The conventional diagnosis challenge: Real-time PCR and nested PCR correlation with the scoring system for individuals at high-risk of Pneumocystis jirovecii pneumonia / Desafío diagnóstico: PCR anidada y en tiempo real frente a un sistema de puntuación en individuos con gran riesgo de neumonía por Pneumocystis jirovecii

Introduction. Pneumocystis jirovecii is an opportunistic fungus that affects mainly people living with HIV (CD4 cell count lower than 200 cells/ml) and other immunosuppressed patients. Since P. jirovecii does not grow on routine mycological media, diagnosis of P. jirovecii pneumonia relies on indirect evidence of its presence in respiratory samples. Objectives. To associate the results of direct immunofluorescence and two molecular methods with a score to predict P. jirovecii pneumonia in patients with AIDS. Materials and methods. A prospective study was conducted with 40 patients. A respiratory sample collected before treatment was subjected to direct immunofluorescence using the Merifluor kit, to nested PCR targeting the mitochondrial large subunit ribosomal RNA, and to the VIASURE real-time PCR kit. Results. These three techniques revealed P. jirovecii in 6, 12, and 15 samples, respectively. All positive samples by direct immunofluorescence were positive by nested PCR, and all positive samples by nested PCR amplified by real-time PCR. There was a statistically significant association between the P. jirovecii pneumonia score and the molecular methods. Two patients were early diagnosed and responded well to treatment. Conclusion. Molecular methods, especially real-time PCR, are recommended for early diagnosis of P. jirovecii pneumonia in AIDS patients.

Introducción. Pneumocystis jirovecii es un hongo oportunista que afecta principalmente a personas con HIV (recuento de CD4 menor de 200 células/ml) y a otros pacientes inmunosuprimidos. Como P. jirovecii no crece en los medios micológicos de rutina, el diagnóstico de neumonía por P. jirovecii se basa en la evidencia presente en muestras respiratorias. Objetivos. Asociar los resultados de la inmunofluorescencia directa y los de dos métodos moleculares con un puntaje para predecir la neumonía causada por P. jirovecii en pacientes con sida. Materiales y métodos. Se realizó un estudio prospectivo de 40 pacientes. Se recolectó una muestra respiratoria antes del inicio de tratamiento y se sometió a una prueba de inmunofluorescencia directa con el kit Merifluor, una PCR anidada para la amplificación de la subunidad larga del ribosoma mitocondrial y una PCR en tiempo real usando el kit VIASURE. Resultados. Estas tres técnicas evidenciaron la presencia de P. jirovecii en 6, 12 y 15 muestras, respectivamente. Todas las muestras positivas por inmunofluorescencia directa fueron positivas en la PCR anidada y todas las muestras positivas en la PCR anidada amplificaron por PCR en tiempo real. Se encontró una asociación estadística entre los valores de la neumonía causada por P. jirovecii y los métodos moleculares. Dos pacientes con diagnóstico temprano respondieron satisfactoriamente al tratamiento. Conclusión. Se recomiendan los métodos moleculares, especialmente la PCR en tiempo real, para el diagnóstico temprano de neumonía causada por P. jirovecii en pacientes con sida.

Immunofluorescence study of the placenta in a case of pemphigoid gestationis: Pathophysiological bases / Estudio de inmunofluorescencia de la placenta a propósito de un caso de penfigoide gestacional: bases fisiopatológicas

Introducción: El herpes gestationis (HG) es una de las principales dermatosis del embarazo que debe ser reconocida y tratada oportunamente ya que se relaciona con un empeoramiento del pronóstico fetal. Aunque se ha investigado la afectación cutánea, hay escasez de estudios morfológicos y funcionales de la placenta en esta patología. Principales síntomas o hallazgos clínicos: Erupción vesicular eritematosa a las 32+1 semanas de gestación. Diagnósticos principales: HG. Intervenciones terapéuticas y resultados: Inmunogammaglobulina en casos graves refractarios a los corticoides por vía oral con desaparición completa de las lesiones. Conclusión: Hasta donde sabemos, este es el primer caso que reporta un análisis detallado de los depósitos de IgG y C3 en la membrana basal de las vellosidades de la placenta mediante un estudio de inmunofluorescencia. Estos hallazgos podrían relacionarse con el ligero mal funcionamiento de la placenta que puede explicar los efectos neonatales adversos.(AU)

Introduction: Pemphigoid gestationis (PG) is one of the main dermatoses of pregnancy that must be recognized and treated promptly, since it is related to worsening of foetal prognosis. Although skin involvement has been investigated, there is a lack of morphological and functional studies of the placenta in this pathology. Main symptoms and/or clinical findings: Erythematous vesicular rash at 32+1 weeks of gestation. Main diagnoses: PG. Therapeutic interventions and results: Immunogammaglobulin in severe cases refractory to oral corticosteroids with complete disappearance of the lesions. Conclusion: To our knowledge, this is the first case to report a detailed analysis of IgG and C3 deposits in the basement membrane of the placental villi by means of an immunofluorescence study. These findings could be related to a slight malfunction of the placenta that may explain the adverse neonatal effects.(AU)

Compromiso renal atípico en amiloidosis de cadena ligera

Introducción la amiloidosis de cadena ligera (AL) es una entidad desencadenada por la proliferación de un clon de células plasmáticas que genera la acumulación de cadenas ligeras, las cuales se depositan en forma de fibrillas amiloides generando una disfunción orgánica. El compromiso renal generalmente se manifiesta como síndrome nefrótico, con un deterioro lento y progresivo de la función renal que puede llevar a un requerimiento de terapia dialítica. Objetivo demostrar el compromiso renal agresivo y subagudo de la amiloidosis sistémica. Presentación del caso paciente masculino de 35 años que consulta por malestar general, visión borrosa, mareos y oliguria con elevación de azoados, y que progresó rápidamente hasta el requerimiento de hemodiálisis en aproximadamente tres meses. Asociado se documenta polineuropatía periférica, infiltración cardiaca y ligera elevación de transaminasas. Ante negatividad de estudios de extensión, se logra documentar la presencia histopatológica de depósitos amiloides con inmunofluorescencia positiva para AL. Actualmente, se encuentra recibiendo esquema de quimioterapia con adecuada estabilidad clínica y tolerancia. Discusión y conclusión la amiloidosis AL es una entidad infrecuente, con compromiso multiorgánico importante y altas tasas de morbilidad y mortalidad. Se recalca en este caso el compromiso subagudo con requerimiento temprano de terapia dialítica y además se enfatiza la importancia de una sospecha y un diagnóstico oportuno en pacientes con compromiso renal y otras manifestaciones sistémicas.

