A rapid homogenous bioassay for detection of thyroid-stimulating antibodies based on a luminescent cyclic AMP biosensor.

Graves' disease (GD) is an autoimmune disease caused by antibodies to the thyroid stimulating hormone receptor (TSHR). The FDA-cleared Thyretain™ TSI bioassay is a highly specific method to detect thyroid stimulating antibodies (TSAb/TSI) in the blood of patients with autoimmune thyroid disease (AITD), particularly GD. To simplify the workflow of this bioassay and to support a semi-quantitative result, we have generated a stable CHO-K1 cell line expressing both a chimeric TSH receptor (TSHR-Mc4) and a luciferase-based homogeneous cAMP biosensor (GS luciferase). Here, we describe a rapid, real-time, homogenous bioassay (Turbo™ TSI Bioassay) to directly assess the functional activity of TSI and produce results in International Units of IU/L. The Turbo™ TSI bioassay works by measuring changes in the intracellular cAMP level induced by a G-protein coupled receptor (G-PCR) signaling cascade which is triggered by the binding of TSI to the TSHR. Upon binding to cAMP, the GS luciferase reporter is activated through conformational changes and generates light that can be measured in intact cells with a luminometer. The LoD and LoQ of the assay were determined to be 0.016 IU/L and 0.03 IU/L, respectively and the preliminary assay cutoff was determined to be 0.024 IU/L by ROC analysis using the Thyretain™ TSI bioassay results as reference. The analytical performance of the Turbo™ TSI bioassay is comparable to the Thyretain™ TSI bioassay as evidenced by similar EC50 values for a TSHR stimulating monoclonal antibody (M22). The specificity of the Turbo™ TSI bioassay was demonstrated by showing no response to a high concentration of a human monoclonal TSHR blocking antibody (K1-70). The precision of the assay was excellent with an overall within-laboratory precision <15% CV. When testing 198 clinical samples, the positive and negative percent agreement between the Turbo™ TSI and the Thyretain™ TSI bioassays were 98.7% and 93.5%, respectively. While both bioassays yield equivalent analytical and clinical performances, the Turbo™ TSI bioassay is much simpler to perform. It does not require cell culture, sample dilution, washing or cell lysis steps, resulting in a dramatically reduced turnaround time from about 21 h to 60 min. In addition, the same cell line showed its capability of detecting thyroid blocking antibodies (TBAb/TBI) in a competitive format. The Turbo™ TSI bioassay is user-friendly and is a very promising advancement to aid the diagnosis of autoimmune thyroid disease (AITD).

Significados y representaciones sociales atribuidos a los miedos de los hijos: Promoción de la parentalidad positiva / Significados e representações sociais atribuídos aos medos dos filhos: promoção para a parentalidade positiva / Meanings and social representations attributed to children’s fears: promotion for positive parenting

Objetivo: Conocer las representaciones sociales y los significados de los padres en lo querespecta a los miedos de los hijos, para definir intervenciones que promuevan la parentalidadpositiva en este área de dificultades del desarrollo infantil. Método: Estudio descriptivo basadoen la Teoría de las Transiciones de Meleis y en la Teoría de las Representaciones Sociales deAbric. La evaluación diagnóstica se realizó entre mayo y junio de 2019, utilizando unarecopilación libre de palabras a través del estímulo inductor – miedo de los niños y adolescentes.La muestra fue constituida por 143 padres de hijos ≤ a 18 años. Resultados: Se identificaron lossignificados cuánto a la posición en el núcleo central y periférico, que ayudaron en lacategorización de seis tipos de parentalidad: parentalidad en sufrimiento, parentalidad condificultades, parentalidad positiva, parentalidad atenta, parentalidad preocupada y parentalidaddesconectada. Conclusiones: El proyecto de promoción para la parentalidad positiva en relaciónal miedo de los hijos basado en el conocimiento de los significados atribuidos permitió no soloentender la dimensión del problema, cómo desarrollar intervenciones y materiales adecuados, asícomo indicar el mejor abordaje para promover la parentalidad positiva en relación a los miedosde los hijos.(AU)

