Comparison of Four Real-Time Polymerase Chain Reaction Assays for the Detection of SARS-CoV-2 in Respiratory Samples from Tunja, Boyacá, Colombia.

Coronavirus disease (COVID-19) is an infectious disease caused by SARS-CoV-2. In Colombia, many commercial methods are now available to perform the RT-qPCR assays, and laboratories must evaluate their diagnostic accuracy to ensure reliable results for patients suspected of being positive for COVID-19. The purpose of this study was to compare four commercial RT-qPCR assays with respect to their ability to detect the SARS-CoV2 virus from nasopharyngeal swab samples referred to Laboratorio Carvajal IPS, SAS in Tunja, Boyacá, Colombia. We utilized 152 respiratory tract samples (Nasopharyngeal Swabs) from patients suspected of having SARS-CoV-2. The diagnostic accuracy of GeneFinderTM COVID-19 Plus RealAmp (In Vitro Diagnostics) (GF-TM), One-Step Real-Time RT-PCR (Vitro Master Diagnostica) (O-S RT-qPCR), and the Berlin modified protocol (BM) were assessed using the gold-standard Berlin protocol (Berlin Charité Probe One-Step RT-qPCR Kit, New England Biolabs) (BR) as a reference. Operational characteristics were estimated in terms of sensitivity, specificity, agreement, and predictive values. Using the gold-standard BR as a reference, the sensitivity/specificity of the diagnostic tests was found to be 100%/92.7% for GF-TM, 92.75%/67.47% for O-S RT-qPCR, and 100%/96.39% for the BM protocol. Using BR as a reference, the sensitivity/specificity for the diagnostic tests were found to be 100%/92.7% for the GF-TM assay, 92.72%/67.47% for the O-S RT-qPCR, and 100%/96.39% for BM. Relative to the BR reference protocol, the GF-TM and BM RT-PCR assays obtained similar results (k = 0.92 and k = 0.96, respectively), whereas the results obtained by O-S-RT-qPCR were only moderately similar. We conclude that the GF-TM and BM protocols offer the best sensitivity and specificity, with similar results in comparison to the gold-standard BR protocol. We recommend evaluating the diagnostic accuracy of the OS-RT-qPCR protocol in future studies with a larger number of samples.

BID regulates AIF-mediated caspase-independent necroptosis by promoting BAX activation.

Alkylating DNA-damage agents such as N-methyl-N'-nitro-N'-nitrosoguanidine (MNNG) trigger necroptosis, a newly defined form of programmed cell death (PCD) managed by receptor interacting protein kinases. This caspase-independent mode of cell death involves the sequential activation of poly(ADP-ribose) polymerase-1 (PARP-1), calpains, BAX and AIF, which redistributes from mitochondria to the nucleus to promote chromatinolysis. We have previously demonstrated that the BAX-mediated mitochondrial release of AIF is a critical step in MNNG-mediated necroptosis. However, the mechanism regulating BAX activation in this PCD is poorly understood. Employing mouse embryonic knockout cells, we reveal that BID controls BAX activation in AIF-mediated necroptosis. Indeed, BID is a link between calpains and BAX in this mode of cell death. Therefore, even if PARP-1 and calpains are activated after MNNG treatment, BID genetic ablation abolishes both BAX activation and necroptosis. These PCD defects are reversed by reintroducing the BID-wt cDNA into the BID(-/-) cells. We also demonstrate that, after MNNG treatment, BID is directly processed into tBID by calpains. In this way, calpain non-cleavable BID proteins (BID-G70A or BID-Δ68-71) are unable to promote BAX activation and necroptosis. Once processed, tBID localizes in the mitochondria of MNNG-treated cells, where it can facilitate BAX activation and PCD. Altogether, our data reveal that, as in caspase-dependent apoptosis, BH3-only proteins are key regulators of caspase-independent necroptosis.

[Importance of water in the hydration of the Spanish population: FESNAD 2010 document]. / Importancia del agua en la hidratación de la población española: documento FESNAD 2010.