Introduction Light chain amyloidosis is an entity triggered by the proliferation of a clone of plasma cells that generates the accumulation of light chains, which are deposited in the form of amyloid fibrils generating organic dysfunction. Renal compromise generally manifests as nephrotic syndrome, with a slow and progressive decline of renal function that can lead to dialysis therapy. Purpose The objective of this case report is to demonstrate the aggressive and subacute renal involvement of systemic amyloidosis. Case presentation We present a case of a 35-year-old male patient who consulted for general malaise, blurred vision, dizziness and oliguria with elevated nitrogen levels that progressed fastly to the requirement of hemodialysis in approximately 3 months. It was also reveal peripheral polyneuropathy, cardiac infiltration, and slight elevation of transaminases. Given the negativity of extension studies, amyloid deposits were documented histopathologically with positive immunofluorescence for LA. He is currently receiving chemotherapy regimen with adequate clinical stability and tolerance. Conclusion and discussion To conclude, AL amyloidosis is a rare entity, with significant multi-organ involvement and high rates of morbidity and mortality. In this case, the subacute involvement with an early requirement for dialysis therapy is emphasized, and the importance of suspicion and timely diagnosis in patients with kidney involvement and other systemic manifestations is emphasized.

Evaluación y caracterización de la sirtuina 2.3 de Giardia duodenalis (CdSir2.3)

La giardiasis es la enfermedad gastrointestinal de mayor incidencia mundial, causada por el protozoario Giardia duodenalis, para la cual no se cuenta con una vacuna o tratamiento eficiente. En aras de buscar nuevos blancos farmacológicos contra este parásito, se han estudiado las enzimas del metabolismo energético, como las sirtuinas, deacetilasas dependientes del dinucleótido de adenina y nicotinamida (NAD). Previamente se identificó a GdSir2.1 y GdSir2.2 como deacetilasas dependientes de NAD, con localizaciones subcelulares diferentes. En este trabajo se estudió otro candidato a sirtuina (GdSir2.3) mediante herramientas bioinformáticas para la identificación de características típicas de la familia sirtuina en la secuencia del candidato, y experimentales como la obtención de la proteína recombinante 6xHis-GdSir2.3 que demostró actividad deacetilasa dependiente de NAD y que sirvió como antígeno en la producción de los IgY - α -6xHis-GdSir2.3 para la localización subcelular de la proteína endógena en G. duodenalis. Lo anterior concuerda con otros estudios donde se señala a GdSir2.3 como un importante regulador de la enquistación, debido a su aumento de expresión durante esta etapa del ciclo de vida, constituyéndola como un blanco farmacológico promisorio para el control de esta parasitemia.

Giardiasis is the gastrointestinal disease with the highest incidence worldwide, caused by the protozoan Giardia duodenalis, for which there is no vaccine or efficient treatment. In order to find new pharmacological targets against this parasite, energy metabolism enzymes such as sirtuins, deacetylases dependent on the nicotinamide adenine dinucleotide (NAD), have been studied. GdSir2.1 and GdSir2.2 were previously identified as NAD-dependent deacetylases, with different subcellular locations. In this work, another candidate for sirtuin (GdSir2.3) was studied using bioinformatic tools for the identification of typical characteristics of the sirtuin family in the sequence of the candidate; and experimental ones such as obtaining the recombinant protein 6xHis-GdSir2.3 that demonstrated NAD-dependent deacetylase activity; and that it served as an antigen in the production of IgY - α - 6xHis-GdSir2.3 for the subcellular localization of the endogenous protein in G. duodenalis. The foregoing is consistent with other studies where GdSir2.3 is indicated as an important regulator of encyst due to its increased expression during this stage of the life cycle, constituting it as a promising drug target for the control of this parasitaemia.

A giardíase é a doença gastrointestinal de maior incidência no mundo, causada pelo protozoário Giardia duodenalis, para a qual não existe vacina ou tratamento eficaz. Com o objetivo de encontrar novos alvos farmacológicos contra esse parasita, têm sido estudadas enzimas do metabolismo energético, como as sirtuínas, desacetilases dependentes do dinucleotídeo adenina nicotinamida (NAD). GdSir2.1 e GdSir2.2 foram previamente identificados como desacetilases dependentes de NAD, com diferentes localizações subcelulares. Neste trabalho, outro candidato a sirtuin (GdSir2.3) foi estudado usando ferramentas de bioinformática para a identificação de características típicas da família sirtuin na sequência do candidato; e experimentais, como a obtenção da proteína recombinante 6xHis-GdSir2.3 que demonstrou atividade desacetilase dependente de NAD; e que serviu como antígeno na produção de IgY - α - 6xHis-GdSir2.3 para a localização subcelular da proteína endógena em G. duodenalis. O exposto é consistente com outros estudos em que o GdSir2.3 é apontado como um importante regulador de encisto devido à sua expressão aumentada durante esta fase do ciclo de vida, constituindo-se como um alvo promissor para o controle dessa parasitemia.

Serologic study of Bartonella sp. infection among human population of Southern Spain.

INTRODUCTION: The purpose of this study was to determine the prevalence of IgG antibodies against Bartonella sp. in a randomly selected sample from the population of the patients of North Sanitary District of Jaén. METHODS: We used a commercially available immunofluorescent test (Focus-Technology IFA Bartonella quintana and B. henselae test). RESULTS: Six hundred five healthy individuals were divided by sex into three age groups. We detected that 13.55% and 11.07% subjects were IgG seropositive to B. henselae and B. quintana, respectively. CONCLUSIONS: Our data show that the prevalence of both Bartonella species in Andalusia (Southern Spain) is relatively high. No statistical difference in the seropositivity was observed among these groups. In both cases, the IgG antibody titers ranged from 1/128 to 1/512.

