RESUMEN
Objective The present study compares immune and endocrine parameters between HIV-infected patients who underwent the Immune Reconstitution Inflammatory Syndrome (IRIS-P) during antiretroviral therapy (ART) and HIV-patients who did not undergo the syndrome (non-IRIS-P). Materials and methods Blood samples were obtained from 31 HIV-infected patients (15 IRIS-P and 16 non-IRIS-P) before ART (BT) and 48 ± 2 weeks after treatment initiation (AT). Plasma Interleukin-6 (IL-6) and Interleukin-18 (IL-18) were determined by ELISA. Cortisol, dehydroepiandrosterone sulfate (DHEA-S) and thyroxin concentrations were measured using chemiluminescence immune methods. Results Concentrations of IL-6 (7.9 ± 1.9 pg/mL) and IL-18 (951.5 ± 233.0 pg/mL) were significantly higher (p < 0.05) in IRIS-P than in non-IRIS-P (3.9 ± 1.0 pg/mL and 461.0 ± 84.4 pg/mL, respectively) BT. Mean T4 plasma level significantly decreased in both groups of patients after treatment (p < 0.05). In both groups cortisol levels were similar before and after ART (p > 0.05). Levels of DHEA-S in IRIS-P decreased AT (1080.5 ± 124.2 vs. 782.5 ± 123.8 ng/mL, p < 0.05) and they were significantly lower than in non-IRIS-P (782.5 ± 123.8 vs. 1203.7 ± 144.0 ng/mL, p < 0.05). IRIS-P showed higher values of IL-6 and IL-18 BT and lower levels of DHEA-S AT than in non-IRIS-P. Conclusion These parameters could contribute to differentiate IRIS-P from non-IRIS-P. The significant decrease in DHEA-S levels in IRIS-P after ART might suggest a different adrenal response in these patients, which may reflect the severity of the disease.
Asunto(s)
Terapia Antirretroviral Altamente Activa/efectos adversos , Biomarcadores/sangre , Infecciones por VIH/sangre , Síndrome Inflamatorio de Reconstitución Inmune/sangre , Relación CD4-CD8 , Sulfato de Deshidroepiandrosterona/sangre , Ensayo de Inmunoadsorción Enzimática , Femenino , Infecciones por VIH/tratamiento farmacológico , Infecciones por VIH/inmunología , Infecciones por VIH/metabolismo , Humanos , Hidrocortisona/sangre , Síndrome Inflamatorio de Reconstitución Inmune/inmunología , Síndrome Inflamatorio de Reconstitución Inmune/metabolismo , Interleucina-18/sangre , Interleucina-6/sangre , Luminiscencia , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Tiroxina/sangre , Carga ViralRESUMEN
El objetivo del trabajo fue estudiar la desialización de eritrocitos que desenmascaran el antígeno T y de los glóbulos que no lo exponen, por contacto con larvas recién nacidas (LRN) de T. spiralis. Se trabajó con 15 suspensiones eritrocitarias en medio enzimático, incubadas con LRN. Los GR control se incubaron con solución salina. Para analizar el efecto del parásito sobre la exposición del antígeno T, se aplicó el Método de Aglutinación anti-T con antígeno T y se estudió la agregación por los métodos de Polibrene y de Análisis Digital de Imágenes, determinando CexpST, CCA y Distribución de los agregados eritrocitarios (DAE). La media de los coeficientes en los eritrocitos que expusieron el antígeno T fueron CexpST=0,22±0,179 y coeficiente de células aisladas (CCA)=0,73±0,108 y en los que no 0,86±0,125 y 0,205±0,163 respectivamente. La DAE mostró que los GR que lo expusieron, presentaron marcada disminución de células aisladas y pronunciado aumento de grandes agregados en relación a los controles, mientras que los GR en los que no hubo exposición, la disminución de células aisladas con respecto a los controles fue menor y el aumento de agregados estuvo homogéneamente distribuido entre todas las categorías. La experiencia realizada concluye que los valores obtenidos de los coeficientes difieren significativamente en los GR que exponen el antígeno T y en los que no, por lo que estas técnicas podrían ser predictivas para detectar desenmascaramiento T.
The aim of this work was to study the desialylation of erythrocytes that expose the T antigen by contact with T. spiralis newborn larvae (NL) and of those that do not. Work was carried out with 15 red cell suspensions in enzymatic medium, which were incubated with NL. The respective Control RBC were incubated with saline solution. To analyze the effect of the parasite on the exposure of the T antigen, the Anti T- antigen T Agglutination Method was applied and the aggregation was studied by Polybrene and Digital Image Analysis Methods determining CexpTS, ICC and Distribution of Erythrocyte Aggregates (DEA). The mean of coefficients in the erythrocytes that exposed the T antigen were CexpTS= 0.22 ± 0.179 and ICC= 0.73 ± 0.108 and of those that did not it was 0.86±0.125 y 0.205±0,163 respectively. DEA showed that the RBCs that exposed it showed a marked decrease of isolated cells and a pronounced increase of large aggregates in relation to the Controls, while the RBCs in which there was no exposure, the decrease of isolated cells with respect to Controls was lower and the increase of aggregates was homogeneously distributed among all categories. The experience concludes that the obtained values of the coefficients differ significantly in the RBCs that expose the T antigen from those that do not, so these techniques could be predictive to detect T unmasking.
O objetivo do trabalho foi estudar a desialização de eritrócitos que desmarcaram o antígeno T e dos glóbulos que não o expõem, por contato com LRN de T. spiralis. Trabalhou-se com 15 suspensões de eritrócitos em meio enzimático, incubadas com LRN. Os GV controle foram incubados com solução salina. Para analisar o efeito do parasita sobre a exposição do antígeno T, foi aplicado o Método de Aglutinação anti T com antígeno T e se estudou a agregação pelos Métodos de Polibrene e de Análise Digital de Imagens, determinando CexpST, CCA e Distribuição dos agregados eritrocitários (DAE). A média dos coeficientes nos eritrócitos que expuseram o antígeno T foram CexpST= 0.22±0.179 e CCA= 0.73±0.108 e naqueles que não o expuseram 0.86±0.125 y 0.205±0,163 respectivamente. Foi demonstrado que os GV que o expuseram, apresentaram marcada diminuição de células isoladas e pronunciado aumento de grandes agregados em relação aos controles, enquanto que os GV nos quais não houve exposição, a diminuição de células isoladas com respeito aos controles foi menor e o aumento de agregados esteve homogeneamente distribuído entre todas as categorias. A experiência realizada conclui que os valores obtidos dos coeficientes diferem significativamente nos GV que expõem o antígeno T e nos que não, portanto estas técnicas poderiam ser preditivas para detectar desmascaramento T.
