[Exposure to service user violence and job satisfaction among nursing staff in public hospitals in Murcia Region]. / Exposición a violencia de los usuarios y satisfacción laboral en el personal de enfermería de los hospitales públicos de la Región de Murcia.

BACKGROUND: Manifestations of physical and psychological violence can affect the levels of job satisfaction. The objective of this study is to determine the frequency of exposure to user violence by the nursing staff in public hospitals in the region of Murcia and its relationship with their level of job satisfaction. METHODS: Cross-sectional descriptive study through a self-administered and anonymous questionnaire sent to the nursing staff of the public hospitals in the region of Murcia (1,489 professionals). The instruments used were the Hospital Aggressive Behaviour Scale-Users (HABS-U) to rate violence and the Overall Job Satisfaction (OJS) to assess job satisfaction. RESULTS: Most workers (71%) reported suffering at least one of the manifestations of non-physical violence and 19.9% one of the physical manifestations with at least an annual frequency. Mean extrinsic satisfaction was 30.1 (SD=6.9), being coworkers the factor with higher frequency of satisfied workers (68.2%) and how the company was managed the factor with more dissatisfied workers (28.8%). Intrinsic satisfaction was lower (25.5, SD=7.2), being variety of tasks the factor with more satisfied workers (51.7%) and promotion opportunity the one with more dissatisfied workers (22.4%). Both extrinsic and intrinsic satisfaction correlated inversely with the exposure to non-physical and physical violence (higher exposure to violence lowered job satisfaction). CONCLUSIONS: In general, the scores for nurses' job satisfaction were low. One of the aspects affecting this dissatisfaction might be exposure to workplace violence; thus reducing exposure to violence could increase job satisfaction that, in turn, would improve patient care.

Practical considerations for clinical use of mesenchymal stem cells: From the laboratory to the horse.

Since the clinical use of mesenchymal stem cells (MSCs) for treating musculoskeletal injuries is gaining popularity, practitioners should be aware of the factors that may affect MSCs from tissue harvesting for MSC isolation to cell delivery into the injury site. This review provides equine practitioners with up-to-date, practical knowledge for the treatment of equine patients using MSCs. A brief overview of laboratory procedures affecting MSCs is provided, but the main focus is on shipping conditions, routes of administration, injection methods, and which commonly used products can be combined with MSCs and which products should be avoided as they have deleterious effects on cells. There are still several knowledge gaps regarding MSC-based therapies in horses. Therefore, it is important to properly manage the factors which are currently known to affect MSCs, to further strengthen the evidence basis of this treatment.

Exposición a violencia de los usuarios y satisfacción laboral en el personal de enfermería de los hospitales públicos de la Región de Murcia / Exposure to service user violence and job satisfaction among nursing staff in public hospitals in Murcia Region

Fundamento: Las manifestaciones de violencia tanto física como psicológica (no física) pueden afectar a los niveles de satisfacción laboral. El objetivo de este estudio fue determinar la frecuencia de exposición del personal de enfermería (incluidos auxiliares) a conductas violentas de baja intensidad de usuarios y comprobar si existe una relación entre esta exposición y el nivel de satisfacción laboral. Material y métodos: Estudio descriptivo transversal a través de un cuestionario anónimo autoaplicado al personal de enfermería de los hospitales públicos de la Región de Murcia (1.489 profesionales). Los instrumentos utilizados fueron la escala HABS-U (Hospital Aggressive Behaviour Scale-Users) para valorar la violencia y la OJS (Overall Job Satisfaction) para valorar la satisfacción laboral. Resultados: La frecuencia de trabajadores que, al menos anualmente, refirieron haber sufrido alguna manifestación de violencia fue superior en el caso de la violencia no física (71,0%) que de la física (19,9%). La satisfacción extrínseca media fue 30,1 (DT=6,9), siendo los compañeros de trabajo el factor con más trabajadores satisfechos (68,2%) frente al modo en el que la empresa está gestionada, factor con mayor porcentaje de insatisfechos (28,8%). La satisfacción intrínseca fue menor (M=25,5 y DT= 7,2), siendo la variedad de tareas que realiza en el trabajo el aspecto con mayor porcentaje de satisfechos (51,7%) y la posibilidad de promoción el factor con más insatisfechos (22,4%). Se observó que la satisfacción, tanto extrínseca como intrínseca, se asoció de forma inversa con la exposición a violencia no física y física: una mayor exposición a violencia disminuyó la satisfacción. Conclusiones: En general, las puntuaciones de satisfacción laboral en el personal de enfermería fueron bajas. Uno de los aspectos que podría afectar es la exposición a violencia laboral, por lo que disminuir la exposición a violencia podría aumentar la satisfacción laboral, lo que probablemente redundaría en una mejor atención al paciente

