Organotypic culture of breast tumor explants as a multicellular system for the screening of natural compounds with antineoplastic potential

Breast cancer is the leading cause of death in women worldwide. The search for novel compounds with antitumor activity, with less adverse effects and higher efficacy, and the development of methods to evaluate their toxicity is an area of intense research. In this study we implemented the preparation and culture of breast tumor explants, which were obtained from precision-cut breast tumor slices. In order to validate the model we are proposing to screen antineoplastic effect of natural compounds, we selected caffeic acid, ursolic acid, and rosmarinic acid. Using the Krumdieck tissue slicer, precision-cut tissue slices were prepared from breast cancer samples; from these slices, 4 mm explants were obtained and incubated with the selected compounds. Viability was assessed by Alamar Blue assay, LDH release, and histopathological criteria. Results showed that the viability of the explants cultured in the presence of paclitaxel (positive control) decreased significantly

Demographic status and genetic tagging of endangered Capercaillie in NW Spain

Counting rare and elusive animals and evaluating their demographic status, are fundamental yet challenging aspects of population ecology and conservation biology. We set out to estimate population size (Nc), genetic effective population size (Ne gen), sex ratio, and movements based on genetic tagging for the threatened Cantabrian capercaillie. We used 9 microsatellite loci to genotype 134 droppings collected at 34 display areas during the breeding season. Using genetic capture-mark-recapture, we estimated 93 individuals (Nc, 95% CI: 70-116) in an area of about 500 km 2, with sex ratio biased towards males (1:1.6). Estimated N e gen (35.5) was 38% of Nc, notably higher than the published average in wild populations. This capercaillie population is small and well within concern in terms of population viability. By genetic tagging, we detected mostly short movements; just a few males were recaptured between contiguous display areas. Non-invasive surveys of endangered populations have a great potential, yet adequate sample size and location are key to obtain reliable information on conservation status. © 2014 Morán-Luis et al.Peer Reviewe

Prevalencia de mastitis subclinica en la región Ciénega del estado de Jalisco

The aim of this study was to evaluate the prevalence and associated with subclinical mastitis pathogens in dairy cows Cienega region of Jalisco. For the study, 5,450 were milk samples from individual quarters of 1,362 Holstein cows, and high crossbreeding, from 37 pens located in the municipalities of Atotonilco el Alto, La Barca, Jamay, Ocotlán, King and Ixtlahuacán Zapotlán of Quinces. We considered producing cows in each pen, except for those with less than 30 days and over 250 days of lactation. The samples were applied California test (CMT) and the indicative use epidemiological called Subclinical Mastitis Index (IMSC) to determine prevalencia.También microbiological analysis was performed a total of 66 milk samples from tanks milk tank (13 samples per study herd). The data obtained were analyzed statistically with the Kruskal-Wallis test. The results showed that 35.64% were indicative of subclinical mastitis and municipalities with the highest prevalence were Barca, Poncitlán and Ocotlán. The rear quarters were the most susceptible to present this type of mastitis (p <0.05) and was obtained 1.93 IMSC, microbiological analysis revealed that 100% of the samples had the presence of Staphylococcus aureus and Salmonella spp. These results show that the municipalities in the región of Jalisco Ciénega has a high prevalence of subclinical mastitis, which may represent losses of up to 16 to 24.5% in the dairy herdEl objetivo del presente estudio fue evaluar la prevalencia y los patógenos asociados a mastitis subclínica en vacas productoras de leche región Ciénega del Estado de Jalisco. Para el estudio se obtuvieron 5,450 muestras de leche de cuartos individuales de 1,362 vacas Holstein, y alto mestizaje, procedentes de 37 corrales ubicados en los municipios de Atotonilco el Alto, La Barca, Jamay, Ocotlán, Zapotlán del Rey e Ixtlahuacán de los Membrillos. Se consideraron las vacas en producción de cada corral, a excepción de aquellas con menos de 30 días y con más de 250 días de lactancia. A las muestras se les aplicó la prueba de California (CMT) y se utilizó el indicativo epidemiológico denominado Índice de Mastitis Subclínica (IMSC) para determinar la prevalencia. También se realizó un análisis microbiológico a un total de 66 muestras de leche provenientes de los tanques de depósito de leche (13 muestras por cada hato del estudio). Los datos obtenidos, se analizaron estadísticamente con la prueba Kruskal-Wallis. Los resultados mostraron que 35.64% fueron indicativas de mastitis subclínica y los municipios con la más alta prevalencia fueron la Barca, Poncitlán y Ocotlán. Los cuartos posteriores fueron los más susceptibles a presentar este tipo de mastitis (p<0.05) y se obtuvo un IMSC de 1.93, el análisis microbiológico reveló que el 100 % de las muestras tuvieron la presencia de Staphylococcus aureus y Salmonella spp. Estos resultados muestran que los municipios de la región Ciénega, del Estado de Jalisco tiene un alta prevalencia de mastitis subclínica, lo que puede representar pérdidas de hasta 16 a 24.5 % en la producción láctea del hat

