Ascaris lumbricoides: Mimetismo molecular por absorción de epitopes P1 / Ascaris lumbricoides: Molecular mimicry by P1 epitope absorption

El mimetismo molecular se ha asociado con la cronicidad de las infecciones parasitarias. Este mecanismo puede deberse a absorción de moléculas del hospedador o a síntesis de moléculas similares. P1 ha sido detectado en diversas especies parásitas. Experiencias previas han demostrado epitopes P1 en ejemplares adultos de Ascaris lumbricoides. El objetivo de este trabajo fue estudiar la absorción de estos epitopes por estadios larvales del nematodo, a los fines de determinar el mecanismo de expresión de estos antígenos sobre la cutícula parasitaria. Las larvas fueron eclosionadas a partir de heces con huevos de este helminto. Fueron cultivadas en medio RPMI con eritrocitos P1. Se realizó la Técnica de Inhibición de la Aglutinación Semicuantitativa usando cuatro anticuerpos monoclonales anti-P1. El sistema revelador fue una suspensión de eritrocitos frescos P1 en medio enzimático de bromelina. Se investigaron por la misma técnica productos de excreción-secreción símil P1 usando larvas cultivadas en medio RPMI en ausencia de eritrocitos P1. Las larvas cultivadas con glóbulos rojos inhibieron la aglutinación de todos los anticuerpos monoclonales con los eritrocitos P1. No se identificaron productos de excreción- secreción símil P1. Se ha demostrado que las larvas de A. lumbricoides absorben epitopes P1 del medio de cultivo. Se concluye que el parásito puede absorber este antígeno durante su ciclo de vida para usarlo en el mimetismo molecular.

Molecular mimicry has been associated with chronicity of parasitic infections. This mechanism can be due to absorption of the host's molecules or to synthesis of similar molecules. P1 has been detected in various parasite species. Previous experiences have shown P1 epitopes in Ascaris lumbricoides's adult parasites. The aim of this study was to analyse absoption of these epitopes by nematode larvae in order to determine the mechanism by which the parasite can express these antigens on their cuticle. Larvae were hatched from the faeces with helminth eggs. Larvae were cultivated in RPMI medium with P1 red cells. The Quantitative Inhibition Agglutination Test was performed using 4 anti- P1 monoclonal antibodies. P1 fresh red cells in bromelin enzymatic medium was used as revealing system. Simil P1 excretory-secretory products were investigated using larvae cultivated in RPMI medium without P1 erythrocytes by the same technique. The larvae which were cultivated with red cells inhibited the agglutination between all anti- P1 monoclonal antibodies and P1 erythrocytes. Simil P1 excretory-secretory products were not identified. It was demonstrated that A. lumbricoides larvae absorb P1 epitopes from the culture medium. It can be concluded that the parasite can absorb this antigen during its life cycle in order to use it in molecular mimicry.

Ascaris lumbricoides: Estudio de la cinética de captación de ácido hialurónico / Ascaris lumbricoides: Study of kinetics of hyaluronic acid capture

El ácido hialurónico (AH) participa de la respuesta inmune porque es un constituyente de las barreras mecánicas naturales, un estimulador de la inflamación y un ligando de receptores involucrados en la inmunidad. En experiencias previas se demostró que adultos y larvas de Ascaris lumbricoides pueden unir AH. El objetivo de este trabajo fue estudiar la cinética de captación de AH por este nematodo. Se trabajó con un extracto de parásito adulto y un concentrado de larvas. Se utilizó la técnica modificada de Inhibición de la Agregación por Adhesión para detección del receptor CD44 soluble de hialuronato. Se evaluó la cinética de captación de AH por el parásito, variando el tiempo de contacto entre ambos desde un tiempo inicial hasta 90 minutos, con intervalos de 10 minutos. Para todos los tiempos se calcularon los valores de CexpAdhE AH e IexpCP AH que fueron analizados por las pruebas de Wilcoxon y de Friedman. Los resultados mostraron que al tiempo inicial de contacto, el parásito adulto y las larvas captan una pequeña cantidad de AH. A los 10 minutos se produjo la mayor captación que se mantuvo sin diferencias significativas durante 50 minutos. A partir de ese tiempo, hubo un leve incremento en la captación que permaneció sin diferencia significativa hasta el tiempo final. Esta experiencia demuestra que in vitro, A. lumbricoides realiza la mayor captación a los 10 minutos de contacto, por lo que se sugiere que in vivo sería capaz de unir AH rápidamente a los fines de modular la respuesta inmune.