For any healthy individual, thirst is an appropriate sign to drink water, except for babies, sportsmen, and most of ill and elderly people. In these instances, it is convenient to schedule appropriate times to drink water since great demands and the physiological mechanisms that determine thirst in these situations may condition water unbalances with important consequences for health and the physical and intellectual performance. The human body has a number of mechanisms that allow keeping constant the water content by adjusting intakes and wastes. Water balance is determined by intake (consumed water, beverages, and water contained in foods) and wastes (urine, stools, the skin, and expired air from the lungs). Failure of these mechanisms and subsequent impairments in water balance may produce severe disarrangements that may threaten somebody's life. In the present document, we analyze the evidences regarding the factors conditioning water needs in the different life stages and physiological situations, as well as the consequences of water unbalance under different situations. A proper hydration may be achieved by feeding and the use of water and other liquids. Although water is the beverage by excellence and represents the ideal way of restoring the losses and get hydrated, we should be aware that, from the very beginning, we have sought other liquid sources with hydration properties. In the last decades we have increased the consumption of different beverages, with a proliferation of sugar-containing beverages. Since excessive sugar consumption has been related to obesity and other chronic conditions, it is evident that the use of these caloric beverages should be rationalized, especially in children. In this document all the considerations regarding hydration are presented and different recommendations are exposed.

[Comparison of dietary reference intakes (DRI) between different countries of the European Union, The United States and the World Health Organization]. / Comparativa de las Ingestas Dietéticas de Referencia (IDR) de los diferentes países de la Unión Europea, de Estados Unidos (EEUU) y de la Organización Mundial de la Salud (OMS).

BACKGROUND: In Spain, there are some organizations and scientific societies that have edited reference values of recommended nutrient intake, but whose data does not always agree in terms of format and content. AIMS: To review the definitions, data and methodology that other countries or groups of countries have followed to obtain and document their own reference values in order to offer basic information to facilitate the establishment of the best reference values for the Spanish population. FIELD: Review of the available information in different countries (or groups of countries) from the European Union, the United States and World Health Organization. The analysed data concerned to healthy populations. CONCLUSIONS: Reference intakes differ among the examined countries according to population groups, included nutrients, methodology and frequency of published reviews. However, most of the countries define major concepts in the same way, although with different names in each country. On the other hand, most of the studied cases represent only a scientific organization in charge of the publication and update of the values of dietary reference intakes, but not in Spain. In that context, it looks convenient to reach a consensus among all Spanish organizations and scientific societies that are involved in this task, in order to establish an acceptable reference values.

Comparativa de las Ingestas Dietéticas de Referencia (IDR) de los diferentes países de la Unión Europea, de Estados Unidos (EEUU) y de la Organización Mundial de la Salud (OMS) / Comparison of dietary reference intakes (DRI) between different countries of the European Union, The United States and the world health organization

Antecedentes: En España, existen varios organismos y sociedades científicas que han editado sus propios de valores de referencia sobre la ingesta recomendada de nutrientes, que no siempre coinciden en los contenidos y formas presentadas. Objetivo: Revisar los conceptos, datos y la metodología que han seguido otros países o grupos de países para obtener y documentar sus propios Valores de Referencia, con objeto de ofrecer una información básica que facilite el establecimiento de los valores de referencia que mejor puedan adaptarse a la población española, en base a la mejor evidencia científica disponible en la actualidad. Ámbito: Revisión de la información disponible en los distintos países (o grupos de países) de la Unión Europea, Estados Unidos y la Organización Mundial de la Salud. Los datos estudiados corresponden a poblaciones sanas. Conclusiones: Las ingestas de referencia difieren notablemente entre los distintos países estudiados en cuanto a grupos de población, tipo de nutrientes incluidos, metodología y periodicidad de las revisiones publicadas. Sin embargo, la mayoría definen de manera semejante los conceptos más importantes, aunque tengan distintas denominaciones en cada país. Por otro lado, en la mayoría de los casos estudiados existe un único organismo científico encargado de la publicación y actualización de los valores de ingestas dietéticas de referencia, no siendo así en España. En este contexto, parece conveniente alcanzar un acuerdo entre todos los organismos y sociedades científicas españolas implicadas en esta tarea, con objeto de establecer unos valores de referencia únicos para todo el país, con el consenso de todos (AU)