Background: Manifestations of physical and psychological violence can affect the levels of job satisfaction. The objective of this study is to determine the frequency of exposure to user violence by the nursing staff in public hospitals in the region of Murcia and its relationship with their level of job satisfaction. Methods: Cross-sectional descriptive study through a selfadministered and anonymous questionnaire sent to the nursing staff of the public hospitals in the region of Murcia (1,489 professionals). The instruments used were the Hospital Aggressive Behaviour Scale-Users (HABS-U) to rate violence and the Overall Job Satisfaction (OJS) to assess job satisfaction. Results: Most workers (71%) reported suffering at least one of the manifestations of non-physical violence and 19.9% one of the physical manifestations with at least an annual frequency. Mean extrinsic satisfaction was 30.1 (SD=6.9), being coworkers the factor with higher frequency of satisfied workers (68.2%) and how the company was managed the factor with more dissatisfied workers 28.8%). Intrinsic satisfaction was lower (25.5, SD=7.2), being variety of tasks the factor with more satisfied workers (51.7%) and promotion opportunity the one with more dissatisfied workers (22.4%). Both extrinsic and intrinsic satisfaction correlated inversely with the exposure to non-physical and physical violence (higher exposure to violence lowered job satisfaction). Conclusions: In general, the scores for nurses’ job satisfaction were low. One of the aspects affecting this dissatisfaction might be exposure to workplace violence; thus reducing exposure to violence could increase job satisfaction that, in turn, would improve patient care

Dissection of ancestral genetic contributions to Creole goat populations.

Goats have played a key role as source of nourishment for humans in their expansion all over the world in long land and sea trips. This has guaranteed a place for this species in the important and rapid episode of livestock expansion triggered by Columbus' arrival in the Americas in the late 1400s. The aims of this study are to provide a comprehensive perspective on genetic diversity in American goat populations and to assess their origins and evolutionary trajectories. This was achieved by combining data from autosomal neutral genetic markers obtained in more than two thousand samples that encompass a wide range of Iberian, African and Creole goat breeds. In general, even though Creole populations differ clearly from each other, they lack a strong geographical pattern of differentiation, such that populations of different admixed ancestry share relatively close locations throughout the large geographical range included in this study. Important Iberian signatures were detected in most Creole populations studied, and many of them, particularly the Cuban Creole, also revealed an important contribution of African breeds. On the other hand, the Brazilian breeds showed a particular genetic structure and were clearly separated from the other Creole populations, with some influence from Cape Verde goats. These results provide a comprehensive characterisation of the present structure of goat genetic diversity, and a dissection of the Iberian and African influences that gave origin to different Creole caprine breeds, disentangling an important part of their evolutionary history. Creole breeds constitute an important reservoir of genetic diversity that justifies the development of appropriate management systems aimed at improving performance without loss of genomic diversity.

Molecular analysis of three Clostridium difficile strain genomes isolated from pig farm-related samples.

Clostridium difficile is an anaerobic spore-forming bacillus that usually causes gastrointestinal disorders in man and other animal species. Most of the strains isolated from animals are toxigenic being the virulent ribotype (RT) 078 predominant in several animal species. Although C. difficile is pathogenic to both humans and animals, there is no direct evidence of zoonosis. Deep genome sequencing provides sufficient resolution to analyse which strains found in animals might be related to human pathogens. So far, there are only a few fully sequenced genomes of C. difficile strains isolated from domestic and wild animals. Using Illumina technology, we have sequenced the genome of three isolates; a strain isolated from the vagina of a sow (5754), one from rat (Rattus spp) intestinal content (RC10) and a third one isolated from environmental rat faeces (RF17). Both, rat and rat faeces were sampled in fattening pig farms. Our study reveals a close genetic relationship of two of these isolates with the virulent strain M120 (RT078) isolated from a human patient. The analysis of the sequences has revealed the presence of antibiotic resistance genes, mobile elements, including the transposon linked with virulence Tn6164, and the similarity of virulence factors between these isolates and human strains. This is the first study focused on the sequencing of C. difficile genomes obtained from wild animals like rats, which can be considered as potential reservoirs for humans and other animal species. This study can help to understand the genome composition and epidemiology of this bacterium species.

