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Introduction: Candidiases is a group of opportunistic infections caused by yeasts belonging to the genus Candida. Candida albicans is the most prevalent species in both superficial and deep infections, however, the clinical importance of non-albicans Candida has increased during the last decade, driving an urgent need for diagnostic tests that allow for species-level resolution and selection of the optimum therapeutic approach.Objective: To design and to optimize a new multiplex PCR assay for the simultaneous identification of the five most relevant species of Candida involved in human candidiasis etiology.Materials and methods: For primers design, the physical and thermodynamic restrictions that affect multiplex PCR performance were analyzed using Gene Runner and Mult-PSOS. As templates, the internal transcribed region 2 (ITR2) was selected for C. albicans (AJ249486.1), and topoisomerase II (TOPII) for C. parasilopsis (AB049144.1), C. krusei (AB049139.1), C. tropicalis (AB049141.1), and C. guillermondii (AB049145.1). We used ATCC strains of all these five species and clinical isolates as templates.Results: We designed ten oligonucleotides for the simultaneous amplification of the Candida species. The electrophoresis band profile was: C. albicans (206 bp), C. guillermondii (244 bp), C. tropicalis (474 bp), C. parasilopsis (558 bp), and C. krusei (419 bp). Conclusion: The new multiplex PCR assay designed in this study allowed a simultaneous and efficient amplification of the amplicons corresponding to the five species of Candida under study, with an adequate resolution in standard agarose gel., Introducción. Las candidiasis son un grupo de infecciones oportunistas causadas por levaduras del género Candida. Candida albicans es la especie de mayor prevalencia en las infecciones superficiales y profundas. Sin embargo, en la última década, la frecuencia de especies diferentes a C. albicans ha aumentado y, por ende, su relevancia clínica, lo cual exige la utilización de técnicas diagnósticas que permitan su detección y el tratamiento adecuado de los pacientes afectados.Objetivo. Diseñar y optimizar una técnica de reacción en cadena de la polimerasa múltiple (PCR múltiple) considerando parámetros termodinámicos para la detección simultánea de cinco especies de Candida relevantes en la etiología de la candidiasis humana.Materiales y métodos. Para el diseño de los cebadores se consideraron restricciones físicas y termodinámicas que afectan la PCR múltiple, usando el programa Gene Runner y la herramienta Mult-PSOS. Como plantillas se utilizaron la región transcrita interna 2 (ITS2) (AJ249486.1) para C. albicans y la topoisomerasa II (TOPII) para C. parasilopsis (AB049144.1), C. krusei (AB049139.1), C. tropicalis (AB049141.1) y C. guillermondii (AB049145.1), y como moldes, extractos de ADN total obtenidos de cepas ATCC y de aislamientos clínicos de las especies de Candida.Resultados. Se diseñaron diez cebadores para la amplificación simultánea de las especies de Candida. Se obtuvo el siguiente patrón de bandas: C. albicans (206 pb), C. guillermondii (244 pb), C. tropicalis (474 pb), C. parasilopsis (558 pb) y C. krusei (419 pb).Conclusión. El ensayo diseñado de PCR múltiple permitió la amplificación simultánea y eficiente de todos los amplicones correspondientes a las especies estudiadas de Candida, así como su adecuada resolución en gel de agarosa al 1,3 %.

Understanding the survival capacity of forest trees to periods of severe water stress could improve knowledge of the adaptive potential of different species under future climatic scenarios. In long lived organisms, like forest trees, the combination of induced osmotic stress treatments and field testing can elucidate the role of drought tolerance during the early stages of establishment, the most critical in the life of the species. We performed a Polyethylene glycolosmotic induced stress experiment and evaluated two common garden experiments (xeric and mesic sites) to test for survival and growth of a wide range clonal collection of Maritime pine. This study demonstrates the importance of additive vs non additive effects for drought tolerance traits in Pinus pinaster, and shows differences in parameters determining the adaptive trajectories of populations and family and clones within populations. The results show that osmotic adjustment plays an important role in population variation, while biomass allocation and hydric content greatly influence survival at population level. Survival in the induced osmotic stress experiment presented significant correlations with survival in the xeric site, and height growth at the mesic site, at population level, indicating constraints of adaptation for those traits, while at the within population level no significant correlation existed. These results demonstrate that population differentiation and within population genetic variation for drought tolerance follow different patterns.