Background: In Spain, there are some organizations and scientific societies that have edited reference values of recommended nutrient intake, but whose data does not always agree in terms of format and content. Aims: To review the definitions, data and methodology that other countries or groups of countries have followed to obtain and document their own reference values in order to offer basic information to facilitate the establishment of the best reference values for the Spanish population. Field: Review of the available information in different countries (or groups of countries) from the European Union, the United States and World Health Organization. The analysed data concerned to healthy populations. Conclusions: Reference intakes differ among the examined countries according to population groups, included nutrients, methodology and frequency of published reviews. However, most of the countries define major concepts in the same way, although with different names in each country. On the other hand, most of the studied cases represent only a scientific organization in charge of the publication and update of the values of dietary reference intakes, but not in Spain. In that context, it looks convenient to reach a consensus among all Spanish organizations and scientific societies that are involved in this task, in order to establish an acceptable reference values (AU)

Structure of the capsular polysaccharide of pneumococcal serotype 11A reveals a novel acetylglycerol that is the structural basis for 11A subtypes.

We have undertaken a structural assessment of Streptococcus pneumoniae 11A polysaccharide as well as two clinical isolates related to 11A. The clinical isolates were labeled 11Aalpha and 11Abeta. The result of our experiments is a revision to the old structure for S. pneumoniae 11A polysaccharide. The new structure differs from the old structure in both the primary connectivities and acetylation pattern. We also show that 11A contains an acetylglycerol-PO4 moiety, a substitution that is heretofore unknown in the bacterial polysaccharide literature. The two clinical isolates were also structurally characterized. 11Aalpha was determined to be identical to 11A. 11Abeta is a new serotype, which differs from 11A in the absence of the acetylation of the glycerol-PO4 moiety and a different acetylation pattern of the saccharides. Thus, we propose that the acetylglycerol is the structural basis for 11Aalpha and 11Abeta subtypes.

Immunohistochemical detection of interleukin-12 and interferon-gamma in pigs experimentally infected with Mycoplasma hyopneumoniae.

The expression of interleukin-12 (IL-12) and interferon-gamma (IFN-gamma) was examined immunohistochemically in the lungs of pigs aged 21 days infected experimentally with Mycoplasma hyopneumoniae (Mh). Ten pigs were inoculated intranasally with Mh and killed in pairs weekly from 7 to 35 days post-infection (dpi). Immunolabelling for IL-12 and IFN-gamma was usually associated with inflammation, particularly in macrophages and lymphocytes in the thickened alveolar septa and in the hyperplastic bronchus-associated lymphoid tissue (BALT). Cells positive for both cytokines were detected at 7 dpi, their numbers increasing at 14 and 21 dpi, and slightly decreasing thereafter. The results suggest that IL-12 and IFN-gamma play a role in pulmonary defence mechanisms against Mh infection.

A reversed phase HPLC assay for the simultaneous quantitation of non-ionic and ionic surfactants in bioprocess intermediates.

This report describes a rapid and accurate reversed phase HPLC method for the simultaneous quantitation of multiple surfactants in various bioprocess solution matrices including cell lysates. Separation and quantitation of a mixture of the cationic detergent domiphen bromide from the non-ionic detergent Triton X-100 in crude cell mixtures can be achieved within 15 min using a TSK-gel C18-NPR reversed phase column and an aqueous mobile phase gradient of acetonitrile:water with the reagent PIC-B8 as ion-pairing modifier. The linear dynamic range for quantitation of domiphen bromide (DB) and Triton in this assay extends from 20 to 2000 microM. Linear regression analyses from the standard curve determinations showed an R2 of > or = 0.990. The assay does not show any interferences from proteins or other cellular contaminants such as nucleic acids. The assay has been used to evaluate clearance of these compounds throughout the purification process of an adenovirus-based vaccine candidate, as well as to determine the effects of process changes on detergent clearance.

Type A-like retroviral particles in a metastatic intestinal adenocarcinoma in an emerald tree boa (Corallus caninus).

A metastatic intestinal papillary adenocarcinoma was diagnosed histologically in an emerald tree boa (Corallus caninus). Metastasis was detected in the liver, both kidneys, lung, and coelomic wall. Ultrastructural examination of the metastatic intestinal epithelial cells in the liver revealed the presence of a moderate number of viral particles that most closely resembled type A retroviral particles and were mainly associated with granular endoplasmic reticulum membranes. This case is the first description of type A-like retroviral particles in a neoplasm of a snake. The role of the virions in the etiology of the intestinal adenocarcinoma is uncertain. In addition, this is the first confirmed report of a metastatic intestinal adenocarcinoma in a snake.