Serologic study of Bartonella sp. infection among human population of Southern Spain / Estudio serológico de la infección por Bartonella sp. en la población humana del sur de España

IntroductionThe purpose of this study was to determine the prevalence of IgG antibodies against Bartonella sp. in a randomly selected sample from the population of the patients of North Sanitary District of Jaén.MethodsWe used a commercially available immunofluorescent test (Focus-Technology IFA Bartonella quintana and B. henselae test).ResultsSix hundred five healthy individuals were divided by sex into three age groups. We detected that 13.55% and 11.07% subjects were IgG seropositive to B. henselae and B. quintana, respectively.ConclusionsOur data show that the prevalence of both Bartonella species in Andalusia (Southern Spain) is relatively high. No statistical difference in the seropositivity was observed among these groups. In both cases, the IgG antibody titers ranged from 1/128 to 1/512.

IntroducciónEl propósito de este estudio fue determinar la prevalencia de anticuerpos IgG frente a Bartonella sp. en una muestra escogida al azar de la población de pacientes del Distrito Sanitario Norte de Jaén.MétodosSe ha utilizado una prueba de inmunofluorescencia disponible comercialmente (Focus-Technology IFA Bartonella quintana y prueba de B. henselae). Seiscientos cinco individuos sanos se dividieron por sexo en 3 grupos de edad.ResultadosDetectamos que el 13,55% y el 11,07% de los sujetos eran IgG seropositivos a B. henselae y B. quintana, respectivamente. En ambos casos, los títulos de anticuerpos IgG variaron de 1/128 a 1/512.ConclusiónNuestros datos muestran que la prevalencia de ambas especies de Bartonella en Andalucía (sur de España) es relativamente alta. No se observaron diferencias estadísticas en la seropositividad entre grupos de edad.

Indocyanine green fluorescence angiography in colorectal surgery. First case series in Mexico.

INTRODUCTION AND AIM: Anastomosis leak occurs in 1-19% of colorrectal surgeries. Our objective was to present the first Mexican case series on colorrectal surgery using indocyanine green fluorescence angiography to evaluate perfusion prior to carrying out the anastomosis. MATERIALS AND METHODS: A retrospective, analytic, descriptive study was conducted. We studied the case records of consecutive patients that underwent colorrectal surgery with indocyanine green angiography performed by the same group of colorrectal surgeons. RESULTS: Twenty-one case records were reviewed. Eleven (52.3%) of the patients were women, mean patient age was 57 years (38-82), and mean body mass index was 25 kg/m2 (17-34). Fifteen (71.4%) patients were diagnosed with malignant disease. Indocyanine green angiography changed our therapeutic decision in three (14.2%) patients. Two colorrectal anastomoses (14.2%) were performed at fewer than 5 cm from the anal verge and 13 (61.9%) were performed at more than 5 cm from the anal verge. Three of the anastomoses were ileocolic (14.2%), two were coloanal (9.5%), and one was ileoanal (4.7%). There were six (28.5%) complications, no cases of anastomotic leak, and no complications associated with the use of indocyanine green. The mortality rate was 0%. CONCLUSION: The present case series is the first on colorrectal surgery conducted in Mexico using indocyanine green fluorescence angiography, with excellent results.

Detección de anticuerpos tipo igg contra borrelia burgdorferi, y factores asociados a la enfermedad de lyme en población canina, de los municipios honda-Tolima, la mesa y Chia-Cundinamarca. detección de anticuerpos tipo igg contra borrelia burgdorferi en población canina / Detection of IgG-type antibodies against Borrelia Burgdorferi, and factors associated with Lyme disease in the canine population, of the municipalities Honda-Tolima, La Mesa and Chia-Cundinamarca

Borrelia burgdorferi, is the spirochete responsible for causing Lyme disease in man and different animals. Objective. Detect specific IgG type antibodies against Borrelia burgdorferi, in canines using the immunofluorescence technique and its correlation with other factors associated with Lyme disease. Methods. Blood was taken for IgG detection against Borrelia burgdorferi sl; Peripheral blood smear of the canines and hemolymph of the ticks to search for spirochetes with Wright staining and finally classification of the ticks using morphometric keys. Results. In the serological test, on average 69.0% of the canines sampled gave positive results in the different titrations. Bacterial structures were spirally visualized both in the peripheral blood of the canines and in the hemolymph of the ticks. The vector found was classified as Rhipicephalus sanguineus, until now not reported in the scientific literature as a carrier of Borrelia, nor associated with the disease.

[The relevance of electron microscopy in kidney biopsies to 21st century pathology]. / Microscopía electrónica en biopsias renales: una evaluación de su utilidad en el siglo XXI.

INTRODUCTION: Electron microscopy (EM) has been used in the study of renal biopsies for more than 5 decades; however, it is expensive and the possibility of restricting it to selected cases has been considered. This study aims to reevaluate the necessity for EM in the diagnosis of renal biopsies today. MATERIAL AND METHODS: All renal biopsies taken between 2016 and 2019 with adequate light microscopy (LM), immunofluorescence (IF) and EM studies were included. The initial diagnosis (without EM) and the final diagnosis (with EM) was recorded. EM was considered necessary in cases in which the initial and final diagnoses did not concur, when diagnosis could not be made with LM and IF only or if the EM study revealed further clinically relevant findings. RESULTS: A total of 621 biopsies were included, 498 (80.2%) of native kidneys and 123 (19.8%) of transplanted kidneys. In 115 cases (18.5%) EM had been deemed necessary for diagnosis; it was required more frequently in hereditary diseases (96.8%) and isolated hematuria (88.9%) but less often in nephrotic syndrome (6.7%) and renal transplant biopsy (5.7%) (p < 0.001). CONCLUSIONS: EM was required in less than a fifth of renal biopsies, being more necessary in isolated hematuria and hereditary diseases and less so in nephrotic syndrome and in renal graft biopsies. These findings may prove useful as a guide to case selection protocols in which EM could be considered as a non-mandatory technique.

