Caracterización morfológica, hematológica y bioquímica clínica en cinco razas asnales españolas para programas de conservación

Consultable des del TDXTítol obtingut de la portada digitalitzadaDeterminadas razas autóctonas de asnos (Andaluza, Catalana, Mallorquina, Asno de las Encartaciones y Zamorano-Leonesa) han sufrido a lo largo del tiempo importantes y frecuentes variaciones cuantitativas, generalmente negativas y particularmente atribuibles al olvido técnico impuesto por la marginación que ha provocado la explotación de otras especies y la industrialización del campo. La disminución de sus efectivos y el cruce indiscriminado con otras razas las ha conducido al estado de razas en peligro de extinción (RD 1682, 1997), y ha conllevado a un gran confusionismo descriptivo de sus características etnológicas. El objetivo último y fundamental del proyecto CICYT AG98-0503 que se está llevando a cabo, es sentar las bases para la conservación, mantenimiento y mejora de los animales que integran cada una de estas razas, centrándose principalmente en las fases I, II y III del protocolo marcado por la FAO para poblaciones en peligro de extinción, es decir, en la conservación «in situ». Esta tesis doctoral se ha centrado en una parte de la fase II del programa de conservación, concretamente en su caracterización racial a nivel morfológico, hematológico y bioquímico clínico. Para ello, se ha llevado a cabo un trabajo de campo centrado en la búsqueda de ejemplares de las 5 razas en distintas comunidades autónomas, con la finalidad de reflejar la situación actual de la población, tomar diferentes medidas zoométricas para realizar su caracterización fenotípica (morfológica), así como extraer muestras sanguíneas, para poder establecer los valores de referencia y normalidad tanto a nivel hematológico, bioquímico y de proteínas plasmáticas. Se efectuaron extracciones sanguíneas a 491 animales de ambos sexos, tanto jóvenes como adultos; y en los animales adultos, o sea, aquéllos mayores de 3 años (317 animales), fueron tomadas un total de 26 medidas corporales (cefálicas, troncales y extremidades) para el análisis biométrico. La descripción y análisis de los parámetros estadísticos de tendencia central y de dispersión, así como el estudio y análisis de los factores: edad, sexo y raza, tanto para las variables hematológicas, bioquímicas, proteínas plasmáticas y morfológicas, nos proporcionaron valores de referencia fiables para ser utilizados tanto en la caracterización racial como en el ámbito clínico. Mediante la caracterización morfológica, aspiramos a la diagnosis racial; esto es, al encuadre del animal objeto de observación y estudio, a un grupo etnológico diferenciado. Los resultados obtenidos a partir de las 26 mediciones biométricas nos proporcionaron datos importantes para diferenciar unos animales de otros, para agruparlos en conjuntos específicos y, fundamentalmente, para deducir proporciones que a su vez indiquen aptitudes funcionales. Estas proporciones las obtuvimos con el análisis de 12 índices zoométricos corporales (etnológicos y funcionales), los cuales evidenciaron las relaciones existentes entre algunos elementos de compacidad, alzada, longitud y peso. El análisis de las correlaciones entre las 26 medidas morfométricas, para cada una de las razas y por sexos, permitieron identificar las interacciones existentes entre y dentro las distintas regiones corporales (tronco, extremidades y cabeza), y el análisis factorial de componentes principales (ACP) permitió observar el grado de relación existente entre dichas razas, así como determinar las variables de mayor importancia en la definición morfoestructural, con la finalidad de reducir el número de variables a emplear en la caracterización práctica y rutinaria de los animales en trabajos posteriores, reduciendo así la complejidad de los mismos. Para ello la matriz de datos de las 26 variables zoométricas estudiadas fue sometida a un análisis multivariante, es decir, sometimos los datos a un análisis canónico, en el que las variables fueron transformadas en variables canónicas (factor I, II y III). Estos tres factores resumieron en total un 99,71 % de la información aportada por las 26 variables de origen, el factor I contribuyó en un 95,85 %, el factor II en un 3,25 % y el factor III en un 0,61 % al total de la varianza explicada. El perímetro torácico, alzada a la cruz, diámetro longitudinal, alzada a las palomillas, alzada al nacimiento de la cola, alzada al dorso, alzada a la pelvis, y la alzada a la grupa, diámetro dorso-esternal y longitud de la cabeza, determinaron principalmente el factor I siendo éstas por tanto, las variables de mayor peso en la caracterización racial Por otro lado, el análisis del nivel de divergencia existente entre las 5 poblaciones de asnos estudiados, basándonos en caracteres cuantitativos, lo realizamos mediante el uso de la Distancia de Mahalanobis, así como con el análisis de componentes principales (ACP). Los resultados mostraron la existencia de tres grupos, sin una evidencia clara de agrupamiento por distancia geográfica. Por ello, debemos suponer que la filogenia y evolución morfológica de los asnos peninsulares ha sido un proceso complejo, englobando distintos patrones de diferenciación para los distintos grupos de caracteres, debido probablemente a la acción medioambiental y a presiones selectivas desiguales. Las mayores diferencias las encontramos entre la raza Andaluza y el Asno de las Encartaciones, mientras que las menores se localizaron entre el asno Zamorano-Leonés y el Mallorquín. Tanto en el ACP como en el cálculo de la distancia de Mahalanobis observamos como la raza Zamorano-Leonesa es la que de algún modo se encontraría en medio de todas ellas, es decir, la que