Immunohistochemical labelling of cytokines in lung lesions of pigs naturally infected with Mycoplasma hyopneumoniae.

Mycoplasma hyopneumoniae (Mh) is the primary agent of porcine enzootic pneumonia (PEN), a chronic respiratory disease endemic to pig farms, and characterized histologically by infiltration of mononuclear cells in airways and prominent hyperplasia of the bronchus-associated lymphoid tissue (BALT). To gain further insight into the pathogenesis of PEN, cytokine expression in the lung, with particular attention to the BALT, was examined immunohistochemically in pigs naturally infected with Mh. An increase (P < 0.05) in proinflammatory and immunoregulatory cytokines (especially interleukin [IL]-2, IL-4 and tumour necrosis factor [TNF]-alpha, and to a lesser extent IL-1 [alpha and beta] and IL-6) was detected in the BALT, which showed intense lymphoid hyperplasia. IL-1beta and TNF-alpha were also detected in the bronchoalveolar exudate of infected pigs, and IL-6 and IL-8 were demonstrated in mononuclear cells of the alveolar septa. The results showed that in Mh infection, macrophage and lymphocyte activation results in the expression of a number of cytokines capable of inducing lung lesions and lymphoreticular hyperplasia of the BALT.

A morphologic and immunohistochemical study of the bronchus-associated lymphoid tissue of pigs naturally infected with Mycoplasma hyopneumoniae.

Porcine enzootic pneumonia (PEN), caused by Mycoplasma hyopneumoniae (Mh), has been described in pigs in all geographic areas. The disease is characterized by high morbidity and low mortality rates in intensive swine production systems. A morphologic and immunohistochemical study was done to determine the cellular populations present in lung parenchyma of infected pigs, with special attention to the bronchus-associated lymphoid tissue (BALT). Polyclonal and monoclonal antibodies were used for the detection of antigens of Mh, T lymphocytes (CD3+, CD4+, and CD8+), IgG+ or IgA+ lymphocytes, and cells containing lysozyme, S-100 protein, major histocompatibility complex class II antigen or myeloid-histiocyte antigen. Findings in lung tissues associated with Mh infection were catarrhal bronchointerstitial pneumonia, with infiltration of inflammatory cells in the lamina propria of bronchi and bronchioles and alveolar septa. Hyperplasia of mononuclear cells in the BALT areas was the most significant histologic change. The BALT showed a high morphologic and cellular organization. Macrophages and B lymphocytes were the main cellular components of germinal centers. T lymphocytes were primarily located in perifollicular areas of the BALT, lamina propria and within the airway epithelium, and plasma cells containing IgG or IgA at the periphery of the BALT, in the lamina propria of bronchi and bronchioles, in alveolar septa, and around bronchial submucosal glands. The hyperplastic BALT in PEN cases consisted of macrophages, dendritic cells, T and B lymphocytes, and IgG+ and IgA+ plasma cells. CD4+ cells predominated over CD8+ cells. Local humoral immunity appears to play an important role in the infection.

Mullerian inhibiting substance inhibits ovarian cell growth through an Rb-independent mechanism.

Müllerian inhibiting substance (MIS), a transforming growth factor-beta family member, causes regression of the Müllerian duct in male embryos. MIS overexpression in transgenic mice ablates the ovary, and MIS inhibits the growth of ovarian cancer cell lines in vitro, suggesting a key role for this hormone in postnatal development of the ovary. This report describes a mechanism for MIS-mediated growth inhibition in both a human epithelial ovarian cancer cell line and a cell line derived from normal ovarian surface epithelium, which is the origin of human epithelial ovarian cancers. MIS-treated cells accumulated in the G(1) phase of the cell cycle and subsequently underwent apoptosis. MIS up-regulated the cyclin-dependent kinase inhibitor p16 through an MIS type II receptor-mediated mechanism and inhibited growth in the absence of detectable or inactive Rb protein. Prolonged treatment with MIS down-regulated the Rb-related protein p130 and increased the Rb family-regulated transcription factor E2F1, overexpression of which inhibited growth. These findings demonstrate that p16 is required for MIS-mediated growth inhibition in ovarian epithelial cells and tumor cells and suggest that up-regulation of E2F1 also plays a role in this process.