Asunto(s)
Eritrocitos , Trichinella spiralis , Interpretación Estadística de DatosRESUMEN
ABSTRACT Objective The present study compares immune and endocrine parameters between HIV-infected patients who underwent the Immune Reconstitution Inflammatory Syndrome (IRIS-P) during antiretroviral therapy (ART) and HIV-patients who did not undergo the syndrome (non-IRIS-P). Materials and methods Blood samples were obtained from 31 HIV-infected patients (15 IRIS-P and 16 non-IRIS-P) before ART (BT) and 48 ± 2 weeks after treatment initiation (AT). Plasma Interleukin-6 (IL-6) and Interleukin-18 (IL-18) were determined by ELISA. Cortisol, dehydroepiandrosterone sulfate (DHEA-S) and thyroxin concentrations were measured using chemiluminescence immune methods. Results Concentrations of IL-6 (7.9 ± 1.9 pg/mL) and IL-18 (951.5 ± 233.0 pg/mL) were significantly higher (p < 0.05) in IRIS-P than in non-IRIS-P (3.9 ± 1.0 pg/mL and 461.0 ± 84.4 pg/mL, respectively) BT. Mean T4 plasma level significantly decreased in both groups of patients after treatment (p < 0.05). In both groups cortisol levels were similar before and after ART (p > 0.05). Levels of DHEA-S in IRIS-P decreased AT (1080.5 ± 124.2 vs. 782.5 ± 123.8 ng/mL, p < 0.05) and they were significantly lower than in non-IRIS-P (782.5 ± 123.8 vs. 1203.7 ± 144.0 ng/mL, p < 0.05). IRIS-P showed higher values of IL-6 and IL-18 BT and lower levels of DHEA-S AT than in non-IRIS-P. Conclusion These parameters could contribute to differentiate IRIS-P from non-IRIS-P. The significant decrease in DHEA-S levels in IRIS-P after ART might suggest a different adrenal response in these patients, which may reflect the severity of the disease.
Asunto(s)
Humanos , Masculino , Femenino , Persona de Mediana Edad , Biomarcadores/sangre , Infecciones por VIH/sangre , Terapia Antirretroviral Altamente Activa/efectos adversos , Síndrome Inflamatorio de Reconstitución Inmune/sangre , Tiroxina/sangre , Ensayo de Inmunoadsorción Enzimática , Hidrocortisona/sangre , Infecciones por VIH/inmunología , Infecciones por VIH/metabolismo , Infecciones por VIH/tratamiento farmacológico , Estudios Prospectivos , Interleucina-6/sangre , Relación CD4-CD8 , Sulfato de Deshidroepiandrosterona/sangre , Carga Viral , Interleucina-18/sangre , Luminiscencia , Síndrome Inflamatorio de Reconstitución Inmune/inmunología , Síndrome Inflamatorio de Reconstitución Inmune/metabolismoRESUMEN
Oral rehydration salt (ORS) treatment in young children with acute diarrhoea (AD) has contributed to decrease mortality associated with dehydration although effective strategies to reduce morbidity associated with this disease are required. The aim of this study was to evaluate the diarrhoea duration when using combined colloidal bismuth hydroxide gel (CBHG) and oral rehydration salt treatment compared with ORS therapy in children with AD. We designed a double-blind, randomised prospective study with treatment and control groups. Patients aged one to 12 years, with no prior pathology and with AD of less than 48 h were included. The Chi-squared and Mann-Whitney tests were used, as well as the Cox proportional hazards model and the Kaplan-Meier estimator. Patients were randomised into an ORS and CBHG treatment group and a control group for ORS plus placebo. (Average age: 3.2 years). The result of the post-treatment evaluation with respect to the average duration of AD was 25.5 h for the treated group vs. 41.5 h for the control group (p = 0.015). The average number of stools was 4.8 in the treated group and 8.2 in the control group (p = 0.032). We conclude that the use of CBHG plus ORS significantly reduced the duration of AD, the number of stools and the percentage of children with persistent AD after 24 h of treatment compared to the control group. AD remitted almost twice as fast in patients treated with CBHG and ORS compared to those who received ORS plus placebo.
RESUMEN
Trichinella spiralis es agente causal de una zoonosis endémica en Argentina. El objetivo fue estudiar la desialización eritrocitaria producida por larvas musculares (LM) de T. spiralis. Se trabajó con concentrados de LM, incubados en partes iguales con glóbulos rojos (GR) Grupo O (37 °C) durante 3 horas (con y sin agitación controlada) durante 150 minutos, a intervalos de 30 minutos, para estudiar el curso de la desialización en el tiempo. Los GR controles fueron incubados de la misma manera, con igual volumen de solución fisiológica. Se aplicó el método de titulación de la agregación, calculando título y coeficiente de puntuación total (CexpST). Se encontró que los eritrocitos incubados con LM presentaron mayor agregación que los controles. El valor medio de CexpST con agitación (0,43) fue significativamente menor que cuando los GR no se agitaron (0,72). El estudio de la desialización eritrocitaria en el tiempo mostró que el título de los GR control disminuyó significativamente a los 90 minutos en 5/10 repeticiones y a los 150 minutos en 9/10. El aumento del tiempo de incubación produjo el incremento de la desialización excepto a los 120 y 150 minutos donde no existieron diferencias significativas en el valor de CexpST. La experiencia realizada in vitro, sugeriría que en la infección in vivo, las LM podrían captar el ácido siálico a partir de los residuos sializados presentes en la célula muscular.
Trichinella spiralis is the cause of an endemic zoonosis in Argentina. The objective was to study the erythrocyte desialylation by T. spiralis muscle larvae (ML) applying an aggregation titulation method. We worked with ML concentrates, which were incubated with an equal volume of O Group erythrocytes (RBCs) at 37° C for 3 hours, (with and without controlled agitation) and for 150 minutes, taking samples at 30 minutes intervals to study the course in time of the desialylation. RBCs control were incubated with an equal volume of physiological saline solution. Aggregation titulation method was applied and the title and total score coefficient (TSexpC) were calculated. The results showed that erythrocytes incubated with ML showed more aggregation than controls. The average TSexpC with agitation (0.43) was significantly lower than when erythrocytes were not stirred (0.72). The course in time of the erythrocyte desialylation showed that the RBCs contol title decreased significantly at 90 minutes in 5/10 repetitions and at 150 minutes in 9/10. Increasing the incubation time produced an increase in erythrocyte desialylation, except at the 120 and 150 minutes interval where no significant differences in TSexpC values were found. The in vitro experience would suggest that in cases of in vivo infection, ML could capture sialic acid from sialylate residues present in the muscle cell.