presenta menores distancias morfológicas con las cuatro restantes. Los resultados de los principales estadísticos descriptivos, analizados en el estudio de 15 variables hematológicas, 11 variables bioquímicas y proteínas plasmáticas, nos proporcionaron los valores de normalidad en cada una de las razas. El análisis de varianza para el factor raza indicó la existencia de diferencias estadísticamente significativas entre las 5 poblaciones de asnos. La edad resultó ser el factor modulador con más peso sobre los parámetros sanguíneos, ya que las diferencias encontradas entre animales jóvenes (3 años) fueron notables (11 de 15 variables hematológicas y 7 de 11 bioquímicas presentaron diferencias significativas). Sin embargo, las diferencias encontradas para el factor sexo reflejaron la existencia de un bajo dimorfismo sexual en lo referente a parámetros hematológicos y bioquímicos, y nulo en cuanto a las proteínas plasmáticas.Certain autochthonous breeds of donkeys (Andaluza, Catalana, Mallorquina, Encartaciones and Zamorano-Leonesa) have suffered important and frequent quantitative variations throughout the time, that generally have been negative, probably due to the intense mechanization of the country-side and the introduction of other species of asses. The decrease of the census of their populations and the indiscriminate mixture with other breeds have lead them to the state of «endangered breeds» (RD 1682, 1997), and have done confuse their ethnic characteristics. The main objective of this thesis is first of all, reflect the present situation of the Spanish donkey breeds and in the other hand, make the biometrical, haematological and biochemical characterization of their endangered populations. This work has followed the rules marked by the FAO Expert Consultation for the identification of possible stocks for conservation and as basic tool for the study, maintenance and conservation of animal genetic biodiversity. For the biometrical study 317 adult animals (older than 3 years) have been sampled and a total of 26 corporal measures (cephalic, trunk and extremities) for each individual were taken. The results obtained from these measurements provided us important information to differ individuals from others, to group them in specific sets and, mainly, to deduce proportions that indicate functional aptitudes. We obtained these proportions analysing 12 corporal indices (ethnological and functional), which demonstrates the existing relationships between some elements of compactness, height, length and weight. For the haematological, biochemical and plasma proteins study, blood samples have taken from 491 animals of both genders and age (young and adult). The description and analysis of the average, variance, and age, sex and race factors provided us guaranteed reference values to be used for the racial characterization and for the clinical scope. The correlation's analyses between the 26 corporal measures, for each one of the breeds and genders, allowed identifying the existing interactions between and within the different corporal regions (trunk, extremities and head). The factorial analysis of principal components (ACP) allowed to observe the existing relationship degree between these breeds, and to determine the most important variables of the morphological definition, with the purpose to reduce the number of variables to be used in the practical and routine characterization of the animals in later studies, reducing then the complexity of them. To do it, all the information of the 26 corporal variables was subject to a multivariate analysis, in fact, subject in a canonical analysis, where the variables were transformed into canonical variables (factor I, II and III). These three factors summarized 99.71 % of the information by the original 26 variables, the factor I contributed in 95.85 %, factor II in 3.25 % and factor III in 0.61 % to the total of the explained variance. The thoracic perimeter, length diameter, withers height, sacrum height, root of tail height, back height, pelvis height, and rump height, the back-sternal diameter and head length, mainly determined factor I, being the heaviest weight variables for the racial characterization. On the other hand, using the Mahalanobis Distance, and also with ACP, we obtained the divergence level between the 5 donkey populations studied, according on quantitative characters. The results showed the existence of three groups, without clear evidence of clustering by geographic distance. For this reason, we must suppose that phylogenetic and morphologic evolution of the peninsular donkeys have been a complex process, including different patterns of differentiation for the different characters groups, probably coming from the environmental action and to unequal selective pressures. The greater differences were found between the Andaluza and Encartaciones breeds, while the minors were located between the Zamorano-Leonesa and Mallorquina. Using the ACP and also in the Mahalanobis Distance, we observed that the Zamorano-Leonesa breed is the one that presents minor morphologic distances with the others. The analysis of breed factor of 15 haematological parameters, 11 biochemical parameters and plasma proteins indicated the existence of statistically significant differences between the 5 populations analyzed. Age had the most influence on blood parameters because the differences found between young and adult animals were remarkable (11 of 15 haematological variables and 7 of 11 biochemical ones presented significant differences). In contrast, less significant differences between genders were found for sex factor so we demonstrate the existence of a low sexual dimorphism for haematological and biochemical parameters