Hyaluronic acid (HA) particípales in the immune response because it is a constituent of natural mechanical barriers, an inflammation stimulator and a ligand for receptors involved in immunity. Previous experiences have shown that adult specimens and larval concéntrales oí A. lumbricoides have hyaluronan binding capacity. The aim ofthis work was to study the kinetics ofhyaluronic acid capture by this parasite. Work was carried out with an extract of adult specimen and a larval concéntrate. The modlfled test of serum soluble CD44 Detectlon by Aggregatlon Inhlbltlon was used. Kinetics hyaluronlc acid capture by the nematode was assessed by varylng the contad time between HA and parasite, from an initial time to 90 minutes, with 10-minute intervals. CexpAdhE HA ana lexpCP HA were calculated for all the times and they were analysed by Wilcoxon and Friedman Tests The results showed that at initial time of contad, the adult paraslte and the larvae had captured a small amount of HA. The major capture was made at 10 minutes and it was maintained for 50 minutes without significad differences. From that time there was a slight increase in capture and it remaíned so untíi the end of the experiment without considerable variations. This experience demonstrates that at 10 minutes of in vitro contad, A. lumbricoides captures most of the HA, whích suggests that in vivo, the parasite would be able to capture HA quickly for the purpose of modulating the immune response.

Preliminary characterization of erythrocytes deformability on the entropy-complexity plane.

We present an application of wavelet-based Information Theory quantifiers (Normalized Total Shannon Entropy, MPR-Statistical Complexity and Entropy-Complexity plane) on red blood cells membrane viscoelasticity characterization. These quantifiers exhibit important localization advantages provided by the Wavelet Theory. The present approach produces a clear characterization of this dynamical system, finding out an evident manifestation of a random process on the red cell samples of healthy individuals, and its sharp reduction of randomness on analyzing a human haematological disease, such as ß-thalassaemia minor.

Estudio de la captación de ácido hialurónico por Ascaris lumbricoides / Study of hyaluronic acid capture by Ascaris lumbricoides

El ácido hialurónico (AH) tiene importantes funciones en la inmunidad. En experiencias previas se demostró que extractos de adultos de A. lumbricoides y concentrados larvarios, tienen capacidad de unión a AH. El objetivo de este trabajo fue estudiar la captación de AH por este helminto. Se trabajó con tres extractos del parásito adulto ([EA]B,C,D) y 3 concentrados de larvas ([CLAL1]: 1100 a 1200 larvas/ mL; [CLAL2]: 400 a 600 larvas/ mL y [CLAL3]: 100 a 200 larvas/ mL). Se empleó la técnica modificada de Inhibición de la Adhesión para detección del Receptor CD44 soluble de hialuronato en suero humano. Se definió CexpAdhE AH como el cociente entre los eritrocitos adheridos por el AH en presencia y ausencia del parásito y se definió IexpCP AH, como la cantidad de eritrocitos que se dejaron de adherir debido a la captación de AH por el parásito, referido al número total de eritrocitos. Los resultados mostraron diferencias significativas en CexpAdhE AH y en IexpCP AH, por efecto de la concentración larvaria y del [EA]. Las medias aritméticas de CexpAdhE AH y de IexpCP AH para los concentrados larvarios fueron 0,636 y 0,21 ([CLAL1]); 0,819 y 0,068 ([CLAL2]); 0,97 y 0,013 ([CLAL3]). Las medianas de CexpAdhE AH y de IexpCP AH para los extractos fueron [EA]C 0,275 y 0,4; [EA]B: 0,20 y 0,43; [EA]D: 0,075 y 0,495. La experiencia permitiría suponer que el parásito puede captar AH para interferir en la respuesta inmune del hospedador.

Hyaluronan Acid (HA) has important functions in immunity. Previous experiences have shown that A. lumbricoides's extracts of adult specimens and larval concentrates have hyaluronan binding capacity. The aim of this study was to analyse the HA capture by this helminth. Three extracts of adult specimens ([AE]B,C,D) and 3 larval concentrates ([ALLC1]: 1100 to 1200 larvae/ mL; [ALLC2]: 400 to 600 larvae/ mL and [ALLC3]: 100 to 200 larvae/ mL) were studied. The modified test of serum soluble CD44 Detection by Aggregation Inhibition was used. CexpAdhE AH was defined as the quotient between erythrocytes adhered by the HA in presence and in absence of the parasite, and IexpCP AH was defined as the amount of erythrocytes that stopped adhering due to the HA capture by the parasite in relation to the total number of erythrocytes. The results showed significant differences in CexpAdhE AH and IexpCP AH as the result of larvae concentration and of the extracts. The arithmetic means of CexpAdhE AH and IexpCP AH for the larvae were 0.636 and 0.21 ([ALLC1]); 0.819 and 0.068 ([ALLC2]); 0.97 and 0.013 ([ALLC3]). Medians of CexpAdhE AH and IexpCP AH for the extracts were [EA]C:0.275 and 0.4; [EA]B: 0.20 and 0.43; [EA]D: 0.075 and 0.495. From the experience, it could be assumed that the parasite can capture the HA to interfere in the host's immune response.

Ascaris lumbricoides: alteration of the erythrocyte superficial charge using the Partition Method in aqueous two-phase system.

