Herramientas biotecnológicas aplicadas a la mejora genética de melocotón y nectarina

[SPA] En la Región de Murcia, la fruticultura presenta una gran importancia económica, destacándose el melocotón como el frutal de hueso al que más extensión se dedica. Éste papel destacado del melocotón, propició la unión entre sector productivo e investigación, dando como resultado el programa de mejora genética IMIDA-NOVAMED de melocotón y nectarina. Desde sus inicios, este programa tuvo como objetivo la obtención de variedades mejoradas genéticamente seleccionadas en la zona de cultivo, evitando el pago de costosos royalties por parte de los agricultores y donde ellos mismos seleccionaran los objetivos de las variedades a obtener. Sin embargo, los resultados obtenidos en programas de mejora genética clásica de melocotón y nectarina, se ven limitados por una metodología que no ha sido renovada desde hace décadas: recolección de polen, hibridación en campo, recolección de frutos, cultivo de embriones y/o germinación de semillas y traspaso de plantas a campo. Además, estos programas se enfrentan a dos factores limitantes, ciclos reproductivos largos y largos períodos de junenilidad. Así, la conjunción de todas éstas técnicas completan un ciclo, elevando el tiempo medio de registro de una variedad a 10 años. Uno de los objetivos primordiales en un programa de mejora genética de frutales es la obtención de variedades que adelanten en su salida al mercado a sus competidoras. Para alcanzar este objetivo se realizan hibridaciones empleando variedades de rápida maduración. En la mayor parte de los casos, cuando el parental de rápida maduración es el femenino, las semillas resultantes no puedan germinar por sí solas. Así, nacieron las técnicas de cultivo in vitro de embriones, que suponen un excelente complemento para los programas de mejora clásica, ya que proporcionan a las semillas inmaduras de un ambiente propicio para su germinación. Por otro lado, hay determinados caracteres que tras años de mejora no han conseguido transmitirse. Este es el caso de la resistencia a Sharka, una enfermedad viral con gran capacidad de infección cuya fuente de resistencia se encuentra en otras especies y en variedades que no están adaptadas a nuestras condiciones de cultivo. En los últimos años, la biotecnología se presenta como alternativa a este tipo de limitaciones de la mejora clásica, mediante la incorporación de genes de interés que, o bien confieren resistencia para determinadas enfermedades, o bien potencian las propiedades beneficiosas de determinadas especies. Para la consecución de estos objetivos mediante técnicas de transformación genética se requiere de una regeneración de novo de plantas a partir de células vegetales individuales en cultivo in vitro. Sin embargo, aunque en algunas especies este fenómeno ocurre de forma casi espontánea en otras como las leñosas, es un proceso bastante dificultoso. El melocotón es una especie especialmente recalcitrante en este sentido, donde solo algunos autores han publicado protocolos de regeneración somática, y en la mayoría de los casos a partir de tejidos zigóticos. El inconveniente de la utilización de estos tejidos para técnicas de transformación genética radica en que las semillas son nuevos genotipos en los que sus cualidades organolépticas están aún por determinar.[ENG] The world population is currently growing in a global food crisis framework, in which science has to rise to the occasion. In the last decades, plant science has offered alternatives to the world, including a deep understanding and improvement of crop quality. Several techniques have been developed in order to increase production or quality by means of adapting horticulture to environmental conditions. Plant breeders have been searching for crop improvements in terms of solutions for different problems, namely diseases, production or postharvest problems. But breeders have also varied objectives such as adaptation of new cultivars to growing areas, disease resistance, ripening fruit, high productivity and organoleptic quality. However, the results obtained in classical breeding programs are limited by a methodology that has not been updated for decades: pollen collection, hybridization, crop harvest, seed germination, and transplanting to the field. Other important factors are seeds containing immature embryos. They are due to a short period between blooming and ripening that leads to low percentages of germination. Immature embryo rescue techniques provide an alternative means to recover seeds from early ripening fruit, which usually fails to completely develop in vivo. But also, fruit breeding programs are facing two limiting factors: long reproductive cycles and long juvenile phases. Thus, the combination of all of them rises the average release time of a new cultivar to a 10-year range. Peach (Prunus persica L. Batsch) is one of the major fruits in the world. Thus, the economic value of this crop is one of the bases of agriculture in Europe. Currently, after decades of breeding, breeding still requires too much time obtaining peach fruits with interesting characteristics because traditional techniques are being used and the genetic basis of economically important traits are unknown. But also, in spite of the presence of some 70 peach breeding programs, peach breeding programs have failed to stabilize certain characters after many cycles of selection. This is the case of resistance to Plum Pox Virus (PPV), widely known as Sharka, a disease with high capacity for viral infection, which affects the productivity and fruit quality of Prunus species. Biotechnology has been presented as a tool to overcome the limitations of classical improvement, through the incorporation of desirable genes that either confer resistance to diseases or boost the beneficial properties of some species. In this sense, genetic transformation is an important tool that can be used in different ways. This technique is very well known as an application to provide protection to crops against certain diseases or insect actions. However, as the genome of different plant species has been sequenced, genetic transformation has played a major role in the elucidation of gene function. To efficiently harness genetic transformation technology, an efficient plant regeneration protocol from plant cells is required. Although in some species this phenomenon occurs almost spontaneously, in others such as woody plants it is quite difficult to achieve. Peach is a particularly recalcitrant species in this sense, in which only some authors have reported somatic regeneration protocols, and in most cases from zygotic tissue. The drawback of using seed-derived material for genetic transformation techniques are that each genotype is unique and not a clone of the parent. The development of plant regeneration protocols from mature tissues is important for the modification of desirable commercial cultivars that have been selected because of economic potential. Since cultivars are typically heterozygous, the seed they produce segregate and the desirable traits may not be present in the progeny that they produce. This research focuses in the development of protocols for both in vitro rescue of embryos and somatic regeneration in peach. On one hand, the searching for a general and improved protocol for embryo rescue in peach can provide a supplementary tool for the peach and nectarine breeding programme developed in IMIDA. On the other hand, developing a series of studies that lead to achieve an efficient protocol for plant regeneration will contribute to take a step forward in genetic transformation in peach. Preliminary field studies of genetic transformation are also included in this present dissertation.Universidad Politécnica de CartagenaPrograma de doctorado en Técnicas Avanzadas en Investigación y Desarrollo Agrario y Alimentari

