Parámetros hemocitarios y química de la hemolinfa en reproductores de la concha prieta Anadara tuberculosa (Arcoida: Arcidae)

Con motivo de conocer el estado de salud de poblaciones silvestres de la concha prieta Anadara tuberculosa se estudiaron los parámetros hemocitarios (viabilidad, conteo total y diferencial, estabilidad lisosomal, fragilidad osmótica) y la química sanguínea (hemoglobina-Hb, proteínas, triglicéridos, glucosa, lactato deshidrogenasa-LDH y catalasa-CAT). Se extrajo hemolinfa en reproductores salvajes aparentemente sanos colectados en la isla Corazón, río Chone, Ecuador. La viabilidad celular fue elevada, con presencia de 5 morfotipos: eritrocitos (74%), granulocitos traslucidos (6%), amebocitos (3%), hialinocitos (12%), blastocitos (5%). Los hemocitos presentaron membranas lisosomales estables al rojo neutro durante 240 min y una fragilidad osmótica media (FO50) de 4.8‰. Las concentraciones de Hb, proteínas, lípidos y glucosa denotan la función respiratoria y reservas energéticas durante los cambios de marea. La actividad de LDH está vinculada al metabolismo anaeróbico y CAT a la capacidad de mantener el equilibrio redox del sistema inmunitario. Los parámetros hemocitarios y química de la hemolinfa pueden servir como índices fisiológicos normales de referencia en reproductores de A. tuberculosa.

In this work we studied the health status of wild populations of the black shell Anadara tuberculosa, haemocyte parameters (viability, total and differential count, lysosomal stability, osmotic fragility) and blood chemistry (haemoglobin-Hb, proteins, triglycerides, glucose, lactate dehydrogenase-LDH and catalase-CAT). Haemolymph was extracted from apparently healthy wild broodstock collected from Corazón Island, Chone River, Ecuador. Cell viability was high, with 5 morphotypes present: erythrocytes (74%), colorless granulocytes (6%), amebocytes (3%), hyalinocytes (12%), blastocytes (5%). Haemocytes showed stable lysosomal membranes to neutral red for 240 min and a mean osmotic fragility (OF50) was 4.8‰. Hb, protein, lipid and glucose concentrations denote respiratory function and energy reserves during tidal changes. LDH activity is linked to anaerobic metabolism and CAT denotes the ability to maintain the redox balance of the immune system. The haemocitary parameters and haemolymph chemistry can serve as normal physiological reference indices in A. tuberculosa broodstock.

Índice neutrófilo-linfocito, relación plaquetas-linfocito y distribución de la anchura del eritrocito en pacientes con preeclampsia / Neutrophil-to-lymphocyte ratio, plateletto- lymphocyte ratio, and erythrocyte width distribution. and erythrocyte width distribution in patients with preeclampsia

Resumen OBJETIVO: Comparar el índice neutrófilo-linfocito, la relación plaquetas-linfocito y la distribución de la anchura del eritrocito de mujeres con preeclampsia con o sin criterios de severidad y los de mujeres sin ésta. MATERIALES Y MÉTODOS: Estudio retrospectivo, de casos y controles, efectuado en mujeres con y sin preeclampsia atendidas entre enero y diciembre de 2019. RESULTADOS: Se estudiaron 70 mujeres con preeclampsia y 70 con embarazo sin esta complicación. El índice neutrófilo-linfocito fue significativamente mayor en las mujeres con preeclampsia (4.11 ± 2.76; IC95%: 3.47-4.75) que en las mujeres sin esta complicación (2.99 ± 1.6; IC95%: 2.62-3.36; p = 0.004), similar a la relación plaquetas-linfocitos (117.61 ± 47.53; IC95%:106.48-128.24 vs 97.64 ± 43.67; IC95%: 87.41-107.87; p = 0.006) y para la distribución de la anchura del eritrocito (14.46 ± 1.9; IC95%: 14.02-14.9 vs 13.56 ± 1.38; IC95%: 13-13.72; p = 0.0002). Ninguno de estos parámetros logró discriminar entre las pacientes con preeclampsia con o sin criterios de severidad. CONCLUSIÓN: Un índice neutrófilo-linfocito ≥ 5.1 y una relación plaquetas-linfocito ≥ 113.1 son capaces de discriminar de manera adecuada entre preeclampsia con o sin criterios de severidad.

Abstract OBJECTIVE: To compare the neutrophil-lymphocyte ratio, platelet-lymphocyte ratio and erythrocyte width distribution, of women with preeclampsia with or without severity criteria and those of women without. MATERIALS AND METHODS: Retrospective case-control study in 70 women with preeclampsia and 70 with normal pregnancy between January and December 2019. RESULTS: Seventy women with preeclampsia and 70 with pregnancy without this complication were studied. The neutrophil-lymphocyte ratio was significantly higher in women with preeclampsia (4.11 ± 2.76; 95%CI: 3.47-4.75), than in women with normal pregnancies (2.99 ± 1.6; 95%CI: 2.62-3.36; p = 0.004); which is similar for the platelet-lymphocyte ratio (117.61 ± 47.53, 95%CI: 106.48-128.24 vs 97.64 ± 43.67; 95%CI: 87.41-107.87; p = 0.006) and for the distribution of the width of the erythrocyte; (14.46 ± 1.9, CI95%: 14.02-14.9 vs 13.56 ± 1.38; CI95%: 13-13.72; p = 0.0002). None of these parameters was able to discriminate between patients with preeclampsia with or without severity criteria. A neutrophil-lymphocyte ratio ≥ 5.1 discriminates between women with a normal pregnancy and those with preeclampsia with or without severity criteria [area under the curve of 0.746, (95%CI: 0.664-0.827)], sensitivity 42%, specificity 91%, positive predictive value 82%, negative predictive value 60% and Odds Ratio 7.1 (95%CI: 2.7-18.6, p = 0.001). The platelet-lymphocyte ratio ≥ 113.4 can discriminate between women with a normal pregnancy and preeclampsia with or without severity criteria, with an area under the curve of 0.617 (95% CI 0.525-0.709). CONCLUSION: A neutrophil-lymphocyte ratio ≥ 5.1, and a platelet-lymphocyte ratio ≥ 113.1 are able to adequately discriminate between patients with normal pregnancy and those with preeclampsia with or without severity criteria.