Objective: Know the social representations and the meanings attributed to children’s fearsby the parents, for delineate interventions that promote positive parenting in this area of childdevelopment difficulties. Method: descriptive study based on Meleis’ Theory of Transitions andAbric’s Theory of Social Representations. A diagnostic evaluation was performed between Mayand June of 2019, using a free word evocation through the stimulus inductor – children’s andadolescents’ fears. The sample consisted of 143 parents of children ≤ 18 years. Results: themeanings were identified by the position in the central and peripheral nucleus, which helped inthe categorization of six types of parenting: parenting in distress, parenting with difficulties,positive parenting, attentive parenting, concerned parenting, and disconnected parenting.Conclusions: The project for positive parenting promotion in relation to children’s fears based onthe knowledge of the meanings attributed allowed not only to understand the dimension of theproblem, but also to develop appropriate interventions and materials, as well as indicate the bestapproach to promote positive parenting in relation to children’s fears.(AU)

Objetivo: conhecer as representações sociais e os significados dos pais no que diz respeitoaos medos dos filhos, para delinear intervenções que promovam a parentalidade positiva nestaárea de dificuldades do desenvolvimento infantil. Método: estudo descritivo baseado na Teoriadas Transições de Meleis e na Teoria das Representações Sociais de Abric. Foi realizada aavaliação diagnóstica entre maio e junho de 2019, com recurso à evocação livre de palavrasatravés do estímulo indutor – medo das crianças e adolescentes. A amostra foi constituída por 143pais de filhos ≤ a 18 anos. Resultados: identificaram-se os significados quanto à posição no núcleocentral e periférico, que auxiliaram na categorização de seis tipos de parentalidade: parentalidadeem sofrimento, parentalidade com dificuldades, parentalidade positiva, parentalidade atenta,parentalidade preocupada e parentalidade desligada. Conclusões: O projeto de promoção para aparentalidade positiva em relação ao medo dos filhos baseado no conhecimento dos significadosatribuídos permitiu não só entender a dimensão do problema, como desenvolver intervenções emateriais adequados, tal como indicar a melhor abordagem para promover a parentalidadepositiva em relação aos medos dos filhos.(AU)

Cuantificación de bacterias diazótrofas aisladas de suelos cacaoteros (Theobroma cacao L.), por la técnica de Número Más Probable (NMP) / Quantification of diazotrophs bacteria isolated from cocoa soils (Theobroma cacao L.), by the technique of Most Probable Number (MPN)

Esta investigación tuvo como objetivo cuantificar bacterias diazótrofas y comparar fisicoquímicamente suelos rizosféricos de tres cacaotales (Theobroma cacao L.) del Departamento Norte de Santander, Colombia; para lo cual se caracterizaron, diferenciándose en área cultivada, manejo agronómico y edad del cultivo. A partir de diluciones seriadas de las muestras y empleando la técnica de Número Más Probable (NMP), en medios de cultivo semiselectivos (NFb, JMV, LGI, JNFb) semisólidos, se cuantificaron las diazótrofas, evaluando como positivo la formación de una película subsuperficial en el medio contenido en viales sellados; muestras pares se enviaron al laboratorio Bioambiental (UNET) para los análisis fisicoquímicos. Como resultado, las muestras evaluadas mostraron deficiencias en el porcentaje de materia orgánica y elementos como Potasio, Fósforo y Magnesio. Se reportaron estadísticamente diferencias altamente significativas en NMP. La mayor cuantificación de diazótrofas se reportó en la finca Florilandia, que se caracterizó por tener riego por goteo. La mayor cuantificación de diazotrófas se registró en los medios NFb y JMV, demostrándose una mayor presencia de los presuntos géneros Azospirillum sp. y Burkholderia sp. los cuales son fácilmente aislados de suelos rizosféricos, a diferencia de los géneros Herbaspirillum sp. y Gluconacetobacter sp. que por su carácter endófito suelen ser menos predominantes en este tipo de muestras. Se concluye además, que las características fisicoquímicas del suelo, la humedad y las relaciones climáticas al momento de la toma de las muestras, condicionan la cantidad de exudados de las raíces y por tanto son factores que condicionaron la presencia de diazótrofas en las muestras.

