Usefulness of a Simple Immunohistochemical Staining Technique to Differentiate Anti-p200 Pemphigoid From Other Autoimmune Blistering Diseases: A Report of 2 Cases. / Descripción de 2 casos de penfigoide anti-p200. Utilidad de una técnica inmunohistoquímica sencilla en el diagnóstico diferencial con otras enfermedades ampollosas autoinmunes.

Anti-p200 pemphigoid is a rare autoimmune subepidermal blistering disease characterized by the presence of circulating immunoglobulin G antibodies directed against laminin gamma-1, a 200-kDa protein located in the lamina lucida of the basement membrane. We review the clinical, histopathological and immunological characteristics of the first 2 cases described in Spain. Anti-p200 pemphigoid shares histopathological and immunopathological findings with epidermolysis bullosa acquisita, the main entity in the differential diagnosis. However, its management follows the same guidelines as those used for bullous pemphigoid. The diagnosis is confirmed by immunoblotting, which is a complex technique available in few centers. We propose the immunohistochemical detection of collagen type IV on the floor of the blister, combined with standard immunofluorescence techniques, as a simple, accessible alternative to differentiate anti-p200 pemphigoid from epidermolysis bullosa acquisita.

Evaluation and comparison of serological methods for the detection of bovine neosporosis in Argentina.

Neospora caninum is a protozoan parasite that causes abortion and important economic losses in cattle worldwide. The accurate diagnosis of neosporosis is essential for management and control measures. The aims of this study were: i) to evaluate the performance of an in-house enzyme-linked immunosorbent assay based on the 38kDa native antigen (p38-ELISA) to diagnose bovine neosporosis in Argentina using a well- characterized local sera panel from experimentally infected and naturally exposed cattle and ii) to compare the diagnostic performance and agreement of three N. caninum serological tests: p38-ELISA, indirect fluorescence antibody test (IFAT) and immunoblotting (IB) using the same sera panel. Serum samples testing either positive or negative by IFAT and IB were considered "Relative Standards of Comparison" (RSC) and used for p38-ELISA evaluation. Receiver operating characteristics analysis revealed that p38-ELISA was highly accurate (area under the curve= 0.982) according to RSC with a cut-off index of 0.0905. Relative sensitivity and specificity of p38-ELISA were 97.8% and 99.5%, respectively and agreement between RSC and p38-ELISA was almost perfect (k= 0.97). The evaluation and performance comparison of serological tests were performed according to the definition of gold standard based on the decision of the "majority of tests". All tests displayed high sensitivity and specificity values (greater than 95%); and excellent agreement. This study describes the accurate performance of p38-ELISA evaluated locally and the highly accurate diagnostic performance of the studied tests for the detection of anti-N. caninum antibodies in cattle from Argentina.

Sensitization to Vitis vinifera Pollen in a Wine Production Area: Identification of the Allergens Involved

Background: Vine cultivation is widely distributed in La Rioja, Spain (37% of all crops) and is associated with exposure of the general population to vine pollen. The aims of this study were to investigate the prevalence of sensitization to Vitis vinifera pollen in persons with respiratory allergy in the general population and to identify the allergens involved. Materials and Methods: The study population comprised patients who came to the hospital between September 2019 and January 2020 with suspected respiratory allergy. All patients underwent skin prick testing with a panel of standardized aeroallergens, profilin, lipid transfer protein (LTP), and V vinifera pollen extract and prick-prick testing with fresh grapes. The in vitro study included specific IgE by ImmunoCap and ELISA, allergenic profile by immunoblot with individual sera from patients positive to V vinifera pollen extract, and 2D immunoblot with a pool of sera. The spots recognized by IgE were identified using mass spectrometry. Results: A total of 151 patients were included. Of these, 124 were positive to some of the allergens tested. Thirty-four (27.4%) were positive to vine pollen in the skin prick tests. The serology study revealed positive results in 20 patients. Five vine pollen allergens were identified, and profilin was the most prevalent (30%). The other 4 allergens could be considered specific to this pollen. Conclusions: Sensitization to vine pollen was frequent in the general population in a vine growing area. The clinical relevance of this finding is unknown owing to sensitization to other pollens in the vine pollen–positive patients. Five new vine pollen allergens were identified (AU)