Comparação das metodologias óptica e de impedância para a determinação de parâmetros hematológicos no analisador Sysmex-XN

Objetivo: Comparar as metodologias de impedância e óptica para contagem de eritrócitos (RBC) e dosagem de hemoglobina (HGB) como forma de corrigir a contagem de eritrócitos em casos de pacientes que apresentam índice de concentração de hemoglobina corpus- cular média (CHCM) acima do ponto de corte. Métodos: Para comparar as metodologias, foram utilizadas amostras com CHCM normal e aumentado. Os parâmetros avaliados foram RBC, HGB e CHCM. Resultados: As amostras com valores de CHCM aumentado demonstraram uma correlação significativa entre RBC e HGB, porém o CHCM apresen- tou uma correlação fraca. Observamos que a metodologia por impedância obteve maior sucesso na correção do CHCM quando as amostras apresentavam aglutinação. Conclusão: A metodologia óptica é uma boa alternativa para correção de valores aumentados de CHCM em situações específicas, mas para casos de aglutinação não se mostrou efetiva.

Objetctive: Compare the impedance and optical methodologies for red blood cells (RBC) count and hemoglobin (HGB) measurement as a way to correct the RBC count in cases of patients who have a MCHC index above the cutoff. Methods: To compare the methodologies, samples with normal and augmented MCHC were used. The parameters evaluated were RBC, HGB and MCHC. Results: Samples with increased MCHC values demonstrated a significant correlation between RBC and HGB, but the MCHC showed a weak correlation. We observed that the impedance methodology was more successful in correcting the MCHC when the samples presented agglutination. Conclusion: Optical methodology is a good alternative for correction of increased CHCM values in specific situations, but for cases of agglutination it has not been shown to be effective.

Evaluación genotóxica de las moléculas con efectos tripanocida y leishmanicida cantin-6-ona y 5-metoxicantin-6-ona en células de médula ósea de ratones / Genotoxic evaluation of the molecules with trypanocidal and leishmanicidal effects, canthin-6-one and 5-methoxycanthin-6-one in mouse bone marrow cells

La tripanosomiasis americana y la leishmaniasis son problemas de salud pública relevantes en Iberoamérica. Las drogas utilizadas actualmente para el tratamiento de estas enfermedades poseen efectos colaterales tóxicos severos. Varios grupos de investigación están abocados a la búsqueda de productos naturales y sintéticos para encontrar nuevos agentes terapéuticos efectivos que no presenten reacciones colaterales adversas. En la evaluación de compuestos de la especie vegetal Zanthoxylum chiloperone (Rutaceae), se demostró que compuestos aislados del extracto presentaban actividad leishmanicida, tripanocida y antifúngica in vivo. Teniendo como antecedentes estos resultados, en el presente estudio se evaluaron los efectos genotóxico y citotóxico del cantín-6-ona y del 5-metoxicantin-6-ona, moléculas aisladas de la planta, en células de médula ósea de animales tratados. El estudio de los efectos genotóxicos se hizo a través del ensayo de modificaciones en la frecuencia de micronúcleos y el efecto citotóxico por modificaciones en la relación entre eritrocitos policromáticos y eritrocitos normocromáticos. Se realizaron 2 ensayos independientes y en cada ensayo los animales fueron divididos en tres grupos de tratamiento: GRUPO I: control negativo que recibió 200 uL de agua y 2.1% de DMSO, vía oral, GRUPO II: compuesto a ser evaluado (canthin-6-ona o 5-methoxicantin-6-ona) con 2.1% de DMSO, y GRUPO III: control positivo que recibió ciclofosfamida 50mg/kg/peso del animal, vía intraperitoneal. El análisis estadístico mostró que ambos compuestos no presentaron efectos genotóxicos ni citotóxicos. Estos resultados permiten proponer a estas moléculas como candidatas a ser sometidas a estudios más detallados como potenciales fármacos contra estas dos enfermedades

American trypanosomiasis and leishmaniasis are relevant public health problems in Latin America. The drugs currently used to treat these diseases have severe toxic side effects. Several research groups are dedicated to the search of natural and synthetic products to find new effective therapeutic agents that do not present adverse collateral reactions. In the evaluation of compounds of the plant species Zanthoxylum chiloperone (Rutaceae), it was shown that isolated compounds of the extract had leishmanicidal, trypanocidal and antifungal in vivo activities. Based on these results, the genotoxic and cytotoxic effects of canthin-6-one and 5-methoxycanthin-6-one, molecules isolated from the plant, on bone marrow cells of treated mice were evaluated in the present study. The study of genotoxic effects was made through the test of modifications in the frequency of micronuclei and the cytotoxic effects by modifications in the relationship between polychromatic erythrocytes and normochromic erythrocytes. Two independent assays were performed and in each assay the animals were divided into three treatment groups: GROUP I: negative control that received 200 µL of water and 2.1% of DMSO, orally, GROUP II: compound to be evaluated (canthin-6 -one or 5-methoxycanthin-6-one) with 2.1% DMSO, and GROUP III: positive control that received cyclophosphamide 50mg /kg animal weight, intraperitoneal. Statistical analysis showed that both compounds had neither genotoxic nor cytotoxic effects. These results allow these molecules to be proposed as candidates to be subjected to more detailed studies as potential drugs against these two diseases