Comparison of autologous bone marrow and adipose tissue derived mesenchymal stem cells, and platelet rich plasma, for treating surgically induced lesions of the equine superficial digital flexor tendon.

Several therapies have been investigated for equine tendinopathies, but satisfactory long term results have not been achieved consistently and a better understanding of the healing mechanism elicited by regenerative therapies is needed. The aim of this study was to assess the separate effects of autologous bone marrow (BM) and adipose tissue (AT) derived mesenchymal stem cells (MSCs), and platelet rich plasma (PRP), for treating lesions induced in the superficial digital flexor tendon (SDFT) of horses. Lesions were created surgically in both SDFTs of the forelimbs of 12 horses and were treated with BM-MSCs (six tendons), AT-MSCs (six tendons) or PRP (six tendons). The remaining six tendons received lactated Ringer's solution as control. Serial ultrasound assessment was performed prior to treatment and at 2, 6, 10, 20 and 45 weeks post-treatment. At 45 weeks, histopathology and gene expression analyses were performed. At week 6, the ultrasound echogenicity score in tendons treated with BM-MSCs suggested earlier improvement, whilst all treatment groups reached the same level at week 10, which was superior to the control group. Collagen orientation scores on histological examination suggested a better outcome in treated tendons. Gene expression was indicative of better tissue regeneration after all treatments, especially for BM-MSCs, as suggested by upregulation of collagen type I, decorin, tenascin and matrix metalloproteinase III mRNA. Considering all findings, a clear beneficial effect was elicited by all treatments compared with the control group. Although differences between treatments were relatively small, BM-MSCs resulted in a better outcome than PRP and AT-MSCs.

The legacy of Columbus in American horse populations assessed by microsatellite markers.

Criollo horse populations descend from horses brought from the Iberian Peninsula over the period of colonization (15th to 17th century). They are spread throughout the Americas and have potentially undergone genetic hybridization with other breeds in the recent past. In this study, 25 autosomal microsatellites were genotyped in 50 horse breeds representing Criollo populations from 12 American countries (27 breeds), breeds from the Iberian Peninsula (19), one breed each from France and Morocco and two cosmopolitan horse breeds (Thoroughbred and Arabian). The genetic relationships among breeds identified five clusters: Celtic; Iberian; North American with Thoroughbred influence; most Colombian breeds; and nearly all other Criollo breeds. The group of "all other Criollo breeds" had the closest genetic relationship with breeds originating from the Iberian Peninsula, specifically with the Celtic group. For the whole set of Criollo breeds analysed, the estimated genetic contribution from other breeds was approximately 50%, 30% and 20% for the Celtic, Iberian and Arab-Thoroughbred groups, respectively. The spatial distribution of genetic diversity indicates that hotspots of genetic diversity are observed in populations from Colombia, Ecuador, Brazil, Paraguay and western United States, possibly indicating points of arrival and dispersion of Criollo horses in the American continent. These results indicate that Criollo breeds share a common ancestry, but that each breed has its own identity.

Inflammatory response to the administration of mesenchymal stem cells in an equine experimental model: effect of autologous, and single and repeat doses of pooled allogeneic cells in healthy joints.