Conservation in the southern edge of Tetrao urogallus distribution: Gene flow despite fragmentation in the stronghold of the Cantabrian capercaillie

The Cantabrian capercaillie (Tetrao urogallus cantabricus) is an endangered subspecies of the Western capercaillie, endemic of northern Spain, inhabiting the south-western limit of the species range. Assessing genetic variability and the factors that determine it is crucial in order to develop an effective conservation strategy. In this work, non-invasive samples were collected in some of the best preserved areas inhabited by Cantabrian capercaillie. Nine microsatellite loci and a sex-specific marker were analysed. We included five zones, separated by valleys with different levels of habitat modifications. No evidence of genetic clustering was found which suggests that fragmentation and development in the area do not act as barriers to gene flow. Nonetheless, significant differences among sampling zones were encountered in terms of their allelic frequencies (global FST = 0.035, p = 0.001). Pairwise FST comparisons showed differences between all sampling zones included, except between the two ones located in the South (Degaña and Alto Sil). These findings, along with the results of individual based genetic differences, indicate that gene flow among sampling zones might be at least slightly compromised, except between the two zones located in the South. Despite this, the sampling zones seem to exchange migrants at a rate that prevents genetic differentiation to the point of creating clusters. Our results show that the capercaillies in the study area constitute a single interbreeding group, which is an important piece of information that provides support to better understand the dynamics of this endangered subspecies.Fil: Fameli, Alberto Francisco. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Museo Argentino de Ciencias Naturales "Bernardino Rivadavia"; ArgentinaFil: Morán Luis, María. Universidad de Oviedo; EspañaFil: Rodríguez Muñoz, Rolando. University of Exeter; Reino UnidoFil: Bañuelos, María José. Universidad de Oviedo; EspañaFil: Quevedo, Mario. Universidad de Oviedo; EspañaFil: Mirol, Patricia Monica. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Museo Argentino de Ciencias Naturales "Bernardino Rivadavia"; Argentin

Population dynamics of an endangered forest bird using mark-recapture models based on DNA-tagging