The objective of this research was to quantify diazotrophic bacteria and compare physicochemically rhizospheric soils of three cocoa plantations (Theobroma cacao L.) in Norte de Santander Department, Colombia; for which they were characterized, differing in cultivated area, agronomic management and crop age. From serial dilutions of the samples and using the technique of Most Probable Number (MPN), In semisolid culture media (NFb, JMV, LGI, JNFb), the diazotrophs were quantified, evaluating as positive the formation of a subsurface film in the medium contained in sealed vials; equal samples were sent to the Bioambiental laboratory (UNET) for physicochemical analyzes. As a result, the evaluated samples showed deficiencies in the percentage of organic matter and elements such as Potassium, Phosphorus and Magnesium. Statistically highly significant differences in MPN were reported. The highest quantification of diazotrophs was reported in the Florilandia farm, which was characterized by drip irrigation. The highest quantification of diazotrophs was recorded in the media NFb and JMV, demonstrating a greater presence of the presumed genera Azospirillum sp. and Burkholderia sp. which are easily isolated from rhizospheric soils, unlike the genera Herbaspirillum sp. and Gluconacetobacter sp. which by their endophytic character tend to be less predominant in this type of samples. It is also concluded that the physicochemical characteristics of the soil, humidity and climatic relationships at the moment of sampling, condition the amount of root exudates and therefore are factors that conditioned the presence of diazotrophs in the samples.

Tratamiento quirúrgico del síndrome de apnea hipopnea obstructiva del sueño en la Clínica Rivas, Bogotá ­ Colombia / Surgical treatment of obstructive sleep apnea hypopnea syndrome in the Rivas Clinic, Bogota ­ Colombia

Introducción: El Síndrome de Apnea Hipopnea Obstructiva del Sueño es un trastorno respiratorio del sueño mayor ampliamente conocido, con importantes implicaciones para los pacientes y cuya incidencia ha venido en aumento durante los últimos años; comprende diversas manifestaciones clínicas que varían desde el ronquido hasta consecuencias cardiovasculares importantes. Objetivo: Describir la experiencia de los procedimientos quirúrgicos más utilizados para el tratamiento de pacientes con Trastornos Respiratorios del Sueño en la Clínica Rivas. Diseño: Estudio observacional descriptivo. Métodos: Revisión de 366 historias clínicas de pacientes con diagnóstico clínico y Polisomnográfico de SAHOS intervenidos quirúrgicamente debido al Trastorno Respiratorio del Sueño por rechazo de terapia de presión positiva en 3 años de observación. Resultados: Se evaluaron diferencias en medianas de los cambios del IAH, índice de Saturación de oxigeno basal y mínima, y el índice de microdespertares nocturnos tanto prequirúrgica como postquirúrgicamente. Como medida de evaluación secundaria se evaluaron las complicaciones quirúrgicas. Conclusión: En nuestra institución, como centro de referencia en apnea del sueño, la cirugía ha demostrado que disminuye de forma significativa gravedad del SAHOS y disminuye el riesgo de los pacientes con trastornos respiratorios del sueño que han rechazado el dispositivo de presión positiva.

Introduction: Obstructive Sleep Apnea Hypopnea Syndrome is a major sleep breathing disorder widely known, with important implications for patients whose incidence has been increasing in recent years; comprises various clinical manifestations ranging from snoring to major cardiovascular consequences. Objective: To describe the experience of surgical procedures commonly used for the treatment of patients with respiratory sleep disorders in Rivas Clinic. Design: Descriptive study. Methods: A review of medical records of 366 patients with clinical and polysomnographic diagnosis of OSAHS and underwent surgically due to Sleep Respiratory Disorder for rejection of positive pressure therapy in 3 years of observation was performed. Results: Differences in median changes in Apnea Hipopnea Index, baseline saturation index and minimum oxygen, and index of nocturnal arousals both preoperative and postoperatively were evaluated. As a secondary assessment of surgical complications were evaluated. Conclusion: At our institution, as a reference center on sleep apnea, surgery has been shown to decrease significantly OSAHS severity and decreases the risk of patients with respiratory sleep disorders that have rejected positive pressure device.

