scholarly journals Assessment of the oxidative metabolism of calves’ bronchoalveolar cells: comparison between NBT and fluorimetric techniques

2018 ◽  
Vol 85 (0) ◽  
Author(s):  
Bruna Artner ◽  
Angela Reck ◽  
Alessandra Mayer Coelho ◽  
Julio José Prediger ◽  
Desiree Vera Pontarolo ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Flow Cytometry ◽  
Alveolar Macrophages ◽  
Oxidative Metabolism ◽  
Therapeutic Dose ◽  
Flow Cytometry Assay ◽  
Colorimetric Test ◽  
Healthy Cattle ◽  
The Difference ◽  
Cattle Husbandry

ABSTRACT: Evaluation of alveolar macrophage functions of cattle is an important tool in order to assess whether measures taken during the cattle husbandry can decrease the respiratory tract defense. The aim of this study was to determine whether dexamethasone used at therapeutic dose can affect the oxidative metabolism of alveolar macrophages of cattle. This was evaluated by two tests, the fluorometric and colorimetric. The similarity of the results was studied, using alveolar macrophages of six healthy cattle, obtained from bronchoalveolar lavage on a basal and an immunosuppressant moment after the application of dexamethasone. For the fluorometric test, alveolar macrophages were incubated with Staphylococcus aureus and 2’-7’dichlorohidroflurescein, and analyzed by flow cytometer. For the colorimetric test, alveolar macrophages were incubated with Phorbol 12- miristate-13 acetate and nitroblue tetrazolium, dissolved and analyzed in a spectrophotometer. It was noted that dexamethasone therapeutic dose (0.05 mg/kg) reduced the functions of alveolar macrophages from healthy bovine. This result was observed by both tests with the difference that the flow cytometry assay was more informative for identifying which specific cellular function has been compromised.

scholarly journals Use of Flow Cytometry to Evaluate Phagocytosis of Staphylococcus aureus by Human Neutrophils

2021 ◽  
Vol 12 ◽  
Author(s):  
Elena Boero ◽  
Iris Brinkman ◽  
Thessely Juliet ◽  
Eline van Yperen ◽  
Jos A. G. van Strijp ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Flow Cytometry ◽  
Immune Responses ◽  
Human Neutrophils ◽  
Host Proteins ◽  
Flow Cytometry Assay ◽  
Immune Therapies ◽  
Human Immune Response ◽  
Human Immune ◽  
Phagocytic Uptake

Neutrophils play a key role in the human immune response to Staphylococcus aureus infections. These professional phagocytes rapidly migrate to the site of infection to engulf bacteria and destroy them via specialized intracellular killing mechanisms. Here we describe a robust and relatively high-throughput flow cytometry assay to quantify phagocytosis of S. aureus by human neutrophils. We show that effective phagocytic uptake of S. aureus is greatly enhanced by opsonization, i.e. the tagging of microbial surfaces with plasma-derived host proteins like antibodies and complement. Our rapid assay to monitor phagocytosis can be used to study neutrophil deficiencies and bacterial evasion, but also provides a powerful tool to assess the opsonic capacity of antibodies, either in the context of natural immune responses or immune therapies.

Whole Spleen Flow Cytometry Assay

BIO-PROTOCOL ◽  
2013 ◽  
Vol 3 (15) ◽  
Author(s):  
Cathy Yam ◽  
Adeline Hajjar
Keyword(s):  
Flow Cytometry ◽  
Flow Cytometry Assay

scholarly journals Identification of pluripotent cells in bovine uterus: in situ and in vitro studies

Reproduction ◽  
2015 ◽  
Vol 149 (4) ◽  
pp. 317-327 ◽  
Author(s):  
Martyna Łupicka ◽  
Gabriel Bodek ◽  
Nahum Shpigel ◽  
Ehud Elnekave ◽  
Anna J Korzekwa
Keyword(s):  
Flow Cytometry ◽  
Protein Expression ◽  
Mrna Expression ◽  
Uterine Horn ◽  
Uterine Tissue ◽  
Pluripotent Cells ◽  
Flow Cytometry Assay ◽  
Myometrial Cells ◽  
Gene And Protein Expression