Antecedentes: El cultivo de la vid está ampliamente distribuido en La Rioja (37% de los cultivos), lo que supone una exposición de la población general al polen de esta planta. El objetivo de este estudio fue investigar la prevalencia de sensibilización al polen de Vitis vinifera en la población general con alergia respiratoria e identificar los alérgenos implicados. Materiales y métodos: Se incluyeron en el estudio pacientes que acudieron al hospital entre septiembre de 2019 y enero de 2020 con sospecha de alergia respiratoria. A todos ellos se les realizó una prueba cutánea con el panel de aeroalérgenos estandarizados, profilina, LTP, extracto de polen de V. vinifera y Prick prick con uva. El estudio in vitro incluyó IgE específica mediante ImmunoCap y ELISA, perfil alergénico por inmunoblot con sueros individuales de pacientes positivos al extracto de polen de V. vinifera e inmunoblot 2D con un pool de sueros. Las proteínas reconocidas por la IgE fueron identificadas por espectrometría de masas. Resultados: Se incluyeron un total de 151 pacientes. De ellos, 124 fueron positivos a algunos de los alérgenos analizados. 34 (27,4%) fueron positivos a polen de vid por prueba cutánea. 20 fueron positivos tras el estudio serológico. Se identificaron 5 alérgenos del polen de la vid, siendo la profilina el más prevalente (30%). Los otros 4 alérgenos podrían considerarse específicos de este polen. Conclusión: Se detectó una alta sensibilización al polen de vid en la población general en una zona de viñedos. Se desconoce la relevancia clínica debido a la sensibilización a otros pólenes en los pacientes positivos a polen de vid. Se identificaron 5 nuevos alérgenos del polen de la vid (AU)

Estudio Poblacional De Prevalencia De Cisticercosis Porcina En Atahualpa, Ecuador. Metodología Y Definiciones Operacionales / Population-Based Study Of Porcine Cysticercosis Prevalence In Atahualpa, Ecuador. Methodology And Operational Definitions

RESUMEN Objetivo: Evaluar la prevalencia de cisticercosis porcina en la parroquia rural Atahualpa, un poblado donde se ha demostrado endemia de neurocisticercosis humana. Diseño: Para el presente estudio, se colectarán muestras de sangre de aproximadamente 600 cerdos, las cuales serán procesadas mediante la prueba de EITB para estimar la tasa actual de sero-prevalencia de cisticercosis porcina. Unicamente se considerará el muestreo en cerdos a partir de los 2 meses de edad. Asimismo, solo se considerarán a aquellos animales sin evidencia de manifestaciones clínicas que hagan riesgosa su manipulación durante la toma de muestras de sangre. El presente estudio no contempla riesgo para la salud de los animales durante la toma de muestras. De acuerdo a las características de endemicidad de cisticercosis humana en Atahualpa, consideramos que la seroprevalencia esperada de cisticercosis porcina mediante EITB sea de al menos 25%. Entre los objetivos secundarios está el determinar los factores relacionados a una mayor o menor seroprevalencia de cisticercosis porcina (de la esperada). Todos los cerdos que resulten positivos (al menos 3 bandas) en el EITB, serán subsecuentemente tratados con oxfendazole, utilizando el régimen de dosis única de 30 mg/kg. Comentario: La importancia de definir la presencia de infección en la población porcina de una zona híper-endémica de neurocisticercosis humana radica en confirmar la existencia de transmisión activa. Si ésta es demostrada, el tratamiento de los cerdos infectados ayudará a interrumpir el ciclo de la Taenia solium y reducirá la prevalencia de esta enfermedad.