Comparative study with dry crude extracts of Ananas comosus L. Merril using LC50 and erythrocyte osmotic fragility as toxicity parameters / Estudio comparativo con extractos crudos secos de Ananas comosus L. Merril utilizando CL50 y fragilidad osmótica eritrocitaria como parámetros de toxicidad

Ananas comosus, conocido popularmente como piña, tiene una anatomía de planta bien definida y numerosos fitoquí­micos farmacológicamente activos. Algunos de éstos son responsables del potencial antimicrobiano de la especie, que ha sido ampliamente estudiado dada la resistencia bacteriana a los antibióticos actualmente utilizados. Teniendo esto en cuenta, se define que la característica principal de un medicamento es la ausencia de efectos tóxicos, por lo tanto, es necesario buscar datos sobre la toxicidad de A. comosus, colaborando para su posible uso como fármaco. Comparando extractos hidroalcohólicos de las hojas de la corona, cáscara y pulpa de la infrutescencia, fue posible determinar que los tres extractos probados no son tóxicos, siendo el de menor toxicidad para Artemia salina Leach (extracto de hojas con CL50 igual a 994 μg/mL) y a sangre de cordero (extracto de pulpa con porcentaje de hemólisis igual a 0,83%). Los estudios sobre principios y metodologías similares a los utilizados aquí han encontrado resultados comparables que indican la baja toxicidad de la planta. Estos resultados aportan a las investigaciónes que promueven el uso de la piña como agente fitoterapéutico y reafirman su presencia en el Sistema Único de Salud de Brasil.

Ananas comosus, popularly known as pineapple, has well defined plant anatomy and numerous pharmacologically active phytochemicals. Some of these are responsible for the species antimicrobial potential, which has been widely studied given the bac­terial resistance to the currently used antibiotics. Considering this, it is defined that the main characteristic of a drug is the absence of toxic effects, thus, there is a need to seek data regarding the toxicity of A. comosus, collaborating for its possible use as a drug. Comparing hydroalcoholic extracts from the crown leaves, skin and pulp of the infructescence, it was possible to determine that the three tested extracts are non-toxic, being the one with the lowest toxicity to Artemia salina Leach (leaves extract with LC50 equal to 994 μg/mL) and to lamb's blood (pulp extract with percentage of haemolysis equal to 0.83%). Studies concerning principles and methodologies similar to those used here have found comparable results stating the low toxicity of the plant. These results contribute to the research that promotes the use of pineapple as a phytotherapeutic agent and reaffirms its presence in the Brazilian Unified Health System.

Exploring the relationship of peripheral total bilirubin, red blood cell, and hemoglobin with blood pressure during childhood and adolescence / Explorando a relação entre os níveis periféricos de bilirrubina total, glóbulos vermelhos e hemoglobina e a pressão arterial na infância e adolescência

Abstract Objective: Total bilirubin is beneficial for protecting cardiovascular diseases in adults. The authors aimed to investigate the association of total bilirubin, red blood cell, and hemoglobin levels with the prevalence of high blood pressure in children and adolescents. Methods: A total of 3776 students (aged from 6 to 16 years old) were examined using cluster sampling. Pre-high blood pressure and high blood pressure were respectively defined as the point of 90th and 95th percentiles based on the Fourth Report on the Diagnosis, Evaluation, and Treatment of High Blood Pressure in Children and Adolescents. Both systolic and diastolic blood pressure were standardized into z-scores. Results: Peripheral total bilirubin, red blood cell and hemoglobin levels were significantly correlated with age, and also varied with gender. Peripheral total bilirubin was negatively correlated with systolic blood pressure in 6- and 9-year-old boys, whilst positively correlated with diastolic blood pressure in the 12-year-old boys and 13- to 15-year-old girls (p < 0.05). Higher levels of red blood cell and hemoglobin were observed in pre-high blood pressure and high blood pressure students when compared with their normotensive peers (p < 0.01). The increases in red blood cell and hemoglobin were significantly associated with high blood pressure after adjusting for confounding factors. The ORs (95% CI) of each of the increases were 2.44 (1.52-3.92) and 1.04 (1.03-1.06), respectively. No statistical association between total bilirubin and high blood pressure was observed (p > 0.05). Conclusion: Total bilirubin could be weakly correlated with both systolic and diastolic blood pressure, as correlations varied with age and gender in children and adolescents; in turn, the increased levels of red blood cell and hemoglobin are proposed to be positively associated with the prevalence of high blood pressure.