Facile solid-phase synthesis of biotinylated alkyl thiols

Biotinylated alkyl thiols with the capacity to graft avidin proteins are in increasing demand for the development of self-assembled monolayers on gold. Here we propose 2-Chlorotrityl Chloride solid-phase resin as a new platform to produce these functionalized alkyl thiols. Biotinylated alkyl thiols of non-obvious solution synthesis were obtained rapidly using this method and without previous purification steps. © 2006 Elsevier Ltd. All rights reserved

Subconjunctival injection of mesenchymal stromal cells protects the cornea in an experimental model of GVHD

Purpose: To evaluate the therapeutic effect of subconjunctival injection of human mesenchymal stromal cells (hMSCs) in the cornea of mice with graft versus host disease (GVHD). Methods: GVHD was induced in mice after hematopoietic stem cell transplantation (HSCT) between MHC-mismatched mouse strains. Subconjunctival injection of hMSCs was applied at day 10 post-HSCT. Infiltration of CD3+ cells in the cornea and epithelial alterations were analyzed by immunofluorescence. Tear was assessed using the PRT test and TearLab Osmolarity System. qPCR was used to evaluate changes in cytokines, Pax6 and Sprr1b expression. To evaluate the effect of irradiation, we analyzed the expression of these genes in TBI mice. Results: Immune cell invasion occurs in mice with GVHD, as shown by the presence of CD3+ cells in the cornea. Interestingly, eyes treated with hMSC did not present CD3+ cells. Tear osmolarity was increased in GVHD eyes, but not in treated eyes. TNFa expression was highly increased in all corneas except in Control and treated eyes. Pax6 in corneal epithelium showed a similar pattern in GVHD and Control mice, and its gene expression was enhanced in GVHD corneas. In contrast, Pax6 was reduced in GVHD + MSC corneas. We also found an increase in SPRR1B staining in GVHD eyes that was lower in GVHD + MSC mice, demonstrating that corneal keratinization is less frequent after treatment with hMSC. Conclusions: The treatment with hMSCs by subconjunctival injection is effective in reducing corneal inflammation and squamous metaplasia in ocular GVHD (oGVHD). Local treatment with hMSCs is a promising strategy for oGVHD.This study was supported by Fund for Health of Spain (FIS) grant PI12/00939 and Red de Terapia Celular de Castilla y León. Rafael Martínez-Carrasco was supported by a grant from Junta de Castilla y León. A. Velasco, J. Aijón and E. Hernández-Galilea belong to UIC.077 and L.I. Sánchez-Abarca and F. Sánchez-Guijo to UIC-116 from Junta de Castilla y León

Versatile Graphene-Based Platform for Robust Nanobiohybrid Interfaces

Technologically useful and robust graphene-based interfaces for devices require the introduction of highly selective, stable, and covalently bonded functionalities on the graphene surface, whilst essentially retaining the electronic properties of the pristine layer. This work demonstrates that highly controlled, ultrahigh vacuum covalent chemical functionalization of graphene sheets with a thiol-terminated molecule provides a robust and tunable platform for the development of hybrid nanostructures in different environments. We employ this facile strategy to covalently couple two representative systems of broad interest: metal nanoparticles, via S-metal bonds, and thiol-modified DNA aptamers, via disulfide bridges. Both systems, which have been characterized by a multi-technique approach, remain firmly anchored to the graphene surface even after several washing cycles. Atomic force microscopy images demonstrate that the conjugated aptamer retains the functionality required to recognize a target protein. This methodology opens a new route to the integration of high-quality graphene layers into diverse technological platforms, including plasmonics, optoelectronics, or biosensing. With respect to the latter, the viability of a thiol-functionalized chemical vapor deposition graphene-based solution-gated field-effect transistor array was assessed