Estudios de genòmica comparativa en especies vegetales de interés

Los microsatélites son marcadores moleculares ampliamente utilizados en estudios de biodiversidad, siendo particularmente útiles en la identificación varietal. Otra de las aplicaciones más recientes de los microsatélites se halla en el campo de la agroalimentación, siendo una herramienta útil en el desarrollo de sistemas de trazabilidad y autentificación. La comparación de secuencias y genomas (genómica comparativa) ha permitido recientemente desarrollar nuevas herramientas como los polimorfismos de un solo nucleótido (SNP; del inglés, “Single Nucleotide Polymorphisms”) e inserciones/deleciones (del inglés, “indels”), que completan la cobertura genómica de los microsatélites. Los principales objetivos de esta tesis son el uso y evaluación de: i) marcadores microsatélite basados en secuencias nucleares, como herramientas en el estudio y caracterización de la agrodiversidad, en especies olvidadas e infrautilizadas (cultivos prometedores), como la higuera; y ii) marcadores microsatélite e inserciones/deleciones basados en la comparación de genomas de cloroplastos de olivo, para la identificación varietal y la aplicabilidad de ellos a la trazabilidad de los aceites de oliva de distintas variedades. Los resultados han demostrado que se trata de herramientas eficaces en la caracterización de este germoplasma local, poniendo de manifiesto la gran variabilidad que éste alberga. Todo ello ha permitido identificar variedades de olivo, tanto a partir de hoja como de aceite. Algunos de los haplotipos identificados pueden ser empleados en la trazabilidad del aceite de oliva.Microsatellites are molecular markers widely used for biodiversity studies, being particularly useful in varietal identification. A more recent application of these markers is in the agrifood field, being a suitable tool for the development of authentication and traceability systems. Comparisons of sequences and genomes (comparative genomics) have recently allowed the development of new tools, such as single nucleotide polymorphisms (SNP) and insertions/deletions (indels), which complete the genomic coverage of microsatellites. The main goals of the present thesis are the use and evaluation of: i) nuclear sequence-based microsatellite markers, as a tool for the study and characterization of the agrodiversity in neglected and underutilized species (promising crops), such as the fig tree; and ii) microsatellite and indel markers, based on comparisons of genome chloroplast of the olive tree, for varietal identification and their application to the traceability of olive oils from different varieties. Nuclear sequence-based microsatellite markers have been selected to evaluate the germplasm agrodiversity in fig tree. The presence of unique alleles has allowed the discrimination amongst local populations. The results have demonstrated that these markers are effective tools in the characterization of this local germplasm, revealing a high level of variability. All this has allowed the identification of olive tree varieties, both from the leaves and from the olive oils. Some haplotypes could be used for olive-oil traceability purposes