Sialic acid is responsible for the negative charge of the erythrocyte. The decrease of sialic acid has hemodynamical and hemorheological importance. The aim was to study the effect of A. lumbricoides on the erythrocyte superficial charge using the Partition Method in aqueous two-phase system in order to indirectly evaluate the alteration of sialic acid in the red cells. We worked with five parasite extracts (AE) and larvae concentrate (LC). Erythrocyte superficial charge was studied by working with non-treated (Controls) and treated erythrocytes. The treatment consisted of incubating the erythrocytes with AE or LC for 30 minutes at 4 degrees C, 20 degrees C and 37 degrees C. The red cells were separated in a sensitive charge two-phase system (Dx/ PEG). The partition coefficient (P) of treated and untreated erythrocytes were calculated. The results showed a P decrease at the three temperatures for red cells treated with four of the AE. The remaining extract did change P values at any of the temperatures studied. The erythrocytes treated with LC showed a decrease in the P value at 37 degrees C and 4 degrees C but no change was observed at 25 degrees C. Statistical analysis concluded that P values were significantly lower in treated erythrocytes than in their corresponding untreated ones (p < 0.05). The Partition Method showed that this parasite alters the erythrocyte superficial charge which may indicate that it can catch sialic acid.

Ascaris lumbricoides: alteration of the erythrocyte superficial charge using the Partition Method in aqueous two-phase system / Ascaris lumbricoides: alteración de la carga superficial eritrocitaria usando el Método de Partición en sistemas bifásicos acuosos

Sialic acid is responsible for the negative charge of the erythrocyte. The decrease of sialic acid has hemodynamical and hemorheological importance. The aim was to study the effect of A. lumbricoides on the erythrocyte superficial charge using the Partition Method in aqueous two-phase system in order to indirectly evaluate the alteration of sialic acid in the red cells. We worked with five parasite extracts (AE) and larvae concentrate (LC). Erythrocyte superficial charge was studied by working with non-treated (Controls) and treated erythrocytes. The treatment consisted of incubating the erythrocytes with AE or LC for 30 minutes at 4 ºC, 20 ºC and 37 ºC. The red cells were separated in a sensitive charge two-phase system (Dx/ PEG). The partition coefficient (P) of treated and untreated erythrocytes were calculated. The results showed a P decrease at the three temperatures for red cells treated with four of the AE. The remaining extract did change P values at any of the temperatures studied. The erythrocytes treated with LC showed a decrease in the P value at 37 ºC and 4 ºC but no change was observed at 25 ºC. Statistical analysis concluded that P values were significantly lower in treated erythrocytes than in their corresponding untreated ones (p < 0.05). The Partition Method showed that this parasite alters the erythrocyte superficial charge which may indicate that it can catch sialic acid.

La disminución de ácido sialico, responsable de la carga negativa del eritrocito, tiene importancia hemodinámica y hemorreológica. El objetivo fue estudiar el efecto de A. lumbricoides sobre la carga superficial eritrocitaria aplicando el método de partición en sistemas bifásicos acuosos, a los fines de evaluar de manera indirecta la alteración de acido sialico de los eritrocitos. Se trabajó con 5 extractos del parásito adulto (EA) y con un concentrado de larvas (500-600 larvas/mL) (CL). Se estudió la carga superficial eritrocitaria, trabajando con eritrocitos no tratados y tratados. El tratamiento consistió en la incubación de los eritrocitos con EA o CL durante 30 minutos a 4 ºC, 25 ºC y 37 ºC. Los eritrocitos fueron sometidos a la separación en un sistema bifásico carga sensible constituido por Dx / PEG. Se calculó el coeficiente de reparto (P), de los eritrocitos sin tratar y tratados. Los resultados mostraron disminución de P a las 3 temperaturas, en hematíes tratados con 4 de los EA. El EA restante no modificó los valores de P a ninguna de las temperaturas estudiadas. CL produjo la disminución de P a 37 ºC y 4 ºC, pero no se observó modificación a 25 ºC. Los análisis estadísticos concluyeron que los valores de P son significativamente menores en los eritrocitos tratados que en los respectivos eritrocitos sin tratar (p < 0.05). El método de partición demostró que A. lumbricoides altera la carga superficial eritrocitaria lo que indicaría que el parásito, tanto en su estado adulto como en sus fases larvales, puede captar acido sialico.

[Serum sialic acid, cellular anionic charge and erythrocyte aggregation in diabetic and hypertensive patients]. / Acido siálico sérico, carga aniónica y agregación eritrocitaria en pacientes diabéticos e hipertensos.