Armonización de la determinación de ANCA por inmunofluorescencia indirecta: primera reunión argentina / Harmonization of the determination of ANCA by indirect immunofluorescence: first argentine meeting / Harmonização da determinação de ANCA por imunofluorescência indireta: primeira reunião argentina

Resumen La determinación de anticuerpos anti-citoplasma de neutrófilos (ANCA) es utilizada en la clínica diaria como una herramienta diagnóstica en distintas formas de vasculitis de pequeños vasos (vasculitis asociadas a ANCA), incluyendo la granulomatosis con poliangeítis (antes: granulomatosis de Wegener), poliangeítis microscópica y granulomatosis eosinofílica con poliangeítis (antes: síndrome de Churg-Strauss), y como apoyo diagnóstico de colitis ulcerosa, colangitis esclerosante primaria y enfermedad de Crohn. Estos anticuerpos están dirigidos contra distintos epítopos antigénicos de diferentes proteínas presentes en el citoplasma del neutrófilo. La determinación de ANCA, habitualmente realizada por la técnica de inmunofluorescencia indirecta (IFI), presenta cierto grado de complejidad en la definición de las imágenes de los diferentes patrones, variabilidad en la forma de trabajo y expresión de resultados. El 24 de octubre de 2018 en el marco del X Congreso Argentino de la Calidad en el Laboratorio Clínico y VIII Jornadas Latinoamericanas de la Calidad en el Laboratorio Clínico, en la Ciudad de Buenos Aires, se realizó una reunión de Armonización de la Determinación de ANCA por IFI con el objeto de presentar, discutir y consensuar los distintos aspectos que se presentan en esta técnica. Las propuestas iniciales fueron discutidas arribándose a recomendaciones generales para proporcionar estándares de trabajo e interpretación de imágenes con el objeto de disminuir la variabilidad de resultados entre los laboratorios clínico-inmunológicos.

Abstract Anti-neutrophil cytoplasmic antibodies (ANCA) tests are widely used in daily clinical practice as a useful tool for the diagnosis of pathologies such as granulomatosis with polyangiitis, microscopic polyangiitis, pauci-immune necrotizing segmental glomerulonephritis and eosinophilic granulomatosis with polyangiitis, ulcerative colitis, primary sclerosing cholangitis, Crohn's disease, etc. These antibodies are directed against different antigenic epitopes of various proteins which are present in the neutrophil cytoplasm. ANCA testing is usually carried out by using indirect immunofluorescence (IIF) method. The determination of ANCA presents some difficulties in the definition of the images of the different patterns, work protocols and result reports uniformity. The "Harmonization Conference on the Determination of ANCA by IIF" was held within the framework of the X Argentine Congress on Quality in the Clinical Laboratory and VIII Latin American Conference on Quality in the Clinical Laboratory, in Buenos Aires, Argentina, on October 24, 2018, in order to present, discuss and agree on different aspects of this method. Some initial proposals were discussed, arriving at general recommendations to provide standards of work and an image interpretation, with the aim of reducing the variability of results among the clinical-immunological laboratories.

Resumo A determinação de anticorpos anti-citoplasma de neutrófilos (ANCA) é utilizada na prática clínica diária como um suporte diagnóstico para várias patologias, como granulomatose com poliangiite, poliangiite microscópica, glomerulonefrite segmentar necrotizante pauci-imune e granulomatose eosinofílica com poliangiite, poliarterite nodosa, colite ulcerosa, colangite esclerosante primária, doença de Crohn. Esses anticorpos são dirigidos contra diferentes epítopos antigênicos de diferentes proteínas presentes no citoplasma do neutrófilo. A determinação de ANCA, normalmente realizada pela técnica de imunofluorescência indireta IFI, apresenta certo grau de complexidade na definição das imagens dos diferentes padrões, variabilidade na forma de trabalho e expressão de resultados. Em 24 de outubro de 2018, no âmbito do X Congreso Argentino de la Calidad en el Laboratorio Clínico (X Congresso Argentino da Qualidade no Laboratório Clínico) e VIII Jornadas Latinoamericanas de la Calidad en el Laboratorio Clínico (VIII Jornadas Latino-americanas da Qualidade no Laboratório), na cidade de Buenos Aires, foi realizada uma Jornada de Harmonização da Determinação de ANCA pelo IFI a fim de apresentar, discutir e concordar sobre os diferentes aspectos que são apresentados nesta técnica. As propostas iniciais foram discutidas, chegando a recomendações gerais para fornecer padrões de trabalho e interpretação de imagens visando reduzir a variabilidade de resultados entre os laboratórios clínico-imunológicos.

Virus respiratorios y características clínico-epidemiológicas en los episodios de infección respiratoria aguda / Respiratory viruses and clinical- epidemiological characteristics in episodes of acute respiratory infection

RESUMEN Con el objetivo de determinar la frecuencia viral y las características clínico-epidemiológicas en los episodios de infección respiratoria aguda de pacientes del Instituto Nacional de Salud del Niño San Borja en Lima, se analizó la información de los episodios de pacientes que requirieron al menos una prueba de inmunofluorescencia directa desde el 1 de enero del 2017 al 31 de diciembre del 2018. Se identificaron 1153 episodios en 707 pacientes. La mediana de la edad fue de 1 año y el 55% fueron del sexo masculino. La frecuencia viral fue del 13,4%; el virus respiratorio sincitial se identificó en el 10,7% de los episodios. La frecuencia viral fue mayor en los menores de 1 año (16,2%); en aquellos con enfermedad congénita respiratoria (38,9%) y durante el otoño (24,2%). Los síntomas más comunes fueron tos (70,3%) y fiebre (53,4%); y los principales diagnósticos fueron neumonía viral (31,8%) y bronquiolitis (23,4%). Se concluye que la frecuencia viral respiratoria estuvo relacionada con la edad, estacionalidad y patología preexistente.

ABSTRACT The aim of the study was to determine the viral frequency and clinical-epidemiological characteristics in the episodes of acute respiratory infection in patients of the Instituto Nacional de Salud del Niño San Borja in Lima, the information of the episodes of patients who required at least one direct Immunofluorescence test from January 1, 2017 to December 31, 2018 was analyzed. 1153 episodes were identified in 707 patients. The median age was 1 year and 55% were male. The viral frequency was 13.4%; respiratory syncytial virus was identified in 10.7% of the episodies. The viral frequency was higher in children under 1 year of age (16.2%); in those with congenital disease respiratory (38.9%) and during the autumn (24.2%). The most common symptoms were cough (70.3%) and fever (53.4%); and the main diagnoses, viral pneumonia (31.8) and bronchiolitis (23.4%). It is concluded that the respiratory viral frequency was related to age, seasonality and pre-existing pathology.

