzoleni, A. Urlacher, M. Falco, M. Vitale, R. W. Karr, and G. B. Ferrara. "ANALYSIS OF HLA SPECIFICITY OF HUMAN MONOCLONAL ANTIBODIES BY CYTOFLUORIMETRY AND CELL ELISA." European Journal of Immunogenetics 18, no. 5-6 (October 1991): 345–53. http://dx.doi.org/10.1111/j.1744-313x.1991.tb00034.x.

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11

Rat, P., K. Bemard-Beaubois, N. Moniot-Ville, Vincen du Laurier, M. Adolphe, and J. M. Warnet. "New quinolones cytotoxicity assessment on living tenocytes in microplates using cold light cytofluorimetry." Toxicology Letters 95 (July 1998): 198–99. http://dx.doi.org/10.1016/s0378-4274(98)80791-1.

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12

Tubiana, N., Z. Mishal, F. le Caer, J. M. Seigneurin, Y. Berthoix, P. M. Martin, and Y. Carcassonne. "Quantification of oestradiol binding at the surface of human lymphocytes by flow cytofluorimetry." British Journal of Cancer 54, no. 3 (September 1986): 501–4. http://dx.doi.org/10.1038/bjc.1986.203.

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13

Rosati, Anna, Luigi Candussio, Enrico Crivellato, Fiora Bartoli Klugmann, Tullio Giraldi, Daniela Damiani, Angela Michelutti, and Giuliana Decorti. "Bodipy-FL-Verapamil: A Fluorescent Probe for the Study of Multidrug Resistance Proteins." Analytical Cellular Pathology 26, no. 1-2 (January 1, 2004): 3–11. http://dx.doi.org/10.1155/2004/576173.

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Most of the substances used as fluorescent probes to study drug transport and the effect of efflux blockers in multidrug resistant cells have many drawbacks, such as toxicity, unspecific background, accumulation in mitochondria. New fluorescent compounds, among which Bodipy‐FL‐verapamil (BV), have been therefore proposed as more useful tools. The uptake of BV has been evaluated by cytofluorimetry and fluorescence microscopy using cell lines that overexpress P‐glycoprotein (P388/ADR and LLC‐PK1/ADR) or MRP (multidrug resistance‐related protein) (PANC‐1) and clinical specimens from patients. The effect of specific inhibitors for P‐glycoprotein (verapamil and vinblastine) or MRP (MK571 and probenecid) has been also studied. BV intracellular concentrations were significantly lower in the two P‐glycoprotein overexpressing cell lines in comparison with the parental lines. In addition, verapamil and vinblastine increased the intracellular concentrations of the dye; MK571 and probenecid, two MRP inhibitors, increased BV levels in PANC‐1 cells, that express this protein. These findings were confirmed in clinical specimens from patients. Fluorescence microscopy revealed a faint fluorescence emission in P‐glycoprotein or MRP expressing cell lines; however, treatment with specific inhibitors significantly increased the fluorescence. BV is a useful tool for studying multidrug resistance proteins with different techniques such as cytofluorimetry and fluorescence microscopy, but does not discriminate between P‐glycoprotein and MRP. In comparison with other classic fluorescent probes, the assay with this dye is extremely rapid, simple, not toxic for cells, devoid of fluorescent background, and can be useful in the clinical settings.
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14

Brickman, M. J., J. M. Cook, and A. E. Balber. "Low temperature reversibly inhibits transport from tubular endosomes to a perinuclear, acidic compartment in African trypanosomes." Journal of Cell Science 108, no. 11 (November 1, 1995): 3611–21. http://dx.doi.org/10.1242/jcs.108.11.3611.

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We have used electron microscopy and flow cytofluorimetry to study endocytosis and intracellular transport of fluid phase bovine serum albumen gold complexes and membrane bound concanavalin A through endosomal compartments of bloodstream forms of Trypanosoma brucei rhodesiense. Both markers were rapidly endocytosed from the flagellar pocket. Within 20 minutes at 37 degrees C the markers reached a large, vesicular, perinuclear compartment that stained heavily with the CB1 monoclonal antibody. Neither marker left the flagellar pocket and entered cells at 4 degrees C. When cells were incubated at 12 degrees C, both markers entered the cell and were transported to collecting tubules, a tubular endosomal compartment that receives endocytosed material from coated endocytic vesicles. However, no material was transported from collecting tubules to the late, perinuclear compartment at 12 degrees C. The morphology of collecting tubule membranes was specifically altered at 12 degrees C; tubules became shorter and were arrayed near the flagellar pocket. The morphological alteration and the block in transport of endocytic markers to the perinuclear compartment seen at 12 degrees C were reversed 10 minutes after cells were returned to 37 degrees C. We also used flow cytofluorimetric measurements of pH dependent fluorescence quenching to measure the pH of the terminal endocytic compartment. Fluoresceinated lectins accumulated in a terminal compartment with a pH of 6.0-6.1, a value considerably higher than that of mammalian lysosomes. Fluorescence from fluoresceinated lectins in this terminal endocytic compartment was dequenched when bloodstream forms were incubated in the presence of chloroquine.
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15

Robins, R. A., R. R. Laxton, M. Garnett, M. R. Price, and R. W. Baldwin. "Measurement of tumour reactive antibody and antibody conjugate by competition, quantitated by flow cytofluorimetry." Journal of Immunological Methods 90, no. 2 (June 1986): 165–72. http://dx.doi.org/10.1016/0022-1759(86)90072-4.

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16

Levi, Marisa, Flavia Tarquini, S. Sgorbati, and E. Sparvoli. "Determination of DNA content by static cytofluorimetry in nuclei released from fixed plant tissue." Protoplasma 132, no. 1-2 (February 1986): 64–68. http://dx.doi.org/10.1007/bf01275791.

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17

JACOB, Marie-Christine, Françoise VACON, and Jean-Claude BENSA. "DOUBLE LABELING WITH c'FDA AND PI IN THE STUDYING OF CELL-CYTOTOXICITY BY CYTOFLUORIMETRY." Biology of the Cell 73, no. 2-3 (January 1991): 37a. http://dx.doi.org/10.1016/0248-4900(91)90213-7.