The aim of this work was to study the relationship between serum sialic acid (SSA) and erythrocyte anionic charge (EAC) with erythrocyte aggregation in two groups: diabetic (DBT, n=20) and hypertensive (HT, n=21) patients, compared to a control group (n=20). We worked with anticoagulated blood with EDTA and serum. The erythrocyte aggregation was studied by microscopically observing and quantifying aggregates using an ASP (Aggregate Shape Parameter). The EAC was determined by binding an Alcian blue dye to the membrane sialic acid and SSA was determined by spectrophotometric method with an Erlich reactant. The values of ASP and SSA increased significantly in HT and DBT patients compared to the control group. The HT and DBT groups showed amorphous aggregates, evident in an alteration in the values of ASP, which were significantly higher ( p < 0.005) than in healthy patients. The EAC values were much lower in HT and DBT patients than in the control group (p < 0.0001). In this work, abnormalities in the erythrocyte aggregation could be detected by the values of ASP, EAC and SSA, which might be involved in vascular disorders of diseases such as hypertension and diabetes.

Ácido siálico sérico, carga aniónica y agregación eritrocitaria en pacientes diabéticos e hipertensos / Serum sialic acid, cellular anionic charge and erythrocyte aggregation in diabetic and hypertensive patients

El objetivo de este trabajo fue estudiar la relación del ácido siálico sérico (AS) y la carga aniónica eritrocitaria (CAE) con la agregación eritrocitaria en dos grupos de pacientes: diabéticos (DBT n= 20) e hipertensos (HTA n= 21), comparados con un grupo control (n= 20). Se trabajó con sangre anticoagulada con EDTA y suero. La agregación eritrocitaria se estudió por observación microscópica de los agregados y cuantificación a través de un parámetro de forma denominado ASP (Aggregation Shape Parameter). La CAE se determinó por unión a colorante alcian blue y el AS por método espectrofotométrico con reactivo de Erlich. Los valores de ASP y AS resultaron significativamente aumentados en los HTA y DBT respecto de los normales. Los HTA y DBT presentaron agregados amorfos, lo que se refleja en los valores alterados de ASP, significativamente mayores (p < 0.005) respecto de los individuos normales. Los valores de CAE resultaron significativamente inferiores en los HTA y DBT respecto del grupo control (p < 0.0001). En este trabajo se demostraron anormalidades en la agregación eritrocitaria, detectadas por los valores de ASP, CAE y AS que podrían estar involucradas en las complicaciones vasculares de vasculopatías como la hipertensión y la diabetes.

The aim of this work was to study the relationship between serum sialic acid (SSA) and erythrocyte anionic charge (EAC) with erythrocyte aggregation in two groups: diabetic (DBT, n=20) and hypertensive (HT, n=21) patients, compared to a control group (n=20). We worked with anticoagulated blood with EDTA and serum. The erythrocyte aggregation was studied by microscopically observing and quantifying aggregates using an ASP (Aggregate Shape Parameter). The EAC was determined by binding an Alcian blue dye to the membrane sialic acid and SSA was determined by spectrophotometric method with an Erlich reactant. The values of ASP and SSA increased significantly in HT and DBT patients compared to the control group. The HT and DBT groups showed amorphous aggregates, evident in an alteration in the values of ASP, which were significantly higher ( p< 0.005) than in healthy patients. The EAC values were much lower in HT and DBT patients than in the control group (p < 0.0001). In this work, abnormalities in the erythrocyte aggregation could be detected by the values of ASP, EAC and SSA, which might be involved in vascular disorders of diseases such as hypertension and diabetes.

[Larval stages of Ascaris lumbricoides: hyaluronan-binding capacity]. / Estados larvales de Ascaris lumbricoides: capacidad de unión a acido hialurónico.

Hyaluronic acid has important functions in inflammatory and tissue reparation processes. Owing to the varied strategies of the parasites to evade the host's immune response, as well as the multiple functions and physiological importance of hyaluronic acid, the aim was to study the hyaluronan binding capacity by Ascaris lumbricoides larval stages. Larval concentrates were prepared by hatching A. lumbricoides eggs. The larvae were collected by the Baermann method. The test of serum soluble CD44 detection by Agregation Inhibition was modified. All the larval concentrates presented hyaluronan binding capacity. The obtained results allow to suppose the existence of an hyaluronic acid specific receptor in A. lumbricoides. This receptor eventually might compete with the usual receptors of the host. The parasite might use this mechanism to evade the immune response.