RESUMEN
El objetivo del trabajo fue estudiar las alteraciones en la agregación eritrocitaria producidas por recién nacidas (LRN) de T. spiralis. Se usaron concentrados de larvas LRN incubados en partes iguales con glóbulos rojos (GR) Grupo O (GR Tratados) durante 2 horas, con y sin agitación controlada, tomando muestras al tiempo inicial, 60 y 120 minutos. Los Controles fueron incubados con igual volumen de solución salina. Se aplicó Análisis Digital de Imágenes para estudiar la distribución de los agregados eritrocitarios y calcular el valor de coeficiente de células aisladas (CCA) y la Técnica de Titulación de la Agregación para determinar el Título y el CexpST. Se utilizó ANOVA bifactorial para analizar el efecto de la agitación y del tiempo de incubación en los valores de CCA. Los resultados mostraron que el aumento del tiempo de tratamiento produjo la disminución de las células aisladas y los pequeños rouleaux, y el aumento de los agregados formados por 5 o más glóbulos, lo cual incrementó significativamente el valor de CCA. El análisis estadístico determinó que la agregación de los GR Tratados a los 60 minutos fue mayor que al tiempo 0, y a los 120 minutos mayor que a los otros dos tiempos. La Titulación de la agregación mostró la disminución del CexpST y del Título de los GR Tratados. Las metodologías empleadas no mostraron diferencias significativas en tratamientos con y sin agitación. Se concluye que la disminución de carga eritrocitaria producida por LRN podría provocar alteraciones hemorreológicas en el hospedador.
The aim of this paper was to study the alterations in the erythrocyte aggregation produced by newborn larvae (NL) of T. spiralis. Work was performed with NL concentrates, which were incubated with an equal volume of O Group RBC (Treated RBC) for 2 hours, with and without controlled agitation, taking samples at the initial time, at 60 and 120 minutes. RBC Controls were incubated with equal volume of saline solution. Digital Image Analysis was applied to study the distribution of erythrocyte aggregates and to calculate ICC values, and the Aggregation Titration Technique was used to determine the Title and TSexpC. Two-factor ANOVA was used to analyze the effect of agitation and incubation time on the ICC values. The results showed that at higher treatment time, there was a decrease of isolated cells and small rouleaux and an increase of the aggregates formed by 5 or more cells, with a significant increase of ICC values. Statistical analysis determined that Treated RBC aggregation at 60 minutes was higher than at initial time and that at 120 minutes it was higher than the other two times. Aggregation Titration showed a decrease in the Treated RBC Title and TSexpC. The methodologies employed showed no significant differences in treatments with and without agitation. It is concluded that the decrease in erythrocyte charge produced by NL could cause hemorrheologic alterations in the host.
O objetivo foi estudar as alterações na agregação de eritrócitos produzidas por larvas recém-nascidas (LRN) da T. spiralis. O trabalho foi feito com concentrados de LRN incubados em partes iguais com glóbulos vermelhos (GV) Grupo O (GV Tratados), durante 2 horas, com e sem agitação controlada, levando as amostras ao tempo inicial, 60 e 120 minutos. Os controles foram incubados com igual volume de solução salina. Análise Digital de Imagens foi aplicada para estudar a distribuição dos agregados de eritrócitos e calcular o valor do Quoficiente de Células Isoladas (QCI) e a Técnica de Titulação da Agregação para determinar o Título e CexpST. Para analisar o efeito da agitação e do tempo de incubação sobre os valores do QCI foi utilizada ANOVA bifatorial. Os resultados mostraram que o aumento do tempo de tratamento produziu a diminuição das células isoladas e os pequenos rouleaux, e o aumento dos agregados formados por 5 ou mais glóbulos, aumentando o valor de QCI significativamente. A análise estatística determinou que a agregação de GV Tratados aos 60 minutos foi maior que no tempo inicial e, aos 120 minutos era maior que durante os outros dois tempos. A Titulação da Agregação mostrou a diminuição do CexpST e do Título de GV Tratados. As metodologias usadas não mostraram diferenças significantes em tratamentos com e sem agitação. Conclui-se que a diminuição de carga de eritrócitos produzida por LRN poderia resultar em alterações hemorreológicas no hospedeiro.
Asunto(s)
Agregación Eritrocitaria , Trichinella spiralis , Alergia e InmunologíaRESUMEN
Experiencias previas comunicaron que los eritrocitos (GR) incubados con larvas recién nacidas (LRN) y larvas musculares (LM) de T. spiralis presentan menor contenido de ácido siálico que los correspondientes GR Controles. El objetivo del trabajo fue estudiar el efecto de LRN y LM sobre la desialización aplicando el Método de Polibrene. Se trabajó con concentrados de LRN y LM incubados en partes iguales con GR Grupo O, en medio salino y enzimático, durante 120 min a intervalos de 30 min. Los Controles fueron incubados con igual volumen de solución salina. Se aplicó el Método de Polibrene y se calculó el valor del coeficiente CexpCASP. Para analizar el efecto del parásito, medio de incubación y tiempo de tratamiento en los valores de CexpCASP, se utilizó un ANOVA multifactorial. Los resultados mostraron que el medio enzimático y el aumento del tiempo de incubación producen la mayor disminución de carga globular, reflejada en menores valores de CexpCASP, sin diferencias significativas con el estadio larval utilizado. La experiencia sugeriría que durante su migración por el torrente circulatorio las LRN podrían captar ácido siálico eritrocitario y alterar el comportamiento hemorreológico, así como también permitiría especular que ambos estadios larvales podrían secuestrarlo de células del hospedador para interferir y/o evadir su respuesta inmune...