Due to the high economic importance of Pinus pinaster Ait.;there is considerable interest in developing, improving and extending the use of its families for mass clonal propagation and in breeding programmes. In the current study, we evaluated shoot growth, rooting ability and mini-cuttings production of P. pinaster in response to nitrogen fertilization and seasons. We compared eight half-sib families of P. pinaster from Asturias and Galicia (Northern Iberian Peninsula), searching for useful parameters and growing conditions to be included in a mass propagation program for clonal family forestry. We fertilized P. pinaster seedling mother plants kept in a greenhouse with three levels of nitrogen high (HN), medium (MN) and low (LN) to evaluate rooting ability of mini-cuttings. In addition, we evaluated the maximal potential production of rooted mini-cuttings considering nine cycles of propagation over 1 year, also using three levels of nitrogen. The HN treatment significantly influenced the rooting process, with length, area and volume of roots all being positively affected. Spring was the most favourable season for mini-cuttings in the HN treatment. This study provides valuable new information to optimize the clonal propagation protocol for P. pinaster and shows that the mini-cuttings technique has great potential in mass scale cloning, providing high quality sprout production and well-formed new plants. © 2012 Springer Science+Business Media B.V.

Maritime pine (Pinus pinaster) is an important commercial species throughout its Atlantic distribution. With the anticipated increase in desiccation of its habitat as a result of climate change, the selection of genotypes with increased survival and growth capability under these conditions for breeding programs is of great interest for this species. We aimed to study the response to a realistic drought stress under controlled conditions, looked for a method to measure dehydration resistance, and analyzed dehydrin expression in drought-resistant and drought-sensitive clones from different ecotypes. We report here the sequence characteristics and the expression patterns of five dehydrins from P. pinaster, along with the physiological characterization of drought stress responses in different genotypes (clonally replicated plants), originating from a broad geographical distribution across France and Spain (provenances). In total, we distinguished five different dehydrin genes in silico, grouped into two types-K 2 and SK n. Three of the dehydrin genes had several sequence variants, differing by multiple or single amino acid substitutions. Only two of the dehydrins (PpinDhn3 and PpinDhn4) showed an increase in transcription with increased drought stress which was dependent on provenance and genotype, suggesting their involvement in drought resistance. The other dehydrins showed decreased expression trends with increased severity of the drought stress. The lack of close association between the drought stress and expression patterns of these dehydrin genes suggest that they could have other functions and not be involved in drought resistance. Our results suggest large differences in function between different dehydrin genes. © 2012 Springer-Verlag.

Mediterranean trees are expected to be exposed to an increased aridity in the near future. The lack of genetic variability of stone pine (Pinus pinea L.) could limit the adaptation potential of the species to more severe droughts. However, information on intra-specific variability in adaptive traits that might confer potential drought tolerance is scant for stone pine. In this study, we explored the adaptation potential of stone pine to water deficit through variability of key functional traits. In particular, inter-clonal variation in whole plant performance in response to water deficit was studied. The role of inter-clonal variation in functional traits and phenotypic plasticity as determinants of whole plant performance under water deficit were also assessed. Relative growth rate (RGR) and key morphological and physiological traits were studied in ramets of 20 different clones of stone pine from a wide range of populations in the Iberian Peninsula. The plants were grown in climatic chamber under controlled environmental conditions and were submitted to an experimental water deficit of 20 days. The results showed significant inter-clonal variation in total dry mass, RGR, biomass partitioning specific leaf area (SLA), leaf area ratio (LAR), net photosynthetic rate (An), stomatal conductance (gs) and intrinsic water use efficiency (WUEi). Significant phenotypic plasticity in response to drought was found for An, gs and WUEi but it did not differ among clones. The clones could be clustered into different groups according to the variability of the studied traits in a principal components analysis. LAR, SLA and net photosynthetic rates explained most of the variation. A combination of traits (high LAR and low WUEi) agreeing with a water spending strategy rather than with a water saving strategy seemed to underlie enhanced RGR under water deficit. Stone pine showed high phenotypic plasticity and significant intra-specific variation in functional traits in response to wa

El software realizado lleva por titulo “Diseño e Implementación de un Software de Historias Clínicas Neurológicas para la Liga Colombiana contra la Epilepsia”, cuyos autores son: Tania Sofía Fernández Velasco y Francisco Javier Palmera Anaya. Director del proyecto Ingeniero Giovanny Vásquez Mendoza y asesor Doctor Jaime Fandiño Franky. El objetivo principal que se tuvo en cuenta fue el de diseñar e implementar un software de historias clínicas neurológicas con el fin de dar soporte en la forma de accesar y manipular la información de las historias clínicas. Para la realización del software se utilizó el lenguaje de programación Visual Basic 6.0 edición empresarial con la base de datos SQL Server 7.0. Para la ayuda se utilizó la herramienta HelpScribble. SOHCLINE permite ingreso, modificación, consulta y eliminación de una historia clínica así como el uso de estadísticas y reportes. Básicamente esta dividido en 5 módulos: Ficha clínica, hojas clínicas, cirugía, evolución y control y estadísticas y reportes.