The aim of this study was to identify uterine pluripotent cells both in bovine uterine tissues as well in epithelial, stromal, and myometrial uterine cell populations. Moreover, the relationship of pluripotent markers expression with age and the uterine horn side was considered. Uterine tissue was collected from ipsilateral and contralateral horns (days 8–10 of the estrous cycle). Immunohistostaining for C-KIT, OCT3/4, NANOG, and SOX2 in uterine tissue was determined. mRNA expression of C-KIT, OCT3/4, NANOG and SOX2 was evaluated in uterine tissue relative to the age of the cow and uterine horn side. Gene and protein expression of these markers in the uterine luminal epithelial, stromal, and myometrial cells was evaluated by real-time PCR and western blotting respectively. The expression of pluripotent cell markers OCT3/4, NANOG, and SOX2 was identified by flow cytometry assay in epithelial, stromal, and myometrial cells. Multilineage differentiation of the bovine uterine cells was performed. mRNA expression of OCT3/4, NANOG, and SOX2 in uterine tissue was higher in the ipsilateral horn than in the contralateral horn. Flow cytometry assay revealed positive fluorescence for OCT3/4, NANOG, and SOX2 in all uterine cell types. Results showed the age-dependent expression of pluripotent markers in uterine tissue. Beside, the different expression of pluripotent cells in each horn of uterus suggests the influence of ovarian hormones on these characteristics. The highest mRNA and protein expression for pluripotent markers was observed in stromal cells among uterine cells, which indicates this population of cells as the main site of pluripotent cells in the cow uterus.

scholarly journals Measurement of Separase Proteolytic Activity in Single Living Cells by a Fluorogenic Flow Cytometry Assay

PLoS ONE ◽  
2015 ◽  
Vol 10 (8) ◽  
pp. e0133769 ◽  
Author(s):  
Wiltrud Haaß ◽  
Helga Kleiner ◽  
Martin C. Müller ◽  
Wolf-Karsten Hofmann ◽  
Alice Fabarius ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Proteolytic Activity ◽  
Living Cells ◽  
Flow Cytometry Assay ◽  
Single Living Cells ◽  
Single Living

scholarly journals Multiparameter Flow Cytometric Analysis of Antibiotic Effects on Membrane Potential, Membrane Permeability, and Bacterial Counts of Staphylococcus aureus andMicrococcus luteus

2000 ◽  
Vol 44 (4) ◽  
pp. 827-834 ◽  
Author(s):  
David J. Novo ◽  
Nancy G. Perlmutter ◽  
Richard H. Hunt ◽  
Howard M. Shapiro
Keyword(s):  
Staphylococcus Aureus ◽  
Flow Cytometry ◽  
Membrane Potential ◽  
Membrane Permeability ◽  
Micrococcus Luteus ◽  
Flow Cytometric Analysis ◽  
Bacterial Counts ◽  
Flow Cytometric ◽  
Permeability Parameter ◽  
Particle Counts

ABSTRACT Although flow cytometry has been used to study antibiotic effects on bacterial membrane potential (MP) and membrane permeability, flow cytometric results are not always well correlated to changes in bacterial counts. Using new, precise techniques, we simultaneously measured MP, membrane permeability, and particle counts of antibiotic-treated and untreated Staphylococcus aureus andMicrococcus luteus cells. MP was calculated from the ratio of red and green fluorescence of diethyloxacarbocyanine [DiOC2(3)]. A normalized permeability parameter was calculated from the ratio of far red fluorescence of the nucleic acid dye TO-PRO-3 and green DiOC2(3) fluorescence. Bacterial counts were calculated by the addition of polystyrene beads to the sample at a known concentration. Amoxicillin increased permeability within 45 min. At concentrations of <1 μg/ml, some organisms showed increased permeability but normal MP; this population disappeared after 4 h, while bacterial counts increased. At amoxicillin concentrations above 1 μg/ml, MP decreased irreversibly and the particle counts did not increase. Tetracycline and erythromycin caused smaller, dose- and time-dependent decreases in MP. Tetracycline concentrations of <1 μg/ml did not change permeability, while a tetracycline concentration of 4 μg/ml permeabilized 50% of the bacteria; 4 μg of erythromycin per ml permeabilized 20% of the bacteria. Streptomycin decreased MP substantially, with no effect on permeability; chloramphenicol did not change either permeability or MP. Erythromycin pretreatment of bacteria prevented streptomycin and amoxicillin effects. Flow cytometry provides a sensitive means of monitoring the dynamic cellular events that occur in bacteria exposed to antibacterial agents; however, it is probably simplistic to expect that changes in a single cellular parameter will suffice to determine the sensitivities of all species to all drugs.