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18

Aguinaga-Barrilero, Ana, Patricia Castro-Sánchez, Ignacio Juárez, Alberto Gutiérrez-Calvo, Noelia Rodríguez-Pérez, Adela Lopez, Remedios Gómez, and José M. Martin-Villa. "Defects at the Posttranscriptional Level Account for the Low TCRζ Chain Expression Detected in Gastric Cancer Independently of Caspase-3 Activity." Journal of Immunology Research 2020 (November 28, 2020): 1–8. http://dx.doi.org/10.1155/2020/1039458.

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Background. Reduced TCRζ chain surface has been reported in T cells from patients with different inflammatory conditions and cancer. However, the causes of this diminished expression in cancer remain elusive. Methods. T cell-enriched populations of blood or tissue (tumoral and nontumoral) origin from 44 patients with gastric adenocarcinoma and 33 healthy subjects were obtained. Samples were subjected to cytofluorimetry, Western blot analysis, TCRζ cDNA sequencing experiments, measurement of TCRζ mRNA levels, and caspase-3 activity assays. Results. Cytofluorimetry revealed a decreased TCRζ expression in T cells of patients, assessed either as percentage of cells expressing this chain (blood: control subjects 99.8 ± 0.1 % , patients 98.8 ± 1.1 % P < 0.001 ; tissue: control subjects 96.7 ± 0.9 % , patients tumoral tissue 67.9 ± 27.0 % , patients nontumoral tissue 82.8 ± 12.6 % , P = 0.019 ) or mean fluorescence intensity (MFI) value (blood: control subjects 102.2 ± 26.0 ; patients 58.0 ± 12.3 , P = 0.001 ; tissue: control subjects 99.4 ± 21.4 ; patients tumoral tissue 41.6 ± 21.4 ; patients nontumoral tissue 62.3 ± 16.6 , P = 0.001 ). Other chains pertaining to the TCR-CD3 complex (CD3ε) showed no significant differences (MFI values). Subsequent TCRζ cDNA sequencing experiments or measurements of TCRζ mRNA levels disclosed no differences between patients and control subjects. Evaluation of caspase-3 activity showed higher levels in T cell extracts of patients, and this activity could be decreased by 70% with the use of the inhibitor Ac-DEVD-FMK, although CD3ζ expression levels did not recover. Conclusions. These results further place the defect responsible for the low TCRζ expression in cancer at the posttranscriptional level and suggests contrary to what has been proposed in other pathologies that elevated caspase-3 activity is not the causative agent.
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19

Nakidkina, Alena, and T. I. KUZMINA. "Evaluation of viability indicators of bovine spermatozoa after exposure to silicon dimethylglycerolate using flow cytofluorimetry." Agrarian Bulletin of the 209, no. 06 (July 15, 2021): 53–60. http://dx.doi.org/10.32417/1997-4868-2021-209-06-53-60.

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Abstract. Silicon and its dioxide (silica) demonstrate good biological compatibility and a wide range of physical and chemical properties, depending on the production and processing method. In particular, silicon dimethylglycerolate (SDMG) has transmucous and transcutaneous drug conductivity, and, as a hydrogel, may be of interest for the oocytes and embryos cultivation medium structuring and/or media for cryopreservation/thawing of gametes. The aim of this study was to examine the effect of SDMG at concentrations of 0.2 % and 0.02 % on the transmembrane potential of mitochondria and cell viability of bovine spermatozoa. Methods. Sperm subpopulations were assessed for (non)viability indicators (disrupted transmembrane potential of mitochondria, externalization of phosphatidylserine and plasma membrane integrity loss) by flow cytometry with two sets of fluorescent probes. Mitochondrial transmembrane potential was measured using 3,3'-dihexyloxacarbocyanine iodide (DiOC6(3))/ethidium bromide, and externalization of phosphatidylserine – using Annexin V-FITC/propidium iodide pair. The results of this work indicate that SDMG in concentrations of 0.2 % and 0.02 % does not affect the transmembrane mitochondrial potential, externalization of phosphatidylserine or necrotic processes in the population of bovine spermatozoa. The scientific novelty. The data is obtained for the first time on the absence of cytotoxicity of SDMG for male gametes. Together with the shown positive effect of this compound on the morphological parameters and the state of nuclear chromatin of porcine oocytes after intrafollicular vitrification, it should be concluded that silicon-containing glycerohydrogels are of interest as a component of sperm cryopreservation/thawing media.
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20

Aussel, Claude, Amir-Hossein Taheri Mahmoudi, Ghislaine Bernard, Jean-Philippe Breittmayer, and Alain Bernard. "Sphingosine, oleylamine and stearylamine inhibit both CD11aCD18-dependent and -independent homotypic aggregation: demonstration by cytofluorimetry." Immunology Letters 47, no. 3 (September 1995): 175–80. http://dx.doi.org/10.1016/0165-2478(95)00073-3.

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21

Nikitayev, V. G., A. N. Pronichev, N. N. Tupitsin, V. Yu Selchuk, V. V. Dmitrieva, A. D. Palladina, E. V. Polyakov, K. A. Liberis, and V. I. Tsyplyak. "Integrated information and measurement system for the diagnosis of acute leukemia and minimal residual disease based on computer microscopy, flow laser cytofluorimetry and artificial intelligence." Journal of Physics: Conference Series 2058, no. 1 (October 1, 2021): 012033. http://dx.doi.org/10.1088/1742-6596/2058/1/012033.

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Abstract The article considers a new integrated information and measurement system for the diagnosis of acute leukemia and minimal residual disease based on computer microscopy and flow laser cytometry. The system is based on combining the results of computer microscopy in the analysis of bone marrow preparations and the results of flow laser cytofluorimetry. A special feature of the system is the use of artificial intelligence technologies in the recognition of images of bone marrow cells in the computer microscopy subsystem. The work was the result of joint work of the Department of Computer Medical Systems of the National Research Nuclear University "MEPhI" and the Laboratory of Hematopoietic Immunology of the National Medical Research Center of Oncology named after N. N. Blokhin.
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22

Dosina, Margarita Khotyanovich. "Culture cells in a model of microgravity." EuroBiotech Journal 1, no. 1 (January 27, 2017): 93–94. http://dx.doi.org/10.24190/issn2564-615x/2017/01.17.