Varietal Tracing of Virgin Olive Oils Based on Plastid DNA Variation Profiling

Olive oil traceability remains a challenge nowadays. DNA analysis is the preferred approach to an effective varietal identification, without any environmental influence. Specifically, olive organelle genomics is the most promising approach for setting up a suitable set of markers as they would not interfere with the pollinator variety DNA traces. Unfortunately, plastid DNA (cpDNA) variation of the cultivated olive has been reported to be low. This feature could be a limitation for the use of cpDNA polymorphisms in forensic analyses or oil traceability, but rare cpDNA haplotypes may be useful as they can help to efficiently discriminate some varieties. Recently, the sequencing of olive plastid genomes has allowed the generation of novel markers. In this study, the performance of cpDNA markers on olive oil matrices, and their applicability on commercial Protected Designation of Origin (PDO) oils were assessed. By using a combination of nine plastid loci (including multi-state microsatellites and short indels), it is possible to fingerprint six haplotypes (in 17 Spanish olive varieties), which can discriminate high-value commercialized cultivars with PDO. In particular, a rare haplotype was detected in genotypes used to produce a regional high-value commercial oil. We conclude that plastid haplotypes can help oil traceability in commercial PDO oils and set up an experimental methodology suitable for organelle polymorphism detection in the complex olive oil matrices

Investigación cuantitativa sobre como afecta el régimen de visitas a menores víctimas de violencia de género

La asociación DEMÉTER por la Igualdad junto con el Instituto Andaluz Interuniversitario de Criminología, Sección de Málaga, están llevando a cabo una investigación cuantitativa sobre cómo afecta el régimen de visitas del padre, impuesto a las hijas e hijos de mujeres víctimas de violencia de género. La investigación se ha llevado a cabo con menores que han sido atendidas/os en el Servicio de “Atención e Intervención a Menores Víctimas de Violencia de Género” ofrecido por la asociación DEMÉTER por la Igualdad en Málaga. El estudio se ha realizado sobre una población de 269 menores de Málaga, hijas e hijos de mujeres víctimas de maltrato de género de entre 0 y 17 años. La muestra corresponde a menores atendidos entre los años 2008 a 2012, con una media de 12 meses de intervención semanal, que no convivían con el agresor en el momento de ingresar en nuestro programa, pero estaban expuestos a un régimen de visitas con el mismo. El objeto de la investigación es valorar cómo afecta a estas y estos menores el seguir expuestos a las arbitrariedades de los agresores/maltratadores. 1186The Association “Demeter por la Igualdad”, together with the Malaga branch of the Instituto Andaluz Interuniversitario de Criminología (Andalucian Inter-University Institute of Criminology) , is undertaking a quantative research into the effects of parental (father) visits on the sons and daughters of women who have been victims of domestic violence. The research has been conducted on underage youngsters who have been dealt with by the department of “DEMÉTER por la Igualdad in Málaga” dedicated to looking after underage victims of domestic violence. The research has been conducted on a group of 269 youngsters, aged between 0-17 years, living in Malaga, all children of women who have suffered domestic violence. The sample covers youngsters who have been dealt with between 2008 and 2012, having an average of 12 months of weekly treatment, who were not living with the agresor at the time of entering our treatment programme, but were receiving regular visits from him. The purpose of the study is to determine the effects on the underage youngsters of being continually subject to the uncertain (uncontrolable?) behaviour of these agresors/wife beaters

Porous Poly(3,4-ethylenedioxythiophene)-Based Electrodesfor Detecting Stress Biomarkers in Artificial Urine and Sweat

When danger is perceived, the human body responds to overcome obstacles and survive a stressful situation; however, sustained levels of stress are associated with health disorders and diminished life quality. Hence, stress biomarkers are monitored to control stress quantitatively. Herein, a porous sensor (4l-COP/p) composed of poly(3,4-ethylenedioxythiophene) (PEDOT) and poly(3,4-ethylenedioxythiophene-co-N-methylpyrrole) (COP), which is prepared in a four-layered fashion to detect dopamine (DA) and serotonin (5-HT), is presented. Specifically, the detection is conducted in phosphate-buffered saline (PBS), as well as artificial urine and sweat, by applying cyclic voltammetry. The limit of detection values obtained are as low as 5.7 × 10−6 and 1.4 × 10−6 m for DA and 5-HT, respectively, when assessed individually in artificial urine. When mixed in PBS, 4l-COP/p detects both biomarkers with a resolution of 0.18 V and a sensitivity of 40 and 30 μA mm−1 for DA and 5-HT, respectively. Additionally, by theoretical calculations, the interaction pattern that each stress biomarker establishes with the PEDOT outer layer is elucidated. Whereas DA interacts with the ������-system of PEDOT, 5-HT forms specific hydrogen bonds with the conducting polymer chains. The resolution value obtained depends upon such interactions. Overall, 4l-COP/p electrodes display potential as stress sensing devices for healthcare technologies.Authors acknowledge MINECO/FEDER (RTI2018-098951-B-I00), the Agència de Gestió d'Ajuts Universitaris i de Recerca (2017SGR359). M.M.P.-M. thanks the Spanish Ministry of Educación y Formación Profesional for the Junior Beatriz Galindo Award (BG20/00216)