Trabalho em equipe: o significado atribuído por profissionais da estratégia de saúde da família / Teamwork and its meaning to professionals working in the family health strategy program

O objetivo com esta pesquisa foi compreender o significado do trabalho em equipe para os profissionais da Estratégia de Saúde da Família(ESF). Trata-se de pesquisa qualitativa, na qual foram utilizadas entrevistas semiestruturadas com oito profissionais da área da saúde. A análiseevidenciou o significado de trabalho em equipe como um trabalho familiar e idealizado, uma relação de ajuda, um trabalho hierárquico e umtrabalho coletivo. Foram apontados, também, dificuldades, agravantes, benefícios e sugestões relacionadas ao trabalho em equipe. Os resultadospossibilitaram reflexões sobre a complexidade do trabalho em equipe e a necessidade de capacitação para o desenvolvimento de competênciasinterpessoais e de trabalho em grupo.

The purpose of this study was to understand the meaning of teamwork to Family Health Strategy Program professionals. It is a qualitative research;semi-structured interviews with eight health professionals were conducted. Study analysis highlighted the teamwork as family work, helpingrelationship, hierarchical and collective work. It also pointed out difficulties, conflicts, benefits and suggestions related to teamwork. The resultsenabled reflections on the complexity of teamwork and the need for training in order to develop interpersonal skills and team work strategies.

El objetivo del presente estudio fue entender el significado del trabajo en equipo para los profesionales de la Estrategia Salud de la Familia. Esta investigacióncualitativa utilizó la entrevista semiestructurada con ocho profesionales de la salud. El análisis realzó la importancia de dicha tarea como trabajo familiar eidealizado, relación de ayuda, trabajo jerárquico y trabajo colectivo. También se mencionan sus dificultades, agravantes y beneficios y se hacen algunas sugerencias.Los resultados permitieron reflexionar sobre su complejidad y necesidad de capacitación para el desarrollo de habilidades interpersonales y del trabajo en equipo.

Subtyping influenza A virus with monoclonal antibodies and an indirect immunofluorescence assay.

The recent association of certain influenza A virus subtypes with clinically relevant phenotypes has led to the increasing importance of subtyping by clinical virology laboratories. To provide clinical laboratories with a definitive immunofluorescence assay for the subtyping of influenza A virus isolates, we generated a panel of monoclonal antibodies (MAbs) against the major circulating influenza A virus subtypes using multiple inactivated H1N1, H3N2, and 2009 H1N1 strains individually as immunogens. Eleven MAbs that target hemagglutinin (HA) of H1N1 and H3N2 subtypes were selected. These MAbs were combined into three subtype-specific reagents, one each for pan-H1 (seasonal and 2009 strains), H3, and 2009 H1, for the subtyping of influenza A virus-positive specimens by indirect immunofluorescence assay (IFA). Each subtype-specific reagent was tested on 21 prototype influenza A virus strains and confirmed to be specific for its intended subtype. In addition, the subtyping reagents did not cross-react with any of 40 other viruses. The clinical performance of the subtyping reagents was evaluated with 75 archived clinical samples collected between 2006 and 2009 using the D(3) Ultra DFA influenza A virus identification reagent (Diagnostic Hybrids, Inc., Athens, OH) and the influenza A virus subtyping reagents by IFA simultaneously. Sixty-four samples grew virus and were subtyped as follows: 30 as H3N2, 9 as seasonal H1N1, and 25 as 2009 H1N1. RT-PCR was used to confirm the influenza A virus subtyping of these samples, and there was 100% agreement with IFA. This subtyping IFA provides clinical laboratories with a cost-effective diagnostic tool for better management of influenza virus infection and surveillance of influenza virus activity.

Monoclonal antibody kit for identification of the novel 2009 H1N1 influenza A virus.