Asunto(s)
Humanos , Trichinella spiralis , Dracunculus , Eritrocitos , LarvaRESUMEN
Se ha comunicado que los eritrocitos (GR) incubados con larvas infectantes de Trichinella spiralis presentan mayor agregación que los GR Controles incubados con solución salina, lo cual indica que el parásito capta el ácido siálico globular. El objetivo fue estudiar la cinética de captación de ácido siálico por larvas musculares de T. spiralis aplicando Análisis Digital de Imágenes. Se trabajó con concentrados larvales de T. spiralis incubados en partes iguales con GR Grupo O, en medio salino y enzimático (GR Tratados), durante 120 minutos a intervalos de 15 minutos. Los Controles fueron incubados con igual volumen de solución salina. Se aplicó Análisis Digital de Imágenes y se calculó el valor del Coeficiente de células aisladas (CCA). Para analizar el efecto del tiempo de incubación en los valores de CCA, se utilizó un análisis de la variancia para un diseño en bloques completos aleatorizados. Los resultados mostraron que el valor promedio de CCA, en ambos medios, varió significativamente con el tiempo de incubación, lo que evidencia que el aumento del contacto de las larvas con los GR produce una disminución del ácido siálico que se refleja en mayores valores de CCA. La experiencia realizada sugeriría que durante su permanencia y viabilidad en el quiste, las larvas de T. spiralis podrían ir captando ácido siálico de la célula muscular con el objeto de interferir y/o evadir la respuesta inmune del hospedador.(AU)
It has been reported that erythrocytes (RBC) which were incubated with infective larvae of T. spiralis exhibit higher aggregation than RBC Controls incubated with saline solution, indicating that the parasite captures erythrocyte sialic acid. The objective of this work was to study the kinetics of sialic acid capture by muscle larvae of T. spiralis using Digital Image Analysis. The work was performed with larvae concentrates of T. spiralis, which were incubated with an equal volume of O Group RBC in saline and enzymatic mediums (Treated RBC), for 120 minutes at 15 minute intervals. Digital Image Analysis was applied and the value of Isolated Cells Coefficient (ICC) was calculated. To analyze the effect of incubation time on the ICC values, an analysis of variance was used to design randomized complete blocks. The results showed that the average value of ICC, both saline and enzymatic mediums, varied significantly with incubation time. It was shown that the increase in the contact of the larvae with erythrocytes produces the decrease of globular sialic acid, which is reflected in higher ICC values. The experience would suggest that during their stay and viability in the trichina cysts, the larvae could be capturing muscle sialic acid in order to interfere and/or evade the host immune response.(AU)
Foi relatado que os eritrócitos (GV) incubados com larvas infectantes da Trichinella spiralis apresentam maior agregaþÒo que os GV Controles incubados com soro fisiológico, indicando que o parasita capta o ácido siálico globular. O objetivo do trabalho foi estudar a cinética de captaþÒo de ácido siálico por larvas musculares da T. spiralis por Análise Digital de Imagens. O trabalho foi feito com concentrados de larvas de T. spiralis incubados em partes iguais com GV Grupo O, em meio salino e enzimático (GV Tratados), durante 120 minutos a intervalos de 15 minutos. Os controles foram incubados com igual volume de soluþÒo salina. Foi aplicada a Análise Digital de Imagens e calculado o Coeficiente de Células Isoladas (CCI). Para analisar o efeito do tempo de incubaþÒo sobre os valores do CCI foi usada uma análise da variÔncia para um desenho em blocos completos casualizados. Os resultados mostraram que o valor médio de CCI, em ambos os meios, variou significativamente com o tempo de incubaþÒo, evidenciando que o aumento do contato das larvas com os GV produz uma diminuiþÒo do acido siálico, que se reflete em valores de CCI mais elevados. A experiÛncia realizada sugeriria que as larvas T. spiralis durante sua permanÛncia e viabilidade no cisto, poderiam ir captando ácido siálico da célula muscular, com o fim de interferir e /ou evitar a resposta imune do hospedeiro.(AU)
RESUMEN
Se ha comunicado que los eritrocitos (GR) incubados con larvas infectantes de Trichinella spiralis presentan mayor agregación que los GR Controles incubados con solución salina, lo cual indica que el parásito capta el ácido siálico globular. El objetivo fue estudiar la cinética de captación de ácido siálico por larvas musculares de T. spiralis aplicando Análisis Digital de Imágenes. Se trabajó con concentrados larvales de T. spiralis incubados en partes iguales con GR Grupo O, en medio salino y enzimático (GR Tratados), durante 120 minutos a intervalos de 15 minutos. Los Controles fueron incubados con igual volumen de solución salina. Se aplicó Análisis Digital de Imágenes y se calculó el valor del Coeficiente de células aisladas (CCA). Para analizar el efecto del tiempo de incubación en los valores de CCA, se utilizó un análisis de la variancia para un diseño en bloques completos aleatorizados. Los resultados mostraron que el valor promedio de CCA, en ambos medios, varió significativamente con el tiempo de incubación, lo que evidencia que el aumento del contacto de las larvas con los GR produce una disminución del ácido siálico que se refleja en mayores valores de CCA. La experiencia realizada sugeriría que durante su permanencia y viabilidad en el quiste, las larvas de T. spiralis podrían ir captando ácido siálico de la célula muscular con el objeto de interferiry /o evadir la respuesta inmune del hospedador...
Asunto(s)
Larva , Trichinella spiralis , Ácido N-Acetilneuramínico , Cinética , Parasitosis Intestinales , TriquinelosisRESUMEN
Debido al pleomorfismo y la variabilidad cultural que presentan las especies del género Trichophyton, los métodos de identificación basados exclusivamente en caracteres morfológicos a menudo no son suficientes para su clasificación. El objetivo de este estudio fue probar un conjunto de métodos fenotípicos para identificar aislamientos fúngicos pertenecientes al género Trichophyton empleando una técnica de taxonomía molecular como método de referencia. Se utilizaron 56 aislamientos clínicos y 6 cepas de referencia, con los que se realizaron estudios macro y micromorfológicos, así como las siguientes pruebas bioquímicas y fisiológicas: perforación del pelo in vitro, requerimientos nutricionales en medios agar Trichophyton, producción de ureasa y crecimiento en el medio agar púrpura de bromocresol-leche-glucosa (APBC-L-G). A su vez se realizó una PCR fingerprinting utilizando el cebador simple (GACA)4. Como resultado de la reacción de PCR se observaron perfiles específicos bien diferenciables para Microsporum canis, Epidermophyton fl occosum, Trichophyton rubrum y Trichophyton interdigitale, pero un mismo perfil para Arthroderma benhamiae y Trichophyton erinacei. Del total de los aislamientos clínicos evaluados, 39 coincidieron con el perfil de T. rubrum, 13 con el de A. benhamiae y 4 con el de T. interdigitale. El ensayo fenotípico más útil para diferenciar T. rubrum del complejo T. mentagrophytes fue la alcalinización del medio APBC-L-G. Con las pruebas fenotípicas se pudo diferenciar T. rubrum de los aislamientos del complejo T. mentagrophytes, pero no las especies pertenecientes a dicho complejo. La técnica molecular permitió caracterizar tanto los aislamientos de T. rubrum como los pertenecientes al complejo T. mentagrophytes, que en nuestro estudio correspondieron a T. interdigitale y Trichophyton sp. anamorfo de A. benhamiae.
Due to the pleomorphism and cultural variability displayed by species of the genus Trichophyton, the identification methods based solely on morphological features are usually insufficient for their classification. The goal of the present work was to test a set of phenotypic methods in order to identify fungal isolates that belong to the aforementioned genus. These methods were based on a molecular taxonomic technique used as standard. Clinical isolates (56) were used as samples along with 6 reference strains. Macro and micromorphological studies were performed as well as biochemical and physiological tests such as in vitro hair perforation, nutritional requirements in Trichophyton agar media, urease production and growth on bromocresol purple-milk solids-glucose (BCP-MS-G) agar. Additionally, PCR fingerprinting using the (GACA)4 primer was employed. As a result of the PCR method, specific profiles were observed for Microsporum canis, Epidermophyton floccosum, Trichophyton rubrum and Trichophyton interdigitale. Identical profiles were obtained for Arthroderma benhamiae y Trichophyton erinacei. Of the total number of clinical isolates, 39 matched the T. rubrum profile while 13 corresponded to A. benhamiae and 4 to T. interdigitale. The most useful phenotypic test to differentiate between T. rubrum and T. mentagrophytes complex strains was alkalinization of the BCP-MS-G medium. Phenotypic tests did not allow differentiation among the T. mentagrophytes complex species. On the other hand, the molecular technique allowed characterization of T. rubrum isolates as well as of those observed in our study and included in the T. mentagrophytes complex: T. interdigitale and Trichophyton sp. , the anamorph of A. benhamiae.