Estados larvales de ascaris lumbricoides: capacidad de unión a ácidos hialurónico / Larval stages of ascaris lumbricoides: hyaluronan binding capacity

El ácido hialurónico tiene importantes funciones en los procesos inflamatorios y de reparación tisular. Debido a la variedad de estrategias utilizadas por los parásitos para evadir la respuesta inmune del hospedador y las múltiples funciones e importancia fisiológica del ácido hialurónico, el objetivo fue estudiar la capacidad de unión a ácido hialurónico de estados larvarios de A. lumbricoides Se trabajó con 4 Concentrados de Larvas obtenidas de la eclosión de huevos de A. lumbricoides y recolectadas por el método de Baermann. Se modificó la técnica de detección de CD44 soluble en suero por Inhibición de la Agregación por Adhesión. Se observó que los 4 Concentrados Larvales unían ácido hialurónico. Los resultados obtenidos permiten suponer la existencia de un receptor con especificidad para ácido hialurónico en A. lumbricoides. Este receptor eventualmente podría competir con los receptores habituales del hospedador. El parásito podría utilizar este mecanismo para evadir la respuesta inmune

Hyaluronic acid has important functions in inflammatory and tissue reparation processes. Owing to the varied strategies of the parasites to evade the host´s immune response, as well as the multiple functions and physiological importance of hyaluronic acid, the aim was to study the hyaluronan binding capacity by Ascaris lumbricoides larval stages. Larval concentrates were prepared by hatching A. lumbricoides eggs. The larvae were collected by the Baermann method. The test of serum soluble CD44 detection by Agregation Inhibition was modified. All the larval concentrates presented hyaluronan binding capacity. The obtained results allow to suppose the existence of an hyaluronic acid specific receptor in A. lumbricoides. This receptor eventually might compete with the usual receptors of the host. The parasite might use this mechanism to evade the immune response

Identificación de epitopes Tipo Grupo Sanguíneo ABO en larvas de Ascaris lumbricoides mantenidas in vitro / Identification of ABO Blood Group like epitopes in vitro maintained Ascaris lumbricoides larvae

En investigaciones previas se identificaron los mismos epitopes del Sistema ABO en extractos de A. lumbricoides y en sus respectivos hospederos. Los objetivos de este trabajo fueron estudiar la absorción de determinantes antigénicas ABO por cultivo in vitro de estadios larvales de este helminto e investigar la presencia de sustancias Tipo Grupo Sanguíneo ABO en los productos de secreción/excreción de las larvas. Se trabajó con larvas obtenidas de la eclosión de huevos de A. lumbricoides y recolectadas por el método de Baermann en cinco tubos con medio RPMI 1640. En tres de los tubos se adicionaron eritrocitos (A, B y O). Los dos tubos restantes se usaron como control negativo de absorción y para investigar los productos de excreción- /secreción de las larvas. Se aplicaron técnicas de Inhibición de la Aglutinación Semicuantitativa usando anticuerpos monoclonales. Los resultados mostraron que las larvas absorben epitopes A, B y H de los eritrocitos agregados al medio de cultivo. Se identificaron sustancias Tipo A y B en los productos de excreción/secreción de las larvas. La experiencia puede contribuir a la comprensión de los mecanismos de evasión de la respuesta inmune del hospedero utilizados por A. lumbricoides.

Previous experiences have demonstrated the same ABO System epitopes in A. lumbricoides extracts and in their hosts. The aims of this work were to study ABO antigenic determiner absorption by in vitro culture of the helminth's larvae and to investigate ABO-Blood-Group-like substances in larvae's excretion/ secretion products. Larvae were obtained by hatching of A. lumbricoides eggs. The larvae were collected in 5 tubes with RPMI 1640 medium by the Baermann method. A, B and O erythrocytes were added in 3 of the tubes. The two remaining tubes were used as absorption Negative Control and to investigate larvae's excretion/secretion products. The Semiquantitative Inhibition Agglutination Test was applied by using monoclonal antibodies. The results showed that the larvae may adsorb A, B and H epitopes from the erythrocytes that were added to the culture medium. A and B-Blood-Group-like substances were identified in the excretion/secretion products. The experience can contribute to understand the host's escape mechanism used by A. lumbricoides.

Hemorheologic changes in type 2 diabetic patients with microangiopathic skin lesions. A linear discriminant categorizing analysis.

The development of diabetic microangiopathy may produce lesions in distinct organic territories (the skin, among others). With an intention of identifying hemorheologic variables for a better characterization of diabetic patients, we studied plasmatic and blood viscosities (P Visc at 2.30 s(-1) and B Visc at 4.60 and 230 s(-1)), fibrinogenemia, and erythrocytic aggregation in 40 type 2 diabetic patients (13 with microangiopathic skin lesions and 27 without) and in 30 healthy controls. Considering its alterations in diabetic patients and applying linear discriminant analysis, two models may be characterized: (a) discriminant function (Disc F)=0.58 aggregate shape parameter (ASP)-0.61 B Visc(230)+0.89 fibrinogenemia for discriminating healthy individuals from diabetic patients with microangiopathic skin lesions and (b) Disc F=6.325 ASP-0.347 B Visc(230)+0.013 fibrinogenemia for discriminating healthy controls from diabetic patients with and without microangiopathic skin lesions. Both models appear to be valid due to the following: (a) Model 1: a coefficient of canonic correlation of 0.924, a highly significant Mahalanobis distance (P<10(-3)), a correct percentage of classification (100%), and the centroids of each group (0.94 and 5.63); (b) Model 2: a coefficient of canonic correlation of 0.898, a highly significant Mahalanobis distance (P<10(-3)), a correct percentage of classification (85.7%), and the centroids of each group (-1.9, 1.9, and 2.4). Just as the alterations in the analyzed hemorheologic variables could be suggesting their possible involvement in the physiopathogenia of diabetic microangiopathic skin lesions, the proposed models could characterize a microcirculatory profile in diabetic patients for preventing irreversible damages.

