Analysis of qPCR reference genes stability determination methods and a practical approach for efficiency calculation on a turbot (Scphthalmus maximus) gonad dataset

Gene expression analysis by reverse transcription quantitative PCR (qPCR) is the most widely used method for analyzing the expression of a moderate number of genes and also for the validation of microarray results. Several issues are crucial for a successful qPCR study, particularly the selection of internal reference genes for normalization and efficiency determination. There is no agreement on which method is the best to detect the most stable genes neither on how to perform efficiency determination. In this study we offer a comprehensive evaluation of the characteristics of reference gene selection methods and how to decide which one is more reliable when they show discordant outcomes. Also, we analyze the current efficiency calculation controversy. Our dataset is composed by gonad samples of turbot at different development times reared at different temperatures. Turbot (Scophthalmus maximus) is a relevant marine aquaculture European species with increasing production in the incoming years. Since females largely outgrow males, identification of genes related to sex determination, gonad development and reproductive behavior, and analysis of their expression profiles are of primary importance for turbot industryVersión del edito

La dependencia: Efectos en la salud familiar

Objetivos: Los objetivos de este trabajo son analizar cómo influye el hecho de que en el hogar haya una persona dependiente en el estado de salud de las personas con quien convive y en los estilos de vida; compararlo con otras situaciones que impliquen dedicación de tiempo y energía, y analizar diferencias de género y edad en cada etapa del ciclo vital. Diseño: Estudio descriptivo transversal analizando datos secundarios. Emplazamiento: El método de recogida de información es el de entrevista personal asistida por ordenador en las viviendas seleccionadas, realizada por el Ministerio de Sanidad, Servicios Sociales e Igualdad. Participantes: Un total de 19.351 individuos mayores de 25 años que realizaron la Encuesta Nacional de Salud 2011-2012 (ENSE 2011/2012). Mediciones principales: Estudio llevado a cabo con datos sobre la sociedad española procedentes de la ENSE 2011/12. Como marco empírico, seleccionamos el modelo Logit, reportando los datos en odds ratio. Las estimaciones se repiten de manera independiente por submuestras de edad y género. Resultados: La salud de las personas que conviven con una persona dependiente es peor que la de aquellas personas que no conviven (hasta 5 veces más riesgo de tener problemas de salud), especialmente si se es mujer, de edad avanzada, con un nivel educativo bajo o no trabaja. También ser cuidador reduce la probabilidad de mantener unos hábitos saludables como son el realizar ejercicio, el descanso o una dieta equilibrada. Conclusiones: Por lo general, convivir con una persona dependiente reduce la probabilidad de mantener estilos de vida saludables y deteriora la salud. Encontramos importantes diferencias de género y edad. Objetives: The purpose of this work is to analyse the effects on informal caregiver‘s health and lifestyle when living with a dependent person at home. A comparison will be made between this situation and other situations involving commitment of time and energy, taking into account gender and age differences in each stage of the life cycle. Design: Cross-sectional study analysing secondary data. Setting: The method used for collecting information is the computer assisted personal interview carried out in selected homes by the Ministry of Health, Social Services and Equality. Participants: The study included 19,351 participants aged over 25 years who completed the 2011-2012 Spanish National Health Survey. Main measurements: This research is based on demographic information obtained from a Spanish National Health Survey (2011/12). Using an empirical framework, the Logit model was select and the data reported as odds ratio. The estimations were repeated independently by sub-groups of age and gender. Results: The study showed that the health of people who share their lives with a dependent person is worse than those who do not have any dependent person at home (they are 5 times at higher risk of developing health problems). The study found that being a woman, advance age, low educational level and does not work, also has an influence. Being a caregiver reduces the likelihood of maintaining a healthy lifestyle through physical exercise, relaxation, or eating a balanced diet. Conclusions: Living with a dependent person reduces the likelihood of maintaining healthy lifestyles and worsens the state of health of family members. Significant differences in gender and age were found

Impact of house dust mite-driven asthma on children's school performance and activity