Resumo Objetivo: A bilirrubina total é benéfica para proteger contra doenças cardiovasculares em adultos. Nosso objetivo foi investigar a associação dos níveis de bilirrubina total, glóbulos vermelhos e hemoglobina com a prevalência de pressão arterial elevada em crianças e adolescentes. Métodos: Um total de 3.776 estudantes (com idade entre 6-16 anos) foram examinados utilizando uma amostra em blocos. A pressão arterial elevada anterior e a pressão arterial elevada foram definidas como o 90° e 95° percentil, respectivamente, com base nos critérios do Quarto Relatório sobre Diagnóstico, Avaliação e Tratamento da Pressão Arterial elevada em Crianças e Adolescentes. A pressão arterial sistólica e pressão arterial diastólica foram padronizadas no escore z. Resultados: Os níveis periféricos de bilirrubina total, glóbulos vermelhos e hemoglobina foram significativamente correlacionados à idade, que também variou de acordo com o sexo. A bilirrubina total periférica apresentou uma correlação negativa com a pressão arterial sistólica em meninos com 6 e 9 anos, ao passo que apresentou uma correlação positiva com a pressão arterial diastólica em meninos de 12 anos e meninas de 13 a 15 anos (p < 0,05). Foram observados níveis mais elevados de glóbulos vermelhos e hemoglobina em estudantes com pressão arterial elevada anterior e pressão arterial elevada em comparação a indivíduos normotensos (p < 0,01). Os aumentos de glóbulos vermelhos e hemoglobina tiveram uma associação significativa com a pressão arterial elevada após ajuste dos fatores de confusão. As RC (IC de 95%) de cada um dos aumentos foram 2,44 (1,52-3,92) e 1,04 (1,03-1,06) respectivamente. Não foi observada nenhuma associação estatística entre o nível de bilirrubina total e a pressão arterial elevada (p > 0,05). Conclusão: A bilirrubina total pode ter correlações fracas com a pressão arterial sistólica e a pressão arterial diastólica, variando de acordo com a idade e o sexo em crianças e adolescentes, enquanto isso, propõe-se que o aumento dos níveis de glóbulos vermelhos e hemoglobina está positivamente associado à prevalência de pressão arterial elevada.

Nueva metodología geométrica para evaluar la morfología del eritrocito normal / New physical and mathematical methodology to evaluate the morphology of normal erythrocyte

Resumen Objetivo. Desarrollar una nueva metodología para caracterizar la estructura del eritrocito normal mediante el espacio ocupado por el anillo del eritrocito normal caracterizado con el método de Box Counting. Método. Se analizaron las imágenes de 20 extendidos de sangre periférica, cuyos eritrocitos fueron evaluados por un experto como normales. Se superpusieron dos rejillas Kp de 5 x 5 pixeles y Kg de 10 x 10 pixeles, para calcular el espacio ocupado por dos regiones del eritrocito estos son, el disco y centro de este, visto de manera frontal mediante el método de Box Counting. Resultados. Los espacios ocupados por la región del disco con la rejilla Kp variaron entre 47 y 56, la región del centro del eritrocito, varió entre 9 y 14. La dimensión fractal de estas dos regiones varió entre 0,941 y 1,115 para el disco, entre 0,652 y 1,222 para el centro. Conclusiones. La estructura del eritrocito normal puede ser caracterizada mediante el espacio ocupado por cada una de las regiones del eritrocito a partir de la geometría fractal.

Abstract Objective. Develop a new methodology to characterize the structure of the normal erythrocyte through the space occupied by the ring of the normal erythrocyte characterized by the method of Box Counting. Method. Images of10 peripheral blood smears were analysed, whose erythrocytes were evaluated by an expert as normal. There were superimposed two Kp grids of 5 x 5 pixels and Kg of 10 x 10 pixels, to calculate the space occupied by two regions of the erythrocyte which are, disc and centre of this, seen of way frontal by the method of Box Counting. Results. The spaces occupied by the disc region with grid Kp varied between 47 and 56, the central region of the erythrocyte, varied between 9 and 14. The fractal dimension of these two regions varied between 0,941 and 1,115 for the disc, between 0.652 and 1,222 for the centre. Conclusions. The normal erythrocyte structure can be characterized by the space occupied by the regions erythrocyte from fractal geometry.

Clasificación automática de glóbulos rojos en frotis de sangre periférica / Healthy lifestyles and risk behavior of medical students

Introducción: El diagnóstico del estado eritrocitario en frotis de sangre periférica es un proceso realizado normalmente de forma manual a partir de observación microscópica, lo cual implica una considerable inversión de tiempo y recursos, además de posibles problemas de subjetividad y dificultad en la reproducibilidad del diagnóstico. Objetivo: Desarrollar una aplicación que permita la clasificación automática de glóbulos rojos en frotis de sangre periférica, de utilidad como herramienta de ayuda diagnóstica. Metodología: Se usaron técnicas de procesamiento de imágenes para segmentar los eritrocitos en las fotografías microscópicas y medir en ellos área, perímetro, solidez, circularidad, excentricidad, textura y dimensión box-counting. Se usó una red neuronal artificial para clasificar los eritrocitos según sus características en siete clases, incluyendo normalidad y seis alteraciones patológicas. La red se entrenó de acuerdo con la clasificación de 262 eritrocitos realizada por un hematólogo experto. Los desarrollos se hicieron en matlab®, una poderosa plataforma de computación científica. Resultados: La red escogida alcanza el 97.3% de aciertos en los datos de validación. Las equivocaciones en la red corresponden a células de dudosa clasificación aún para un experto, por presentar características correspondientes a varias clasificaciones patológicas. Conclusiones: La aplicación desarrollada clasifica de manera rápida y acertada los diferentes tipos de glóbulos rojos presentes en una muestra microscópica de frotis de sangre periférica, siendo de utilidad como herramienta de apoyo diagnóstico.