Optimization and application of electrophoretic mobility analysis of human red blood cells to study their in vitro stability, interaction with polycations and proteolytic enzymes.

Electrophoretic light scattering method has been considered to determine both the mean and polydispersity of electrophoretic mobility of normal human red blood cells. The final goal of our study was to optimize an in vitro test allowing us to investigate the interaction of xenobiotics, in particular polyelectrolytes, with blood cells. The feasibility of our method has been evaluated based on the reproducibility of our technique to analyze the native electrophoretic mobility of human RBCs, as well as their evolution in the presence of polycationic compounds, i.e. a natural polyamine, spermine, and a synthetic polycation, a polyethylenimine (PEI). Additionally, the follow-up of the human RBCs electrophoretic mobility has been controlled after their incubation with neuraminidase.

Ascaris lumbricoides: Capacidad de unión a hialuronato / Ascaris lumbricoides: Hyaluronan binding capacity

El ácido hialurónico tiene importantes funciones en los procesos inflamatorios y de reparación tisular. Sus principales receptores son CD44, RHAMM e ICAM-I. Debido a la variedad de estrategias utilizadas por los parásitos para evadir la respuesta inmune del hospedador y considerando las múltiples funciones e importancia fisiológica del ácido hialurónico, el objetivo de este trabajo fue determinar si Ascaris lumbricoides tiene capacidad de unión a hialuronato. Se trabajó con 36 extractos de A. lumbricoides obtenidos por remoción quirúrgica de la cutícula y ruptura mecánica refrigerada. Se modificó la técnica de detección de CD44 soluble en suero por Inhibición de la Agregación por adhesión. Los resultados demostraron que 23 de los 36 extractos estudiados tenían capacidad de unión a hialuronato. Este hecho podría deberse a la existencia de algún receptor en el parásito que une hialuronato y que eventualmente competiría con los receptores habituales del hospedador. A. lumbricoides podría utilizar este mecanismo para evadir la respuesta inmune.

Hyaluronan acid has important functions in inflammatory and tissue reparation processes. Its main receptors are CD44, RHAMM and ICAM-I. Owing to the varied strategies of the parasites to evade the host´s immune response, and also considering the multiple functions and physiological importance of hyaluronan acid, the aim was to study if Ascaris lumbricoides has hyaluronan binding capacity. Extracts of A. lumbricoides were prepared by surgical remotion of the cuticle and refrigerated mechanical rupture. The study was done on 36 parasite extracts. The test of serum soluble CD44 Detection by Aggregation Inhibition was modified. Of the 36 extracts studied, 23 presented hyaluronan binding capacity. This fact can possibly be due to the existence of a receptor with hyaluronan acid binding capacity in the parasite, which eventually might compete with the usual receptors of the host. A. lumbricoides might use this mechanism to evade the immune response.

Ascaris lumbricoides: Capacidad inhibitoria relacionada al antígeno P1 / Ascaris lumbricoides: Inhibitory capacity in relation to P1 antigen

El Sistema de Grupo Sanguíneo P tiene un único antígeno: P1 que fue identificado en varias especies parásitas. El objetivo de este trabajo fue determinar la capacidad inhibitoria de A. lumbricoides en relación al antígeno P1. Se trabajó con 11 extractos parasitarios (EA). Para evaluar la capacidad inhibitoria de los extractos en relación a los epitopes P1, se realizó la técnica semicuantitativa de inhibición de la aglutinación. Se prepararon dos series de diluciones de anticuerpo monoclonal anti- P1; a la primera se le agregó PBS a todos los tubos y a la otra, igual volumen de EA. El sistema revelador fue una suspensión de eritrocitos P1. Se determinó el título del anticuerpo monoclonal en ambas series. Se consideró significativa la aglutinación cuando hubo una diferencia de 2 o más diluciones entre ambos títulos. De los 11 EA estudiados, 8 presentaron epitopes P1. A estos 8 extractos se les calculó el Poder Inhibitorio y se lo relacionó con su contenido proteico a los fines de poder comparar la capacidad inhibitoria entre los extractos. La técnica aplicada permitió evidenciar la presencia de epitopes del Sistema P en extractos de A. lumbricoides y comparar la capacidad inhibitoria de los extractos en relación a epitopes P1.

