Determination of osmotic resistance of erythrocytes using flow cytometry in patients with hereditary spherocytosis

Long-term high fat-carbohydrates diet (HF-CD) contributes to the formation of irreversible changes in the organism that lead to the emergence of civilization diseases. In this study, the impact of three-month high-fat diet on the physical properties of erythrocytes (RBCs) was studied. Furthermore, the biological activity of Cistus incanus L. extracts, plant known with high pro-health potential, in relation to normal and HF-CD RBCs, was determined. Obtained results have shown that, applied HF-CD modified shape, membrane potential and osmotic resistance of erythrocytes causing changes in membrane lipid composition and the distribution of lipids. The impact of HF-CD on physical properties of RBCs along with atherosclerotic lesions of the artery was visible, despite the lack of statistically significant changes in blood morphology and plasma lipid profile. This suggests that erythrocytes may be good markers of obesity-related diseases. The studies of biological activity of Cistus incanus L. extracts have demonstrated that they may ameliorate the effect of HF-CD on erythrocytes through the membrane-modifying and antioxidant activity.

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The Past, Present and Future of Flow Cytometry in Central Nervous System Malignancies

Methods and Protocols ◽

10.3390/mps4010011 ◽

2021 ◽

Vol 4 (1) ◽

pp. 11

Author(s):

Evrysthenis Vartholomatos ◽

George Vartholomatos ◽

George A. Alexiou ◽

Georgios S. Markopoulos

Keyword(s):

Central Nervous System ◽

Flow Cytometry ◽

Nervous System ◽

Therapeutic Agents ◽

Phenotypic Characterization ◽

Cell Phenotype ◽

Analytical Technique ◽

The Past ◽

Current Review

Central nervous system malignancies (CNSMs) are categorized among the most aggressive and deadly types of cancer. The low median survival in patients with CNSMs is partly explained by the objective difficulties of brain surgeries as well as by the acquired chemoresistance of CNSM cells. Flow Cytometry is an analytical technique with the ability to quantify cell phenotype and to categorize cell populations on the basis of their characteristics. In the current review, we summarize the Flow Cytometry methodologies that have been used to study different phenotypic aspects of CNSMs. These include DNA content analysis for the determination of malignancy status and phenotypic characterization, as well as the methodologies used during the development of novel therapeutic agents. We conclude with the historical and current utility of Flow Cytometry in the field, and we propose how we can exploit current and possible future methodologies in the battle against this dreadful type of malignancy.

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KD determination from time-resolved experiments on live cells with LigandTracer and reconciliation with end-point flow cytometry measurements

European Biophysics Journal ◽

10.1007/s00249-021-01560-2 ◽

2021 ◽

Author(s):

Diana Spiegelberg ◽

Jonas Stenberg ◽

Pascale Richalet ◽

Marc Vanhove

Keyword(s):

Flow Cytometry ◽

Live Cells ◽

Time Resolved ◽

Surface Receptors ◽

Full Characterization ◽

End Point ◽

Titration Curves ◽

Kinetics Of

AbstractDesign of next-generation therapeutics comes with new challenges and emulates technology and methods to meet them. Characterizing the binding of either natural ligands or therapeutic proteins to cell-surface receptors, for which relevant recombinant versions may not exist, represents one of these challenges. Here we report the characterization of the interaction of five different antibody therapeutics (Trastuzumab, Rituximab, Panitumumab, Pertuzumab, and Cetuximab) with their cognate target receptors using LigandTracer. The method offers the advantage of being performed on live cells, alleviating the need for a recombinant source of the receptor. Furthermore, time-resolved measurements, in addition to allowing the determination of the affinity of the studied drug to its target, give access to the binding kinetics thereby providing a full characterization of the system. In this study, we also compared time-resolved LigandTracer data with end-point KD determination from flow cytometry experiments and hypothesize that discrepancies between these two approaches, when they exist, generally come from flow cytometry titration curves being acquired prior to full equilibration of the system. Our data, however, show that knowledge of the kinetics of the interaction allows to reconcile the data obtained by flow cytometry and LigandTracer and demonstrate the complementarity of these two methods.

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Development of multiparametric analysis of human PBMC by flow cytometry: Determination of inflammatory and calcic profile

Archives of Cardiovascular Diseases Supplements ◽

10.1016/j.acvdsp.2021.04.126 ◽

2021 ◽

Vol 13 (2) ◽

pp. 199-200

Author(s):

C. Brun ◽

A. Paccalet ◽

T. Bochaton ◽

M. Paillard ◽

C. Crola Da Silva

Keyword(s):

Flow Cytometry ◽

Human Pbmc ◽

Multiparametric Analysis

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Flow Cytometry-Based Determination of Ploidy from Dried Leaf Specimens in Genomically Complex Collections of the Tropical Forage Grass Urochloa s. l.