Asunto(s)
Humanos , Trichophyton/clasificación , Argentina , Técnicas de Tipificación Micológica/métodos , Trichophyton/aislamiento & purificaciónRESUMEN
Debido al pleomorfismo y la variabilidad cultural que presentan las especies del género Trichophyton, los métodos de identificación basados exclusivamente en caracteres morfológicos a menudo no son suficientes para su clasificación. El objetivo de este estudio fue probar un conjunto de métodos fenotípicos para identificar aislamientos fúngicos pertenecientes al género Trichophyton empleando una técnica de taxonomía molecular como método de referencia. Se utilizaron 56 aislamientos clínicos y 6 cepas de referencia, con los que se realizaron estudios macro y micromorfológicos, así como las siguientes pruebas bioquímicas y fisiológicas: perforación del pelo in vitro, requerimientos nutricionales en medios agar Trichophyton, producción de ureasa y crecimiento en el medio agar púrpura de bromocresol-leche-glucosa (APBC-L-G). A su vez se realizó una PCR fingerprinting utilizando el cebador simple (GACA)4. Como resultado de la reacción de PCR se observaron perfiles específicos bien diferenciables para Microsporum canis, Epidermophyton fl occosum, Trichophyton rubrum y Trichophyton interdigitale, pero un mismo perfil para Arthroderma benhamiae y Trichophyton erinacei. Del total de los aislamientos clínicos evaluados, 39 coincidieron con el perfil de T. rubrum, 13 con el de A. benhamiae y 4 con el de T. interdigitale. El ensayo fenotípico más útil para diferenciar T. rubrum del complejo T. mentagrophytes fue la alcalinización del medio APBC-L-G. Con las pruebas fenotípicas se pudo diferenciar T. rubrum de los aislamientos del complejo T. mentagrophytes, pero no las especies pertenecientes a dicho complejo. La técnica molecular permitió caracterizar tanto los aislamientos de T. rubrum como los pertenecientes al complejo T. mentagrophytes, que en nuestro estudio correspondieron a T. interdigitale y Trichophyton sp. anamorfo de A. benhamiae.(AU)
Due to the pleomorphism and cultural variability displayed by species of the genus Trichophyton, the identification methods based solely on morphological features are usually insufficient for their classification. The goal of the present work was to test a set of phenotypic methods in order to identify fungal isolates that belong to the aforementioned genus. These methods were based on a molecular taxonomic technique used as standard. Clinical isolates (56) were used as samples along with 6 reference strains. Macro and micromorphological studies were performed as well as biochemical and physiological tests such as in vitro hair perforation, nutritional requirements in Trichophyton agar media, urease production and growth on bromocresol purple-milk solids-glucose (BCP-MS-G) agar. Additionally, PCR fingerprinting using the (GACA)4 primer was employed. As a result of the PCR method, specific profiles were observed for Microsporum canis, Epidermophyton floccosum, Trichophyton rubrum and Trichophyton interdigitale. Identical profiles were obtained for Arthroderma benhamiae y Trichophyton erinacei. Of the total number of clinical isolates, 39 matched the T. rubrum profile while 13 corresponded to A. benhamiae and 4 to T. interdigitale. The most useful phenotypic test to differentiate between T. rubrum and T. mentagrophytes complex strains was alkalinization of the BCP-MS-G medium. Phenotypic tests did not allow differentiation among the T. mentagrophytes complex species. On the other hand, the molecular technique allowed characterization of T. rubrum isolates as well as of those observed in our study and included in the T. mentagrophytes complex: T. interdigitale and Trichophyton sp. , the anamorph of A. benhamiae.(AU)
Asunto(s)
Humanos , Trichophyton/clasificación , Argentina , Técnicas de Tipificación Micológica/métodos , Trichophyton/aislamiento & purificaciónRESUMEN
Due to the pleomorphism and cultural variability displayed by species of the genus Trichophyton, the identification methods based solely on morphological features are usually insufficient for their classification. The goal of the present work was to test a set of phenotypic methods in order to identify fungal isolates that belong to the aforementioned genus. These methods were based on a molecular taxonomic technique used as standard. Clinical isolates (56) were used as samples along with 6 reference strains. Macro and micromorphological studies were performed as well as biochemical and physiological tests such as in vitro hair perforation, nutritional requirements in Trichophyton agar media, urease production and growth on bromocresol purple-milk. solids-glucose (BCP-MS-G) agar. Additionally, PCR fingerprinting using the (GACA)4 primer was employed. As a result of the PCR method, specific profiles were observed for Microsporum canis, Epidermophyton floccosum, Trichophyton rubrum and Trichophyton interdigitale. Identical profiles were obtained for Arthroderma benhamiae y Trichophyton erinacei. Of the total number of clinical isolates, 39 matched the T. rubrum profile while 13 corresponded to A. benhamiae and 4 to T. interdigitale. The most useful phenotypic test to differentiate between T. rubrum and T. mentagrophytes complex strains was alkalinization of the BCP-MS-G medium. Phenotypic tests did not allow differentiation among the T. mentagrophytes complex species. On the other hand, the molecular technique allowed characterization of T. rubrum isolates as well as of those observed in our study and included in the T. mentagrophytes complex: T. interdigitale and Trichophyton sp., the anamorph of A. benhamiae.
Asunto(s)
Trichophyton/clasificación , Argentina , Humanos , Técnicas de Tipificación Micológica/métodos , Trichophyton/aislamiento & purificaciónRESUMEN
Due to the pleomorphism and cultural variability displayed by species of the genus Trichophyton, the identification methods based solely on morphological features are usually insufficient for their classification. The goal of the present work was to test a set of phenotypic methods in order to identify fungal isolates that belong to the aforementioned genus. These methods were based on a molecular taxonomic technique used as standard. Clinical isolates (56) were used as samples along with 6 reference strains. Macro and micromorphological studies were performed as well as biochemical and physiological tests such as in vitro hair perforation, nutritional requirements in Trichophyton agar media, urease production and growth on bromocresol purple-milk. solids-glucose (BCP-MS-G) agar. Additionally, PCR fingerprinting using the (GACA)4 primer was employed. As a result of the PCR method, specific profiles were observed for Microsporum canis, Epidermophyton floccosum, Trichophyton rubrum and Trichophyton interdigitale. Identical profiles were obtained for Arthroderma benhamiae y Trichophyton erinacei. Of the total number of clinical isolates, 39 matched the T. rubrum profile while 13 corresponded to A. benhamiae and 4 to T. interdigitale. The most useful phenotypic test to differentiate between T. rubrum and T. mentagrophytes complex strains was alkalinization of the BCP-MS-G medium. Phenotypic tests did not allow differentiation among the T. mentagrophytes complex species. On the other hand, the molecular technique allowed characterization of T. rubrum isolates as well as of those observed in our study and included in the T. mentagrophytes complex: T. interdigitale and Trichophyton sp., the anamorph of A. benhamiae.