Absenteeism; Allergic asthma; Subcutaneous allergen immunotherapyAbsentisme; Asma al·lèrgica; Immunoteràpia subcutània amb al·lèrgensAbsentismo; Asma alérgica; Inmunoterapia subcutánea con alérgenosEvidence regarding asthma's impact on children's daily lives is limited. This prospective and cross-sectional, observational, multicenter study assessed school/work and activity impairment in children and adolescents with allergic asthma and their caregivers and allergen immunotherapy (AIT) effects. Included patients were schooled children and adolescents (5 to 17 years) with allergic asthma due to house dust mites (HDM). Impairment of school/work (i.e., absenteeism and presenteeism) and activity was measured in patients and their caregivers using the Work Productivity Impairment Questionnaire plus Classroom Impairment Questions: Allergy Specific (WPAI + CIQ:AS). HDM allergic patients with school impairment received subcutaneous AIT with a MicroCrystalline Tyrosine-associated allergoid. WPAI + CIQ:AS and effectiveness variables were compared between baseline and 1-year post-AIT. Of the 113 patients included, 59 (52.2%) and 51 (45.1%) showed school and activity impairment, respectively, missing a mean (SD) of 37.6 (24.4) % and 42.6 (25.6) % of school and activity time, respectively. Twenty-six (23%) caregivers reported activity impairment and, of the 79 (69.9%) employed, 30 (38%) reported work impairment. Of the 65 patients with school/activities impairment, 41 (63.1%) received AIT, of which 21 (51.2%) completed 1 year of treatment. Effectiveness variables and WPAI + CIQ:AS significantly improved: Mean (SD) school impairment decreased from 39.7 (26.7) to 2.1 (7.1) % (p < 0.001) and activity impairment from 46.2 (34.6) to 1.4 (3.6) % (p < 0.001). Conclusion: Allergic asthma due to HDMs results in school/work and activity impairment in children and adolescents and their caregivers. One year of AIT provided clinical benefits and reduced school and activity impairment. What is known: • Allergic asthma impairs children's school performance and daily activities. • Allergen immunotherapy modifies allergic disease course and ameliorates its symptoms. What is new: • Asthma symptoms due to allergy to house dust mites impair children's school attendance and productivity and daily activity and their caregivers' work performance and daily lives. • Allergen immunotherapy with a house dust mite MicroCrystalline Tyrosine (MCT)-associated allergoid seems to provide clinical benefits, associated with decreased school and activity impairment, supporting it as an effective treatment option

The mouse deafness locus (dn) is associated with an inversion on chromosome 19

Recombination data for the mouse deafness locus (dn) on chromosome 19 are consistent with the presence of an inversion for which one of the breakpoints is between D19Mit14 and D19Mit96, a distance of less than 226 kb. Fluorescence in situ hybridization studies using a bacterial artificial chromosome on interphase (G1) nuclei provide additional support for the presence of an inversion. The dn gene is probably the orthologue of the human DFNB7/DFNB11 gene on chromosome 9. Copyright (C) 1998 Elsevier Science B.V

Malaria falciparum y síndrome nefrótico: nuestras experiencias. Falciparum malaria and nephrotic Síndrome: Our experience

Las especies de Plasmodium que infectan al hombre son: P. vivax, P. Malariae, P. Ovale y P. Falciparum. En Mozambique, como en la mayor parte de la llamada África Subsahariana, la especie predominante es P. falciparum cloroquina resistente. La infección por P. falciparum es potencialmente mortal, tiende a manifestarse como una enfermedad febril sin signos localizados o específicos. En los casos más graves, sin embargo puede presentarse asociada a variados síndromes clínicos que plantean serios retos terapéuticos.Es reconocido que la malaria o paludismo puede asociarse a síndrome nefrótico y se han dado explicaciones de esta relación patogénica. En Mozambique, en un período de seis meses, tuvimos la oportunidad de tratar tres casos de Malaria falciparum grave, asociado a síndrome nefrótico.Divulgar y trasmitir las experiencias prácticas y consideraciones teóricas a propósito de uno de estos casos es la motivación de los autores de este trabajo. Palabras clave: PALUDISMO- MALARIA - Síndrome nefrÓtico ABSTRACT Plasmodiumspecies infecting man are the following: P.vivax, P. Malariae, P. Ovale and P. Falciparum. In Mozambique, like the biggest area from the so called Subsaharian Africa,the resistent-chloroquine P. Falciparum is the predominating specie in this area. The P. Falciparum infection is potentially fatal, with a trend to show as a febrile condition with no localized or specific signs. In more severe cases, however, it may be presented in association with different clinical syndromes which represent serious therapeutic challenges. It is not unknown that Malaria or Paludism may be associated with the Nephrotic Syndrome, and many explanations have been given on this pathogenic relatioship. In a sixth month's period in Mozambique we had the chance to test three severe cases of Falciparum Malaria associated with a Nephrotic Syndrome. Spreading the practical experience and theoretic considerations on one of these cases is the aim of this work. Key words: MALARIA, NEPHROTIC SYNDROM

Resultados preliminares de la influencia de la temperatura de cultivo sobre la proporción de sexos en el rodaballo (Scophthalmus maximus L.)

