Bibiana D. Riquelme

Complex viscoelasticity of normal and lectin treated erythrocytes using laser diffractometry

A new method to find directly complex viscoelastic parameters (CVP) of red blood cells (RBC) is presented in this paper. Experimental determinations were carried out in an Erythrodeformeter (Rasia et al., 1986) operating in oscillating mode (0.5 to 3.5 Hz). The Erythrodeformeter performs direct determination of CVP of erythrocytes undergoing sinusoidal shear stresses by laser diffractometry. The measurements lead to the determination of mean values of the four components of erythrocyte complex viscoelasticity. The influence of the alterations induced on erythrocyte membrane by vegetable lactins (Ulex europaeus, wheat germ agglutinin and Enterolobium contorticilicum seeds) was analyzed to verify the sensitivity of this method. Differences observed between the CVP parameters of treated cells and the ones corresponding to control samples (non treated cells) are analyzed. Results obtained from cells treated with wheat germ agglutinin agree with observations published by Smith and Hochmuth (1982). Determinations of RBC complex viscoelasticity carried out by laser diffractometry could become an important tool to understand the influence of the factors associated with alterations of the rheologic properties of RBC membrane, which can affect the in vivo blood flow.

Quantification of glycophorin A and glycophorin B on normal human RBCs by flow cytometry

BACKGROUND: The quantification of antigens and proteins on RBCs has been achieved by different approaches. Flow cytometry allows the results of the earliest studies to be to reappraised because it offers the possibility of measuring the immunofluorescence intensity of single cells and integrating the individual data of a large number of cells within a very short time.

Kinetic study of antibody adhesion on a silicon wafer by laser reflectometry

Abstract Antibody adhesion kinetic in real time has been studied by laser reflectometry technique. An ellipsometer is used to measure the light intensity reflected by a silicon wafer. Light intensity reflected by the wafer presents a minimum at the pseudo-Brewster angle. Then, the reflectance increases as the antibodies (monoclonal anti- AB ) adhere on interface. Mathematical analysis of reflectance curves versus time verifies that the antibody adhesion at the interface follows Langmuir kinetics (Prog. Biomed. Opt. Imaging 1(5) (2000) 19) for low antibody concentrations. Parameters obtained allow to carry out a detailed study of the antibody adsorption and the antigen–antibody interaction. This conduces to development of an optical immunosensor for detection and quantification of soluble antigens, and a novel method for commercial antiserum quality control. This technique does not require labeled antibodies, being also independent of cellular factors. Also, this technique is quicker and sensible than the conventional immunohematology methods.

Study of polycation effects on erythrocyte agglutination mediated by anti-glycophorins using microscopic image digital analysis

The aim of this work was to study synthetic polycation effects on erythrocyte agglutination mediated by anti-glycophorin using image digital analysis. Polycations are oligomers or polymers of natural or synthetic origin, which bear a great number of positive charges at pH 7.4. Several of these polycations are nowadays used in clinic for human and veterinary purposes. New applications of polycations to the development of new drug delivery systems are investigated, in order to promote the drug absorption through the gastro-intestinal and blood brain barriers. However, up to now, there are no clear relationships between macromolecular features of polycations (molecular weight, mean charge density, charge repartition, etc.) and their interactions with blood elements (which bear superficial negative charges). The interaction on the red blood cell membrane with synthetic polycations having well-controlled macromolecular features and functionalized with pendent polyethylene glycol segments was investigated. The alterations over stationary and dynamic viscoelastic properties of erythrocyte membranes were analyzed through laser diffractometry. Image digital analysis was used to study erythrocyte agglutination mediated by anti-glycophorin. Results show different reactivities of the polycations on the erythrocyte membrane. These findings could provide more information about the mechanisms of polycation interaction on erythrocyte membranes. We consider that this work could provide useful tools to understand and improve the haemocompatibility of polycations and enlarge their potential in clinic.