El software realizado lleva por titulo “Diseño e Implementación de un Software de Historias Clínicas Neurológicas para la Liga Colombiana contra la Epilepsia”, cuyos autores son: Tania Sofía Fernández Velasco y Francisco Javier Palmera Anaya. Director del proyecto Ingeniero Giovanny Vásquez Mendoza y asesor Doctor Jaime Fandiño Franky. El objetivo principal que se tuvo en cuenta fue el de diseñar e implementar un software de historias clínicas neurológicas con el fin de dar soporte en la forma de accesar y manipular la información de las historias clínicas. Para la realización del software se utilizó el lenguaje de programación Visual Basic 6.0 edición empresarial con la base de datos SQL Server 7.0. Para la ayuda se utilizó la herramienta HelpScribble. SOHCLINE permite ingreso, modificación, consulta y eliminación de una historia clínica así como el uso de estadísticas y reportes. Básicamente esta dividido en 5 módulos: Ficha clínica, hojas clínicas, cirugía, evolución y control y estadísticas y reportes.

El software realizado lleva por titulo “Diseño e Implementación de un Software de Historias Clínicas Neurológicas para la Liga Colombiana contra la Epilepsia”, cuyos autores son: Tania Sofía Fernández Velasco y Francisco Javier Palmera Anaya. Director del proyecto Ingeniero Giovanny Vásquez Mendoza y asesor Doctor Jaime Fandiño Franky. El objetivo principal que se tuvo en cuenta fue el de diseñar e implementar un software de historias clínicas neurológicas con el fin de dar soporte en la forma de accesar y manipular la información de las historias clínicas. Para la realización del software se utilizó el lenguaje de programación Visual Basic 6.0 edición empresarial con la base de datos SQL Server 7.0. Para la ayuda se utilizó la herramienta HelpScribble. SOHCLINE permite ingreso, modificación, consulta y eliminación de una historia clínica así como el uso de estadísticas y reportes. Básicamente esta dividido en 5 módulos: Ficha clínica, hojas clínicas, cirugía, evolución y control y estadísticas y reportes.

Three generations of restricted selection index involving pupal weight at 21 days (X1) and adult weight at 31 days (X2) were carried out in each of three replicates of Tribolium castaneum. The efficiency of two restricted indexes-optimum (O) and desired gains (G)-was evaluated experimentally. The optimum index (-0.0260X1 + 0.1551X2) was designed to maximize adult weight while setting the response of pupal weight by a fixed amount The desired-gains index (4.71X1 - 6.26X2) was intended to achieve genetic gains in pupal weight and adult weight at a prechosen ratio of 31. Both methods of selection were considered examples of antagonistic selection, with the positive genetic correlation between pupal and adult weight being very large. The results confirmed theoretical expectations. The optimum index set the response of pupal weight by a fixed amount while increasing adult weight; observed responses in the O line were significant for pupal weight and adult weight. The results from the G line showed that responses approximated a ratio of 61 for pupal weight and adult weight. In this line, the response for pupal weight was very similar to the expected value, and it did not change adult weight significantly. © 1989 The American Genetic Association.

Effects of errors in estimates of the genetic correlation on the accuracy of unrestricted, optimum, and desired gains selection indices were examined experimentally in Tribolium castaneum. Three lines were selected for three generations for pupal weight at 21 days and adult weight at 31 days, using unrestricted (I9), optimum (O9), and desired gains (G9) index selection methods. The genetic correlation between pupal and adult weights in the base population was 0.95. The optimum index was designed to set the response of pupal weight by a fixed amount, while in the desired gains index the responses of pupal and adult weights were specified as being equal to 31. Three other indices were constructed using a deliberately incorrect genetic correlation (0.25), i.e.;unrestricted (I2), optimum (O2), and desired gains (G2). Responses observed in unrestricted index lines (I9 versus I2) and optimum index lines (O9 versus O2) did not differ significantly, even though lines I9 and I2 differed in a practical sense. Responses in desired gains index lines (G9 versus G2) differed significantly. Responses obtained for aggregate genotype (pupal weight + adult weight) and for the component traits were greater in line I9 than those obtained in line I2. Responses obtained in the O9 and O2 lines for pupal and adult weights were similar, while those obtained in the G9 and G2 lines were similar for pupal weight but not (P<0.05) for adult weight. Therefore, underestimation of the genetic correlation seems to affect the efficiency of a desired gains index more than that of unrestricted or optimum indices. © 1989 Springer-Verlag.