P Blood Group has a unique antigen: P1. It was identified in various parasite species. The aim of this work was to determine A. lumbricoides´s Inhibitory Capacity in relation to P1 antigen. The work was based on 11 parasite extracts (AE). The semiquantitative inhibition agglutination technique was used to assess the extracts´ Inhibitory Capacity in relation to P1 antigen. Two series of anti- P1 monoclonal antibody dilutions were prepared. PBS was added to the first series and an equal volume of AE to the second. Suspensions of P1 red cells were used as a revealing system. Monoclonal antibody titres were determined in both series. The agglutination was considered significative when there was a difference of two or more dilutions between the titres. Eight of the 11 extracts studied presented P1 epithopes. Inhibitory power of these 8 extracts was calculated and it was related with their protein concentrations to compare the Inhibitory Capacity among the extracts. The technique used made it possible to prove P System epithopes presence and to compare the extracts´ Inhibitory Capacity in relation to P1 epithopes.

Ascaris lumbricoides: epitopes del Sistema P por método de cinética de la hemaglutinación / Ascaris lumbricoides: P system epithopes by haemogglutination kinetics method

Experiencias previas demostraron epitopes P1 en Ascaris lumbricoides por inhibición de la aglutinación. La cinética de la hemaglutinación aplica la extinción óptica relativa producida por un haz de luz trasmitido a través de una suspensión de pequeñas partículas, y permite obtener una estimación paramétrica de la tasa de hemaglutinación. El objetivo de este trabajo fue utilizar este método para demostrar la presencia de epitopes del Sistema P en extractos de A. lumbricoides. La técnica de inhibición de la aglutinación permitió seleccionar 10 extractos: 3 con y 7 sin epitopes P1. Se registró la cinética de la reacción Control: anti- P1- eritrocitos - P1 y la cinética de la misma reacción, previo contacto del anticuerpo con el extracto. Se calcularon y se compararon los valores de ³ EOR % (variación en el porcentaje de extinción óptica relativa) para ambas reacciones. Los resultados evidenciaron la presencia de epitopes P1, que no habían sido detectados por inhibición de la aglutinación, en 3 extractos. Para los extractos restantes, hubo coincidencia entre los resultados obtenidos por los dos métodos. La cinética de la hemaglutinación es sencilla, rápida y fácilmente adaptable para las experiencias en que se necesite evaluar una reacción antígeno-anticuerpo a través de la hemaglutinación.

Summary Previous experiences have demonstrated P1 epithopes in Ascaris lumbricoides by inhibition agglutination. Haemogglutination kinetics applies the relative optical extinction produced on a light beam transmitted through a suspension of small particles, giving a parametric estimate of the haemagglutination rate.The aim was to use this method for the demonstration of P System epithopes presence in A. lumbricoides extracts. inhibition agglutination test enabled the selection of 10 extracts: 3 with and 7 without P1 epithopes. Kinetics of anti- P1 - P1 erythrocyte control reaction and kinectics of the same reaction with a previous contact between antibody and extract were registered. ³ EOR % values (relative optical extinction variation) for both reactions were calculated and compared between themselves. The results proved P1 epithopes presence in 3 extracts, which had not been detected by inhibition agglutination. The results coincided for the remaining extracts by both methods. Haemogglutination kinetics is simple and fast and it is easily adapted for experiences in which the investigator needs to assess the antibody-antigen reaction by haemogglutination.

[Ascaris lumbricoides: heterogeneity in ABO epithopes expression]. / Ascaris lumbricoides: heterogeneidad en la expresión de epitopes ABO.

Ascaris lumbricoides and its hosts can present the same ABO System epithopes. The aim was to study ABO epithopes expression in A. lumbricoides by Inhibition Agglutination Test using different monoclonal antibodies and policlonal sera. We worked with 27 parasite extracts (14 extracts from A Group patients, 2 AB Group 1, B Group and 10 from unknown ABO Group patients). Inhibition Agglutination Test was made facing the extracts from A, AB and unknown ABO Group patients against monoclonal antibodies and policlonal serum of A specificity and the extracts from B, AB and unknown ABO Group patients against monoclonal antibodies and policlonal serum of B specificity. The Semiquantitative Test was made with the extracts which inhibited the agglutination of any antibody. The 50% of the parasite extracts faced against A specificity antibodies and the 46% of the parasite extracts faced against B specificity antibodies significantly inhibited the agglutination in the Semiquantitative Test. No extract inhibited the totality of the antibodies. All the antibodies reacted with some extract, except one antibody of B specificity. The ABO activity in the extracts was independent of the antibody's immunoglobulin class, titre and concentration and it only was dependent on the antibody union with the epithope at which the antibody is directed. The heterogeneity in the ABO epithopes expression of A.lumbricoides might be involved in the escape of the host's immune response. The used Inhibition Tests are sensitive, simple, rapid and economic. We conclude that the use of different antibodies alows the best definition of the antigen-antibody specificity.