Anatomical Site, Typing, Virulence Gene Profiling, Antimicrobial Susceptibility and Resistance Genes of Streptococcus suis Isolates Recovered from Pigs in Spain

Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/).[EN] A set of 207 Streptococcus suis isolates were collected from ten autonomous communities from Spain in 2019 to 2020 from pigs with meningitis, pneumonic lungs, arthritic joints or other swollen viscera, to a lesser extent. Thirteen capsular types were detected being the most prevalent serotype 2 (21.7%), followed by serotypes 1 (21.3%), 9 (19.3%) and 3 (6.3%). Serotypes 2 and 9 were recovered mainly from the central nervous system (CNS), while serotype 1 was isolated mostly from swollen joints and serotype 3 from the lungs. Twenty-five isolates (12.1%) could not be typed. The most prevalent pathotype was epf + mrp + sly + luxS (49 isolates, 23.8%), and it was related mainly to serotypes 1 and 2. Serotypes 1–3 and 9 were significantly associated with anatomical sites of isolation and virulence factors, serotype 9 (CNS) and serotypes 3 and 9 (lungs) being associated with virulence profiles without the epf gene. S. suis isolates showed globally high antimicrobial resistances, but ampicillin followed by spectinomycin and tiamulin resulted in the highest activities, while the greatest resistances were detected for sulphadimethoxine, tetracyclines, neomycin, clindamycin and macrolides. A total of 87.4% isolates were positive to the tetO gene, 62.4% to the ermB gene and 25.2% to the fexA gene, while 14.6% were positive to all three genes simultaneously. A significative association between isolate resistances to tetracyclines and macrolides and the resistance genes tested was established, except for phenicol resistance and the fexA gene. A set of 14 multiresistance patterns were obtained according to the number of antimicrobials to which the isolates were resistant, the resistances to 12 or more agents being the most prevalent ones. A remarkable amount of multiresistance profiles could be seen among the S. suis serotype 9 isolates.SIWe acknowledge Laboratorios SYVA for the assignment of the 207 S. suis isolates because they are property of this Spanish laboratory.This research was funded by the contract project called “Obtención y caracterización de aislados de Hameophilus parasuis y Streptococcus suis en gando porcino de Castilla y León para la realización de una autovacuna eficaz” (C-309), financed by Laboratorios SYVA S.A.U., León. M.P.R. was a recipient of a contract grant from this Spanish laboratory

ROS-Scavenging Enzymes as an Antioxidant Response to High Concentration of Anthracene in the Liverwort <i>Marchantia polymorpha</i> L.

Marchantia polymorpha L. responds to environmental changes using a myriad set of physiological responses, some unique to the lineage related to the lack of a vascular- and root-system. This study investigates the physiological response of M. polymorpha to high doses of anthracene analysing the antioxidant enzymes and their relationship with the photosynthetic processes, as well as their transcriptomic response. We found an anthracene dose-dependent response reducing plant biomass and associated to an alteration of the ultrastructure of a 23.6% of chloroplasts. Despite a reduction in total thallus-chlorophyll of 31.6% of Chl a and 38.4% of Chl b, this was not accompanied by a significant change in the net photosynthesis rate and maximum quantum efficiency (Fv/Fm). However, we found an increase in the activity of main ROS-detoxifying enzymes of 34.09% of peroxidase and 692% of ascorbate peroxidase, supported at transcriptional level with the upregulation of ROS-related detoxifying responses. Finally, we found that M. polymorpha tolerated anthracene-stress under the lowest concentration used and can suffer physiological alterations under higher concentrations tested related to the accumulation of anthracene within plant tissues. Our results show that M. polymorpha under PAH stress condition activated two complementary physiological responses including the activation of antioxidant mechanisms and the accumulation of the pollutant within plant tissues to mitigate the damage to the photosynthetic apparatus

Circulating Cell Biomarkers in Pulmonary Arterial Hypertension: Relationship with Clinical Heterogeneity and Therapeutic Response