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Abstract The main objective of the work was to clarify the question - how will cell cultures functional state change after microgravity simulation when the shift in full strength direction takes place? Proliferation processes and apoptosis intensity in cell lines of rat glioma and human fibroblasts were compared in changing the position of flasks with cell culture in relation to the horizon. The detection of apoptosis and necrosis processes was carried out using flow cytofluorimetry. It was found that the change in full strength direction provides an inhibitory effect on tumor glial cells and fibroblasts’ proliferative activity enhances along with inhibition of apoptotic processes. Intensification of apoptotic processes in glioma cells and attenuation of cell death processes in normal cells - fibroblasts - are the result of cell cooperation disturbance.
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23

Galassi, Leone. "A gas-proof temperature-controlled microscope chamber that accepts oil or glycerol-immersion objectives for cytofluorimetry." Technical Tips Online 5, no. 1 (January 2000): 7–9. http://dx.doi.org/10.1016/s1366-2120(08)70148-x.

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24

Movchan, L. V., and T. V. Shman. "ANALYSIS OF THE NUMBER OF CELLS WITH CD34+CD38- AND CD34+CD38-CD19+ PHENOTYPES AS POTENTIAL LEUKEMIC STEM CELLS IN ACUTE LYMPHOBLASTIC LEUKEMIA IN CHILDREN." Health and Ecology Issues, no. 2S (December 28, 2011): 66–69. http://dx.doi.org/10.51523/2708-6011.2011-8-2s-22.

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The number of the supposed leukemic stem cells in marrow samples of 54 patients with primary B-linear acute lymphoblastic leukemia was detected by the method of multiparametric flow cytofluorimetry in leukemia diagnosis (zero day). The level of minimal residual disease was estimated on zero and on the fifteenth days of induction therapy. In the course of the research it was found out that leukemic B-cell precursors with СD34+СD38-CD19+ phenotype prevailed among the cells with СD34+СD38-phenotype. The high percentage of both СD34+СD38-, СD34+СD38-, and СD34+СD38-CD19+ among the general population leukemic cells was associated with a worse response to the therapy. Therefore, the initial number of such cells can be considered as a prognostic marker in acute lymphoblastic leukemia in children.
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25

King, S. L., G. St J. Whitley, S. Page, S. S. Nussey, and A. P. Johnstone. "Immune induction of major histocompatibility complex antigens on a human thyroid cell line (SGHTL-34)." Journal of Molecular Endocrinology 2, no. 3 (May 1989): 183–87. http://dx.doi.org/10.1677/jme.0.0020183.

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ABSTRACT Expression of major histocompatibility complex antigens by epithelial cells may play a role in the aetiology of autoimmune disorders. We have studied the effect of γ-interferon on SGHTL-34, a human thyroid cell line which constitutively expresses class I but not class II antigens. γ-Interferon induced the expression of class II and increased the expression of class I molecules (assessed by flow cytofluorimetry) in a dose-dependent manner. Thyrotrophin or phytohaemagglutinin had no effect on either class I or class II expression. However, a supernatant from phytohaemagglutinin-stimulated peripheral blood mononuclear cells, containing 6400 U γ-interferon/ml, was an effective inducer of both class I and class II antigens. These data clarify earlier studies using primary thyroid cultures, which are contaminated with cells of the immune system.
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26

Plaksin, D., K. Polakova, P. McPhie, and D. H. Margulies. "A three-domain T cell receptor is biologically active and specifically stains cell surface MHC/peptide complexes." Journal of Immunology 158, no. 5 (March 1, 1997): 2218–27. http://dx.doi.org/10.4049/jimmunol.158.5.2218.

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Abstract We have expressed in bacteria a single-chain T cell receptor (scTCR) with specificity for an HIV gp120-derived peptide bound to the murine MHC-I molecule, H-2Dd. This scTCR consists of V alpha covalently linked to the VbetaCbeta domains that was solubilized, refolded, and purified in high yield. Specific binding of the scTCR to MHC/peptide complexes was demonstrated by surface plasmon resonance, with a Kd of 2 to 8 x 10(-6) M. This scTCR specifically inhibited T cell activation, and stained cell surface MHC/peptide complexes as measured by cytofluorimetry. The preservation of binding specificity by such a three-domain scTCR suggests that this structure is sufficient for specific MHC/peptide recognition and that this strategy will be of general use as applied to other TCR.
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27

Donati, Francesco, Roberta Acciarini, Ilenia De Benedittis, Xavier de la Torre, Daniela Pirri, Mariangela Prete, Alessandra Stampella, Enza Vernucci, and Francesco Botre. "Detecting Autologous Blood Transfusion in Doping Control: Biomarkers of Blood Aging and Storage Measured by Flow Cytofluorimetry." Current Pharmaceutical Biotechnology 19, no. 2 (June 28, 2018): 124–35. http://dx.doi.org/10.2174/1389201019666180405165118.

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BHAUD, YVONNE, JEAN-MARIE SALMON, and MARIE-ODILE SOYER-GOBILLARD. "The Complex Cell Cycle of the Dinoflagellate Protoctist Crypthecodinium Cohnii as Studied In Vivo and by Cytofluorimetry." Journal of Cell Science 100, no. 3 (November 1, 1991): 675–82. http://dx.doi.org/10.1242/jcs.100.3.675.

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The complete cell cycle of the dinoflagellate Crypthecodinium cohnii Biecheler 1938 was observed in vivo in a synchronized heterogeneous population, after DAPI staining of DNA. In a given population, the relative nuclear DNA content in each class of cell was measured using a new numerical image-analysis method that takes into account the total fluorescence intensity (FI), area (A) and shape factor (SF). The visible degree of synchronization of the population was determined from the number of cells with a nuclear content of 1C DNA at ‘synchronization’, time 0. One method of synchronization (method 1), based on the adhesiveness of the cysts, gave no better than 50% synchronization of the population; method 2, based on swimming cells released from cysts cultured on solid medium, gave 73% of cells with the same nuclear DNA content. A scatter plot of data for FI versus A in the first few hours after time 0 showed that the actual degree of synchronization of the population was lower. The length of the C. cohnii cell cycle determined in vivo by light microscopy was 10, 16 or 24 h for vegetative cells giving two, four or eight daughter cells, respectively. Histograms based on the FI measurements showed that in an initially synchronized population observed for 20 h, the times for the first cell cycle were: G1 phase, 6 h; S phase, 1 h 30 min; G2+M, 1h 30 min, with the release of vegetative cells occurring 1 or 2h after the end of cytokinesis. The times for the second cell cycle were G1+S, 3h; G2+M, 2h. FI and A taken together, suggested that the S phase is clearly restricted, as in higher eukaryotes. A and SF, taken together, showed that the large nuclear areas were always in cysts with two or four daughter cells. FI and SF, taken together, showed that the second S phase always occurred after completion of the first nuclear division. Our data concerning the course of the cell cycle in C. cohnii are compared with those from earlier studies, and the control of the number of daughter cells is discussed; this does not depend on the ploidy of the mother cell.
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29