Asunto(s)
Trichophyton/clasificación , Argentina , Humanos , Técnicas de Tipificación Micológica/métodos , Trichophyton/aislamiento & purificaciónRESUMEN
El objetivo del presente trabajo fue estudiar el efecto de A. lumbricoides sobre la carga aniónica de eritrocitos y eritrocitos desializados. Se trabajó con 30 extractos ([EA]) y 2 concentrados larvales parasitarios ([CLAL] 1: 500- 600/ [CLAL]2: 200- 300 larvas/ mL) y eritrocitos Grupo "O" en medio salino (GR) y enzimático (GRb). Se incubó el sedimento globular con igual volumen de parásito y el Control (GRc) con solución fisiológica, a 37 ºC. Se aplicaron los Métodos de Polibrene y Azul Alcian y se implementó una Titulación de la Agregación a partir del Polibrene. Se definieron coeficientes para cada método que fueron analizados estadísticamente. Los resultados mostraron que a mayor tiempo de tratamiento y a mayor concentración larvaria, aumentó la captación de ácido siálico (AS) disminuyendo la agregación de GR y GRb. El efecto fue mayor en la incubación del parásito con GRb en relación de GR. El tratamiento enzimático redujo la carga porcentual en los glóbulos incubados con ambos [CLAL] de forma similar (16%-17% de la inicial) y la Diferencia de Carga de GR y GRb debida a ambos fue aproximadamente del 10% indicando que la pérdida de carga producida por [CLAL]1 fue significativa con respecto a [CLAL]2. Se concluye que A. lumbricoides puede captar AS eritrocitario. Este fenómeno contribuiría a explicar la anemia y trombos producidos en ascariosis, destacando que en pacientes con patologías que cursen con déficit de AS estas complicaciones serían más relevantes.
The aim of this work was to study the A. lumbricoides' effect on the anionic charge of erythrocytes and desyalated erythrocytes. Work was carried out on 30 extracts ([AE]) and 2 larval concentrates ([ALLC] 1: 500-600; ([ALLC] 2: 200-300 larvae/ mL) of the parasite and Group O erythrocytes in saline (RC) and enzymatic medium (RCb). The red cell sediment was incubated with an equal volume of parasite and the Control (RCc) with saline solution, at 37C. Polybrene and Alcian Blue Methods were applied and Aggregation Titulation from Polybrene was implemented. The coefficients for each method were defined,which were analyzed statistically. The results showed that a longer treatment and also at higher larval concentration,increased the capture of sialic acid (AS),while decreasing the RC and RCb aggregation. The effect was greater on the incubation of the parasite with RCb in relation to RC. The enzymatic treatment reduced similarly the percentage charge in the globules incubated with both [ALLC] (16% -17% of initial) and the variation of RC and RCb charge due to both was about 10%,indicating that the charge loss caused by [ALLC]1 was significant with regard to [ALLC]2. We concluded that A. lumbricoides can capture erythroctyte SA. This phenomenon would contribute to explain the anaemia and thrombi produced in ascariosis,noting that in patients with AS deficit pathologies,these complications would be more relevant.
O objetivo foi estudar o efeito de A. lumbricoides sobre a carga aniônica de eritrócitos e eritrócitos desializados. Trabalhou-se com 30 extratos ([EA]) e 2 concentrados larvais parasitários ([CLAL]1: 500- 600/ [CLAL]2: 200- 300 larvas/ mL) e eritrócitos Grupo "O" em meio salino (GV) e enzimático (GVb). Foi incubado o sedimento globular com igual volume de parasita e o Testemunha (GVc) com solução fisiológica,a 37ºC. Foram aplicados os Métodos de Polybrene e Azul Alciano e se implementou uma Titulação da Agregação a partir do Polybrene. Definiram-se coeficientes para cada método que foram analisados estatisticamente. Os resultados mostraram que a maior tempo de tratamento e a maior concentração larvária,aumentou a captação de ácido siálico (AS) diminuindo a agregação de GV e GVb. O efeito foi maior na incubação da parasita com GVb em relação de GV. O tratamento enzimático reduziu a carga percentual nos glóbulos incubados com ambos [CLAL] de forma similar (16%-17% da inicial) e a Diferença de Carga de GV e GVb devida a ambos foi aproximadamente de 10% indicando que a perda de carga produzida por [CLAL]1 foi significativa com relação a [CLAL]2. A conclusão é que A. lumbricoides pode captar AS eritrocitário. Este fenômeno contribuiria a explicar a anemia e trombos produzidos em ascaridíase,destacando que em pacientes com patologias que cursem com déficit de AS estas complicações seriam mais relevantes.
RESUMEN
El objetivo del presente trabajo fue estudiar el efecto de A. lumbricoides sobre la carga aniónica de eritrocitos y eritrocitos desializados. Se trabajó con 30 extractos ([EA]) y 2 concentrados larvales parasitarios ([CLAL] 1: 500- 600/ [CLAL]2: 200- 300 larvas/ mL) y eritrocitos Grupo "O" en medio salino (GR) y enzimático (GRb). Se incubó el sedimento globular con igual volumen de parásito y el Control (GRc) con solución fisiológica, a 37 ºC. Se aplicaron los Métodos de Polibrene y Azul Alcian y se implementó una Titulación de la Agregación a partir del Polibrene. Se definieron coeficientes para cada método que fueron analizados estadísticamente. Los resultados mostraron que a mayor tiempo de tratamiento y a mayor concentración larvaria, aumentó la captación de ácido siálico (AS) disminuyendo la agregación de GR y GRb. El efecto fue mayor en la incubación del parásito con GRb en relación de GR. El tratamiento enzimático redujo la carga porcentual en los glóbulos incubados con ambos [CLAL] de forma similar (16%-17% de la inicial) y la Diferencia de Carga de GR y GRb debida a ambos fue aproximadamente del 10% indicando que la pérdida de carga producida por [CLAL]1 fue significativa con respecto a [CLAL]2. Se concluye que A. lumbricoides puede captar AS eritrocitario. Este fenómeno contribuiría a explicar la anemia y trombos producidos en ascariosis, destacando que en pacientes con patologías que cursen con déficit de AS estas complicaciones serían más relevantes.(AU)