BACKGROUND: Mesenchymal stem cells (MSCs) transplantation has become a promising therapeutic choice for musculoskeletal injuries. Joint-related disorders are highly prevalent in horses. Therefore, these animals are considered as suitable models for testing MSC-based therapies for these diseases. The aim of this study was to investigate the clinical and inflammatory responses to intra-articular single and repeat dose administration of autologous or of pooled allogeneic MSCs in healthy equine healthy joints. Six horses were intra-articularly injected with a single autologous dose of bone marrow derived MSCs (BM-MSCs) and two separate doses of allogeneic BM-MSCs pooled from several donors. All contralateral joints were injected with Lactated Ringer's Solution (LRS) as the control vehicle. Signs of synovitis and lameness were evaluated at days 0, 1, 2, 3, 5 and 10 after injection. Total protein (TP), white blood cell count (WBC) and neutrophil count (NC) in synovial fluid were also measured at the same time-points. RESULTS: A mild synovial effusion without associated lameness was observed after all BM-MSCs injections. The second allogeneic injection caused the lowest signs of synovitis. Local temperature slightly increased after all BM-MSCs treatments compared to the controls. TP, WBC and NC in synovial fluids also increased during days 1 to 5 after all BM-MSCs injections. Both, clinical and synovial parameters were progressively normalized and by day 10 post-inoculation appeared indistinguishable from controls. CONCLUSIONS: Intra-articular administration of an allogeneic pool of BM-MSCs represents a safe therapeutic strategy to enhance MSCs availability. Importantly, the absence of hypersensitivity response to the second allogeneic BM-MSCs injection validates the use of repeat dose treatments to potentiate the therapeutic benefit of these cells. These results notably contribute to the development of stem cell based therapies for equine and human joint diseases.

Conservation priorities of Iberoamerican pig breeds and their ancestors based on microsatellite information.

Criollo pig breeds are descendants from pigs brought to the American continent starting with Columbus second trip in 1493. Pigs currently play a key role in social economy and community cultural identity in Latin America. The aim of this study was to establish conservation priorities among a comprehensive group of Criollo pig breeds based on a set of 24 microsatellite markers and using different criteria. Spain and Portugal pig breeds, wild boar populations of different European geographic origins and commercial pig breeds were included in the analysis as potential genetic influences in the development of Criollo pig breeds. Different methods, differing in the weight given to within- and between-breed genetic variability, were used in order to estimate the contribution of each breed to global genetic diversity. As expected, the partial contribution to total heterozygosity gave high priority to Criollo pig breeds, whereas Weitzman procedures prioritized Iberian Peninsula breeds. With the combined within- and between-breed approaches, different conservation priorities were achieved. The Core Set methodologies highly prioritized Criollo pig breeds (Cr. Boliviano, Cr. Pacifico, Cr. Cubano and Cr. Guadalupe). However, weighing the between- and within-breed components with FST and 1-FST, respectively, resulted in higher contributions of Iberian breeds. In spite of the different conservation priorities according to the methodology used, other factors in addition to genetic information also need to be considered in conservation programmes, such as the economic, cultural or historical value of the breeds involved.

Effect of inflammatory environment on equine bone marrow derived mesenchymal stem cells immunogenicity and immunomodulatory properties.

Mesenchymal stem cells (MSCs) are being investigated for the treatment of equine joint diseases because of their regenerative potential. Recently, the focus mainly has addressed to their immunomodulatory capacities. Inflammation plays a central role in joint pathologies, since the release of proinflammatory mediators to the synovial fluid (SF) leads to the activation of enzymatic degradation of the cartilage. MSCs can modulate the local immune environment through direct or paracrine interaction with immune cells, suppressing their proliferation and re-addressing their functions. Proinflammatory molecules can induce MSC immunoregulatory potential, but they could also increase the expression of immunogenic molecules. Studying the effect of inflammatory environment on MSC immunomodulation and immunogenicity profiles is mandatory to improve cellular therapies. The aim of this study was to analyse the response of equine bone marrow MSCs (eBM-MSCs) to three inflammatory conditions. Equine BM-MSCs from three animals were exposed to: (a) 20% allogeneic inflammatory SF (SF); (b) 50 ng/ml of TNFα and IFNγ (CK50) and (c) 20 ng/ml of TNFα and IFNγ (CK20). After 72 h of exposure, expression of immunogenic and immunomodulation-related molecules, including cell-to-cell contact and paracrine signalling molecules, were analysed by RT-qPCR and flow cytometry. The gene expression of adhesion molecules was upregulated whereas MSC migration-related genes were downregulated by all inflammatory conditions tested. CK culture conditions significantly upregulated the expression of COX-2, iNOS, IDO and IL-6. MHC-I gene expression was upregulated by all conditions, whereas MHC-II was upregulated only after CK priming. The expression of CD40 did not significantly change, whereas the ligand, CD40L, was downregulated in CK conditions. Flow cytometry showed an increase in the percentage of positive cells and mean fluorescence intensity (MFI) of the MHC-I and MHC-II molecules at CK50 conditions, supporting the gene expression results. These outcomes reinforce the change of the immunophenotype of the eBM-MSCs according to the surrounding conditions. Inflammatory synovial environment did not lead to significant changes, so the environment found by eBM-MSCs when they are intraarticular administered may not be enough to activate their immunomodulatory potential. CK priming at tested doses enhances the immunoregulatory profile of eBM-MSCs, which may promote a therapeutic benefit. Even if CK priming induced an upregulation of MHC expression, costimulatory molecule expression however was not upregulated, suggesting that immunogenicity might not be increased. This study provides a better understanding about the behaviour of eBM-MSCs inside the inflamed joint and constitutes a first step to improve MSC-based therapies for equine joint diseases.