Correlation between permanent tooth eruption pattern and the predominance of the motor function laterality

Objectives: To asses whether dental eruption order can play a role in the early diagnosis of crossed laterality. Study Design: Dental eruption pattern along with eye, ear, hand and foot lateralism were examined on 131 chil - dren between 6 to 8 years old from public schools from a multietnic population area of Barcelona city. Statistic methods (Statgraphics Plus 5.1 program) were used to evaluate data recollected. Results: Only foot and dentition lateralities behave as independent variables regarding hand laterality. So dental eruption laterality (along with the foot one) would be one of the parameters more related to hand laterality given that dentition variable relationship is greater that the foot one. This suggests that tooth eruption could be more clinically relevant. Crossed laterality hand-foot is significantly more predominant in men (13%) than in women (1,6%). Meanwhile, the relationship between hand and dentition didn't show any influence of sex. Conclusions: Dental eruption order, can be used as a good parameter in the determination of the patient's laterality

Instruments for assessing the risk of falls in acute hospitalized patients: a systematic review and meta-analysis

Background Falls are a serious problem for hospitalized patients, reducing the duration and quality of life. It is estimated that over 84% of all adverse events in hospitalized patients are related to falls. Some fall risk assessment tools have been developed and tested in environments other than those for which they were developed with serious validity discrepancies. The aim of this review is to determine the accuracy of instruments for detecting fall risk and predicting falls in acute hospitalized patients. Methods Systematic review and meta-analysis. Main databases, related websites and grey literature were searched. Two blinded reviewers evaluated title and abstracts of the selected articles and, if they met inclusion criteria, methodological quality was assessed in a new blinded process. Meta-analyses of diagnostic ORs (DOR) and likelihood (LH) coefficients were performed with the random effects method. Forest plots were calculated for sensitivity and specificity, DOR and LH. Additionally, summary ROC (SROC) curves were calculated for every analysis. Results Fourteen studies were selected for the review. The meta-analysis was performed with the Morse (MFS), STRATIFY and Hendrich II Fall Risk Model scales. The STRATIFY tool provided greater diagnostic validity, with a DOR value of 7.64 (4.86 - 12.00). A meta-regression was performed to assess the effect of average patient age over 65 years and the performance or otherwise of risk reassessments during the patient’s stay. The reassessment showed a significant reduction in the DOR on the MFS (rDOR 0.75, 95% CI: 0.64 - 0.89, p = 0.017). Conclusions The STRATIFY scale was found to be the best tool for assessing the risk of falls by hospitalized acutely-ill adults. However, the behaviour of these instruments varies considerably depending on the population and the environment, and so their operation should be tested prior to implementation. Further studies are needed to investigate the effect of the reassessment of these instruments with respect to hospitalized adult patients, and to consider the real compliance by healthcare personnel with procedures related to patient safety, and in particular concerning the prevention of falls

Análisis económico-financiero de una unidad básica de producción cooperativa

An economic an financial appraisal of a typical livestock cooperative collective venture in Valle del Cauto, Granma province, Cuba, was performed. A data-base was built taking into account the information collected from surveys to the venture administra- tion board and the statistical and financial registers from 1996, 1997, and 1998. Technical indexes such as soil use and herd struc- ture, and financial indexes like solvency and debt level at a short run were discussed. The average guideline for animal current production inventory and bills to be collected and paid were also analyzed. This last analysis showed the venture financial status. There was just a 9,6% and 9% solvency to pay debts in the years under study. They invested 1,31; 1,28 and 1,16 pesos (national currency) from foreing resources for each own financing peso.  Este trabajo se realizó en una unidad básica de producción cooperativa (UBPC) ganadera, representativa del Valle del Cauto, provincia de Granma, Cuba, con el objetivo de analizar el comportamiento de los indicadores económico-financieros que inciden en su actividad económica. La base de datos se confeccionó con encuestas realizadas a los miembros de la junta directiva de la UBPC, y con la información proveniente de los registros estadísticos y los estados financieros correspondientes a tres años de los últimos ejercicios aprobados (1996, 1997 y 1998). Se analizan de forma particular indicadores técnicos como el uso del suelo y estructura del rebaño; también se estudiaron indicadores financieros como liquidez y grado de endeudamiento a corto plazo; así como algunas razones de actividad, como plazo promedio de inventario de producción animal en proceso y cuentas por cobrar y pagar. El análisis de las razones muestra la situación financiera que presenta la UBPC. El índice de prueba ácida en el período demuestra que sólo el 9, 6 y 9 % respectivamente de sus activos a liquidar a corto plazo cubren sus deudas para el período anali- zado. Por cada peso de financiación propia utilizan 1,31; 1,28 y 1,16 pesos de recursos ajenos para estos años, respectivamente