The aim of this work was to study the A. lumbricoides effect on the anionic charge of erythrocytes and desyalated erythrocytes. Work was carried out on 30 extracts ([AE]) and 2 larval concentrates ([ALLC] 1: 500-600; ([ALLC] 2: 200-300 larvae/ mL) of the parasite and Group O erythrocytes in saline (RC) and enzymatic medium (RCb). The red cell sediment was incubated with an equal volume of parasite and the Control (RCc) with saline solution, at 37C. Polybrene and Alcian Blue Methods were applied and Aggregation Titulation from Polybrene was implemented. The coefficients for each method were defined,which were analyzed statistically. The results showed that a longer treatment and also at higher larval concentration,increased the capture of sialic acid (AS),while decreasing the RC and RCb aggregation. The effect was greater on the incubation of the parasite with RCb in relation to RC. The enzymatic treatment reduced similarly the percentage charge in the globules incubated with both [ALLC] (16% -17% of initial) and the variation of RC and RCb charge due to both was about 10%,indicating that the charge loss caused by [ALLC]1 was significant with regard to [ALLC]2. We concluded that A. lumbricoides can capture erythroctyte SA. This phenomenon would contribute to explain the anaemia and thrombi produced in ascariosis,noting that in patients with AS deficit pathologies,these complications would be more relevant.(AU)
O objetivo foi estudar o efeito de A. lumbricoides sobre a carga ani¶nica de eritrócitos e eritrócitos desializados. Trabalhou-se com 30 extratos ([EA]) e 2 concentrados larvais parasitários ([CLAL]1: 500- 600/ [CLAL]2: 200- 300 larvas/ mL) e eritrócitos Grupo "O" em meio salino (GV) e enzimático (GVb). Foi incubado o sedimento globular com igual volume de parasita e o Testemunha (GVc) com soluþÒo fisiológica,a 37ºC. Foram aplicados os Métodos de Polybrene e Azul Alciano e se implementou uma TitulaþÒo da AgregaþÒo a partir do Polybrene. Definiram-se coeficientes para cada método que foram analisados estatisticamente. Os resultados mostraram que a maior tempo de tratamento e a maior concentraþÒo larvária,aumentou a captaþÒo de ácido siálico (AS) diminuindo a agregaþÒo de GV e GVb. O efeito foi maior na incubaþÒo da parasita com GVb em relaþÒo de GV. O tratamento enzimático reduziu a carga percentual nos glóbulos incubados com ambos [CLAL] de forma similar (16%-17% da inicial) e a Diferenþa de Carga de GV e GVb devida a ambos foi aproximadamente de 10% indicando que a perda de carga produzida por [CLAL]1 foi significativa com relaþÒo a [CLAL]2. A conclusÒo é que A. lumbricoides pode captar AS eritrocitário. Este fen¶meno contribuiria a explicar a anemia e trombos produzidos em ascaridíase,destacando que em pacientes com patologias que cursem com déficit de AS estas complicaþ§es seriam mais relevantes.(AU)
RESUMEN
OBJECTIVE: The aim of this work was to evaluate the expression of FUT2 gene in saliva and histo ABH antigens of patients with oral lesions. STUDY DESIGN: In total 178 subjects were examined, half of whom suffered from oral pre-cancerous and cancerous lesions, while the other half were the healthy control group We analyzed the FUT 2 polymorphism by ASO-PCR (allele specific oligonucleotid - polymerase chain reaction) with specific primers for G428 allele and the wild type allele of FUT2 gene. To reveal A, B and H antigens in tissue sections of the patients (n= 89) we used a modified specific red cell adherence technique. RESULTS: We found a high intensity of oral disease in the non-secretor group (OR = 2.43). A total of 58% of the patients with oral pre-cancerous and cancerous lesions was non secretors (se_/_), in contrast with the healthy population (21.5%). A strongly positive reaction was defined as a sheet of indicator erythrocytes adhered to the epithelial cells. In 31 of the 54 samples analyzed the test showed slightly positive results on atypical areas, and there was a complete antigen deletion in areas affected by neoplasia. Nineteen samples showed a total absence of ABH antigens in both histologically normal and pathological areas. Blood group antigens were expressed at a high level in benign and highly differentiated malignant tumors. In poorly differentiated malignant tumors, they were mostly absent. CONCLUSION: Considering these results we suggest the use of this method to monitor probable preneoplastic lesions in risk population, especially in those with no secretor status (absence of FUT2 gene).
Asunto(s)
Sistema del Grupo Sanguíneo ABO/biosíntesis , Fucosiltransferasas/biosíntesis , Neoplasias de la Boca/sangre , Lesiones Precancerosas/sangre , Fucosiltransferasas/análisis , Expresión Génica , Humanos , Neoplasias de la Boca/genética , Lesiones Precancerosas/genética , Saliva/química , Galactósido 2-alfa-L-FucosiltransferasaRESUMEN
Introducción: el estudio de las interacciones hospedador-parásito es un nuevo desafío para comprender aspectos del metabolismo parasitario, los mecanismos de invasión, escape inmunológico y daño. Ascaris lumbricoides puede provocar anemia y trombosis. Previamente se demostró que este parásito altera la carga superficial eritrocitaria, lo cual indica que puede captar ácido siálico del glóbulo rojo. Objetivo: estudiar el efecto producido por extractos del parásito adulto sobre la carga eritrocitaria, utilizando el método de azul Alcian y comparar su sensibilidad con el método de Polibrene. Métodos: se trabajó con 55 extractos parasitarios [EA] y con suspensiones de eritrocitos grupo O. Se realizó el tratamiento de los glóbulos incubando el sedimento con igual volumen de [EA] a 37 ºC durante 1 h. El Control (eritrocitos sin contacto con [EA]) fue incubado con tampón fosfato pH 7,4. Se aplicó el método de azul Alcian y se determinó la carga aniónica eritrocitaria porcentual (CAE por ciento) en el Control y en los glóbulos tratados. Se definió el coeficiente experimental de carga aniónica eritrocitaria (Cexp CAE), como el cociente entre CAE por ciento final e inicial. Resultados: se observó que 27 de los 55 [EA] (49,1 por ciento) modificaron la carga de los glóbulos rojos, el Cexp CAE para estos eritrocitos resultó 0,75 ± 0,1144 y para los que no mostraron variación de carga de 0,94 ± 0,04450. El análisis estadístico concluyó que los métodos de Polibrene y de azul Alcian tienen sensibilidades comparables (p> 0,20). Conclusiones: Ascaris lumbricoides es capaz de captar ácido siálico del eritrocito, lo que contribuiría a explicar la trombosis atribuida al parásito, pero también sugeriría que el nematodo lo podría utilizar en sus rutas metabólicas o en sus estrategias de evasión inmunológica.