Genetic diversity, structure and individual assignment of Casta Navarra cattle: a well-differentiated fighting bull population.

The Casta Navarra lineage was one of the populations used to establish the fighting bull (FB) breed, and it has also been reproductively isolated from the others FBs. A total of 1284 individuals from two generations of 16 Casta Navarra herds were sampled to analyse their diversity, their genetic structure and the ability of 28 microsatellite markers to assign individuals to closely related populations. These animals were compared with closely related phylogenetic (FB) or geographical (Pirenaica and Monchina) populations. Hardy-Weinberg equilibrium analysis showed that 82% of the loci had a significant heterozygote deficit as a consequence of the Wahlund effect. The average proportion of genetic variation explained by farm differences was 9% by Wright's FST index. A phylogenetic tree constructed with a neighbour-joining method based on Reynolds genetic distances and a Bayesian Markov chain Monte Carlo clustering approach revealed clear differences between farm groups that generally corresponded to historical information and could unambiguously differentiate Casta Navarra cattle from the other populations. The percentage of animals correctly assigned to the Casta Navarra population was 91.78% for a q threshold of >0.9. Admixture was only detected in 4.45% (q < 0.8) of the cattle. These results are relevant for the maintenance and development of diversity and conservation in the Casta Navarra population.

Dacriocistorrinostomía endonasal: participación del oftalmólogo / Endoscopic dacryocystorhinostomy: Role of the ophthalmologist

OBJETIVO: Evaluar los resultados de la dacriocistorrinostomía (DCR) endoscópica con o sin apoyo del oftalmólogo. MATERIAL Y MÉTODOS: Estudio retrospectivo de 100 casos operados de DCR endoscópica por un otorrinolaringólogo entre junio de 2008 y diciembre de 2009. De las 100, 50 fueron intervenidas con apoyo quirúrgico de un oftalmólogo, que canalizaba los canalículos superior e inferior con sondas de Bowman, mientras que en los otros 50 casos fue el propio otorrinolaringólogo quién realizó esta canalización, sin apoyo del oftalmólogo. La evaluación de los resultados a 2 años incluyó la percepción subjetiva, la permeabilidad de la vía lagrimal tras lavado y la permeabilidad funcional de la vía lagrimal tras test de Jones modificado. RESULTADOS: De las 100 DCR revisadas, en más del 50% se realizó algún tratamiento complementario por el otorrinolaringólogo, sobre todo septoplastias. En cuanto a la resolución de la epífora, cuando no hubo oftalmólogo presente en el quirófano, los pacientes referían mejoría subjetiva total en 75%; sin embargo, en el grupo cuya canalización quirúrgica era realizada por el oftalmólogo, los resultados satisfactorios llegaban al 92% de forma subjetiva, diferencia estadísticamente significativa. CONCLUSIONES: La DCR endoscópica es efectiva en el tratamiento de la epífora, pero sus resultados mejoran cuando el oftalmólogo colabora en el procedimiento quirúrgico, canalizando la vía lagrimal