¿Es el antyecedente de tumores previos un factor de mal pronóstico en el cváncer superficial de vejiga? / Is the antecedent of former tumors, a bad pronostic factor in superficial bladder cancer?

Objetivo: Establecer si la presencia de recurrencias tumorales es un factor de pronóstico en el carcinoma transicional de vejiga en estadio t1. Material y métodos: Análisis retrospectivo de los carcinomas transicionales en estadio T1 operados por miembros del C.U.S.P.B.A. Se consignaron: la edad, sexo, características del tumor (número, grado, tamaño, etc.), la aparición y tipo de recidiva, así como la presencia de progresión tumoral en éstas. Se registro en cada ficha si el paciente había sido operado previamente de tumores vesicales transicionales. De este modo se conformaron dos grupos a estudiar: Aquéllos sin antecedentes de tumores previos (grupo 1), y aquéllos con antecedentes de los mismos (grupo 2). Resultados: Se analizaron 371 pacientes con diagnóstico de carcinoma transicional de vejiga en estadio T1, operados por miembros del C.U.S.P.B.A. durante el período 1993-1998. Se halló que 331 pacientes no habían presentado nunca cirugías por carcinomas transicionales anteriores (grupo 1), mientras que 40 casos sí habían sido operados previamente por este tipo de tumores (grupo 2). De esta manera se constituyeron los dos grupos a estudiar. Dentro del grupo 1 el 35,6 por ciento presentó recidiva, mientras que este porcentaje fue del 60 por ciento en el grupo 2 (p=0,002). Akl evaluar la aparición de progresión en lasd recidivas se observó que ésta estuvo presente en el 12,6 por ciento de los pacientes en el grupo 1 y el 30 por ciento de los pacientes en el grupo 2 (p=0,003). Comparando aquellos pacientyes que sólo progresaron en el estadio (T) se encontró progresión en el 5,1 por ciento dentro del grupo 1...(AU)

¿Es el antyecedente de tumores previos un factor de mal pronóstico en el cváncer superficial de vejiga? / Is the antecedent of former tumors, a bad pronostic factor in superficial bladder cancer?

Objetivo: Establecer si la presencia de recurrencias tumorales es un factor de pronóstico en el carcinoma transicional de vejiga en estadio t1. Material y métodos: Análisis retrospectivo de los carcinomas transicionales en estadio T1 operados por miembros del C.U.S.P.B.A. Se consignaron: la edad, sexo, características del tumor (número, grado, tamaño, etc.), la aparición y tipo de recidiva, así como la presencia de progresión tumoral en éstas. Se registro en cada ficha si el paciente había sido operado previamente de tumores vesicales transicionales. De este modo se conformaron dos grupos a estudiar: Aquéllos sin antecedentes de tumores previos (grupo 1), y aquéllos con antecedentes de los mismos (grupo 2). Resultados: Se analizaron 371 pacientes con diagnóstico de carcinoma transicional de vejiga en estadio T1, operados por miembros del C.U.S.P.B.A. durante el período 1993-1998. Se halló que 331 pacientes no habían presentado nunca cirugías por carcinomas transicionales anteriores (grupo 1), mientras que 40 casos sí habían sido operados previamente por este tipo de tumores (grupo 2). De esta manera se constituyeron los dos grupos a estudiar. Dentro del grupo 1 el 35,6 por ciento presentó recidiva, mientras que este porcentaje fue del 60 por ciento en el grupo 2 (p=0,002). Akl evaluar la aparición de progresión en lasd recidivas se observó que ésta estuvo presente en el 12,6 por ciento de los pacientes en el grupo 1 y el 30 por ciento de los pacientes en el grupo 2 (p=0,003). Comparando aquellos pacientyes que sólo progresaron en el estadio (T) se encontró progresión en el 5,1 por ciento dentro del grupo 1...