Introduction: The process of erythrocyte classification in peripheral blood smear is normally done manually from microscopic observation. This implies not only a considerable investment of time and resources but also brings potential problems of subjectivity and difficulty in the reproducibility of diagnosis. Objective: To develop an application that allows the automatic classification of red blood cells in peripheral blood smears, as a diagnostic aid tool. Methodology: Image processing techniques were used in order to segment erythrocytes in the microscopic photographs and to measure characteristics as area, perimeter, solidity, circularity, eccentricity, texture and boxcounting dimension. An artificial neural network was used to classify the red blood cells in the images in seven classes, including normal and six pathological changes, according to their characteristics. The network was trained according to the classification of 262 erythrocytes by an expert hematologist. The developments were made in matlab®, a powerful scientific computing platform. Results: The chosen network reaches 97.3% correct in the validation data. Mistakes in the network correspond to cells with various pathological classifications features, which make them difficult to classify even for an expert. Conclusions: The developed application classifies quickly and accurately the different types of red blood cells in a microscopic sample of peripheral blood smear, so it could be useful as a diagnostic support tool.

Caracterización matemática de la unión de la proteína PfNBP-1 al eritrocito mediante la aplicación de la probabilidad y entropía / Mathematical characterization of binding between PfNBP-1 protein and erythrocyite by applying probability and entropy

Objetivo: Estudiar la proteína Plasmodium falciparum Normocyte Binding Protein-1 (PfNBP-1), partiendo de un método de caracterización físico-matemática desarrollado previamente para péptidos de alta unión del merozoito de malaria al eritrocito. Materiales y métodos: Se tomaron 21 péptidos con tamaño de 20 aminoácidos no sobrelapados de los cuales dos son de alta unión, se cuantificó la frecuencia de aparición de los 20 aminoácidos esenciales en cada posición y se calculó la probabilidad, sumatoria de probabilidad y la entropía con el objetivo de diferenciar matemáticamente los péptidos de alta y baja unión. Posteriormente se calcularon los mismos valores para péptidos teóricos análogos, en los que fueron cambiados por glicinas los aminoácidos críticos confirmados experimentalmente. Resultados: Los péptidos de PfNBP-1 comprobados experimentalmente de alta unión, presentaron valores de probabilidad, sumatoria de probabilidad y entropía ubicados dentro del macroestado de unión y sus péptidos teóricos análogos presentaron resultados que se diferenciaban cada vez más del macroestado de unión a medida que se reemplazaban aminoácidos críticos por glicinas. En cuanto a las secuencias de no unión de PfNBP-1, se encontró que los valores calculados son diferentes a los asociados al macroestado de unión, comprobando que en el 100 % de casos estudiados es posible diferenciar los péptidos de no unión y alta unión matemáticamente. Conclusiones: La probabilidad y la entropía permiten caracterizar adecuadamente los péptidos de alta unión de PfNBP-1, y evidenciar el orden matemático subyacente al proceso de unión de proteínas de malaria.

Objective: To study the Plasmodium falciparum Normocyte Binding Protein-1 (PfNBP-1) based on a of physicalmathematical characterization method previously developed for high binding peptides of malaria merozoite to erythrocyte. Materials and methods: 21 non overlapped peptides with size of 20 amino acids, including two of high binding were taken; the frequency of occurrence of the 20 essential amino acids in each position was quantified and probability, summation of probability and entropy were calculated in order to mathematically differentiate high and low binding peptides. Later the same values were calculated for theoretical analogs peptides, where the critical amino acids confirmed experimentally were changed by glycine. Results: The experimentally validated high binding peptides of PfNBP-1 showed values of probability, summation of probability and entropy located within the binding macrostate peptides and their theoretical analogues peptides presented results that differed increasingly of the binding macrostate as critical amino acids were replaced by glycine. For the PfNBP-1 sequences of non-binding, it was found that the calculated values are different from those associated with the macrostate of binding and it was verified that in 100% of studied cases it is possible to mathematically differentiate binding and non-binding peptides. Conclusions: The probability and entropy allow to adequately characterize the high-binding peptides of PfNBP-1 and show the mathematical order underlying the process of protein binding of malaria to the erythrocyte.

Avaliação da genotoxicidade da Ilex paraguariensis (erva mate) pelo teste do micronúcleo / Evaluation of the genotoxicity of Ilex paraguariensis (yerba mate) by micronucleus test

A Ilex paraguariensis é espécie nativa da América do Sul. O consumo de erva mate tem sido associado ao aumento nas taxas de câncer oral, de orofaringe, esôfago e laringe. O objetivo deste estudo foi investigar o potencial genotóxico da exposição a dose única de Ilex paraguariensis através do teste do micronúcleo. Para este estudo, foram utilizados 32 ratos Wistar albinos machos e adultos, divididos em 4 grupos: A - Composto por 8 ratos que receberam infusão de chá preparado na concentração de 5% de erva mate (concentração usualmente encontrada no chá de consumo humano); B - Composto por 8 ratos que receberam chá preparado por imersão em água fria na concentração de 5% de erva mate; C - Composto por 8 ratos, os quais receberam ciclofosfamida em dose única subcutânea (50mg/kg) no primeiro dia do experimento (grupo controle positivo); D - Composto por 8 ratos, os quais receberam somente água (grupo controle negativo). Todos os animais receberam ração ad libitum. Os animais dos grupos A, B e D foram submetidos à eutanásia 48 horas após o início do experimento e os do grupo C, 24 horas após. Foi coletado material da medula óssea de cada rato após a eutanásia para realização do teste do micronúcleo em eritrócito policromático, para avaliação do grau de genotoxicidade. A mediana de micronúcleos para o grupo A (chá mate preparado com infusão) foi de 0,00, do grupo B (chá mate em imersão em água fria) foi de 0,00, do grupo C (ciclofosfamida - controle positivo) foi de 9,00, e no grupo D (controle negativo) foi de 0,00. Não se observou genotoxicidade da Ilex paraguariensis, em ambas as formas de preparo do chá, através do teste de micronúcleo, ao nível de significância de 5%.