Las larvas de tres familias de rodaballos se cultivaron a temperaturas de 15'C, 18'C y 22'C desde el día dos hasta día 90 de vida y a temperatura ambiente hasta el dia 210. En las familias 1 y 2 las proporciones sexuales, determinadas por el fenotipo, oscilaron en los tres grupos entre el 40%- 60%,y no se observaron diferencias siqníñcaffvas entre los diferentes grupos de temperatura (p>0.05). En la familia 3, el porcentaje de hembras fue mayor que el de machos en los tres grupos de temperatura, y además se observó diferencia significativa (p<0.05) entre el grupo de peces cultivados a temperatura fría con respecto a los cultivados en agua ambiente. En tas familias 1 y 2 el sexo genético coincidió en gran medida con~ sexo fenolípico, siendo la discrepancia menor del 10%. Sin embargo, en la familia 3 se observó que el 36,5% de los machos genéticos eran hembras a 15'C, el 29% a 18ºC y el 18% a 23'C. Los resultados sugieren la interacción temperatura-familia en la determinación sexual del rodaballo, que debe ser confirmada en un mayor número de familias.Larvae from three families of turbot were cultured at 15'C, 18'C and 22'C from 2 to 90 days old, and then at ambient temperature until210 daysold. Regarding families 1 and 2, \he sexual proportions determined by \he phenotype varied between 40%·60%, regardless of \he cunure temperature (p>O.05). Gontrary, in family 3, \he percentage of females was higher than for males in all three cunure temperatures. Furthermore, \here were differences (¡¡<O.05) between families cultured at 15'C and 18'C. For families 1 and 2, sex determined by genotype was similarto \hat detennined by the phenotype, wi\h differences <1 0%. Gontrary, for family 3,\he \he percentage of males determined by \he genotype \hat were phenotipycatt¡ females was 36, 29 and 18% for animals cunured at 15'C, 18'C and 22'C, respectively. Results suggest an interaction temperature-family in \he turbot sex determination which should be checked on a higher numberoffamilies

Characterization of the SNAG and SLUG Domains of Snail2 in the Repression of E-Cadherin and EMT Induction: Modulation by Serine 4 Phosphorylation

Snail1 and Snail2, two highly related members of the Snail superfamily, are direct transcriptional repressors of E-cadherin and EMT inducers. Previous comparative gene profiling analyses have revealed important differences in the gene expression pattern regulated by Snail1 and Snail2, indicating functional differences between both factors. The molecular mechanism of Snail1-mediated repression has been elucidated to some extent, but very little is presently known on the repression mediated by Snail2. In the present work, we report on the characterization of Snail2 repression of E-cadherin and its regulation by phosphorylation. Both the N-terminal SNAG and the central SLUG domains of Snail2 are required for efficient repression of the E-cadherin promoter. The co-repressor NCoR interacts with Snail2 through the SNAG domain, while CtBP1 is recruited through the SLUG domain. Interestingly, the SNAG domain is absolutely required for EMT induction while the SLUG domain plays a negative modulation of Snail2 mediated EMT. Additionally, we identify here novel in vivo phosphorylation sites at serine 4 and serine 88 of Snail2 and demonstrate the functional implication of serine 4 in the regulation of Snail2-mediated repressor activity of E-cadherin and in Snail2 induction of EMT

Sexual transmission of American trypanosomiasis in humans : a new potential pandemic route for Chagas parasites