OBJECTIVE: To evaluate the results of endoscopic dacryocystorhinostomy (DCR) with or without support of the ophthalmologist. MATERIAL AND METHODS: A retrospective study of 100 cases of endoscopic DCR surgery conducted by an otolaryngologist between June 2008 and December 2009. Of the 100 cases, 50 were operated with surgical support of the ophthalmologist, who inserted Bowman probes in the upper and lower canaliculi, while in the other 50 cases it was the otolaryngologist who performed this, without support of the ophthalmologist. The evaluation of the results after 2 years included the subjective perception, the lacrimal patency after lacrimal syringing, and lacrimal functional test after modified Jones test. RESULTS: Of the 100 DCR reviewed, more than 50% required complementary treatment by the otolaryngologist, mainly septoplasty. As for the resolution of epiphora, without support of the ophthalmologist, 75% the patients reported an overall subjective improvement, but this reached 92% in the surgical group with support of the ophthalmologist, which was a statistically significant difference. CONCLUSIONS: Endoscopic DCR is effective in the treatment of epiphora, but its results improve when the ophthalmologist inserts the probes in the lacrimal canaliculi during the surgical procedure

[Endoscopic dacryocystorhinostomy: role of the ophthalmologist]. / Dacriocistorrinostomía endonasal: participación del oftalmólogo.

OBJECTIVE: To evaluate the results of endoscopic dacryocystorhinostomy (DCR) with or without support of the ophthalmologist. MATERIAL AND METHODS: A retrospective study of 100 cases of endoscopic DCR surgery conducted by an otolaryngologist between June 2008 and December 2009. Of the 100 cases, 50 were operated with surgical support of the ophthalmologist, who inserted Bowman probes in the upper and lower canaliculi, while in the other 50 cases it was the otolaryngologist who performed this, without support of the ophthalmologist. The evaluation of the results after 2 years included the subjective perception, the lacrimal patency after lacrimal syringing, and lacrimal functional test after modified Jones test. RESULTS: Of the 100 DCR reviewed, more than 50% required complementary treatment by the otolaryngologist, mainly septoplasty. As for the resolution of epiphora, without support of the ophthalmologist, 75% the patients reported an overall subjective improvement, but this reached 92% in the surgical group with support of the ophthalmologist, which was a statistically significant difference. CONCLUSIONS: Endoscopic DCR is effective in the treatment of epiphora, but its results improve when the ophthalmologist inserts the probes in the lacrimal canaliculi during the surgical procedure.

Genetic diversity and differentiation of five Cuban cattle breeds using 30 microsatellite loci.

Conservation and improvement strategies in farm animals should be based on a combination of genetic and phenotypic characteristics. Genotype data from 30 microsatellites were used to assess the genetic diversity and relationships among five Cuban cattle breeds (Siboney de Cuba, Criollo Cubano, Cebú Cubano, Mambí de Cuba and Taíno de Cuba). All microsatellite markers were highly polymorphic in all the breeds. The expected heterozygosity ranged from 0.67 ± 0.02 in the Taíno de Cuba breed to 0.75 ± 0.02 in the Mambí de Cuba breed, and the observed heterozygosity ranged from 0.66 ± 0.03 in the Cebú Cubano breed to 0.73 ± 0.02 in the Siboney de Cuba breed. The genetic differentiation between the breeds was significant (p < 0.01) based on the infinitesimal model (F(ST)). The exact test for Hardy-Weinberg equilibrium within breeds showed a significant deviation in each breed (p < 0.0003) for one or more loci. The genetic distance and structure analysis showed that a significant amount of genetic variation is maintained in the local cattle population and that all breeds studied could be considered genetically distinct. The Siboney de Cuba and Mambí de Cuba breeds seem to be the most genetically related among the studied five breeds.

Genetic characterization of Latin-American Creole cattle using microsatellite markers.

Genetic diversity in and relationships among 26 Creole cattle breeds from 10 American countries were assessed using 19 microsatellites. Heterozygosities, F-statistics estimates, genetic distances, multivariate analyses and assignment tests were performed. The levels of within-breed diversity detected in Creole cattle were considerable and higher than those previously reported for European breeds, but similar to those found in other Latin American breeds. Differences among breeds accounted for 8.4% of the total genetic variability. Most breeds clustered separately when the number of pre-defined populations was 21 (the most probable K value), with the exception of some closely related breeds that shared the same cluster and others that were admixed. Despite the high genetic diversity detected, significant inbreeding was also observed within some breeds, and heterozygote excess was detected in others. These results indicate that Creoles represent important reservoirs of cattle genetic diversity and that appropriate conservation measures should be implemented for these native breeds in order to minimize inbreeding and uncontrolled crossbreeding.

