RESUMO
Fundamento: la depresión es una de las complicaciones no neurológicas más frecuentes en la enfermedad cerebrovascular isquémica. Objetivo: determinar la asociación de marcadores inflamatorios y de disfunción endotelial con la depresión en pacientes con enfermedad cerebrovascular isquémica. Métodos: se realizó un estudio analítico, prospectivo de corte transversal en pacientes con enfermedad cerebrovascular isquémica en fase aguda (N=22) y no aguda (N=37); atendidos en el Instituto de Neurología y Neurocirugía y el Hospital Manuel Fajardo, de La Habana, Cuba. Se recogieron variables demográficas, factores de riesgo, etiología y localización del infarto, deficiencia neurológica, discapacidad para las actividades de la vida diaria (índice de Barthel), neuropsicológicas (depresión por inventario de Beck y test de Hamilton). Se determinó proteína C-reactiva, alfa-1-antitripsina, complementos C3 y C4 y microalbuminuria. Resultados: las puntuaciones de las pruebas neuropsicológicas no tuvieron diferencias significativas entre la fase aguda y no aguda, pero hubo un aumento estadístico de la frecuencia de pacientes sin depresión y con ligera depresión en la fase no aguda. En la fase aguda, el complemento C4 y en la fase no aguda el complemento C3, la proteína C-reactiva y el alfa-1-antitripsina se correlacionaron directamente con la puntuación del inventario de Beck. La proteína C-reactiva y C3 se correlacionaron estadísticamente con la puntuación del test de Hamilton. En el análisis multivariado, la proteína C-reactiva mostró asociación independiente con el grado de depresión por el test de Hamilton. Conclusiones: la proteína C-reactiva pudiera estar relacionada con la severidad de la depresión, quizás por asociación con la discapacidad para las actividades de vida diaria.
Foundation: depression in ischemic cerebrovascular disease is one of the most frequent non-neurological complications. Objective: to determine the association of inflammatory markers and endothelial dysfunction with depression in patients with ischemic cerebrovascular disease. Methods: an analytical, prospective, cross-sectional study was carried out in patients with acute (N=22) and non-acute (N=37) ischemic cerebrovascular disease; treated at the Institute of Neurology and Neurosurgery; and the Manuel Fajardo Hospital, in Havana, Cuba. Demographic variables, risk factors, etiology and location of the infarction, neurological deficiency, disability for activities of daily living (Barthel index), neuropsychological (depression by Beck inventory and Hamilton test) were collected. C-reactive protein, alpha-1-antitrypsin, C3 and C4 complements, and microalbuminuria were determined. Results: the scores of the neuropsychological tests did not have significant differences between the acute and non-acute phase, but there was a statistical increase in the frequency of patients without depression and with slight depression in the non-acute phase. In the acute phase, C4, and in the non-acute phase, C3, C-reactive protein and alpha-1-antitrypsin were directly correlated with the Beck inventory score. C-reactive protein and C3 were statistically correlated with the Hamilton test score. In the multivariate analysis, C-reactive protein showed an independent association with the degree of depression by the Hamilton test. Conclusions: C-reactive protein could be related to the severity of depression, perhaps by association with the disability for activities of daily living.
RESUMO
RESUMEN Objetivos . Comparar diferentes métodos de extracción de ADN a partir de quistes y trofozoítos de Giardia spp. mediante la técnica de reacción en cadena de la polimerasa (PCR) convencional. Materiales y métodos. Se aislaron quistes de Giardia spp. a partir de 65 muestras coprológicas procedentes de hospitales de referencia nacional, obteniéndose una carga promedio de 5x104 parásitos. Asimismo, se cultivaron trofozoítos de Giardia intestinalis (ATCC® 30957™) obteniéndose una carga parasitaria de 5x106. Se compararon once métodos de extracción para quistes y seis para trofozoítos. La concentración y pureza del ADN extraído se determinó por espectrofotometría y el rendimiento de la extracción se evaluó mediante la amplificación de los genes beta giardina (bg) y glutamato deshidrogenasa (gdh) por PCR semi-anidada. Resultados. Se observó que el método I mostró la mayor concentración de ADN a partir de quistes (12,24 ng/µL), pureza (1,4) y mejor rendimiento (100% amplificación bg, 60% gdh) en comparación con los otros métodos evaluados. En el caso de los trofozoítos el método que no tuvo pretratamientos presentó la mayor concentración de ADN, pureza y rendimiento (26,56 ng/µL; 1,85; 100% amplificación bg y gdh). Conclusiones. Los pretratamientos mecánicos, de choque térmico y enzimáticos son necesarios para la ruptura de la pared quística de Giardia spp., siendo el marcador molecular bg de mayor rendimiento para detección de ADN de quistes. Los trofozoítos no requieren pretratamientos para lograr resultados satisfactorios. Se cuenta con una metodología reproducible para la extracción de ADN de Giardia spp. a partir de cualquier estadio evolutivo.