Christen, M. O., R. Osseni, C. Debbasch, J. Maugard, P. Rat, and J. M. Warnet. "Cytoprotective effect of anethole dithiole thione (sulfarlem®) as a free radical scavenger-evaluation by microplate cytofluorimetry." Toxicology Letters 95 (July 1998): 50. http://dx.doi.org/10.1016/s0378-4274(98)80199-9.

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30

Nikolaeva, M. A., I. A. Gukasyan, R. D. Filippova, I. V. Korotkova, D. V. Kuyavskaya, and G. T. Sukhikh. "Adaptation of flow cytofluorimetry for detection of antispermal antibodies in the serum and peritonaal fluid of women." Bulletin of Experimental Biology and Medicine 124, no. 3 (September 1997): 921–23. http://dx.doi.org/10.1007/bf02447003.

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31

Schumann, RR, J. van der Bosch, S. Ruller, M. Ernst, and M. Schlaak. "Monocyte long-term cultivation on microvascular endothelial cell monolayers: morphologic and phenotypic characterization and comparison with monocytes cultured on tissue culture plastic." Blood 73, no. 3 (February 15, 1989): 818–26. http://dx.doi.org/10.1182/blood.v73.3.818.bloodjournal733818.

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Human monocytes were cultured on monolayers of a newly established microvascular endothelial cell strain. As compared to monocytes cultured on plastic, these endothelium-“derived” monocytes (EDM) showed distinct morphology, higher motility, and different antigen-expression pattern for several surface markers, as detected by cytofluorimetry. The MO-1- and the Leu-M1-marker were maintained on EDMs while they were lost on plastic-cultured cells. The MAX 1–26-termed markers failed to increase on EDMs, in contrast to plastic-cultured monocytes. For seven additional markers, expression after two weeks in vitro was higher on EDMs than on plastic-cultured monocytes. Functionally EDMs showed typical monocyte/macrophage behavior and were easily removable from the culture system for further experimentation. Our data suggest that monocytes cultured on microvascular endothelial cells are maintained for several weeks in a more physiologic state than monocytes cultured on plastic.
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32

Schumann, RR, J. van der Bosch, S. Ruller, M. Ernst, and M. Schlaak. "Monocyte long-term cultivation on microvascular endothelial cell monolayers: morphologic and phenotypic characterization and comparison with monocytes cultured on tissue culture plastic." Blood 73, no. 3 (February 15, 1989): 818–26. http://dx.doi.org/10.1182/blood.v73.3.818.818.

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Abstract Human monocytes were cultured on monolayers of a newly established microvascular endothelial cell strain. As compared to monocytes cultured on plastic, these endothelium-“derived” monocytes (EDM) showed distinct morphology, higher motility, and different antigen-expression pattern for several surface markers, as detected by cytofluorimetry. The MO-1- and the Leu-M1-marker were maintained on EDMs while they were lost on plastic-cultured cells. The MAX 1–26-termed markers failed to increase on EDMs, in contrast to plastic-cultured monocytes. For seven additional markers, expression after two weeks in vitro was higher on EDMs than on plastic-cultured monocytes. Functionally EDMs showed typical monocyte/macrophage behavior and were easily removable from the culture system for further experimentation. Our data suggest that monocytes cultured on microvascular endothelial cells are maintained for several weeks in a more physiologic state than monocytes cultured on plastic.
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33

Esposito, Simone, Sonia Colicchia, Xavier de la Torre, Francesco Donati, Monica Mazzarino, and Francesco Botrè. "Liposomes as potential masking agents in sport doping. Part 2: Detection of liposome-entrapped haemoglobin by flow cytofluorimetry." Drug Testing and Analysis 9, no. 2 (February 23, 2016): 208–15. http://dx.doi.org/10.1002/dta.1956.

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34

Vashkevich, E. P., and T. V. Shman. "TESTING OF CD107A MARKER EXPRESSION FOR THE ASSESSMENT OF ANTITUMOR ACTIVITY OF CYTOTOXIC LYMPHOCYTES." Health and Ecology Issues, no. 2S (December 28, 2011): 22–24. http://dx.doi.org/10.51523/2708-6011.2011-8-2s-4.

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The assessment of antitumor activity of cytotoxic lymphocytes is an important indicator of their functional status in cellular immunotherapy. Testing of CD107a marker expression by the method of flow cytofluorimetry has been widely lately used for these purposes. The research that we carried out to determine the antitumor activity of mononuclear lymphocytes in peripheral blood including those stimulated by interleukine-2 showed that incubation of mononuclear lymphocytes in peripheral blood in the presence of К-562 tumor line was conducive to the reliable increase in rates of natural killer and killer-like cells expressing CD107a. A straight correlation between CD107a+killer cells percentage and the number of the dead targets in the test with the use of CFSE fluorescent label was detected. The assessment method of CD107a expression makes it possible to measure the cytotoxic activity in a cell in a concrete population of effectory cells and may be used alongside with standard methods for target-cell lysis.
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35

Grande, Fedora, Francesca Giordano, Maria Antonietta Occhiuzzi, Carmine Rocca, Giuseppina Ioele, Michele De Luca, Gaetano Ragno, Maria Luisa Panno, Bruno Rizzuti, and Antonio Garofalo. "Toward Multitasking Pharmacological COX-Targeting Agents: Non-Steroidal Anti-Inflammatory Prodrugs with Antiproliferative Effects." Molecules 26, no. 13 (June 28, 2021): 3940. http://dx.doi.org/10.3390/molecules26133940.