Comparative study of equine bone marrow and adipose tissue-derived mesenchymal stromal cells.

REASONS FOR PERFORMING STUDY: Mesenchymal stromal cells (MSCs) represent an attractive source for regenerative medicine. However, prior to their application, fundamental questions regarding molecular characterisation, growth and differentiation of MSCs must be resolved. OBJECTIVES: To compare and better understand the behaviour of equine MSCs obtained from bone marrow (BM) and adipose tissue (AT) in culture. METHODS: Five horses were included in this study. Proliferation rate was measured using MTT assay and cell viability; apoptosis, necrosis and late apoptosis and necrosis were evaluated by flow cytometry. The mRNA expression levels of 7 surface marker genes were quantified using RT-qPCR and CD90 was also analysed by flow cytometry. Differentiation was evaluated using specific staining, measurement of alkaline phosphatase activity and analysis of the mRNA expression. RESULTS: High interindividual differences were observed in proliferation in both cell types, particularly during the final days. Statistically significant differences in viability and early apoptosis of cultured AT- and BM-MSCs were found. The highest values of early apoptosis were observed during the first days of culture, while the highest percentage of necrosis and late apoptosis and lowest viability was observed in the last days. Surface marker expression pattern observed is in accordance to other studies in horse and other species. Osteogenic differentiation was evident after 7 days, with an increasing of ALP activity and mRNA expression of osteogenic markers. Adipogenic differentiation was achieved in BM-MSCs from 2 donors with one of the 16 media tested. Chondrogenic differentiation was also observed. CONCLUSIONS: Proliferation ability is different in AT-MSCs and BM-MSCs. Differences in viability and early apoptosis were observed between both sources and CD34 was only found in AT-MSCs. Differences in their osteogenic and adipogenic potential were detected by staining and quantification of specific tissue markers. POTENTIAL RELEVANCE: To provide data to better understand AT-MSCs and BM-MSCs behaviour in vitro.

Genetic diversity, structure, and breed relationships in Iberian cattle.

In Iberia there are 51 officially recognized cattle breeds of which 15 are found in Portugal and 38 in Spain. We present here a comprehensive analysis of the genetic diversity and structure of Iberian cattle. Forty of these breeds were genotyped with 19 highly polymorphic microsatellite markers. Asturiana de los Valles displayed the greatest allelic diversity and Mallorquina the least. Unbiased heterozygosity values ranged from 0.596 to 0.787. The network based on Reynolds distances was star-shaped with few pairs of interrelated breeds and a clear cluster of 4 breeds (Alistana/Arouquesa/Marinhoa/Mirandesa). The analysis of the genetic structure of Iberian cattle indicated that the most probable number of population clusters included in the study would be 36. Distance results were supported by the STRUCTURE software indicating a relatively recent origin or possible crossbreeding or both between pairs or small groups of breeds. Five clusters included 2 different breeds (Betizu/Pirenaica, Morucha/Avileña, Parda de Montaña/Bruna de los Pirineos, Barrosã/Cachena, and Toro de Lidia/Brava de Lide), 3 breeds (Berrenda en Negro, Negra Andaluza, and Mertolenga) were divided in 2 independent clusters each, and 2 breeds were considered admixed (Asturiana de los Valles and Berrenda en Colorado). Individual assignation to breeds was not possible in the 2 admixed breeds and the pair Parda de Montaña/Bruna de los Pirineos. The relationship between Iberian cattle reflects their geographical origin rather than their morphotypes. Exceptions to this geographic clustering are most probably a consequence of crossbreeding with foreign breeds. The relative genetic isolation within their geographical origin, the consequent genetic drift, the adaptation to specific environment and production systems, and the influence of African and European cattle have contributed to the current genetic status of Iberian cattle, which are grouped according to their geographical origin. The greater degree of admixture observed in some breeds should be taken into account before using molecular markers for genetic assignment of individuals to breeds.