Virus respiratorios y características clínico-epidemiológicas en los episodios de infección respiratoria aguda / Respiratory viruses and clinical- epidemiological characteristics in episodes of acute respiratory infection

RESUMEN Con el objetivo de determinar la frecuencia viral y las características clínico-epidemiológicas en los episodios de infección respiratoria aguda de pacientes del Instituto Nacional de Salud del Niño San Borja en Lima, se analizó la información de los episodios de pacientes que requirieron al menos una prueba de inmunofluorescencia directa desde el 1 de enero del 2017 al 31 de diciembre del 2018. Se identificaron 1153 episodios en 707 pacientes. La mediana de la edad fue de 1 año y el 55% fueron del sexo masculino. La frecuencia viral fue del 13,4%; el virus respiratorio sincitial se identificó en el 10,7% de los episodios. La frecuencia viral fue mayor en los menores de 1 año (16,2%); en aquellos con enfermedad congénita respiratoria (38,9%) y durante el otoño (24,2%). Los síntomas más comunes fueron tos (70,3%) y fiebre (53,4%); y los principales diagnósticos fueron neumonía viral (31,8%) y bronquiolitis (23,4%). Se concluye que la frecuencia viral respiratoria estuvo relacionada con la edad, estacionalidad y patología preexistente.

ABSTRACT The aim of the study was to determine the viral frequency and clinical-epidemiological characteristics in the episodes of acute respiratory infection in patients of the Instituto Nacional de Salud del Niño San Borja in Lima, the information of the episodes of patients who required at least one direct Immunofluorescence test from January 1, 2017 to December 31, 2018 was analyzed. 1153 episodes were identified in 707 patients. The median age was 1 year and 55% were male. The viral frequency was 13.4%; respiratory syncytial virus was identified in 10.7% of the episodies. The viral frequency was higher in children under 1 year of age (16.2%); in those with congenital disease respiratory (38.9%) and during the autumn (24.2%). The most common symptoms were cough (70.3%) and fever (53.4%); and the main diagnoses, viral pneumonia (31.8) and bronchiolitis (23.4%). It is concluded that the respiratory viral frequency was related to age, seasonality and pre-existing pathology.

Determinación de anticuerpos anti-dsDNA: evaluación del desempeño de un inmunoensayo quimioluminiscente y correlación con el método de inmunofluorescencia indirecta / Detection of anti-dsDNA antibodies: evaluation of performance of a chemiluminescent immunoassay and correlation to the indirect immunofluorescence assay / Determinação de anticorpos anti-dsDNA: avaliação do desempenho de um imunoensaio quimioluminiscente e correlação com o método de imunoflourescência indireta

Resumen La determinación de anticuerpos anti-dsDNA es de utilidad para el diagnóstico y seguimiento clínico de pacientes con lupus eritematoso sistémico (LES) y es uno de los criterios de clasificación del SLICC-2012. El objetivo del estudio fue verificar el desempeño del inmunoensayo quimioluminiscente (CLIA) QUANTA Flash dsDNA y compararlo con el método en uso de inmunofluorescencia indirecta en Crithidia luciliae (CLIFT). Se analizaron con ambos métodos 195 pacientes con diagnóstico de enfermedades del tejido conectivo y solicitud de anticuerpos anti-dsDNA. Se obtuvieron 38 sueros positivos, 133 negativos y 24 (12,3%) discordantes. Entre estos resultados discordantes, hubo 17 que correspondieron a pacientes con LES y se agruparon en 16 CLIA+/CLIFT- y 1 CLIA-/CLIFT+. Se verificó el desempeño para precisión siguiendo el protocolo EP15-A2 y la linealidad. Se estudió la concordancia mediante el coeficiente Kappa y la correlación con el coeficiente Rho de Spearman. Se observó mayor sensibilidad diagnóstica para CLIA. El grado de acuerdo fue moderado y se obtuvo buena correlación entre los valores cuantitativos de CLIA y los títulos de CLIFT. De acuerdo al buen desempeño encontrado y a los resultados discordantes analizados, la mejor estrategia para la implementación de CLIA sería utilizarla en combinación con CLIFT, lo que aumentaría la sensibilidad diagnóstica sin perder especificidad.

Abstract The detection of anti-dsDNA antibodies is useful for diagnosis and clinical monitoring of patients with systemic lupus erythematosus (SLE) and it is part of the classification criteria according to SLICC 2012. The purpose of this study was to verify the performance of the chemiluminescent immunoassay (CLIA) QUANTA Flash dsDNA and to compare it with the method currently in use, i.e the indirect immunofluorescence in Crithidia luciliae (CLIFT). One hundred and ninety five patients, who presented connective tissue diseases and required the study of anti-dsDNA antibodies, were analyzed. Thirty eight positive serum samples were obtained, 133 were negative and 24 (12.3%) in disagreement. Within the discordant results, there were 17 that corresponded to patients with SLE and they were grouped in 16 CLIA+/CLIFT- and 1 CLIA-/CLIFT+. The accuracy performance was assessed according to the EP15-A2 protocol and linearity. Concordance and correlation were calculated with the Kappa and Spearman's Rho coefficient, respectively. Based on the good performance observed and the discordant results analyzed, the best strategy for CLIA implementation would be to combine it with CLIFT, which would increase the diagnostic sensitivity without losing specificity.

Resumo A determinação dos anticorpos anti-dsDNA é de utilidade para o diagnóstico e seguimento clinico de pacientes com lúpus eritematoso sistêmico (LES) e é um dos critérios de classificação do SLICC 2012. O objetivo do estudo foi verificar o desempenho do imunoensaio quimioluminescente (CLIA) QUANTA Flash dsDNA e compará-lo com o método em uso imunofluorescência indireta em Crithidia luciliae (CLIFT). Foram analisados com os dois métodos, 195 pacientes com diagnóstico de doenças do tecido conjuntivo e solicitude de anticorpos anti-dsDNA. Os resultados foram agrupados em 38 soros positivos, 133 negativos e 24 (12,3%) discordantes. Entre esses resultados discordantes, 17 corresponderam a pacientes com LES e se agruparam em 16 CLIA+/CLIFT- e 1 CLIA-/CLIFT+. Foi verificado o desempenho para precisão seguindo o protocolo EP15-A2 e a linearidade. Foi estudada a concordância mediante o coeficiente Kappa e correlação com o coeficiente Rho de Spearman. Observou-se maior sensibilidade diagnóstica para CLIA. O grau de acordo foi moderado e boa correlação foi observada entre os valores quantitativos de CLIA e os títulos de CLIFT. Com base no bom desempenho encontrado e nos resultados discordantes analisados, a melhor estratégia para implementar o CLIA seria utilizá-lo em combinação com o CLIFT, o que aumentaria a sensibilidade do diagnóstico sem perder a especificidade.