Genes ◽

10.3390/genes12070957 ◽

2021 ◽

Vol 12 (7) ◽

pp. 957

Author(s):

Paulina Tomaszewska ◽

Till K. Pellny ◽

Luis M. Hernández ◽

Rowan A. C. Mitchell ◽

Valheria Castiblanco ◽

...

Keyword(s):

Flow Cytometry ◽

Germplasm Collection ◽

Forage Grass ◽

Breeding Programs ◽

Robust Identification ◽

Ploidy Levels ◽

Sustainable Improvement ◽

Tropical Forage ◽

Relative Differences

Urochloa (including Brachiaria, Megathyrus and some Panicum) tropical grasses are native to Africa and are now, after selection and breeding, planted worldwide, particularly in South America, as important forages with huge potential for further sustainable improvement and conservation of grasslands. We aimed to develop an optimized approach to determine ploidy of germplasm collection of this tropical forage grass group using dried leaf material, including approaches to collect, dry and preserve plant samples for flow cytometry analysis. Our methods enable robust identification of ploidy levels (coefficient of variation of G0/G1 peaks, CV, typically <5%). Ploidy of some 348 forage grass accessions (ploidy range from 2x to 9x), from international genetic resource collections, showing variation in basic chromosome numbers and reproduction modes (apomixis and sexual), were determined using our defined standard protocol. Two major Urochloa agamic complexes are used in the current breeding programs at CIAT and EMBRAPA: the ’brizantha’ and ’humidicola’ agamic complexes are variable, with multiple ploidy levels. Some U. brizantha accessions have odd level of ploidy (5x), and the relative differences in fluorescence values of the peak positions between adjacent cytotypes is reduced, thus more precise examination of this species is required. Ploidy measurement of U. humidicola revealed aneuploidy.

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Determination of testicular function after torsion by DNA flow cytometry of serial fine-needle aspirates

BJU International ◽

10.1046/j.1464-410x.1997.09825.x ◽

1997 ◽

Vol 79 (3) ◽

pp. 449-454 ◽

Cited By ~ 3

Author(s):

T.A. Carroll ◽

M.C. Regan ◽

R. Alyusuf ◽

D. Greene ◽

B. Curran ◽

...

Keyword(s):

Flow Cytometry ◽

Dna Flow Cytometry ◽

Testicular Function ◽

Fine Needle ◽

Fine Needle Aspirates

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Determination of antiviral efficacy against lymphotropic herpesviruses utilizing flow cytometry

Antiviral Research ◽

10.1016/s0166-3542(02)00210-3 ◽

2003 ◽

Vol 58 (2) ◽

pp. 149-157 ◽

Cited By ~ 20

Author(s):

Mary C. Long ◽

Deborah J. Bidanset ◽

Stephanie L. Williams ◽

Nicole L. Kushner ◽

Earl R. Kern

Keyword(s):

Flow Cytometry ◽

Antiviral Efficacy

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Increased Fraction of Circulating Activated Platelets in Acute and Previous Cerebrovascular Ischemia

Thrombosis and Haemostasis ◽

10.1055/s-0037-1615191 ◽

1998 ◽

Vol 80 (08) ◽

pp. 298-301 ◽

Cited By ~ 62

Author(s):

Andreas Ruf ◽

Axel Vogt ◽

Christoph Lichy ◽

Florian Buggle ◽

Heinrich Patscheke ◽

...

Keyword(s):

Flow Cytometry ◽

Phase Contrast ◽

Matched Control ◽

Shape Change ◽

Antigen Expression ◽

Phase Contrast Microscopy ◽

Activated Platelets ◽

Contrast Microscopy ◽

Age And Sex

SummaryDetermination of circulating activated platelets may be helpful to estimate the prognosis and to stratify therapies in arterial vascular disorders including stroke. We used flow cytometry and phase contrast microscopy to study whether the fraction of platelets expressing p-selectin and CD63 and the fraction of platelets with shape change are increased in patients with acute and previous cerebrovascular ischemia.The proportion of platelets expressing activation dependent antigens was higher in patients with acute (n = 24; p-selectin: 8.23 ± 4.21%; CD63: 3.53 ± 2.53%) and with previous cerebrovascular ischemia (n = 46; 3.86 ± 1.98%; 2.80 ± 1.79%) as compared to age- and sex-matched control subjects (n = 35; 2.17 ± 0.96%; 1.79 ± 0.75%; p ≤0.005, respectively). In patients with previous ischemia, there was no difference between treatment with aspirin (n = 25) or phenprocoumon (n = 21). Hypertension, diabetes mellitus and smoking were not associated with increased antigen expression (analysis of variance). The fraction of discoid platelets and platelet counts were not significantly different between groups.Our results indicate increased expression of platelet neoantigens in acute and to a less degree in previous cerebrovascular ischemia. Ongoing platelet activation after cerebrovascular ischemia despite therapy with aspirin or phenprocoumon indicates that new anti-platelet drugs may be of benefit for these patients. Flow cytometry appears to be a useful tool to assess platelet function in cerebrovascular ischemia.

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