ABSTRACT Objective: To assess sero-prevalence of porcine cysticercosis in Atahualpa, a rural Ecuadorian village endemic for human neurocysticercosis. Design: We aim to collect at least 600 blood samples of local pigs, to quantify the number of positive bands in the EITB, and to estimate crude prevalence rates of porcine cysticercosis in the village. Sampling will only include pigs older than 2 months of age. Only healthy pigs will be sampled, to avoid risks inherent to manipulation of sick animals. There will be no risk for pigs as the result of the procedure. According to the estimated endemicity of human cysticercosis, we expect to find about 25% of infected pigs in this study. In the event of prevalence rates above or below the expected percentage, we will attempt to assess the causes of such discrepancies. All pig positive to 3 bands of more in the EITB will receive treatment with oxfendazole at a single dose of 30 mg/kg. Comment: The importance of defining the prevalence of porcine cysticercosis in a village endemic for human neurocysticercosis is to confirm the existence of active transmission. If this is true, treatment of infected pigs will help to interrupt the life cycle of Taenia solium, thus reducing the prevalence of this disease in humans.

Evaluación y comparación de pruebas serológicas para la detección de la neosporosis bovina en Argentina / Evaluation and comparison of serological methods for the detection of bovine neosporosis in Argentina

Neospora caninum is a protozoan parasite that causes abortion and important economic losses in cattle worldwide. The accurate diagnosis of neosporosis is essential for management and control measures. The aims of this study were: i) to evaluate the performance of an in-house enzyme-linked immunosorbent assay based on the 38 kDa native antigen (p38-ELISA) to diagnose bovine neosporosis in Argentina using a well- characterized local sera panel from experimentally infected and naturally exposed cattle and ii) to compare the diagnostic performance and agreement of three N. caninum serological tests: p38-ELISA, indirect fluorescence antibody test (IFAT) and immunoblotting (IB) using the same sera panel. Serum samples testing either positive or negative by IFAT and IB were considered "Relative Standards of Comparison" (RSC) and used for p38-ELISA evaluation. Receiver operating characteristics analysis revealed that p38-ELISA was highly accurate (area under the curve= 0.982) according to RSC with a cut-off index of 0.0905. Relative sensitivity and specificity of p38-ELISA were 97.8 % and 99.5 %, respectively and agreement between RSC and p38-ELISA was almost perfect (k= 0.97). The evaluation and performance comparison of serological tests were performed according to the definition of gold standard based on the decision of the "majority of tests". All tests displayed high sensitivity and specificity values (greater than 95 %); and excellent agreement. This study describes the accurate performance of p38-ELISA evaluated locally and the highly accurate diagnostic performance of the studied tests for the detection of anti-N. caninum antibodies in cattle from Argentina.

Neospora caninum es un parásito protozoo responsable de abortos y pérdidas económicas en bovinos. La realización de un diagnóstico serológico preciso y con resultados comparables obtenidos por diferentes pruebas contribuye al manejo de este problema y a encarar medidas de control. Los objetivos del presente trabajo fueron los siguientes: 1) evaluar en Argentina una prueba de enzimoinmunoensayo in-house con el antígeno nativo de 38 kDa de N. caninum (ELISA-p38) para el diagnóstico de la neosporosis bovina, utilizando un panel de sueros locales bien caracterizados, procedentes de bovinos infectados de modo experimental o naturalmente expuestos; 2) comparar el desempeño y establecer el nivel de concordancia de tres pruebas serológicas para la detección de N. caninum, ELISA-p38, inmunofluorescencia indirecta (IFI) e inmunoblot (IB), con el mismo panel de sueros. Los sueros que resultaron positivos o negativos a IFI e IB fueron considerados como estándares relativos de comparación (ERC) para evaluar la prueba de ELISA-p38. El análisis de característica operativa del receptor determinó que la prueba de ELISA-p38 fue altamente precisa (área bajo la curva= 0,982) usando el punto de corte 0,0905. La sensibilidad y especificidad relativa del ELISA-p38 fue 97,8 % y 99,5 %, respectivamente, con una concordancia casi perfecta (k= 0,97) respecto del ERC. La comparación del desempeño de las pruebas se realizó usando como gold standard el criterio de la decisión de la "mayoría de las pruebas". Las pruebas exhibieron altos valores de sensibilidad y especificidad (mayores del 95 %) y excelente concordancia. Este trabajo describe un buen desempeño de la prueba de ELISA-p38 evaluada localmente y adecuada performance diagnóstica de las pruebas serológicas analizadas para la detección de anticuerpos anti N. caninum en bovinos de Argentina.