Diagnóstico de Leishmaniosis canina mediante la técnica de reacción en cadena de la polimerasa (PCR) : un procedimiento simple para uso en la clínica

Para detectar la presencia de Leishmania ssp, en distintas muestras clínicas de perros sospechosos de padecer esta enfermedad, se ha utilizado la técnica PCR, amplificando para ello un fragmento del gen SSU rRNA, repetido más de 100 veces en el genoma del parásito. El método se ha optimizado para utilizarlo como método de rutina en la clínica. El procesado de las muestras es rápido y simple. El diagnóstico se ha realizado por presencia/ausencia del parásito, utilizando para ello muestras de sangre, médula ósea y ganglio linfático principalmente. En el caso de existir el parásito en el huésped se visualiza una banda nítida de un tamaño de 603 bp y en el caso de que el parásito esté ausente, no se detecta la presencia de esta banda. Los mejores resultados se obtuvieron cuando la muestra de partida fue médula o ganglio linfático. El método presenta la ventaja adicional de detectar portadores asintomáticos, incluidos los titulas de IFI dudosos. La técnica PCR se presenta como test de diagnóstico rutinario, siendo más rápida, eficaz y económica que los métodos de diagnóstico clásicos

Epigenetics modifications and Subclinical Atherosclerosis in Obstructive Sleep Apnea: The EPIOSA study.

Background Obstructive sleep apnea (OSA) is associated with increased risk for cardiovascular morbidity and mortality. Epidemiological and animal models studies generate hypotheses for innovative strategies in OSA management by interferig intermediates mechanisms associated with cardiovascular complications. We have thus initiated the Epigenetics modification in Obstructive Sleep Apnea (EPIOSA) study (ClinicalTrials.gov identifier: NCT02131610). Methods/design EPIOSA is a prospective cohort study aiming to recruit 350 participants of caucasian ethnicity and free of other chronic or inflammatory diseases: 300 patients with prevalent OSA and 50 non-OSA subjects. All of them will be follow-up for at least 5 years. Recruitment and study visits are performed in single University-based sleep clinic using standard operating procedures. At baseline and at each one year follow-up examination, patients are subjected to a core phenotyping protocol. This includes a standardized questionnaire and physical examination to determine incident comorbidities and health resources utilization, with a primary focus on cardiovascular events. Confirmatory outcomes information is requested from patient records and the regional Department of Health Services. Every year, OSA status will be assessed by full sleep study and blood samples will be obtained for immediate standard biochemistry, hematology, inflammatory cytokines and cytometry analysis. For biobanking, aliquots of serum, plasma, urine, mRNA and DNA are also obtained. Bilateral carotid echography will be performed to assess subclinical atherosclerosis and atherosclerosis progression. OSA patients are treated according with national guidelines. Discussion EPIOSA will enable the prospective evaluation of inflammatory and epigenetics mechanism involved in cardiovascular complication of treated and non-treated patients with OSA compared with non OSA subjects

Cosmid-derived markers anchoring the bovine genetic map to the physical map

The mapping strategy for the bovine genome described in this paper uses large insert clones as a tool for physical mapping and as a source of highly polymorphic microsatellites for genetic typing, and was one objective of the BovMap Project funded by the European Union (UE). Eight-three cosmid and phage clones were characterized and used to physically anchor the linkage groups defining all the bovine autosomes and the X Chromosome (Chr). By combining physical and genetic mapping, clones described in this paper have led to the identification of the linkage groups corresponding to Chr 9, 12, 16, and 25. In addition, anchored loci from this study were used to orient the linkage groups corresponding to Chr 3, 7, 8, 9, 13, 16, 18, 19, and 28 as identified in previously published maps. Comparison of the estimated size of the physical and linkage maps suggests that the genetic length of the bovine genome may be around 4000 c

The EASL–Lancet Liver Commission: protecting the next generation of Europeans against liver disease complications and premature mortality

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Genetic landscape of 6089 inherited retinal dystrophies affected cases in Spain and their therapeutic and extended epidemiological implications

Inherited retinal diseases (IRDs), defined by dysfunction or progressive loss of photoreceptors, are disorders characterized by elevated heterogeneity, both at the clinical and genetic levels. Our main goal was to address the genetic landscape of IRD in the largest cohort of Spanish patients reported to date. A retrospective hospital-based cross-sectional study was carried out on 6089 IRD affected individuals (from 4403 unrelated families), referred for genetic testing from all the Spanish autonomous communities. Clinical, demographic and familiar data were collected from each patient, including family pedigree, age of appearance of visual symptoms, presence of any systemic findings and geographical origin. Genetic studies were performed to the 3951 families with available DNA using different molecular techniques. Overall, 53.2% (2100/3951) of the studied families were genetically characterized, and 1549 different likely causative variants in 142 genes were identified. The most common phenotype encountered is retinitis pigmentosa (RP) (55.6% of families, 2447/4403). The most recurrently mutated genes were PRPH2, ABCA4 and RS1 in autosomal dominant (AD), autosomal recessive (AR) and X-linked (XL) NON-RP cases, respectively; RHO, USH2A and RPGR in AD, AR and XL for non-syndromic RP; and USH2A and MYO7A in syndromic IRD. Pathogenic variants c.3386G > T (p.Arg1129Leu) in ABCA4 and c.2276G > T (p.Cys759Phe) in USH2A were the most frequent variants identified. Our study provides the general landscape for IRD in Spain, reporting the largest cohort ever presented. Our results have important implications for genetic diagnosis, counselling and new therapeutic strategies to both the Spanish population and other related populations.This work was supported by the Instituto de Salud Carlos III (ISCIII) of the Spanish Ministry of Health (FIS; PI16/00425 and PI19/00321), Centro de Investigación Biomédica en Red Enfermedades Raras (CIBERER, 06/07/0036), IIS-FJD BioBank (PT13/0010/0012), Comunidad de Madrid (CAM, RAREGenomics Project, B2017/BMD-3721), European Regional Development Fund (FEDER), the Organización Nacional de Ciegos Españoles (ONCE), Fundación Ramón Areces, Fundación Conchita Rábago and the University Chair UAM-IIS-FJD of Genomic Medicine. Irene Perea-Romero is supported by a PhD fellowship from the predoctoral Program from ISCIII (FI17/00192). Ionut F. Iancu is supported by a grant from the Comunidad de Madrid (CAM, PEJ-2017-AI/BMD7256). Marta del Pozo-Valero is supported by a PhD grant from the Fundación Conchita Rábago. Berta Almoguera is supported by a Juan Rodes program from ISCIII (JR17/00020). Pablo Minguez is supported by a Miguel Servet program from ISCIII (CP16/00116). Marta Corton is supported by a Miguel Servet program from ISCIII (CPII17/00006). The funders played no role in study design, data collection, data analysis, manuscript preparation and/or publication decisions