Introduction: the study of the host-parasite interactions is a new challenge to understanding some aspects of the parasitic metabolism and the mechanisms of invasion, immunological evasion and damage. Ascaris lumbricoides may cause anemia and thrombosis. It was previously shown that Ascaris lumbricoides modified the superficial charge of erythrocytes, which means that the parasite can capture sialic acid from the red blood cell. Objective: to study the effect of adult parasite extracts on the erythrocyte charge using the Alcian Blue method and to compare its sensitivity with the Polybrene method. Methods: fifty five adult parasite extracts and Group O erythrocyte suspensions were used. The erythrocytes were treated by incubating the sediment with an equal volume of parasite extracts for one hour at 37 ºC. The control group (erythrocytes without any contact with the parasite extracts) was incubated with pH 7.4phosphate buffer solution. Alcian Blue method was applied and the percentage erythrocyte anionic charge was determined in the control group and in the treated red cells. The experimental coefficient of erythrocyte anionic charge was defined as the quotient between the initial and the final percentage erythrocyte anionic charge. Results: it was shown that 27 out of 55 parasite extracts (49.1 percent) modified the charge of the red blood cells, being their experimental coefficient of the erythrocyte anionic charge 0.75 ± 0.1144 whereas the same coefficient amounted to 0,94 ± 0.0445 for those which did not show any charge variation. The statistical analysis concluded that the Polybrene and Alcian Blue Methods had comparable sensitivities (p>0.20). Conclusions: A. lumbricoides is able to capture sialic acid from the erythrocyte, which would not only explain the thrombosis attributed to the parasite, but also suggest that the nematode could use this acid either in its metabolic routes or for its strategies of immunological evasion.
Asunto(s)
Animales , Azul Alcián , Ascaris lumbricoides , Mezclas Complejas/farmacología , Eritrocitos/fisiología , Fenómenos ElectrofisiológicosRESUMEN
INTRODUCTION: the study of the host-parasite interactions is a new challenge to understanding some aspects of the parasitic metabolism and the mechanisms of invasion, immunological evasion and damage. Ascaris lumbricoides may cause anemia and thrombosis. It was previously shown that Ascaris lumbricoides modified the superficial charge of erythrocytes, which means that the parasite can capture sialic acid from the red blood cell. OBJECTIVE: to study the effect of adult parasite extracts on the erythrocyte charge using the Alcian Blue method and to compare its sensitivity with the Polybrene method: METHODS: fifty five adult parasite extracts and Group O erythrocyte suspensions were used. The erythrocytes were treated by incubating the sediment with an equal volume of parasite extracts for one hour at 37 degrees C. The control group (erythrocytes without any contact with the parasite extracts) was incubated with pH 7.4 phosphate buffer solution. Alcian Blue method was applied and the percentage erythrocyte anionic charge was determined in the control group and in the treated red cells. The experimental coefficient of erythrocyte anionic charge was defined as the quotient between the initial and the final percentage erythrocyte anionic charge. RESULTS: it was shown that 27 out of 55 parasite extracts (49.1 %) modified the charge of the red blood cells, being their experimental coefficient of the erythrocyte anionic charge 0.75 +/- 0.1144 whereas the same coefficient amounted to 0,94 +/- 0.0445 for those which did not show any charge variation. The statistical analysis concluded that the Polybrene and Alcian Blue Methods had comparable sensitivities (p>0.20). CONCLUSIONS: A. lumbricoides is able to capture sialic acid from the erythrocyte, which would not only explain the thrombosis attributed to the parasite, but also suggest that the nematode could use this acid either in its metabolic routes or for its strategies of immunological evasion.
Asunto(s)
Azul Alcián , Ascaris lumbricoides , Mezclas Complejas/farmacología , Eritrocitos/fisiología , Animales , Fenómenos ElectrofisiológicosRESUMEN
OBJECTIVES: To study membrane proteins modifications in Senescent Red Blood Cells (SeRBC). DESIGN ANDMETHODS: SeRBC were obtained on Percoll gradients. Membrane proteins were analyzed by SDS-PAGE, band 3 by immunoblotting, and protein oxidation by measuring the carbonyl groups. RESULTS: Densitometric analysis showed no change in SeRBC while an increase in band 3 and its degradation products was found. An increase of protein oxidation level was found in SeRBC. CONCLUSIONS: These findings provide further experimental evidence about protein modifications occurring during the RBC lifespan.
Asunto(s)
Proteína 1 de Intercambio de Anión de Eritrocito/metabolismo , Envejecimiento Eritrocítico , Eritrocitos/química , Proteínas de la Membrana/metabolismo , Electroforesis en Gel de Poliacrilamida , Eritrocitos/metabolismo , Humanos , Immunoblotting , Oxidación-Reducción , Procesamiento Proteico-PostraduccionalRESUMEN
En el desarrollo de la respuesta inmune a patógenos intracelulares participa el elevado polimorfismo de las moléculas HLA de clase II. El objetivo de este trabajo fue establecer la participación de los alelos HLA-DRB1 en personas con infección con Trypanosoma cruzi (T. cruzi) o con Mycobacterium leprae (M. leprae). Se estudiaron 252 individuos de la ciudad de Rosario, divididos en: 86 personas seropositivas para T cruzi (sin compromiso cardiológico de relevancia), 85 pacientes con diagnóstico de Lepra y 81 individuos controles, sin evidencia de patologías. El ADN genómico fue extraído de sangre periférica utilizando el método de salting out y empleado como templado para amplificar por PCR el segundo exón polimórfico de HLA-DRB1. Los alelos fueron tipificados mediante la técnica de PCR-SSOP. La comparación de frecuencias mostró prevalencia de los alelos DRB1 *0409 y DRB1 *1503 en los individuos seropositivos para T. cruzi con respecto al grupo control. Por otra parte, el análisis estadístico indicó una disminución significativa del alelo DRB1 *1103 en pacientes con esta tripanosomiasis. Al examinar las frecuencias observamos en el grupo de pacientes con Lepra un aumento significativo de los alelos DRB1 *1401 y DRB1 *1406. Además observamos que las proporciones de los alelos DRB1 *0808 y DRB1 *1103 en los enfermos son significativamente inferiores con respecto al grupo control. Los alelos HLA DRB1 podrían actuar solos o en combinación con otros genes para conferir susceptibilidad o resistencia a estas infecciones en la población de Rosario, Argentina.
In the development of the immune response to intracellular pathogens implicated the high polymorphism of HLA class II molecules. The aim of this study was to establish the involvement of the HLA-DRB1 alleles in infected subjects with T. cruzi or leprosy patients in Rosario, Argentina. We studied 252 individuals who divided into: 86 positive people for T. cruzi without cardiac damage, 85 patients diagnosed with leprosy and controls 81 individuals without evidence of disease. Genomic DNA was extracted from peripheral blood using the standard salting out method and used as a template to amplify by the PCR the polymorphic second exon of the HLA-DRB1. PCR products were hybridized separately with sequence-specifics oligonucleotides (SSOP). Statistical analysis indicated that of increased frequencies of DRB1 *0409, and DRB1 *1503 in individuals with Chagas' disease. DRB1 *1103 allele was prevalence in the group control and could be associated with resistance to the presence of trypanosomiasis. DRB1 *1401 and DRB1 *1406 alleles were significantly more prevalent in leprosy patients, whereas a decreased frequency of DRB1 *0808 and DRB1 *1103 alleles was found, by comparison with the group control. The HLA-DRB1 alleles could act alone or in combination with other genes to confer differential susceptibility and also protection to these diseases in Rosario, Argentina.