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The antitumor activity of certain anti-inflammatory drugs is often attributed to an indirect effect based on the inhibition of COX enzymes. In the case of anti-inflammatory prodrugs, this property could be attributed to the parent molecules with mechanism other than COX inhibition, particularly through formulations capable of slowing down their metabolic conversion. In this work, a pilot docking study aimed at comparing the interaction of two prodrugs, nabumetone (NB) and its tricyclic analog 7-methoxy-2,3-dihydro-1H-cyclopenta[b]naphthalen-1-one (MC), and their common active metabolite 6-methoxy-2-naphthylacetic acid (MNA) with the COX binding site, was carried out. Cytotoxicity, cytofluorimetry, and protein expression assays on prodrugs were also performed to assess their potential as antiproliferative agents that could help hypothesize an effective use as anticancer therapeutics. Encouraging results suggest that the studied compounds could act not only as precursors of the anti-inflammatory metabolite, but also as direct antiproliferative agents.
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36

Barilo, A. A., and S. V. Smirnova. "PRURITUS AS A MARKER OF IMMUNE DISTURBANCES IN PSORIASIS." Medical academic journal 19, no. 1S (December 15, 2019): 60–61. http://dx.doi.org/10.17816/maj191s160-61.

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Objective: Based on the study of some indicators of cellular and humoral immunity, evaluate the features of immunoreactivity in patients with psoriasis with concomitant pruritus. Materials and methods. Patients were examined with vulgar PS (n = 97), which were divided into 2 groups: group 1 - PS with pruritus (n = 73, mean age 40.0 ± 1.5 years), group 2 (comparison group) - PS without pruritus (n = 24, mean age 41.5 ± 2.7 years). The cellular immunity indicators was carried out by flow cytofluorimetry. Phagocytic activity of peripheral blood neutrophils was studied microscopically by the absorption of latex particles. Concentrations of immunoglobulins, cytokines, circulating immune complexes in the blood serum were evaluated by the method of enzyme-linked immunosorbent assay. Statistica 6.0 was used for statistical analysis. Our studies allowed us to identify features of PS with concomitant pruritus: severe clinical course of the disease with damage to the musculoskeletal system, associated with selective deficiency of class M immunoglobulins and increased activity of phagocytic neutrophils.
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37

Kurtasova, L. M., N. A. Shakina, and T. V. Lubnina. "Studies on correlations between immunophenotype and the indices of metabolic enzyme activity of blood lymphocytes in children with hypertrophy of the pharyngeal tonsils." Medical Immunology (Russia) 22, no. 1 (January 31, 2020): 165–70. http://dx.doi.org/10.15789/1563-0625-soc-1806.

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The objective of our study was to evaluate correlation between the immune pheno-type and activity indices of NAD (P)-dependent dehydrogenases in peripheral blood lymphocytes in young children with hypertrophy of the pharyngeal tonsil (HPT). We have examined 57 children, 1-3 years of age, with hypertrophy of the pharyngeal tonsils (HPT). The control group consisted of 35 healthy children of the same age. The numbers of CD3+, CD4+, CD8+, CD19+, CD16+/56+ lymphoid cells in peripheral blood were determined by flow cytofluorimetry technique. Activity of NAD (P)-dependant dehydrogenases in peripheral blood lymphocytes was studied using bioluminescent method as described elsewhere (А. Savchenko, L. Suntsova, 1989). Correlation analysis has revealed an increase of positive correlations, a decrease of the correlation strength, and emergence of new connections between phenotype and activity indices of NAD (P)-dependent dehydrogenases in peripheral blood lymphocytes in children with hypertrophy of pharyngeal tonsils (HPT). Specific correlation patterns between the phenotype and activity indices of NAD (P)-dependent dehydrogenases in peripheral blood lymphocytes have been revealed in children with hypertrophy of pharyngeal tonsils (HPT).
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38

Armanini, D., P. Spinella, M. Simoncini, A. Basso, S. Zovato, GB Pozzan, CB De Palo, G. Bucciante, and I. Karbowiak. "Regulation of corticosteroid receptors in patients with anorexia nervosa and Cushing's syndrome." Journal of Endocrinology 158, no. 3 (September 1, 1998): 435–39. http://dx.doi.org/10.1677/joe.0.1580435.

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We have studied 16 patients with anorexia nervosa (11 with a stabilised weight loss and 5 in the weight-losing phase), 11 healthy controls, and 10 patients with Cushing's syndrome, by measuring plasma cortisol (by enzyme-immunoassay), ACTH (by RIA), corticosteroid (Type I-mineralocorticoid and Type II-glucocorticoid) receptors in mononuclear leukocytes (by radio-receptor assay), and lymphocyte subpopulations (by cytofluorimetry). In anorexic patients with a stabilised weight loss and in Cushing's syndrome the mean value of both Type I and Type II corticosteroid receptors in mononuclear leukocytes was significantly lower than in controls. The correlation between Type II receptors and plasma cortisol was inverse in stabilised anorexia nervosa and in Cushing's syndrome, and direct in healthy controls. Anorexic patients in the weight-losing phase showed a significant increase in plasma cortisol levels and a normal number of Type II receptors. From these results we hypothesise that in anorexia nervosa there is a progression from an increase in plasma cortisol in the weight-losing phase, to a concomitant decrease in Type II receptors when the disease is stabilised.
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39

Антонюк, Marina Antonyuk, Виткина, Tatyana Vitkina, Новгородцева, Tatyana Novgorodtseva, Денисенко, Yuliya Denisenko, Жукова, and Natalya Zhukova. "MITOCHONDRIAL DYSFUNCTION IN CHRONIC OBSTRUCTIVE PULMONARY DISEASE." Bulletin physiology and pathology of respiration 1, no. 60 (June 15, 2016): 28–33. http://dx.doi.org/10.12737/20048.