The Ilex paraguariensis is a native species of South America. Yerba mate consumption has been associated with increased rates of oral, oropharynx, esophagus and larynx cancer. The aim of this study was to investigate the genotoxic potential of the exposure to a single dose of Ilex paraguariensis by the micronucleus test. For this study, 32 male, adult Wistar rats were used, divided into 4 groups: A - Consisting of 8 rats, which received an infusion of tea prepared at a concentration of 5% of mate (concentration usually found in human consumption of tea); B - Consisting of 8 rats, which received tea prepared immersed in cold water at a concentration of 5% of mate; C - Consisting of 8 rats, which received a single subcutaneous dose of cyclophosphamide (50mg/kg) on the first day of the experiment (positive control group); D - Consisting of 8 rats, which received only water (negative control group). All animals received food ad libitum. The animals in groups A, B and D were sacrificed 48 hours after the beginning of the experiment and group C, 24 hours after. Material from the bone marrow of each rat was collected after euthanasia to perform the micronucleus test in polychromatic erythrocyte to assess the degree of genotoxicity. The median of micronuclei for group A (mate tea prepared with infusion) was 0.00, for group B (mate tea immersed in cold water) was 0.00, for group C (cyclophosphamide - positive control) was 9.00 and for group D (negative control) was 0.00. No genotoxicity of Ilex paraguariensis was observed in both tea preparation methods by the micronucleus test at a significance level of 5%.

Daño oxidativo en un modelo experimental de hiperglicemia e hiperlipidemia inducida por sacarosa en ratas wistar / Oxidative damage in an experimental model of saccharose-induced hyperglycemia and hyperlipidemia in Wistar rats

Se realizó un estudio experimental de casos y controles en el período comprendido entre noviembre de 2010 y mayo de 2011, con el objetivo de caracterizar el comportamiento del daño oxidativo en un modelo experimental de hiperglicemia e hiperlipidemia inducida por sacarosa en ratas wistar en la Unidad de Toxicología Experimental de la Universidad de Ciencias Médicas de Villa Clara. Para ello se utilizaron ratas machos como modelo biológico, con el fin de evaluar la actividad oxidativa inducida por sacarosa. El modelo fue establecido a partir de la administración de una dieta rica en sacarosa, para lo cual se determinaron los por cientos de hemólisis, fotohemólisis y la susceptibilidad a sustancias con efectos hemolíticos. Se encontró aumento en los valores de glucemia y triacilglicéridos de las ratas del grupo estudio, con diferencias significativas con respecto al grupo control. Asimismo, se obtuvieron diferencias significativas entre ambos grupos estudiados en los test de susceptibilidad a la hemólisis y la fotohemólisis. Se produjo incremento significativo en los niveles de glicemia y triacilglicéridos relacionados con la dieta rica en sacarosa. Se observaron incrementos significativos en los valores de los por cientos de hemólisis en el ensayo de susceptibilidad a la hemólisis y la fotohemólisis y en el primer caso se correspondió con el grupo sometido a la dieta rica en sacarosa. Se observó una fuerte correlación entre los niveles de triglicéridos del grupo estudio en el último mes de experimentación y los valores obtenidos en el test de fotohemólisis.

We carried out an experimental research of cases and controls in the period from November 2010 to May 2011, with the objective of characterizing the behavior of the oxidative damage in an experimental model of saccharose-induced hyperglycemia and hyperlipidemia in Wistar rats in the Unit of Experimental Toxicology of the Medical Sciences University of Villa Clara. We used male rats as biological model, with the objective of evaluating the saccharose- induced oxidative activity. The model was set up on the basis of a diet rich in saccharose, and for that there were calculated the hemolysis, photohemolysis percentages and the susceptibility to substances with hemolytic effects. We found an increase in the glycemia and triacilglyceride values of the rats in the studied group, with significant differences in those of the control group. There were also found significant differences between the both studied groups in the tests of susceptibility to the hemolysis and photohemolysis. There it was a significant increase in the glycemia and triacilglycerides levels related with the saccharose-enriched diet. We observed significant increases in the percentages values of the hemolysis in the tests of susceptibility to the hemolysis and photohemolysis, and in the first case it corresponded with the group receiving a saccharose- enriched diet. There it was a strong correlation between the triglycerides level of the studied group in the last month of the research and the values obtained in the photohemolysis test.