Acid-Induced Aggregation and Gelation of Sodium Caseinate-Guar Gum Mixtures

The aim of this work was to study the formation of bovine sodium caseinate (NaCAS) acid gels induced by addition of glucono-δ-lactone (GDL) in the presence of guar gum (GG). At low biopolymer’s concentrations, a one-phase system was observed, whereas at higher mixture concentrations two-phase systems were formed. Aggregation (at low NaCAS concentrations) and gelation (at high NaCAS concentrations) processes were analyzed through the use of full and fractional factorial experiment designs, using turbidimetric and rheological techniques. Finally, the gel images were obtained by confocal laser scanning microscopy and the images were analyzed. Results showed that at low NaCAS concentrations, the presence of GG affects the pH at which aggregation begins but was not significant for the time at which aggregation begins. On the other hand, at high NaCAS concentrations, the concentration of GG only affected significantly the elastic character of acid gels. As polysaccharide concentration increases, the gels obtained were weaker and with larger pores. Also, the formation of NaCAS droplet-shaped structures at certain biopolymer ratio was observed. The presence of GG affects both the rate of gelation and phase separation, which, in turn, determine the type of gel microstructure. Phase separation seems to occur prior to protein gelation because the protein gel network is discontinued, hindering the gel compactness and reducing gel strength. In summary, GG modifies NaCAS stabilization (self-association and phase separation) and the viscoelasticity and microstructure of NaCAS acid gels. The control of such processes and properties would allow obtaining mixture gels with different textures.

Acid-Induced Aggregation and Gelation of Bovine Sodium Caseinate-Carboxymethylcellulose Mixtures

Maria Eugenia Hidalgo1, Bibiana D. Riquelme1,2, Estela M. Alvarez1, Jorge R. Wagner3 and Patricia H. Risso1,2,4 1Facultad de Ciencias Bioquimicas y Farmaceuticas, Universidad Nacional de Rosario 2Instituto de Fisica Rosario (IFIR), CONICET-UNR, (2000), Rosario, 3Departamento de Ciencia y Tecnologia, 4Facultad de Ciencias Veterinarias,Universidad Nacional de Rosario, Rosario Universidad Nacional de Quilmes, Buenos Aires, Argentina

Hemocompatibility and biofunctionality of two poly(2-(dimethylamino)ethyl methacrylate-co-poly(ethyleneglycol) copolymers.

To mask the antigenic sites of cells for cell therapies, especially for blood transfusion, we investigated the hemocompatibility of two poly(2-(dimethylamino)ethyl methacrylate-co-poly(ethyleneglycol) compared with that of the homopolymer without PEG. Our strategy relies on the potential ability of these copolymers to self-assemble at the erythrocyte surface. The cationic sequence of the copolymer should be able to interact with the glycocalyx by ionic interaction. The other sequence, based on a polyethyleneglycol moiety, should prevent both nonspecific interactions and specific recognition of the biological surface. The hemocompatibility of these copolymers was assessed by analyzing alterations in human erythrocyte membrane viscoelasticity, morphology, granularity, and aggregation. Their properties to mask ABO system and three erythrocyte glycophorin sites were investigated. No alterations in the erythrocyte morphology were observed by confocal microscopy. On the other hand, a partial masking of different specific glycophorin sites leads to future optimization of the macromolecular structures of these functionalized copolymers.

Impairment of Erythrocytes Incubated in Glucose Medium: A Wavelet-Information Theory Analysis

This study investigates the effects produced by an increased concentration of glucose in a suspending medium on the erythrocytes Information Theory quantifiers. Erythrocytes, which were obtained from eight healthy volunteers, were washed and incubated in vitro with glucose solutions at different concentrations. The measured Wavelet-based Information Theory quantifiers include the Relative Wavelet Energy (RWE), the Normalized Total Wavelet Shannon Entropy (NTWS), MPR-Statistical Complexity Measure (SCM) and entropy–complexity plane. The results show that the increase in glucose concentration does not produce significant changes on the RWE, while significant ones on the NTSE, which combined with SCM values allow to identify different behaviour for all the different populations in the entropy–complexity plane. Modification in the hemorheological properties of cells could be clearly detected with these Wavelet-based Information Theory quantifiers.

Determination of the complex viscoelastic parameters of human red blood cells by laser diffractometry

A new method to find directly complex viscoelastic parameters (CVP) of human red blood cells is presented in this paper. Experimental determinations were carried out in an Erythrodeformeter operating in oscillating mode (0.5 to 3.5 Hz). The Erythrodeformeter performs direct determination of CVP of erythrocytes undergoing sinusoidal shear stresses by laser diffractometry.

Simplified variant of an optical chip to evaluate aggregation of red blood cells

Traditional techniques to evaluate the aggregation of red blood cells by optical methods require large sample volume and provide parameters that vary significantly from one method to another. A simplified variant of a chip system previously developed by Shin et al. (2009)1 based on light transmission for measuring erythrocyte aggregation is presented. Through a detailed analysis of intensity versus time curves, relevant information about erythrocyte aggregation and its variables is obtained. Parameters that provide more accuracy for the diagnosis of patients in order to have an immediate application in Clinical Medicine are proposed.