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The aim is to study structural and functional condition of cell mitochondrial apparatus for evaluating the mitochondrial membrane potential (MMР) and the fatty acids (FA) composition of mitochondrial membranes of blood cells in patients with COPD; to establish the role of mitochondrial dysfunction in the mechanism of COPD. The study involved 27 patients with mild COPD, 21 with moderate COPD of a stable course, and 20 healthy people. MMР of leukocytes was assessed with cytofluorimetry. The composition of FA of mitochondrial membranes was studied with gas-liquid chromatography. As a result of the study it was found out that the worsening of the disease (moderate COPD) leads to the increase of the number of white blood cells with reduced MMP and appearance of deficit of the saturated monoenic and n-3 polyunsaturated fatty acids in the membrane of mitochondria. The revealed changes in the structural and functional state of mitochondria show a violation in energetic activity, membrane permeability and transport of substances, which is a sign of the formation of mitochondrial dysfunction and cell hypoxia development at respiratory diseases.
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40

Bédouet, Laurent, Marie-Thérèse Bousser, Natacha Frison, Claire Boccaccio, Jean-Pierre Abastado, Philippe Marceau, Roger Mayer, Michel Monsigny, and Annie-Claude Roche. "Uptake of Dimannoside Clusters and Oligomannosides by Human Dendritic Cells." Bioscience Reports 21, no. 6 (December 1, 2001): 839–55. http://dx.doi.org/10.1023/a:1015592926051.

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Knowing that human blood monocyte-derived dendritic cells express cell-surface mannose-specific lectins, we prepared various mannoses containing glycoconjugates with the aim of developing highly specific synthetic carriers of oligonucleotides and genes. Conjugates were prepared from oligosaccharides obtained by hydrazinolysis of Saccharomyces cerevisiae invertase glycopeptides. The reducing saccharides were converted into glycosynthons, i.e., into glyco-amino acids. Fluorescein derivatives were obtained by coupling the free carboxyl group of oligosaccharyl-pyroglutamate to the α-amino group of ∊-fluoresceinyl-thiocarbamyl lysine methyl ester. It has been shown by others that glycosylated linear oligolysines containing up to six α-D-mannopyranosylphenylthiocarbamyl units have a high affinity for the human mannose receptor. In order to obtain fully biodegradable clusters and to improve both the specificity and the selectivity, disaccharides transformed into glycosynthons were coupled to pentalysine carriers (Lys5-Ala-Cys-NH2). Glycosylated pentalysyl cysteine conjugates were made fluorescent upon substitution of the cysteine thiol group with fluorescein iodoacetamide. As shown by flow cytofluorimetry, both the dimannoside clusters and yeast oligomannosides were very efficiently taken up by DC, conversely lactoside clusters were not.
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41

Rat, P., R. Osseni, M. O. Christen, M. Thevenin, J. M. Warnet, and M. Adolphe. "53 Microplate cytofluorimetry (MCM) and MiFAAC Tests (Microtitration Fluorimetric Assays on Alive Cells) : new tools for screening in cellular pharmacotoxicology." Cell Biology and Toxicology 12, no. 4-6 (December 1996): 393. http://dx.doi.org/10.1007/bf00438225.

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42

Saggia, C., A. Santagostino, G. Forti, G. Biaggi, G. Angeli, M. Dacorsi, D. Cerrato, G. Loi, G. Porcile, and O. Alabiso. "Prospective study on prognostic significance of DNA ploidy and Ki-67 expression in colorectal cancer." Journal of Clinical Oncology 25, no. 18_suppl (June 20, 2007): 21184. http://dx.doi.org/10.1200/jco.2007.25.18_suppl.21184.

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21184 Background: The prognostic value of flow cytometry DNA ploidy and Ki-67 expression in colorectal carcinomas has not been defined yet. The study tries to correlate tumoral DNA ploidy and proliferative activity (Ki-67) with therapy response, Overall Survival (OS), Disesase Specific Survival (DSS) and Disease Free Survival (DFS). Methods: From 01/09/02 to 30/06/05, 3 samples of cancer tissue and 1 sample of normal mucosa has been collected from all operating pieces of colorectal cancer. These samples were frosen and later disgregated, treated and coloured with Propidium Iodide. DNA ploidy was evaluated by FACSCalibur cytofluorimetry. Normal mucosa tissue was our internal control. Ki-67 was evaluated by immuno-histochemistry in all tumoral samples. All the patients were cured with chemo- and/or radiotherapy in our divisions. Results: 67 patients (M/F 35/32); median age was 70; staging: 19% I, 33% II, 30% III, 18% IV. We found aneuploidy in 65,7% of carcinoma and Ki-67 median expression was 55%. DNA tumoral heterogeneity was present in 27% of patients. DNA aneuploidy correlates with advanced disease stage at diagnosis (p<0,01), with high number of metastatic lymphnodes (p<0,005) and with serological markers positivity (p<0,04). After surgery and chemotherapy 35% of the patients with aneuploid carcinoma and high proliferative activity (Ki-67>55%) do not show evident disease versus 100% of patients with DNA diploidy and lower Ki-67. Tumoral aneuploidy correlates in a significative way with lower OS (48% vs 89% of diploid patients), lower DSS (tumor death happened just in patients with aneuploid DNA in every disease stages), with lower DFS (18% vs 86% of diploid patients). Univariated analysis stated that aneuploidy determinates an Odd Ratio=5,7 to develop disease progression (p=0,033). At the moment Ki-67 expression with 55% cut-off does not seem to correlate with OS, DSS and DFS. Conclusions: Preliminar results (the study is still in progress) seem to suggest that cytofluorimetric DNA-ploidy has a prognostic and predictive significance in colo-rectal carcinomas. Ki-67 expression (immuno-histochemistry) has an uncertain significance. The small number of patients and the short follow-up do not allow us to reach any definitive conclusions, but the study is worth to go on. No significant financial relationships to disclose.
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43

Sivaccumar, Jwala Priyadarsini, Antonio Leonardi, Emanuela Iaccarino, Giusy Corvino, Luca Sanguigno, Angela Chambery, Rosita Russo, et al. "Development of a New Highly Selective Monoclonal Antibody against Preferentially Expressed Antigen in Melanoma (PRAME) and Identification of the Target Epitope by Bio-Layer Interferometry." International Journal of Molecular Sciences 22, no. 6 (March 20, 2021): 3166. http://dx.doi.org/10.3390/ijms22063166.