[Localization of Arylsulfatase A during in vitro incubation of human spermatozoa in capacitation media]. / Localización de arilsulfatasa A durante la incubación in vitro de espermatozoides humanos en medio de capacitación.

OBJECTIVE: To determine the localization and distribution of the ArylsulfataseA receptor (ARSA) in human spermatozoa before and after their incubation in capacitation medium for 1 and 4hours. MATERIAL AND METHODS: Semen samples were obtained from five normozoospermic donors. Capacitation was by swim-up technique using capacitation medium for 1 and 4hours. Localization of the ARSA receptor was assessed by indirect immunofluorescence using confocal microscopy. A minimum of 200cells were observed in each physiological condition. RESULTS: Before incubation, no representative pattern was observed among the cells positive for this biomarker (8.61%). This percentage increased significantly after incubation in the capacitation medium for 1 and 4hours (61.86% and 63.38% respectively). A majority pattern was observed among the capacitated cells, with intense labelling in the acrosomal region (27.11% and 28.20% after 1 and 4hours respectively). It should be noted that the pattern corresponding to fluorescence at the level of the periacrosomal region was not observed in the spermatozoa prior to incubation. Only after incubation in capacitation medium for 1 and 4hours, 9.13% and 12.78% of cells with such distribution were detected. CONCLUSIONS: In vitro capacitation, regardless of time, favours the immunolocalization of ARSA in the cephalic region of the spermatozoa. The most representative subpopulation after this process was the one in which ARSA was intensely and homogeneously distributed in the acrosome region, involved in primary gamete recognition.

Diferencias fenotípicas de fibroblastos gingivales en sujetos con hiperplasia gingival idiopática frente a sujetos periodontalmente sanos: estudio piloto / Phenotypic differences of gingival fibroblasts in subjects with idiopathic gingival hyperplasia versus periodontally healthy subjects: a pilot study

Objetivo: identificar, describir y diferenciar las características fenotípicas de los fibroblastos gingivales (FGs) en pacientes con hiperplasia gingival idiopática (HGI) e individuos periodontalmente sanos. Métodos: los FGs fueron aislados a partir de tejido gingival de individuos periodontalmente sanos (n=2) y pacientes con HGI (n=2). Los FGs se cultivaron en el medio DMEM (Dulbecco's Modified of Eagle Medium) a 37°C con 5% de CO2. La identificación y localización de la actina, vimentina y mitocondrias en FGs fue realizada y evaluada microscópicamente mediante inmunofluorescencia con anticuerpos monoclonales. La capacidad de migración de los FGs en los pacientes con HGI e individuos sanos también fue estudiada. Resultados: todos los FGs fueron mononucleares, fusiformes y con prolongaciones citoplasmáticas visibles. La faloidina permitió identificar una densa red de actina en los FGs de pacientes con HGI, contrariamente a los FGs de individuos periodontalmente sanos. La vimentina y mitocondrias fueron identificadas en los FGs de individuos sanos y pacientes con HGI sin ninguna alteración en su expresión y localización. La migración de la monocapa de los FGs indicó una actividad de migración celular importante en los FGs de los pacientes con HGI, en relación a los FGs de los individuos periodontalmente sanos. Conclusión: los FGs de pacientes con HGI conservan características fenotípicas celulares similares a los FGs de individuos periodontalmente sanos. Sin embargo, los FGs de pacientes con HGI simulan tener una mayor capacidad migratoria que amerita ser explorada en futuros trabajos de investigación.

Objective: To identify and to describe the phenotypic characteristics of gingival fibroblasts from patients with idiopathic gingival hyperplasia (IGH) and periodontally healthy individuals. Methods: Gingival fibroblasts (GFs) were isolated from gingival tissue from periodontally healthy individuals (n=2) and patients with IGH (n=2). The GFs were grown in DMEM (Dulbecco's Modified of Eagle Medium) at 37°C with 5% CO2. The identification and location of actin, vimentin and mitochondria in GFs were performed and evaluated microscopically by immunofluorescence with monoclonal antibodies. The migration capacity of GFs from IGH and healthy individuals was also studied. Results: All the GFs were mononuclear, fusiform and with visible cytoplasmic extensions. The phalloidin allowed to identify a dense actin network in the GFs of patients with IGH, contrary to the GFs of periodontally healthy individuals. Vimentin and mitochondria were identified in the GFs of healthy individuals and patients with IGH without any alteration in their expression and location. Monolayer migration of GFs indicates significant cell migration activity in the GFs of patients with IGH in relation to the GFs of periodontally healthy individuals. Conclusion: GFs from patients with IGH retain cellular phenotypic characteristic similar to GFs from periodontally healthy individuals. However, the GFs of patients with IGH simulate having a greater migratory capacity that deserves to be explored in future research works.

Serologic study of Bartonella sp. infection among human population of Southern Spain.

INTRODUCTION: The purpose of this study was to determine the prevalence of IgG antibodies against Bartonella sp. in a randomly selected sample from the population of the patients of North Sanitary District of Jaén. METHODS: We used a commercially available immunofluorescent test (Focus-Technology IFA Bartonella quintana and B. henselae test). RESULTS: Six hundred five healthy individuals were divided by sex into three age groups. We detected that 13.55% and 11.07% subjects were IgG seropositive to B. henselae and B. quintana, respectively. CONCLUSIONS: Our data show that the prevalence of both Bartonella species in Andalusia (Southern Spain) is relatively high. No statistical difference in the seropositivity was observed among these groups. In both cases, the IgG antibody titers ranged from 1/128 to 1/512.