ABSTRACT Objectives. To compare different methods of DNA extraction from cysts and trophozoites of Giardia spp. using the conventional polymerase chain reaction (PCR) technique. Materials and Methods. Cysts of Giardia spp. were isolated from 65 coprological samples from national reference hospitals, obtaining an average load of 5x104 parasites. In addition, Giardia intestinalis trophozoites (ATCC® 30957™) were cultured obtaining a 5x106 parasitic load. Eleven extraction methods for cysts and six for trophozoites were compared. The concentration and purity of the extracted DNA were determined by spectrophotometry and the extraction yield was assessed by amplification of the ß-giardin (bg) and glutamate dehydrogenase (gdh) genes with a semi nested PCR assay. Results. It was observed that method 1 showed the highest concentration of DNA from cysts (12.24 ng/µL), purity (1.4) and best performance (bg: 100% amplification; gdh: 60% amplification) compared to the other methods evaluated. In the case of trophozoites, the method without pre treatment showed the highest level of DNA concentration, purity, and yield (26.56 ng/µL; 1.85; 100% amplification of bg and gdh, respectively). Conclusions . Mechanical, thermal shock, and enzymatic pre-treatments are necessary for the rupture of the cystic wall of Giardia spp. making it the highest-yielding bg molecular marker for detecting cyst DNA. Trophozoites do not require pre-treatment to achieve satisfactory results. A reproducible methodology for the extraction of DNA from Giardia spp. from any evolutionary stage is available.
Assuntos
Humanos , DNA/isolamento & purificação , Reação em Cadeia da Polimerase , Trofozoítos/genética , Giardia/genética , Parasitologia/métodos , Reação em Cadeia da Polimerase/métodos , Técnicas GenéticasRESUMO
Objective: Leishmaniasis is a parasitic disease causing major public health problem in form of visceral and cutaneous types. The cutanoue leishmaniasis is caused by L. tropica, in low-land areas without reservoir; Arthroponatic leishmaniasis (ACL), Zoonotic Cutaneous Leishmaniasis ( ZCL), in high-land. This case report involved; 25 years old Egyptian active young single male adult, stayed in Utama (75 Km far from El-Madina Manowra on the road to Makkah). He presented with three skin lesions on his arms occurred within the last 1-3 months. on examination revealed; volcano- like indurated ulcers which clinically suspected as leishmania lesions. Materials and Methods: Laboratory investigations were involved; skin smear using Giemsa stain, Leishmanin test (LST), polymerase chain reaction (PCR), sequencing and phylogenitic analysis BLAST (NCBI). Results: Microscopy positive LDB (leishmanin donovani bodies), Leishmanin test (LST) was negative. PCR positive L. major. Sequence alignment were 100% with nine Iranian isolates and one Tunisian isolate. After one month of treatment with Pentostam (Sodium stibogluconate) local injections at the site of lesions the lesion progressed from ulcer to scar. Conclusion: L. major is a major species causing cutaneous leishmaniasis in Al-Medina Manowra region in Saudi Arabia. The usage of the polymerase chain reaction (PCR) is a useful diagnostic tool and help to identify the causative species.
RESUMO
OBJECTIVE: In order to evaluate the prevalence of cytomegalovirus infection to terminally failing heart, cytomegaloviral DNA was detected in the explanted hearts of transplantation recipients. METHODS: DNA extractions were performed from explanted failing hearts(N=22) and normal hearts (N=5) and polymerase chain reactions(PCRs) were done for detection of late gene sequence coded pp150 phosphoprotein. The products were confirmed with electrophoresis on 1% agarose gel. In order to improve the sensitivity of detection in cytomegaloviral genome, nested PCRs were executed with the primers designed for the original 607 bp products. RESULTS: All patients had IgG anti-cytomegalovirus antibody and did not have IgM anti-cytomegalovirus antibody. Cytomegaloviral genomes in myocardium were detected by polymerase chain reaction. The 607bp products by PCRs were found in both explanted failing hearts(3 cases/22, 13.5%) and normal hearts(1 case/5, 20.0%). In nested PCRs, 186bp products were found in both failing hearts(LV 4/22, LA 3/20, RV 5/22, HA 0/17) and normal hearts(LV 2/5, LA 1/4, RV 1/5, RA 2/5). There was no significant change in the presence of cytomegaloviral DNA between failing and normal hearts. Total positivity of cytomegaloviral genome in explanted hearts was 44.4% according to nested PCR results. CONCLUSION: Cytomegalovirus was rarely observed in explanted hearts of terminal heart failure and nested PCR could enhance the sensitivity of cytomegaloviral genome detection. But cytomegalovirus might have no direct causal relationship in the development of terminal heart failure.