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Background: Monoclonal antibodies (mAbs) against cancer biomarkers are key reagents in diagnosis and therapy. One such relevant biomarker is a preferentially expressed antigen in melanoma (PRAME) that is selectively expressed in many tumors. Knowing mAb’s epitope is of utmost importance for understanding the potential activity and therapeutic prospective of the reagents. Methods: We generated a mAb against PRAME immunizing mice with PRAME fragment 161–415; the affinity of the antibody for the protein was evaluated by ELISA and SPR, and its ability to detect the protein in cells was probed by cytofluorimetry and Western blotting experiments. The antibody epitope was identified immobilizing the mAb on bio-layer interferometry (BLI) sensor chip, capturing protein fragments obtained following trypsin digestion and performing mass spectrometry analyses. Results: A mAb against PRAME with an affinity of 35 pM was obtained and characterized. Its epitope on PRAME was localized on residues 202–212, taking advantage of the low volumes and lack of fluidics underlying the BLI settings. Conclusions: The new anti-PRAME mAb recognizes the folded protein on the surface of cell membranes suggesting that the antibody’s epitope is well exposed. BLI sensor chips can be used to identify antibody epitopes.
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44

Ivanov, M. F., I. P. Balmasova, and A. V. Zhestkov. "Immunopathogenetic features and prognostic criteria for severe hemorrhagic fever with renal syndrome." RUDN Journal of Medicine 24, no. 3 (December 15, 2020): 207–17. http://dx.doi.org/10.22363/2313-0245-2020-24-3-207-217.

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Aim. Assessment of the features of cellular immunological mechanisms at the early stage of HFRS of varying severity and development of prognostic criteria for the risk of a severe course of the infectious process. Materials and methods. An immunological blood test (flow cytofluorimetry method) was performed in 12 patients with severe HFRS and 53 patients with moderate course in the dynamics of the disease. Statistical data processing was performed based on the SPSS software package. Results. At the initial stages of HFRS, immunological features of the severe course of the disease were established in the form of a higher content of T-helper and regulatory T-cells in the blood and a reduced number of CTL, including their activated pool. Based on these changes, an immunological prognostic coefficient of HFRS was developed, which allows determining the risk of severe course in the early days (febrile period) of the disease with high prognostic accuracy. Conclusion. The results obtained allowed us to identify previously unknown features of the immune process at the initial stages of HFRS development, which allowed us to propose a new approach to predicting the severe course of the disease.
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45

Carlini, Lavinia, Gabriele Tancreda, Valeria Iobbi, Federico Caicci, Silvia Bruno, Alfonso Esposito, Daniela Calzia, et al. "The Flavone Cirsiliol from Salvia x jamensis Binds the F1 Moiety of ATP Synthase, Modulating Free Radical Production." Cells 11, no. 19 (October 9, 2022): 3169. http://dx.doi.org/10.3390/cells11193169.

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Several studies have shown that mammalian retinal rod outer segments (OS) are peculiar structures devoid of mitochondria, characterized by ectopic expression of the molecular machinery for oxidative phosphorylation. Such ectopic aerobic metabolism would provide the chemical energy for the phototransduction taking place in the OS. Natural polyphenols include a large variety of molecules having pleiotropic effects, ranging from anti-inflammatory to antioxidant and others. Our goal in the present study was to investigate the potential of the flavonoid cirsiliol, a trihydroxy-6,7-dimethoxyflavone extracted from Salvia x jamensis, in modulating reactive oxygen species production by the ectopic oxidative phosphorylation taking place in the OS. Our molecular docking analysis identified cirsiliol binding sites inside the F1 moiety of the nanomotor F1Fo-ATP synthase. The experimental approach was based on luminometry, spectrophotometry and cytofluorimetry to evaluate ATP synthesis, respiratory chain complex activity and H2O2 production, respectively. The results showed significant dose-dependent inhibition of ATP production by cirsiliol. Moreover, cirsiliol was effective in reducing the free radical production by the OS exposed to ambient light. We report a considerable protective effect of cirsiliol on the structural stability of rod OS, suggesting it may be considered a promising compound against oxidative stress.
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46

Andreola, Giovanna, Licia Rivoltini, Chiara Castelli, Veronica Huber, Paola Perego, Paola Deho, Paola Squarcina, et al. "Induction of Lymphocyte Apoptosis by Tumor Cell Secretion of FasL-bearing Microvesicles." Journal of Experimental Medicine 195, no. 10 (May 20, 2002): 1303–16. http://dx.doi.org/10.1084/jem.20011624.

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The hypothesis that FasL expression by tumor cells may impair the in vivo efficacy of antitumor immune responses, through a mechanism known as ‘Fas tumor counterattack,’ has been recently questioned, becoming the object of an intense debate based on conflicting results. Here we definitely show that FasL is indeed detectable in the cytoplasm of melanoma cells and its expression is confined to multivesicular bodies that contain melanosomes. In these structures FasL colocalizes with both melanosomal (i.e., gp100) and lysosomal (i.e., CD63) antigens. Isolated melanosomes express FasL, as detected by Western blot and cytofluorimetry, and they can exert Fas-mediated apoptosis in Jurkat cells. We additionally show that melanosome-containing multivesicular bodies degranulate extracellularly and release FasL-bearing microvesicles, that coexpress both gp100 and CD63 and retain their functional activity in triggering Fas-dependent apoptosis of lymphoid cells. Hence our data provide evidence for a novel mechanism potentially operating in Fas tumor counterattack through the secretion of subcellular particles expressing functional FasL. Such vesicles may form a sort of front line hindering lymphocytes and other immunocompetent cells from entering neoplastic lesions and exert their antitumor activity.
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47

Feldman, Tatiana, Dmitriy Ostrovskiy, Marina Yakovleva, Alexander Dontsov, Sergey Borzenok, and Mikhail Ostrovsky. "Lipofuscin-Mediated Photic Stress Induces a Dark Toxic Effect on ARPE-19 Cells." International Journal of Molecular Sciences 23, no. 20 (October 13, 2022): 12234. http://dx.doi.org/10.3390/ijms232012234.