Background: Endothelial dysfunction is central to PAH. In this study, we simultaneously analysed circulating levels of endothelial microvesicles (EMVs) and progenitor cells (PCs) in PAH and in controls, as biomarkers of pulmonary endothelial integrity and evaluated differences among PAH subtypes and as a response to treatment. Methods: Forty-seven controls and 144 patients with PAH (52 idiopathic, 9 heritable, 31 associated with systemic sclerosis, 15 associated with other connective tissue diseases, 20 associated with HIV and 17 associated with portal hypertension) were evaluated. Forty-four patients with scleroderma and 22 with HIV infection, but without PAH, were also studied. Circulating levels of EMVs, total (CD31+CD42b-) and activated (CD31+CD42b-CD62E+), as well as circulating PCs (CD34+CD133+CD45low) were measured by flow cytometry and the EMVs/PCs ratio was computed. In treatment-naïve patients, measurements were repeated after 3 months of PAH therapy. Results: Patients with PAH showed higher numbers of EMVs and a lower percentage of PCs, compared with healthy controls. The EMV/PC ratio was increased in PAH patients, and in patients with SSc or HIV without PAH. After starting PAH therapy, individual changes in EMVs and PCs were variable, without significant differences being observed as a group. Conclusion: PAH patients present disturbed vascular homeostasis, reflected in changes in circulating EMV and PC levels, which are not restored with PAH targeted therapy. Combined measurement of circulating EMVs and PCs could be foreseen as a potential biomarker of endothelial dysfunction in PAH

The Influence of Genetic and Environmental Factors among MDMA Users in Cognitive Performance

This study is aimed to clarify the association between MDMA cumulative use and cognitive dysfunction, and the potential role of candidate genetic polymorphisms in explaining individual differences in the cognitive effects of MDMA. Gene polymorphisms related to reduced serotonin function, poor competency of executive control and memory consolidation systems, and high enzymatic activity linked to bioactivation of MDMA to neurotoxic metabolites may contribute to explain variations in the cognitive impact of MDMA across regular users of this drug. Sixty ecstasy polydrug users, 110 cannabis users and 93 non-drug users were assessed using cognitive measures of Verbal Memory (California Verbal Learning Test, CVLT), Visual Memory (Rey-Osterrieth Complex Figure Test, ROCFT), Semantic Fluency, and Perceptual Attention (Symbol Digit Modalities Test, SDMT). Participants were also genotyped for polymorphisms within the 5HTT, 5HTR2A, COMT, CYP2D6, BDNF, and GRIN2B genes using polymerase chain reaction and TaqMan polymerase assays. Lifetime cumulative MDMA use was significantly associated with poorer performance on visuospatial memory and perceptual attention. Heavy MDMA users (>100 tablets lifetime use) interacted with candidate gene polymorphisms in explaining individual differences in cognitive performance between MDMA users and controls. MDMA users carrying COMT val/val and SERT s/s had poorer performance than paired controls on visuospatial attention and memory, and MDMA users with CYP2D6 ultra-rapid metabolizers performed worse than controls on semantic fluency. Both MDMA lifetime use and gene-related individual differences influence cognitive dysfunction in ecstasy users

How can I assess my patients with Parkinson's disease during a busy clinic day?

[Background] The evaluation of motor impairment in Parkinson's disease (PD) is mainly assessed with the motor subdomain of the Unified Parkinson's Disease Rating scale (UPDRS part III) and, lately, with the MDS-UPDRS part III. To optimize efforts and special needs during specific circumstances in clinical practice, we sought to identify the most sensitive items to assess motor impairment in PD.[Methods] We included the COPPADIS-PD cohort and collected the UPDRS part III at baseline (V0), 12 months (V1), and 24 months (V2). Factor analysis and effect size using Cohen's d formula were performed in the Off and On states at V0, V1, and V2.[Results] We included 667 patients with PD, mean age of 62.59 ± 8.91 years, 410 (60.2%) males, with a median HY stage of 2.00 (1.00; 4.00) at baseline. Over time, the most discriminating items were postural stability and body bradykinesia (“arise from chair” and “gait”) in the Off state, right and left upper extremity bradykinesia ("finger tap", "hand movements" and "prono/supination") in the On state. Body bradykinesia and right-left finger tapping were the items with the largest effect size (0.93, 0.84, 0.83, respectively) to assess motor improvement after receiving antiparkinsonian medications over time.[Conclusion] Under specific circumstances, selecting a few items of the UPDRS part III, including postural stability, body bradykinesia, and upper extremity bradykinesia, could be used to create a quick clinical judgment of motor status and improvement in PD.This work was supported by the Instituto de Salud Carlos III-Fondo Europeo de Desarrollo Regional (ISCIII-FEDER) [PI14/01823, PI16/01575, PI18/01898, PI19/01576, PI20/00613], the Consejería de Economía, Innovación, Ciencia y Empleo de la Junta de Andalucía [CVI-02526, CTS-7685], the Consejería de Salud y Bienestar Social de la Junta de Andalucía [PI-0471-2013, PE-0210-2018, PI-0459-2018, PE-0186-2019], the Fundación Alicia Koplowitz and the Fundación "Curemos el Parkinson".Peer reviewe