Frecuencia de patrones de anticuerpos antinucleares, en pacientes con sospecha de enfermedades sistémicas reumáticas autoinmunes / Frequency of antinuclear antibodies patterns in patients with suspected of systemic autoimmune rheumatic diseases / Frequência de Padrões de anticorpo antinucleares em pacientes com suspeita de doenças sistêmicas reumáticas autoimunes / Fréquence des profils d'anticorps antinucléaires dans les cas de suspicion de maladies rhumatismales auto-immunes systémiques

Resumen Objetivo: El objetivo de este trabajo es describir la frecuencia de los patrones de tinción de Anticuerpos antinucleares (ANA), en pacientes con sospecha de enfermedades autoinmunes, en el sureste mexicano. Materiales y métodos: Se emplearon células Hep-2 y anticuerpo anti-IgG acoplado a FITC (EuroimmunTM) para el análisis de las muestras a través de inmunofluorescencia indirecta. Resultados: Del total de los pacientes, 89 fueron mujeres (87.2%) y 13 hombres (12.7%), en edades de 2 a 88 años. Se observó que 85 muestras (70.6 %) correspondieron al patrón nuclear, 10 (9.8 %) al patrón citoplasmático y 7 (6.8 %) al patrón mitótico. De los patrones nucleares, 37 (36.8%), 17 (16.7%) y 12 (11.8%) correspondieron a patrones homogéneo, granular fino y granular grueso respectivamente. Conclusiones: En este trabajo se observó, que la mayor frecuencia de anticuerpos antinucleares se encontró en pacientes en edad productiva. Los patrones más observados fueron el homogéneo, granular fino y granular grueso. El patrón homogéneo se asocia a LES cuando se presenta en títulos altos.

Abstract Objective: The objective of this work was to describe the frequency of antinuclear antibody (ANA) staining patterns in patients with suspected autoimmune diseases in southeastern Mexico. Materials and methods: Hep2 cells and Anti-Human IgG coupled to FITC (EuroimmunTM) were used for analyzing samples through indirect immunofluorescence. Results: 87.2 % per cent (89) of the total number of patients were women and 12.7 % were men (13) aged from 2 to 88 years. It was observed that 85 samples (70.6%) corresponded to the nucleolar pattern, 10 (9.8%) to the cytoplasmic pattern and 7 (6.8%) to the mitotic pattern. Of the nuclear patterns, 37 (36.8%), 17 (16.7%) and 12 samples (11.8%) corresponded to homogeneous, nuclear fine speckled and nuclear coarse speckled patterns respectively. Conclusions: In this work, it was observed that the highest frequency of antinuclear antibodies was found in patients of productive age. The most observed patterns were the homogeneous, nuclear fine speckled and nuclear coarse speckled. The homogeneous pattern is associated with SLE when presented in high titers.

Resumo Objetivo: deste trabalho é descrever a frequência dos padrões de coloração de anticorpos antinucleares (ANA) em pacientes com suspeita de doenças autoimunes no sudeste mexicano. Materiais e métodos: células Hep-2 e anticorpo anti-IgG acoplado a FITC (EuroimmunTM) foram utilizados para a análise das amostras por imunofluorescência indireta. Resultados: Do total de pacientes, 89 eram mulheres (87,2%) e 13 homens (12,7%), com idade entre 2 e 88 anos. Observou-se que 85 amostras (70,6%) corresponderam ao padrão nuclear, 10 (9,8%) ao padrão citoplasmático e 7 (6,8%) ao padrão mitótico. Dos padrões nucleares, 37 (36,8%), 17 (16,7%) e 12 (11,8%) corresponderam aos padrões homogêneo, granular fino e granular grosso, respectivamente. Conclusões: Neste trabalho observou-se que a maior frequência de anticorpos antinucleares foi encontrada em pacientes em idade produtiva. Os padrões mais observados foram homogêneo, granular fino e granular grosso. O padrão homogêneo está associado ao LES quando ocorre em altos títulos.

Résumé Objectif : L'objectif de ce travail est de décrire la fréquence des profils de coloration des Anticorps Antinucléaires (ANA), dans les cas de suspicion de maladies auto-immunes, dans le sud-est du Mexique. Matériels et méthodes : Des cellules Hep-2 et un anticorps anti-IgG couplé au FITC (EuroimmunTM) ont été utilisés pour l'analyse desé chantillons par immunofluorescence indirecte. Résultats : Sur le total des patients, 89 étaient des femmes (87,2 %) et 13 des hommes (12,7 %), tous âgés de 2 à 88 ans. Il a été observé que 85 échantillons (70,6 %) correspondaient au patron nucléaire, 10 (9,8 %) au patron cytoplasmique et 7 (6,8 %) au patron mitotique. Parmi es patrons nucléaires, 37 (36,8%), 17 (16,7%) et 12 (11,8%) avaient respectivement un aspect homogène, moucheté à grains fins et moucheté à gros grains. Conclusions : Dans travail, il a été observé que la fréquence la plus élevée d'anticorps antinucléaires a été trouvée chez les patients en âge productif. Les aspects les plus observés ont été de type homogène, moucheté à grains fins et moucheté à gros grains. L'aspect homogène est associé à l'ELS lorsqu'il est présenté en titres élevés.

Perspectivas en investigación: células HEp-2 infectadas con EB's de Chlamydia trachomatis serovar 2 (VR-902B) / Chlamydia trachomatis: Research Perspectives. HEp-2 Cells Infected with EB's from Chlamydia trachomatis serovar 2 (VR-902B): Research Perspectives

Resumen Chlamydia trachomatis (C. trachomatis) es una bacteria Gram negativa inmóvil, caracterizada por ser un microorganismo intracelular obligado y por poseer un ciclo reproductivo en el que puede distinguirse una forma infecciosa extracelular metabólicamente inerte (cuerpo elemental - EB's), y una forma no infecciosa intracelular y activa (cuerpo reticulado - RB's). C trachomatis se caracteriza por causar infección en humanos, está relacionada con enfermedades de transmisión sexual e infecciones oculares; por lo que puede conllevar a secuelas de interés, si no se da un tratamiento oportuno. El objetivo de este estudio fue optimizar el modelo de infección de C. trachomatis en células HEp-2 con cuerpos elementales (EB's) de C. trachomatis serovar L2. Inicialmente, se establecieron las condiciones para el crecimiento adecuado de las células HEp-2 en tiempo y con una confluencia del 90%, para continuar con la optimización de un protocolo de infección. La infección fue confirmada a partir de la coloración con Giemsa permitiendo evaluar características morfológicas tanto de las células HEp-2 sin infectar e infectadas, y así mismo, de los cuerpos elementales de C. trachomatis. Finalmente, se corroboró la infección con la técnica de inmunofluorescencia directa que detecta la proteína de membrana MOMP de C. trachomatis. Tras los ensayos realizados se evidenció la presencia de cuerpos elementales próximos y dentro del citoplasma celular, así como células vacuoladas y daño celular causado por la infección.