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Lipofuscin granules from retinal pigment epithelium (RPE) cells contain bisretinoid fluorophores, which are photosensitizers and are phototoxic to cells. In the presence of oxygen, bisretinoids are oxidized to form various products, containing aldehydes and ketones, which are also potentially cytotoxic. In a prior study, we identified that bisretinoid oxidation and degradation products have both hydrophilic and amphiphilic properties, allowing their diffusion through the lipofuscin granule membrane into the RPE cell cytoplasm, and are thiobarbituric acid (TBA)-active. The purpose of the present study was to determine if these products exhibit a toxic effect to the RPE cell also in the absence of light. The experiments were performed using the lipofuscin-fed ARPE-19 cell culture. The RPE cell viability analysis was performed with the use of flow cytofluorimetry and laser scanning confocal microscopy. The results obtained indicated that the cell viability of the lipofuscin-fed ARPE-19 sample was clearly reduced not immediately after visible light irradiation for 18 h, but after 4 days maintaining in the dark. Consequently, we could conclude that bisretinoid oxidation products have a damaging effect on the RPE cell in the dark and can be considered as an aggravating factor in age-related macular degeneration progression.
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48

Moiseeva, Natalia I., Lidia A. Laletina, Timur I. Fetisov, Leyla F. Makhmudova, Angelika E. Manikaylo, Liliya Y. Fomina, Denis A. Burov, et al. "Analysis of Multiple Drug Resistance Mechanism in Different Types of Soft Tissue Sarcomas: Assessment of the Expression of ABC-Transporters, MVP, YB-1, and Analysis of Their Correlation with Chemosensitivity of Cancer Cells." International Journal of Molecular Sciences 23, no. 6 (March 16, 2022): 3183. http://dx.doi.org/10.3390/ijms23063183.

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Chemotherapy of soft tissue sarcomas (STS) is restricted by low chemosensitivity and multiple drug resistance (MDR). The purpose of our study was the analysis of MDR mechanism in different types of STS. We assessed the expression of ABC-transporters, MVP, YB-1, and analyzed their correlation with chemosensitivity of cancer cells. STS specimens were obtained from 70 patients without metastatic disease (2018–2020). Expression level of MDR-associated genes was estimated by qRT-PCR and cytofluorimetry. Mutations in ABC-transporter genes were captured by exome sequencing. Chemosensitivity (SI) of STS to doxorubicin (Dox), ifosfamide (Ifo), gemcitabine (Gem), and docetaxel (Doc) was analyzed in vitro. We found strong correlation in ABCB1, ABCC1, and ABCG2 expression. We demonstrated strong negative correlations in ABCB1 and ABCG2 expression with SI (Doc) and SI (Doc + Gem), and positive correlation of MVP expression with SI (Doc) and SI (Doc + Gem) in undifferentiated pleomorphic sarcoma. Pgp expression was shown in 5 out of 44 STS samples with prevalence of synovial sarcoma relapses and it is strongly correlated with SI (Gem). Mutations in MDR-associated genes were rarely found. Overall, STS demonstrated high heterogeneity in chemosensitivity that makes reasonable in vitro chemosensitivity testing to improve personalized STS therapy, and classic ABC-transporters are not obviously involved in MDR appearance.
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49

Limongi, Tania, Francesca Susa, Bianca Dumontel, Luisa Racca, Michela Perrone Donnorso, Doriana Debellis, and Valentina Cauda. "Extracellular Vesicles Tropism: A Comparative Study between Passive Innate Tropism and the Active Engineered Targeting Capability of Lymphocyte-Derived EVs." Membranes 11, no. 11 (November 18, 2021): 886. http://dx.doi.org/10.3390/membranes11110886.

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Cellular communications take place thanks to a well-connected network of chemical–physical signals, biomolecules, growth factors, and vesicular messengers that travel inside or between cells. A deep knowledge of the extracellular vesicle (EV) system allows for a better understanding of the whole series of phenomena responsible for cell proliferation and death. To this purpose, here, a thorough immuno-phenotypic characterization of B-cell EV membranes is presented. Furthermore, the cellular membrane of B lymphocytes, Burkitt lymphoma, and human myeloid leukemic cells were characterized through cytofluorimetry assays and fluorescent microscopy analysis. Through cytotoxicity and internalization tests, the tropism of B lymphocyte-derived EVs was investigated toward the parental cell line and two different cancer cell lines. In this study, an innate capability of passive targeting of the native EVs was distinguished from the active targeting capability of monoclonal antibody-engineered EVs, able to selectively drive the vesicles, enhancing their internalization into the target cancer cells. In particular, the specific targeting ability of anti-CD20 engineered EVs towards Daudi cells, highly expressing CD20 marker on their cell membrane, was proved, while almost no internalization events were observed in HL60 cells, since they did not express an appreciable amount of the CD20 marker on their plasma membranes.
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50

Belvedere, Raffaella, Elva Morretta, Nunzia Novizio, Silvana Morello, Olga Bruno, Chiara Brullo, and Antonello Petrella. "The Pyrazolyl-Urea Gege3 Inhibits the Activity of ANXA1 in the Angiogenesis Induced by the Pancreatic Cancer Derived EVs." Biomolecules 11, no. 12 (November 24, 2021): 1758. http://dx.doi.org/10.3390/biom11121758.

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The pyrazolyl-urea Gege3 molecule has shown interesting antiangiogenic effects in the tumor contest. Here, we have studied the role of this compound as interfering with endothelial cells activation in response to the paracrine effects of annexin A1 (ANXA1), known to be involved in promoting tumor progression. ANXA1 has been analyzed in the extracellular environment once secreted through microvesicles (EVs) by pancreatic cancer (PC) cells. Particularly, Gege3 has been able to notably prevent the effects of Ac2-26, the ANXA1 mimetic peptide, and of PC-derived EVs on endothelial cells motility, angiogenesis, and calcium release. Furthermore, this compound also inhibited the translocation of ANXA1 to the plasma membrane, otherwise induced by the same ANXA1-dependent extracellular stimuli. Moreover, these effects have been mediated by the indirect inhibition of protein kinase Cα (PKCα), which generally promotes the phosphorylation of ANXA1 on serine 27. Indeed, by the subtraction of intracellular calcium levels, the pathway triggered by PKCα underwent a strong inhibition leading to the following impediment to the ANXA1 localization at the plasma membrane, as revealed by confocal and cytofluorimetry analysis. Thus, Gege3 appeared an attractive molecule able to prevent the paracrine effects of PC cells deriving ANXA1 in the tumor microenvironment.
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