Ascaris lumbricoides: heterogeneidad en la expresión de epitopes ABO / Ascaris lumbricoides: heterogeneity in ABO epithopes expression

Ascaris lumbricoides puede presentar los mismos epitopes del Sistema ABO que sus hospederos. El objetivo fue estudiar la expresión de epitopes ABO en el parásito por la técnica de Inhibición de la Aglutinación usando diferentes anticuerpos monoclonales y sueros policlonales. Se trabajó con 27 extractos parasitarios (14 de pacientes A, 2 AB, 1 B y 10 de Grupo ABO desconocido). Los extractos de pacientes A, AB y Grupo desconocido fueron enfrentados a anticuerpos monoclonales y suero policlonal de especificidad A; y los de pacientes B, AB y Grupo desconocido a anticuerpos monoclonales y suero policlonal de especificidad B. Se hizo la Inhibición Semicuantitativa a los extractos que inhibieron la aglutinación de algún anticuerpo. Se observó que el 50 por ciento de los extractos enfrentados a anticuerpos de especificidad A y el 46 por ciento de los enfrentados a anticuerpos de especificidad B inhibieron la aglutinación de forma significativa. Ningún extracto inhibió a la totalidad de los anticuerpos. Todos los anticuerpos reaccionaron con algún extracto, excepto un anticuerpo de especificidad B. La actividad ABO del extracto fue independiente de la clase de inmunoglobulina, concentración y título del anticuerpo; Se observó que sólo depende de la unión del anticuerpo al epitope al cual está dirigido. La heterogeneidad en la expresión de epitopes ABO de A. lumbricoides podría estar involucrada en la evasión de la respuesta inmune del hospedero. Las técnicas de Inhibición utilizadas son sensibles, sencillas, rápidas y económicas. Se concluye que el uso de diferentes anticuerpos monoclonales permite la mejor definición de la especificidad antígeno-anticuerpo

Laser diffractometry technique: clinical applications to vascular pathologies.

Hypertension (HTA) and dyslipidemia (DLP) represent major risk factors for cardiovascular and cerebral-vascular ischemic disease. The mechanisms through which they can induce vascular damage are both metabolic and mechanical. Hemorheological alterations in HTA are result of changes affecting both red cell intrinsic structure and their interactions with the plasmatic components. Several hemorheological determinants (biochemical, ionic, metabolic and rheologic) could influence and produce an impaired erythrocyte deformability determining an increased flow resistance in the microcirculation. The "Erythrodeformeter" allows obtaining the stationary and dynamical linear parameters of erythrocyte membrane by laser diffractometry. Stationary and oscillatory shear-induced elongation of cells leads to an elliptical diffraction pattern, its geometric characteristics being directly related to those of deformed RBC. Erythrocyte stationary parameters (Deformability Index, surface viscosity and elastic modulus) were obtained in stationary regime. Complex viscoelastic parameters (dynamic elasticity, dynamic loss, viscous and elastic components of the complex viscosity) were obtained when operating in oscillating mode. The diffractometric method is sensitive to detect pathological or induced alterations on RBC membrane, which can affect blood flow in vivo. The rheological parameters obtained give important information about the erythrocyte membrane and allow to detect and characterize erythrocyte alterations in vascular pathologies.

[Hemorheological alterations in hypertensive patients]. / Evaluación de alteraciones hemorreológicas en pacientes hipertensos.

The aim of this study was to investigate the blood viscosity profile and to evaluate the influence of plasmatic (fibrinogen) and cellular (erythrocyte aggregation) factors in a group of hypertensive patients, compared with a normotensive group. We worked with anticoagulated blood of both non diabetic hypertensive patients (n=31), and healthy individuals (n=40). The plasmatic viscosity and whole blood determination were obtained with a cone-plate viscometer. Erythrocyte aggregation was studied by microscopical observation and quantified by an Aggregate Shape Parameter (ASP), defined as the relation projected area/perimeter. Fibrinogen was determined by the Clauss method with a coagulometer. A comparison between these groups led us to assert that whole blood viscosity was significantly higher in hypertensive patients than in the controls at all shear rates. Plasma viscosity values only showed significant differences between both groups at low shear rate (1.15 a 11.56 seg(-1)). The hypertensive patients showed irregular and amorphous aggregates so that ASP appeared significantly higher (p< 0.001) in patients with hypertension (0.69 +/- 0.11) than in healthy subjects (0.25 +/- 0.12). Fibrinogen appeared slightly higher (p<0.01) in the hypertensive group than in the normal group. Several hemorheological parameters play important roles in the pathogenesis of hypertension. Among these factors, several hemorheological parameters could be altered in hypertension (hematocrit, plasma fibrinogen level, erythrocyte deformability and aggregability, plasma and whole blood viscosity). An increased RBC aggregation has been identified as an important factor responsible for disturbing blood rheological behavior in the microcirculation. The present study demonstrates an abnormal erythrocyte aggregation, which was detected by increased ASP values that could be responsible for vascular complications in hypertension.