Deficiencia de glucosa 6 fosfato deshidrogenasa: analisis enzimatico y molecular en una población de Bogota / Glucose-6-phosphate dehydrogenase deficiency: enzimatic and molecular analysis in a Bogotß population

Objetivo: Determinar qué tan frecuente es la deficiencia de glucosa 6 fosfato deshidrogenasa (G6PD) y realizar analisis molecular para identificar las variantes A+, A- y mediterrßnea en una población de residentes en Bogota. Métodos: Se analizaron 348 personas que residen en Bogotß, pertenecientes a la Policía Nacional y a la Universidad del Rosario. La actividad enzimßtica se determinó en muestras de sangre mediante espectrofotometría con el kit Trinity Biotech (Cat 345-B). Los valores de hemoglobina (Hg) y hematócrito (Hto) se determinaron con el método de Drabkin y sedimentación, respectivamente. La determinación de las variantes moleculares se realizó mediante amplificación por reacción en cadena de la polimerasa (PCR) y analisis de fragmentos de restricción de longitud polimórfica (RFLP) con las enzimas NlaIII, Fok I y MboII para A+, A- y mediterrßnea, respectivamente. Se hicieron analisis estadísticos para comparar la concentración de Hg en personas sanas y deficientes, la actividad de G6PD por géneros y los datos de frecuencia a nivel mundial. Resultados y conclusiones: La frecuencia de deficiencia de G6PD para la población en estudio fue 3.1%. En 1.4% de los casos se observó actividad deficiente, en 1.7% actividad intermedia y en 0.6% actividad aumentada. No se encontraron las variantes moleculares A+, A- y mediterranea en ningún afectado. La actividad de G6PD no tuvo diferencias por género. Se encontró diferencia significativa en el valor de hemoglobina entre las personas sanas y deficientes de G6PD. Los individuos deficientes eran asintomßticos lo que indica mecanismos de compensación de estrés oxidativo. Las mujeres deficientes son heterocigotos con una inactivación preferencial del cromosoma X anormal y al ser portadoras tienen riesgo de 50% de tener hijos afectados con la enfermedad. La identificación de mujeres y hombres deficientes permite establecer medidas preventivas ante posibles crisis hemolíticas desencadenadas por infecciones y drogas.

Objective: To determine the frequency of G-6PD and molecular analysis for identification of A+, A- and Mediterranean in healthy persons in Bogota. Methods: Quantitative spectrophotometric assays for enzyme activity of G-6PD were carried out on the red cells of 348 asymptomatic and healthy adult males and females. Through molecular analysis of DNA from G-6PD deficients the relevant exons were amplified for PCR and then analysed with the restriction enzymes NlaIII, Fok I and MboII, for the detection of A+, A- and Mediterranean variants. Results and conclusions: Among 348 samples, 1.4% exhibited total deficiency and 1.7% had intermediate deficiency while 96.3% were normal. The combined prevalence was 3.7%. In enzymatic activity no statistically significance was seen between males and females. No variant was found among these patients and any of the subjects studied displayed any sign of hemolysis and other clinical manifestations. Although it is not yet clearly understood other mechanisms must exist to offer protection from the oxidative stresses. The finding of severe enzyme deficiency in some heterozygote females is due to extreme degree of X inactivation of the normal chromosome.

Trypanosoma vivax: Bometría y eritrometría en infecciones experimentales

La anemia y la poiquilocitosis son manifestaciones clínicas características de la tripanosomosis por Trypanosoma vivax. No obstante, poco se sabe acerca de la multicausalidad de las alteraciones de forma y tamaño que experimentan los eritrocitos durante la infección. La parasitemia, las dimensiones tripomastigóticas y el área eritrocítica fueron registradas a lo largo de las infecciones. El análisis estadístico reveló cambios significativos en las variables consideradas. El estudio demostró relación entre las dimensiones tripomastigóticas y el área eritrocítica. Además, probó que el patrón de cambio morfométrico experimentado por tripomastigotes de aislados diferentes en distintos hospedadores, pudiera ser un evento hospedador-dependiente.

Anemia and poikilocytosis are clinical manifestations characteristic to tripanosomosis by Trypanosoma vivax. Nevertheless, little is known about the multicausality of the form and size alterations that experience the erythrocytes during the infection. The parasitemia, the trypomastigotic dimensions and the erythrocytic area were registered throughout the infections. The statistical analysis reveals significant changes in the considered variables. The study demonstrated relationship between the trypomastigotic dimensions and the erythrocytic area. In addition, it proved that the pattern of trypomastigote morphometric change of different isolates experienced in different hosts could be a host-dependent event.

Erythrocyte membrane, plasma and atherosclerotic plaque lipid pattern in coronary heart disease / Perfil lipídico de membrana de eritrocito, plasma y placa ateromatosa en la enfermedad coronaria

The objective was to analyze the lipid composition of the atherosclerotic plaque (AP), plasma and erythrocyte membrane (EM) in patients with advanced coronary heart disease (CHD). AP were obtained through endarterectomy in 18 patients. Ten normolipemic healthy subjects were selected to obtain the normal lipid pattern profile. Total lipids of AP and EM were determined by HPTLC, and the fatty acid profile from AP, EM and plasma using TLC-FID. The relative amount of the lipid species analyzed in AP was in line with the data in the literature [phospholipids: 23.5 mol% more or less 3.5; total cholesterol 68.9 mol% more or less 7.9; triglyceride 7.6 mol% more or less 3.4]. Plasma and EM from CHD patients compared to controls, showed a decrease in polyunsaturated fatty acids and an increase in saturated fatty acids leading to a decrease in the unsaturation index (plasma: 1.67 more or less 0.06 vs. 1.28 more or less 0.03, P less than 0.05; EM: 2.28 more or less 0.04 vs. 1.25 more or less 0.010, P less than 0.05) and an enhancement in the saturated/unsaturated ratio (plasma: 0.35 more or less 0.02 vs. 0.52 more or less 0.02, P less than 0.05; EM: 0.45 more or less 0.01 vs. 0.83 more or less 0.04, P less than 0.05). These data are consistent with an essential fatty acid deficiency. Total cholesterol was increased in the CHD's EM (32.3 more or less 0.8 vs. 40.6 more or less 2.5, P less than 0.05) with a decrease in phospholipid percentage (67.7 more or less 0.7 vs. 59.4 more or less 2.6, P less than 0.05) indicating an alteration in membrane fluidity. These findings suggest changes in EM lipids in CHD patients in spite of different pathological conditions such as age, smoking status and diabetes. The analysis of the lipid composition of EM could provide a useful tool to monitor the evolution of the CHD.