Tumores incidentales de rñón y tumores menores de cinco centímetros / Incidental renal tumours and tumours less than five cm. in diameter

Objetivo: Establecer la experiencia de un grupo cooperativo sobre tumores renales incidentales y los menores de 5 centímetros. Material y métodos: Se realizó un análisis retrospectivo de los adenocarcinomas renales operados con nefrectomía radical en el período 1986-1996. Se seleccionaron los tumores incidentales, los menores de 5 centímetros, y se realizó un análisis de la función renal posoperatoria de los pacientes. Resultados: Se analizaron 231 tumores renales; 32 por ciento fueron dianóstico incidental (74 pacientes); 35,5 poor cientoi fueron tumores menores de 5 cm (82 pacientes). Se halló que los tumores incidentales se poresentaron en estadios más bajos (62 por ciento Edios 1 y 2), y con mayor porcentaje de tumores menores de 5 cm (51,3 por cientoi) (p=<0,01). La sobrevida de los tumores menores de 5 cm fue estadísticamente superior a los mayores a esa cifra (p=<0,001). Se halló un 7,8 por ciento de detioro de la función renal posoperatoria (3,9 transitoriay 3,9 definitiva), con cifras de creatinina superiores a 1,5 mg/dl. Ningún paciente con tumor incidental o menor de 5 cm requirió hemodiálisis. Conclusiones: Los tumores renalrs incidentales de 5 cm son una entidad cada vez m,ás frecuente, y suelen coincidir con estadios bajos. La nefrectomía radical parece ser una técnica adecuada para su tratamiento, con escaso deterioro de la función renal posoperatoria, y 0 por ciento de necesidad de hemodiálisis(AU)

Tumores incidentales de rñón y tumores menores de cinco centímetros / Incidental renal tumours and tumours less than five cm. in diameter

Objetivo: Establecer la experiencia de un grupo cooperativo sobre tumores renales incidentales y los menores de 5 centímetros. Material y métodos: Se realizó un análisis retrospectivo de los adenocarcinomas renales operados con nefrectomía radical en el período 1986-1996. Se seleccionaron los tumores incidentales, los menores de 5 centímetros, y se realizó un análisis de la función renal posoperatoria de los pacientes. Resultados: Se analizaron 231 tumores renales; 32 por ciento fueron dianóstico incidental (74 pacientes); 35,5 poor cientoi fueron tumores menores de 5 cm (82 pacientes). Se halló que los tumores incidentales se poresentaron en estadios más bajos (62 por ciento Edios 1 y 2), y con mayor porcentaje de tumores menores de 5 cm (51,3 por cientoi) (p=<0,01). La sobrevida de los tumores menores de 5 cm fue estadísticamente superior a los mayores a esa cifra (p=<0,001). Se halló un 7,8 por ciento de detioro de la función renal posoperatoria (3,9 transitoriay 3,9 definitiva), con cifras de creatinina superiores a 1,5 mg/dl. Ningún paciente con tumor incidental o menor de 5 cm requirió hemodiálisis. Conclusiones: Los tumores renalrs incidentales de 5 cm son una entidad cada vez m,ás frecuente, y suelen coincidir con estadios bajos. La nefrectomía radical parece ser una técnica adecuada para su tratamiento, con escaso deterioro de la función renal posoperatoria, y 0 por ciento de necesidad de hemodiálisis

Adenocarcinoma renal: factores pronósticos: Experiencia de 10 años del club urológico del sur de la Provincia de Buenos Aires (C.U.S.P.B.A) / Renal adenocarcinoma: prognostic factors: 10 years experience of the Provincia Buenos Aires urologic Club

Objetivo: Establecer factors pronósticos en el adenocarcinoma renal.Material y método: Se realizó un estudio retrospectivo de todos los casos de adenocarcinoma renal operados por los miembros del Club urológico del sur de la provincia de Buenos Aires que participaron en este estudio, entre los años 1986 y 1996. Resultados: Se revisaron 231 casos de carcinoma renal operados en ese período. La edad fue de 62+= 11,63 por ciento fueron hombres. La sobrevida a 5 años fue : 100 por ciento estadio I, 77 por ciento estadio II, 51 por ciento estadio III y, 0 por ciento estadio IV; 32 por ciento fueron incidentales. Los pacientes < de 60 años y los tumores > de 5 cm presentaronmenor sobrevida. Conclusiones: Los tumores de bajo estadio (I y II), < de 5 cm, y en pascientes mayores de 60 años tienen mejor pronóstico en cuanto a la sobrevida global