Abstract Chlamydia trachomatis (C. Trachomatis) is a Gram negative unmoving bacterium, characterized by being an obligate intracellular microorganism and having a reproductive cycle in which a metabolically inactive extracellular infectious form (elementary body - EB's) can be distinguished from an intracellular active and non-infectious form (reticulated body - RB's). C trachomatis is characterized by causing infection in humans, is related to sexually transmitted diseases and eye infections, so it can lead to sequelae of interest if timely treatment is not given. The objective of this study was to optimize the infection model of C. trachomatis in HEp-2 cells with elementary bodies (EB's) of C. trachomatis serovar L2. Initially, the conditions for the adequate growth of HEp-2 cells were established in time and with a confluence of 90%, to continue with the optimization of an infection protocol. The infection was confirmed from the staining with Giemsa allowing to evaluate morphological characteristics of both uninfected and infected HEp-2 cells and also of the elementary bodies of C. trachomatis. Finally, the infection was corroborated with the direct immunofluorescence technique, that detects the C. trachomatis MOMP membrane protein. After the tests were performed, the presence of elementary bodies nearby and within the cellular cytoplasm was evidenced, as well as vacuolated cells and cellular damage caused by the infection.

Dinámica de anticuerpos e infección activa por Anaplasma marginale en becerras / Dynamics of Anaplasma marginale Active Infection and Antibodies in Calves

Resumen Históricamente, la especie bovina ha sido de vital importancia para el hombre como animal de abasto, por lo que el cuidado y protección a esta especie es muy importante. A nivel mundial existen diversas enfermedades que atacan el ganado bovino, entre las cuales se encuentra anaplasmosis bovina. La presente investigación no experimental de campo tiene como objetivo evaluar la dinámica de anticuerpos anti-Anaplasma y determinar en qué momento el animal presenta infección activa. Para ello, se escogió una población de 35 becerras ubicadas en la unidad de producción del municipio Crespo, en el estado de Lara, Venezuela. A las becerras, de 0 a 7 días de nacidas, se le extrajo la sangre en tubos con anticoagulante, para realizar frotis de capa blanca y determinación del hematocrito, y sin anticoagulante, para la inmunofluorescencia indirecta. Posteriormente, fueron muestreadas cada 15 días hasta obtener un total de 12 muestras por cada becerra. Para analizar los datos, fue utilizada la prueba estadística X2 del programa estadístico Epi Info versión 3.5.4. Se obtuvo un 79,05 % de positividad y 100 % de seropositividad para Anaplasma marginale. Se concluyó que los altos valores obtenidos con las pruebas serológicas son indicativos de animales con memoria inmunológica, lo que quiere decir que no necesariamente estarían infectados al momento del muestreo. Asimismo, se confirma la capacidad de la especie bovina de crear un estado de premunición ante la anaplasmosis a edades tempranas y mantenerla por un periodo prolongado, de modo que se encuentra una estabilidad enzoótica en la unidad de producción.

Abstract Bovine species has been historically of high importance to the human beings as a provision animal. Therefore, it is highly important to care and protect this species. There are different diseases attacking the bovines around the world, such as the bovine anaplasmosis. This non-experimental research in the field aims to evaluate the dynamics of anti-Anaplasma antibodies and to determine when the animal has an active infection. To do so, a population of 35 female calves was used from the production unit in the town Crespo, Lara State, Venezuela. Female calves, from 0 to 7 days born, were taken blood samples in test tubes with anticoagulant. Then a white smear test was carried out and the hematocrit was determined. Samples without anticoagulant were taken to do an indirect immunofluorescence test. The calves were taken blood samples every 15 days in order to complete 12 samples from each individual calf. For the data analysis, the statistical test X2 from the software Epi Info v.3.5.4 was used. Positive tests accounted for 79.05 % of the samples and seropositiveness for Anaplasma marginale reached 100%. It is concluded that the high values resulting from the serologic tests indicate that these animals have an immune memory, which means, in turn, that they were not necessarily infected at the time of being taken the samples. Likewise, this study confirms that this species is able to create a pre-immunity condition to the anaplasmosis at an early age and to keep it for a long term. This way, they reach an enzootic balance in the production unit.

Indocyanine green fluorescence angiography in colorrectal surgery. First case series in Mexico. / Angiografía por fluorescencia con verde de indocianina para cirugía de colon y recto. Primera serie reportada en México.

INTRODUCTION AND AIM: Anastomotic leak occurs in 1-19% of colorrectal surgeries. Our objective was to present the first Mexican case series on colorrectal surgery using indocyanine green fluorescence angiography to evaluate perfusion prior to carrying out the anastomosis. MATERIALS AND METHODS: A retrospective, analytic, descriptive study was conducted. We studied the case records of consecutive patients that underwent colorrectal surgery with indocyanine green angiography performed by the same group of colorrectal surgeons. RESULTS: Twenty-one case records were reviewed. Eleven (52.3%) of the patients were women, mean patient age was 57 years (38-82), and mean body mass index was 25 kg/m2 (17-34). Fifteen (71.4%) patients were diagnosed with malignant disease. Indocyanine green angiography changed our therapeutic decision in three (14.2%) patients. Two colorrectal anastomoses (14.2%) were performed at fewer than 5 cm from the anal verge and 13 (61.9%) were performed at more than 5 cm from the anal verge. Three of the anastomoses were ileocolic (14.2%), two were coloanal (9.5%), and one was ileoanal (4.7%). There were six (28.5%) complications, no cases of anastomotic leak, and no complications associated with the use of indocyanine green. The mortality rate was 0%. CONCLUSION: The present case series is the first on colorrectal surgery conducted in Mexico using indocyanine green fluorescence angiography, with excellent results.