Background: the Trypanosoma cruzi infection endemic in Latin America has now spread to several countries across four continents; this endemic involves triatomine vector-free protists. We hypothesised that the sexual transmission of T. cruzi contributes to the ongoing spread of Chagas disease. Objectives: a short-term longitudinal study was conducted to evaluate this hypothesis. Methods: the study population comprised 109 subjects from four families, among whom 21 had been diagnosed with acute Chagas disease by direct parasitological analysis. Blood mononuclear cells and serum samples were obtained from each study subject once per year for three consecutive years. Enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescence serological examinations were used to detect specific T. cruzi antibodies. Polymerase chain reaction of T. cruzi DNA revealed 188-nucleotide bands, which hybridised to a specific radiolabelled probe and were confirmed by cloning and sequencing. Results: three independent assessments at different time points revealed T. cruzi nuclear DNA footprints in 76% (83/109) of the study population with active infection. In contrast, the ELISA and indirect immunofluorescence assays detected the T. cruzi antibody in 28.4% (31/109) of the study samples. Moreover, the semen from 82.6% (19/23) of subjects people revealed harboured the 188- bp base pair T. cruzi footprint. Interestingly, the ejaculates of nuclear DNA-positive Chagas patient transmitted the T. cruzi upon peritoneal injection or infusion in the vagina of mice, and amastigotes were detected in the skeletal muscle, myocardium, vas deferens, and uterine tube. Main conclusions: T. cruzi infections can be transmitted from females or males to naïve mates through intercourse, and progeny showed discrepancies between the ratios of nuclear DNA footprints and specific antibody that can be explained by the tolerance attained during early embryo growth. Additional studies are needed to develop drugs to eradicate the infections. Additionally, the importance of a vigorous education, information, and communication program to prevent sexually transmitted Chagas disease in humans cannot be underemphasised

Minimal information for studies of extracellular vesicles 2018 (MISEV2018):a position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines

The last decade has seen a sharp increase in the number of scientific publications describing physiological and pathological functions of extracellular vesicles (EVs), a collective term covering various subtypes of cell-released, membranous structures, called exosomes, microvesicles, microparticles, ectosomes, oncosomes, apoptotic bodies, and many other names. However, specific issues arise when working with these entities, whose size and amount often make them difficult to obtain as relatively pure preparations, and to characterize properly. The International Society for Extracellular Vesicles (ISEV) proposed Minimal Information for Studies of Extracellular Vesicles (“MISEV”) guidelines for the field in 2014. We now update these “MISEV2014” guidelines based on evolution of the collective knowledge in the last four years. An important point to consider is that ascribing a specific function to EVs in general, or to subtypes of EVs, requires reporting of specific information beyond mere description of function in a crude, potentially contaminated, and heterogeneous preparation. For example, claims that exosomes are endowed with exquisite and specific activities remain difficult to support experimentally, given our still limited knowledge of their specific molecular machineries of biogenesis and release, as compared with other biophysically similar EVs. The MISEV2018 guidelines include tables and outlines of suggested protocols and steps to follow to document specific EV-associated functional activities. Finally, a checklist is provided with summaries of key points

Whole genome sequencing of turbot (Scophthalmus maximus; Pleuronectiformes):a fish adapted to demersal life

12 páginas, 5 figuras.-- Antonio Figueras ... et al.-- This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly citedThe turbot is a flatfish (Pleuronectiformes) with increasing commercial value, which has prompted active genomic research aimed at more efficient selection. Here we present the sequence and annotation of the turbot genome, which represents a milestone for both boosting breeding programmes and ascertaining the origin and diversification of flatfish. We compare the turbot genome with model fish genomes to investigate teleost chromosome evolution. We observe a conserved macrosyntenic pattern within Percomorpha and identify large syntenic blocks within the turbot genome related to the teleost genome duplication. We identify gene family expansions and positive selection of genes associated with vision and metabolism of membrane lipids, which suggests adaptation to demersal lifestyle and to cold temperatures, respectively. Our data indicate a quick evolution and diversification of flatfish to adapt to benthic life and provide clues for understanding their controversial origin. Moreover, we investigate the genomic architecture of growth, sex determination and disease resistance, key traits for understanding local adaptation and boosting turbot production, by mapping candidate genes and previously reported quantitative trait loci. The genomic architecture of these productive traits has allowed the identification of candidate genes and enriched pathways that may represent useful information for future marker-assisted selection in turbotThis work was funded by the Spanish Government: projects Consolider Ingenio: Aquagenomics (CSD2007-00002) and Metagenoma de la Península Ibérica (CSD2007-00005), Ministerio de Economía y Competitividad and European Regional Development Funds (AGL2012-35904), and Ministerio de Economía y Competitividad (AGL2014-51773 and AGL2014-57065-R); and Local Government Xunta de Galicia (GRC2014/010). P.P. and D.R. gratefully acknowledge the Spanish Ministerio de Educación for their FPU fellowships (AP2010-2408, AP2012-0254). Funding to pay the Open Access publication charges for this article was provided by the Ministerio de Economía y Competitividad (AGL2014-51773) and Xunta de Galicia (GRC2014/010)Peer reviewe