Relative breed contributions to neutral genetic diversity of a comprehensive representation of Iberian native cattle.

This study is aimed at establishing priorities for the optimal conservation of genetic diversity among a comprehensive group of 40 cattle breeds from the Iberian Peninsula. Different sets of breed contributions to diversity were obtained with several methods that differ in the relative weight attributed to the within- and between-breed components of the genetic variation. The contributions to the Weitzman diversity and the expected heterozygosity (He) account for between- and within-breed variation only, respectively. Contributions to the core set obtained for several kinship matrices, incorporate both sources of variation, as well as the combined contributions of Ollivier and Foulley and those of Caballero and Toro. In general, breeds that ranked high in the different core set applications also ranked high in the contribution to the global He, for example, Sayaguesa, Retinta, Monchina, Berrenda en Colorado or Marismeña. As expected, the Weitzman method prioritised breeds with low contributions to the He, like Mallorquina, Menorquina, Berrenda en Negro, Mostrenca, Vaca Palmera or Mirandesa, all showing highly negative contributions to He - that is, their removal would significantly increase the average He. Weighing the within- and between-breed components with the FST produced a balanced set of contributions in which all the breeds ranking high in both approaches show up. Unlike the other methods, the contributions to the diversity proposed by Caballero and Toro prioritised a good number of Portuguese breeds (Arouquesa, Barrosã, Mertolenga and Preta ranking highest), but this might be caused by a sample size effect. Only Sayaguesa ranked high in all the methods tested. Considerations with regard to the conservation scheme should be made before adopting any of these approaches: in situ v. cryoconservation, selection and adaptation within the breeds v. crossbreeding or the creation of synthetic breeds. There is no general consensus with regard to balancing within- and between-breed diversity and the decision of which source to favour will depend on the particular scenario. In addition to the genetic information, other factors, such as geographical, historical, economic, cultural, etc., also need to be considered in the formulation of a conservation plan. All these aspects will ultimately influence the distribution of resources by the decision-makers.

A single nucleotide polymorphism in the coding region of bovine transferrin is associated with milk fat yield.

The bovine transferrin gene (TF) is located at 125 cM on bovine chromosome 1 (BTA1); it codes for transferrin, a glycoprotein that is highly conserved in many species and that is responsible for iron transport. The TF gene has been located in several QTL regions, and some transferrin classes have been associated with fat and milk yields. We analyzed by means of allele-specific oligonucleotide real-time PCR the c.1455A>G SNP in exon 12 of the TF cDNA sequence (accession number U02564), which induces an Asp/Gly substitution at position 469 of the peptide. The c.1455A>G SNP was assayed in eight Spanish cattle breeds, as well as in two groups of Holstein-Friesian animals that had the highest and lowest estimated breeding values for milk fat yield. Analysis of the cSNP showed balanced frequencies in all breeds, with a mean of 0.44. Evaluation of a potential association between the cSNP and the groups of Holstein-Friesian animals selected for milk fat yield showed a significant association (P < 0.0006); the G allele was associated with high fat production. Significant differences in genotypic frequencies between the groups were also detected (P < 0.0028). These results lead us to suggest that the TF gene has an effect on milk fat yield.

The bovine annexin 9 gene (ANXA9) is significantly associated with milk-fat yield in a Spanish Holstein-Friesian population.

On the basis of QTL studies for milk-fat yield trait on BTA3, annexin 9 protein (ANXA9), fatty acid transport protein type 3 (SLC27A3) and diacylglycerol O-acyltransferase 1 (DGAT1) were selected as candidate genes. Three different single nucleotide polymorphisms (SNPs) of bovine ANXA9, SLC27A3 and DGAT1 genes have been tested in a selective genotyping design for milk-fat yield. Significant allele frequency differences were found for ANXA9 (p=0.02), in Holstein-Friesian animals with high and low breeding values for milk-fat yield. Regression analysis also showed a significant effect (p=0.0207) between estimated breeding values (EBVs) for fat milk content and ANXA9 polymorphism. So ANXA9 gene falls into a significant quantitative trait loci interval for milk-fat yield that was previously reported on bovine chromosome 3 in other dairy populations. Our results suggest that the ANXA9 gene polymorphism or a linked segregating QTL contributes to variation in milk-fat yield.