Propuesta de un sistema de evaluación y acreditación de la Especialidad de Reumatología basada en las variables de calidad. Una visión a tener en cuenta por la Sociedad Cubana de Reumatología

The quality of postgraduate education in Cuba is seen as the integration of social relevance and academic excellence. The quality achieved by a postgraduate program, as well as the measures and actions to provide training and continuous improvement, are evident when subjected to processes of self-assessment or external evaluation. As part of the control system of the Ministry of Higher Education, the University System Accreditation Program was established. This system has considered the requirements of the international context and the methodologies, methods and procedures used to create a culture of quality in the university community, by strengthening the self-evaluation and external evaluation for accreditation of programs and institutions. Accreditation is the result of applying a system of internal and external evaluation, aimed at acknowledging publicly that an institution or program meets certain quality requirements, previously defined by collegiate bodies of academic prestige. In Cuba, the National Accreditation Board.La calidad de la educación posgraduada en Cuba se concibe como la integración de la pertinencia social y la excelencia académica. La calidad alcanzada por un programa de posgrado, así como las medidas y acciones de perfeccionamiento y mejora continua se evidencian al someterse a procesos de autoevaluación o evaluación externa. Como parte del sistema de control el Ministerio de Educación Superior estableció el Sistema Universitario de Programas de Acreditación, que ha considerado los requerimientos del contexto internacional y las metodologías, los métodos y los procedimientos empleados para crear una cultura de calidad en la comunidad universitaria a partir del fortalecimiento de la auto evaluación y la evaluación externa con vistas a la acreditación de programas e instituciones. La acreditación es el resultado de la aplicación de un sistema de evaluación interna y externa dirigido a reconocer públicamente que una institución o programa reúne determinados requisitos de calidad, definidos previamente por órganos colegiados de reconocido prestigio académico; en Cuba es la Junta de Acreditación Nacional

Evaluación económica de dos sistemas de pastoreo para la producción de leche con bajos insumos en suelo vertisol

An economic an financial appraisal of a typical livestock cooperative collective venture in Valle del Cauto, Granma province, Cuba, was performed. A data-base was built taking into account the information collected from surveys to the venture administra- tion board and the statistical and financial registers from 1996, 1997, and 1998. Technical indexes such as soil use and herd struc- ture, and financial indexes like solvency and debt level at a short run were discussed. The average guideline for animal current production inventory and bills to be collected and paid were also analyzed. This last analysis showed the venture financial status. There was just a 9,6% and 9% solvency to pay debts in the years under study. They invested 1,31; 1,28 and 1,16 pesos (national currency) from foreing resources for each own financing peso.Se evaluó económicamente el empleo de dos sistemas de pastoreo en una vaquería comercial, en condiciones de secano. Se uti- lizaron los métodos de pastoreo racional Voisin (PRV) y el pastoreo porcionado, cada uno con 37 vacas 5/8 Holstein-3/8 Cebú. Para valorar la eficiencia económica, se tuvieron en cuenta indicadores del volumen, valor y costo de producción, ganancia económica, costo por peso, relación beneficio costo, costo del kg de leche y se calculó el punto de equilibrio y período de recu- peración de la inversión. Los resultados mostraron que los indicadores del volumen de producción (producción de leche), valor de la producción (leche y carne), los costos de producción (gastos fijos y variables) y la ganancia económica revelaron un mejor comportamiento en el método porcionado, donde el costo por peso producido promedio (moneda nacional) es inferior en $0,06, la relación beneficio costo se incrementó en$ 0,11 y el costo del kg de la leche disminuyó en $0,03, con respecto al PRV. Además, se determinó que solo es necesario invertir entre$ 0,08 y $0,12 USD por cada peso en moneda nacional, en el método porcionado, mientras que en el PRV se necesita invertir$ 0,02 adicionales. El costo para producir un kilogramo de leche fue bajo en sentido general y osciló entre $0,05 y$ 0,07 en el PRV y entre $0,04 y$ 0,05 en el porcionado