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)1.

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field

The genetic ancestry of american creole cattle inferred from uniparental and autosomal genetic markers.

Cattle imported from the Iberian Peninsula spread throughout America in the early years of discovery and colonization to originate Creole breeds, which adapted to a wide diversity of environments and later received influences from other origins, including zebu cattle in more recent years. We analyzed uniparental genetic markers and autosomal microsatellites in DNA samples from 114 cattle breeds distributed worldwide, including 40 Creole breeds representing the whole American continent, and samples from the Iberian Peninsula, British islands, Continental Europe, Africa and American zebu. We show that Creole breeds differ considerably from each other, and most have their own identity or group with others from neighboring regions. Results with mtDNA indicate that T1c-lineages are rare in Iberia but common in Africa and are well represented in Creoles from Brazil and Colombia, lending support to a direct African influence on Creoles. This is reinforced by the sharing of a unique Y-haplotype between cattle from Mozambique and Creoles from Argentina. Autosomal microsatellites indicate that Creoles occupy an intermediate position between African and European breeds, and some Creoles show a clear Iberian signature. Our results confirm the mixed ancestry of American Creole cattle and the role that African cattle have played in their development

GENETIC CHARACTERISTICS AND DISTANCES AMONGST SPANISH AND FRENCH RABBIT POPULATIONS

[EN] A total of 990 rabbits were tested using 18 blood electrophoretic markers. These animals belong to 6 breed populations presently breed in Spain (Spanish Common, Spanish Giant, Butterfly, Burgandy Fawn, New Zealand White and Californian), one Spanish cross-breed population, three French crossbreed populations originated from the INRA selected lines (1077, 2066 and 9077) and one Wild Spanish population. 12 markers were found to be polymorphic (Dia-2, 6-Pgd, Es-1, Es-2, Es-3, Es-7, Ada, Hp, Tf, Ca-2 and Hb), each one being controlled by one locus and showing autosomal co-dominant Mendelian inheritance. We have found large differences in gene frequencies of Dia-2, 6-Pgd, Ada, Es-1, Es-2 and Es-3 loci in French populations. The genetic variabiliy estimated using the average degree of heterozygosity was lower in French than in Spanish populations. The results obtained from genetic distance analysis showed differences between French and Spanish populations. France 3 was the most divergent population. The genetic distance values obtained revealed that French populations are as different amongst themselves as they are between Spanish populations (the value obtained was corresponding to subspecie, NEI, 1987). These differences could have originated from the high selectiva pressure applied to parental lines. In the Spanish group, Spanish Giant was the most divergent population.[FR] Un total de 990 lapins appartenant a 11 populations élevés en France et en Espagne (Commun espagnol, Géant espagnol, Papil/on, Fauve de Bourgogne, Néo-Zélandais Blanc et Califomien, une lignée hybride espagnole et tros lignées hybrides fram;aises -obtenues pour croisement d'une lignée mere Néo-Zélandais Blanc et une lignée pere Califomien , toutes les deux ferrnées pendants 25 ans- et une population de lapins de garenne espagnole) ont été étudiés pour 11 marqueurs électrophorétiques sannguins polymorphes, avec une hérédité Mendélienne codominante (Dia-2, 6-Pgd, Es-1, Es-2, Es-3, Es-7, Ada, Hp, Tf, Ca-2 et Hb). On a trouvé de grandes différences pour les fréquences al/éliques des loci Dia-2, 6-Pgd, Ada, Es-1, Es-2 et Es-3 dans les lignées franr;aises. La variabilité génétique, estimée par le degré moyen d'hétérozygote, était plus faible dans les lignées franr;aises que dans les lignées espagnoles. Les résultats des distances génétiques séparent les populations franr;aises des populations espagnoles. La population France 3 se distingue nettement des autres. Les populations franr;aises sont aussi éloignées les unes des autres qu' elles le sont des populations espagnoles (la valeur obtenue correspond a la distance entre 2 sous especes). Ces différences sont peut-4tre dues 8 la grande pression de sélection réalisées dans les lignées parentales franr;aises. Le Géant espagnol étais le plus different des populations espagnoles.Martin-Burriel, I.; Marcos, S.; Osta, R.; García-Muro, E.; Zaragoza, P. (1996). GENETIC CHARACTERISTICS AND DISTANCES AMONGST SPANISH AND FRENCH RABBIT POPULATIONS. World Rabbit Science. 04(3). doi:10.4995/wrs.1996.282SWORD04