Populations of vertebrates are built of individuals of different sex, age class or stage, which often affect distinctly the population dynamics. Such intrapopulation partitioning of vital rates needs to be identified to develop efficient conservation actions. Using DNA extracted from feces and feathers we combined DNA-tagging and mark–recapture analyses to evaluate sex-specific population dynamics of an endangered population of capercaillie (Tetrao urogallus; Phasianidae). We built encounter histories for 120 individuals in the mating seasons of 2009–2011, in a study area of about 424 km2. Minimum number of individuals per mating season and estimates of population size ranged 56–67 and 76–115, respectively. Estimates of population size were consistently lower in multiple-season, open-population models than in single-season closed-population models. The super-population in the study area was 149 individuals for the whole study period. Sex-ratio was notably male-biased. Probability of recapture p ranged 0.62–0.70, and was similar for males and females. Female apparent survival φ was lower than expected, and much lower than male apparent survival. It includes however movements in and out of the sampled population, thus comparison with previously reported values based on conventional tagging should be cautious. Females showed higher turnover, indicated by higher probability β of entering the sampled population, and higher number of entries from the super-population, Bgross. Realized population growth rate ʎ was > 1 for both females and males. The combination of non-intrusive DNA-tagging and the analytical framework of mark–recapture models provided inferences on population dynamics that would have been hardly feasible with conventional methods. Male-biased sex ratios, higher female turnover and seemingly low female apparent survival were our key findings. While the whole population needs continuous monitoring, we believe that adult females deserve priority attention in evaluation and design of conservation actions.Fil: Bañuelos Martínez, María José. Universidad de Oviedo; EspañaFil: Blanco Fontao, Beatriz. Universidad de Oviedo; EspañaFil: Fameli, Alberto Francisco. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Museo Argentino de Ciencias Naturales "Bernardino Rivadavia"; ArgentinaFil: Fernández Gil, Alberto. Consejo Superior de Investigaciones Científicas. Estación Biológica de Doñana; EspañaFil: Mirol, Patricia Monica. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Museo Argentino de Ciencias Naturales "Bernardino Rivadavia"; ArgentinaFil: Morán, Luis María. Universidad de Oviedo; EspañaFil: Rodriguez Muñoz, Rolando. University of Exeter; Reino UnidoFil: Quevedo, Mario. Universidad de Oviedo; Españ

Organotypic Culture of Breast Tumor Explants as a Multicellular System for the Screening of Natural Compounds with Antineoplastic Potential

Breast cancer is the leading cause of death in women worldwide. The search for novel compounds with antitumor activity, with less adverse effects and higher efficacy, and the development of methods to evaluate their toxicity is an area of ​​intense research. In this study we implemented the preparation and culture of breast tumor explants, which were obtained from precision-cut breast tumor slices. In order to validate the model we are proposing to screen antineoplastic effect of natural compounds, we selected caffeic acid, ursolic acid, and rosmarinic acid. Using the Krumdieck tissue slicer, precision-cut tissue slices were prepared from breast cancer samples; from these slices, 4 mm explants were obtained and incubated with the selected compounds. Viability was assessed by Alamar Blue assay, LDH release, and histopathological criteria. Results showed that the viability of the explants cultured in the presence of paclitaxel (positive control) decreased significantly (P<0.05); however, tumor samples responded differently to each compound. When the explants were coincubated with paclitaxel and compounds, a synergic effect was observed. This study shows that ex vivo culture of breast cancer explants offers a suitable alternative model for evaluating natural or synthetic compounds with antitumor properties within the complex microenvironment of the tumor

Minimum (<i>N_min</i>) and estimated (<i>N_c</i>) number of capercaillies in the study area.

<p>Population size (Nc) was estimated both for the whole study area and for Muniellos, the best represented zone in the samples. Separate estimates for each sex (Nc male, Nc female) are indicated for the whole study area. Numbers in parentheses indicate individuals that could not be sexed.</p

Multilocus probability of identity for unrelated (P_ID) and sibling (P_ID-sib) capercaillies.

<p>Probabilities were estimated with all the identified individuals (N = 56). Microsatellites are ordered from least to most informative. Arrows indicate the first value of P_ID and P_ID-sib below 0.01.</p

Distribution of Cantabrian capercaillie (<i>Tetrao urogallus cantabricus</i>) and location of the study area.

<p>A) Distribution of capercaillie in central and southwestern Europe (dark grey). B) Location of the study sites (filled dots) over an index of habitat suitability <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0099799#pone.0099799-Rudnick1" target="_blank">[31]</a>. C) Detail of the study area; different symbols show display areas where samples were collected in spring 2009 in each of five watersheds: Muniellos (stars), Hermo (triangles), Degaña (squares), Leitariegos (diamonds) and Alto Sil (dots). The legend shows habitat suitability according to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0099799#pone.0099799-Rudnick1" target="_blank">[31]</a>.</p