El objetivo fue analizar la composición lipídica de las membranas de eritrocitos (ME), plasma y placas ateromatosas (PA) en pacientes con enfermedad coronaria avanzada (ECV). Las PA fueron obtenidas de endarterectomías coronarias de 18 pacientes. Fueron seleccionados 10 sujetos sanos, normolipémicos, como grupo control. Los lípidos totales de PA y ME se determinaron utilizando HPTLC, y el perfil de ácidos grasos de las PA, ME y plasma mediante TLC-FID. La cantidad relativa de las especies lipídicas obtenidas de las PA coinciden con la literatura [fosfolípidos 23.5 mol% más o menos 3.5; colesterol total 68.9 mol% más o menos 7.9; triglicéridos 7.6 mol% más o menos 3.4]. En el plasma y en las ME de los pacientes con ECV se observó, comparando con los pacientes controles, una disminución de los ácidos grasos poli-no saturados acompañado de un aumento de los ácidos grasos saturados que provocó el descenso del índice de instauración (plasma: 1.67 más o menos 0.06 vs. 1.28 más o menos 0.03, P menor que 0.05; ME: 2.28 más o menos 0.04 vs. 1.25 más o menos 0.010, P menor que 0.05) y el incremento del cociente AG saturados/insaturados (plasma: 0.35 más o menso 0.02 vs. 0.52 más o menos 0.02, P menor que 0.05; ME: 0.45 más o menos 0.01 vs. 0.83 más o menos 0.04, P menor que 0.05). Estos datos indicarían una deficiencia de ácidos grasos esenciales. Se observó una elevación en los valores de colesterol total (32.3 más o menos 0.8 vs. 40.6 más o menos 2.5, P menor que 0.05) y una disminución de los valores de fosfolípidos (67.7 más o menos 0.7 vs. 59.4 más o menos 2.6, P menor que 0.05) en las ME de los pacientes con ECV. Estos hallazgos sugieren cambios en los lípidos de las ME en los pacientes con ECV a pesar de presentar diferencias con respecto a edad, tabaquismo y diabetes. El conocimiento del perfil lipídico de las ME podría constituirse en una herramienta útil para monitorear la evolución de la enfermedad.

La senescencia del Eritrocito humano y su relación con el trasporte de calcio y potasio / The senesces of human erythrocyte ant its relation with Calcium and Potassium transport

Los eritrocitos humanos poseen una vida media de 120 días, y es ampliamente aceptado que su concentración de Ca2+ aumenta con el proceso de la senescencia. El control de este mecanismo es desconocido en estas células no nucleadas. Así, la investigación en busca del reloj biológico de estas células tiene más de cuarenta años y se ha propuesto que los cambios cíclicos asociados al paso por el lecho capilar son buenos candidatos, en especial el estrés mecánico que se genera sobre sus membranas. En este ensayo se rediscuten antiguos datos de otros investigadores y resultados de nuestro laboratorio al respecto de dos nuevos mecanismos de transporte, que permiten una reinterpretación de la hipótesis del reloj biológico. Así, proponemos una nueva hipótesis en la que se plantea que es el potasio y no el calcio y sus permeabilidades, el factor principal que guía el proceso de senescencia.

Human erythrocyte has a mean life time of 120 days, and it is generally accepted that Ca2+ concentration rice withthe senesces process. The control of this mechanism it is not known in these non nucleated cells. The search for a mechanism beneath the biological clock of erythrocytes has more than forty years. It has been proposed that cyclic changes because of the pass thru the capillary can be involved, specially the mechanical stress on the membrane. Here we revisited data from other researchers and from our laboratory regarding two novels transport mechanisms, which permits a reinterpretation of the biological clock. We propose a novel hypothesis in which is potassium instead of calcium and there permeability, the one who leads the senescence process.

Polimorfismo del gen de la banda 3 eritrocítica en grupos étnicos de Costa Rica / Polymorphism of the erythrocyte band 3 gene (EPB3) in ethnic groups of Costa Rica

Band 3 (AE1) is one of the most abundant proteins in the membrane of the human erythrocyte. This protein works as an anionic CI - and HCO3- exchanger and it also functions as an anchor for several proteins of the erythrocyte's cytoesqueleton. Several mutations have been described and many polymorphic variants have been associated to hereditary spherocytosis. The identification of a genetic marker at position 5' of the AEl gene could be associated to several molecular defects of the erythrocyte. This genetic marker is a restriction fragment length polymorphism (RFLP) produced by restriction enzime Pst I. For this polymorphism a total of 216 individuals belonging to seven different populations were analyzed: one from the Central Valley, two African descendants (Lim6n and Guanacaste) and four Amerindians (Bribri, Cabecar, Maleku and Guaymi). The most frequent allele in the Amerindian population was no 1. No significant differences were found with respect to the Hardy-Weinberg equilibrium in six of the populations, although the Guaymi group does present significative differences.