Faster Detection of Staphylococcus aureus in Milk and Milk Powder by Flow Cytometry

2021 ◽  
Author(s):  
Siyuan Liu ◽  
Bin Wang ◽  
Zhiwei Sui ◽  
Ziquan Wang ◽  
Longquan Li ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Flow Cytometry ◽  
Milk Powder

scholarly journals Single-Tube Flow Cytometry Assay for the Detection of Mature Lymphoid Neoplasms in Paucicellular Samples

2016 ◽  
Vol 60 (4) ◽  
pp. 385-394
Author(s):  
Alessandra Stacchini ◽  
Anna Demurtas ◽  
Sabrina Aliberti ◽  
Antonella Barreca ◽  
Domenico Novero ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Hodgkin Lymphoma ◽  
Final Diagnosis ◽  
Molecular Data ◽  
Malignant Lymphomas ◽  
Flow Cytometry Assay ◽  
Single Tube ◽  
Fine Needle Aspirates ◽  
Non Hodgkin Lymphoma ◽  
A Cell

Objectives: Flow cytometry (FC) has become a useful support for cytomorphologic evaluation (CM) of fine-needle aspirates (FNA) and serous cavity effusions (SCE) in cases of suspected non-Hodgkin lymphoma (NHL). FC results may be hampered by the scarce viability and low cellularity of the specimens. Study Design: We developed a single-tube FC assay (STA) that included 10 antibodies cocktailed in 8-color labeling, a cell viability dye, and a logical gating strategy to detect NHL in hypocellular samples. The results were correlated with CM and confirmed by histologic or molecular data when available. Results: Using the STA, we detected B-type NHL in 31 out of 103 hypocellular samples (81 FNA and 22 SCE). Of these, 8 were not confirmed by CM and 2 were considered to be only suspicious. The FC-negative samples had a final diagnosis of benign/reactive process (42/72), carcinoma (27/72), or Hodgkin lymphoma (3/72). Conclusions: The STA approach allowed obtainment of maximum immunophenotyping data in specimens containing a low number of cells and a large amount of debris. The information obtained by STA can help cytomorphologists not only to recognize but also to exclude malignant lymphomas.

scholarly journals A flow cytometry assay to quantify intercellular exchange of membrane components

2017 ◽  
Vol 8 (8) ◽  
pp. 5585-5590 ◽  
Author(s):  
Dimitrios Poulcharidis ◽  
Kimberley Belfor ◽  
Alexander Kros ◽  
Sander I. van Kasteren
Keyword(s):  
Flow Cytometry ◽  
Flow Cytometry Assay ◽  
Membrane Components ◽  
Intercellular Exchange

A technically simple and broadly deployable FACS-based assay to determine intercellular exchange of membrane components.

Lyso(bis)phosphatidic acid: A novel source of arachidonic acid for oxidative metabolism by rabbit alveolar macrophages

1985 ◽  
Vol 130 (2) ◽  
pp. 800-806 ◽  
Author(s):  
Felicia R. Cochran ◽  
Janice R. Connor ◽  
Vicki L. Roddick ◽  
B.Moseley Waite
Keyword(s):  
Arachidonic Acid ◽  
Phosphatidic Acid ◽  
Alveolar Macrophages ◽  
Oxidative Metabolism
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