To develop an immunofluorescence assay for identification of the 2009 H1N1 influenza A virus, we generated a number of monoclonal antibodies (MAbs) by using inactivated H1N1 2009 virus (A/California/07/2009) as the immunogen. Two MAbs that target two different epitopes of the 2009 H1N1 hemagglutinin (HA) were selected to make the D(3) Ultra 2009 H1N1 Influenza A ID kit (2009 H1N1 ID kit; Diagnostic Hybrids, Inc., Athens, OH), which is intended for the identification of the 2009 H1N1 virus by indirect immunofluorescence assay (IFA). The kit does not detect any of 14 seasonal H1N1 or H3N2 prototype influenza virus strains and is also not reactive with seven other major respiratory viruses. Clinical respiratory specimens were evaluated using both the 2009 H1N1 ID kit and the CDC human influenza virus real-time reverse transcription-PCR swine flu panel (CDC rRT-PCR) and showed 100% agreement between the two assays. Four of these clinical specimens, however, were positive by the 2009 H1N1 ID kit but were identified as presumptively positive by the CDC rRT-PCR by virtue of showing threshold cycle (C(T)) values only with universal InfA and swInfA primers, not with swH1 primers. Sequence analysis of the HA genes of these four specimens revealed point mutations in both the primer and probe regions. In addition, unlike the CDC rRT-PCR, the 2009 H1N1 ID kit can differentiate the 2009 H1N1 virus from a swine-derived H1 influenza A virus (A/New Jersey/8/76). The 2009 H1N1 ID kit offers clinical laboratories an alternative to RT-PCR for the identification of the 2009 H1N1 influenza A virus.

Inyección intratimpánica con metilprednisolona como terapia primaria en sordera súbita / Intratympanic therapy with metilprednisolone as an initial therapy in sudden sensorial hearing loss

Objetivo: Evaluar la respuesta de la inyección intratimpánica (IIT) con Metilprednisolona como manejo primario en la Hipoacusia Neurosensorial Súbita Idiopática (HNSSI).Materiales y métodos: Estudio experimental en 16 pacientes y 17 oídos, con HNSSI sometidos a IIT con Metilprednisolona como terapia primariaSe realizó una comparación estadística, posterior a la intervención. Se utilizó prueba de rangos de Wilcoxon y análisis de varianza de Kruskal – Wallis entre los valores de PTA y el porcentaje de discriminación con las variables de sexo, edad, tipo de curva y tiempo transcurrido hasta la consulta.Resultados: Mejoría global del 58,80%, cambios significativos para mejoría del PTA (p=0.01) y para porcentaje de discriminación (p=0.03). Influencia significativa en la recuperación del PTA dependiendo de la clasificación del grado de pérdida inicial (p=0.005), y de la clasificación del tipo de curva (p=0.039). En la discriminación, se encontró efecto significativo de la clasificación del grado de pérdida inicial (p = 0,004).Conclusión: Se demuestra utilidad de la IIT con Metilprednisolona en la sordera súbita como terapia primaria.

Objective: To evaluate the response to intra-tympanic therapy (IIT) with Metilprednisolone as an initial therapy in Sudden Sensorial Hearing loss (SSHL). Materials and methods: 16 patients and 17 ears, with SSHL under ITT with Metilprednisolone as initial therapy were analyzed. A statistical comparison was conducted after the surgery. Wilcoxon range test and Kruskal-Wallis variance analysis were used between the PTA values and the discrimination percentage with the following variables: sex, age, type of curve of the audiogram and time elapsed up to the consultation. Results: A global improvement of 58, 80% of patients was found. The improvement in PTA and discrimination was significant (p=0.01, p=0.03), respectively. There was a significant influence in the recovery of PTA depending on the classification of the initial level of loss (p= 0.005) and on the classification of the type of curve (p=0.039). In discrimination, the only significant variable that influenced the gain was the severity of the initial loss. Conclusion: The usefulness of IIT with Metilprednisolone in sudden sensorial hearing loss has been proved as a primary therapy.