Virtopsy: forensic traumatology of the subcutaneous fatty tissue; multislice computed tomography (MSCT) and magnetic resonance imaging (MRI) as diagnostic tools

Traumatic lesions of the subcutaneous fatty tissue provide important clues for forensic reconstruction. The interpretation of these patterns requires a precise description and recording of the position and extent of each lesion. During conventional autopsy, this evaluation is performed by dissecting the skin and subcutaneous tissues in successive layers. In this way, depending on the force and type of impact (right angle or tangent), several morphologically distinct stages of fatty tissue damage can be differentiated: perilobular hemorrhage (I), contusion (II), or disintegration (III) of the fat lobuli, and disintegration with development of a subcutaneous cavity (IV). In examples of virtopsy cases showing blunt trauma to the skin and fatty tissue, we analyzed whether these lesions can also be recorded and classified using multislice computed tomography (MSCT) and magnetic resonance imaging (MRI). MSCT has proven to be a valuable screening method to detect the lesions, but MRI is necessary in order to properly differentiate and classify the grade of damage. These noninvasive radiological diagnostic tools can be further developed to play an important role in forensic examinations, in particular when it comes to evaluating living trauma victims.

Prevalence and Clinical Significance of DSM-5-Attenuated Psychosis Syndrome in Adolescents and Young Adults in the General Population: The Bern Epidemiological At-Risk (BEAR) Study

Objective: Section III of the Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition (DSM-5) lists attenuated psychosis syndrome as a condition for further study. One important question is its prevalence and clinical significance in the general population. Method: Analyses involved 1229 participants (age 16-40 years) from the general population of Canton Bern, Switzerland, enrolled from June 2011 to July 2012. "Symptom," "onset/worsening," "frequency," and "distress/disability" criteria of attenuated psychosis syndrome were assessed using the structured interview for psychosis-risk syndromes. Furthermore, help-seeking, psychosocial functioning, and current nonpsychotic axis I disorders were surveyed. Well-trained psychologists performed assessments using the computer-assisted telephone interviewing technique. Results: The symptom criterion was met by 12.9% of participants, onset/worsening by 1.1%, frequency by 3.8%, and distress/disability by 7.0%. Symptom, frequency, and distress/disability were met by 3.2%. Excluding trait-like attenuated psychotic symptoms (APS) decreased the prevalence to 2.6%, while adding onset/worsening reduced it to 0.3%. APS were associated with functional impairments, current mental disorders, and help-seeking although they were not a reason for help-seeking. These associations were weaker for attenuated psychosis syndrome. Conclusions: At the population level, only 0.3% met current attenuated psychosis syndrome criteria. Particularly, the onset/worsening criterion, originally included to increase the likelihood of progression to psychosis, lowered its prevalence. Because progression is not required for a self-contained syndrome, a revision of the restrictive onset criterion is proposed to avoid the exclusion of 2.3% of persons who experience and are distressed by APS from mental health care. Secondary analyses suggest that a revised syndrome would also possess higher clinical significance than the current syndrome.

Mapping and characterization of the Xlsirt P11 RNA cis-acting localization element

In many organisms, polarity of the oocyte is established post-transcriptionally via subcellular RNA localization. Many RNAs are localized during oogenesis in Xenopus laevis, including Xlsirts ( Xenopus laevis short interspersed repeat transcripts) [Kloc, 1993]. Xlsirts constitute a large family defined by highly homologous repeat units 79–81 nucleotides in length. Endogenous Xlsirt RNAs use the METRO (Message Transport Organizer) pathway of localization, where RNAs are transported from the nucleus to the mitochondrial cloud in stage I oocytes. Secondly, RNAs anchor at the vegetal pole in stage II oocytes. Exogenous Xlsirt RNAs can also utilize the Late pathway of localization, which involves localization to the vegetal cortex during stage III of oogenesis and results in RNAs anchored in the cortex of the entire vegetal hemisphere. ^ The Xlsirts localization signal is contained within the repeat region. This study was designed to test the hypothesis that there are cis -acting localization elements in Xlsirts, and that higher order structure plays a role. Results of experiments on Xlsirt P11, a 1700 basepair (bp) family member, led to the conclusion that a 137-bp fragment of the repetitive region is necessary and sufficient for METRO and Late pathway localization. This analysis definitively demonstrates that the Xlsirt localization signal for the METRO and Late pathways reside within the repetitive region and not within the flanking regions. Analysis of Xlsirt linker scanning mutations revealed two METRO-pathway specific subelements, and one Late-pathway specific subelement. Functional, computer, and biochemical evidence relates the higher order structure of this element to its ability to function as a localization element. ^ Xlsirt 137 is 99% identical to the Xlsirt consensus sequence identified in this study, suggesting that it is the localization element for all localized Xlsirt family members. The repeat unit was reframed based on function, rather than arbitrarily based on sequence. This work supports the hypothesis presented in 1981 by George Spohr, who originally isolated the Xlsirts, which stated that the highly conserved repetitive elements must be constrained from variability due to some unknown function of the repeats themselves. These studies shed light on the mechanism of RNA localization, linking structure and function. ^

Life, wanderings and labours in Eastern Africa : with an account of the first successful ascent of the equatorial snow mountain, Kilima Njaro, and remarks upon East African slavery

by Charles New, of the late Livingstone search and relief expedition

Policy analysis of the Americans with Disabilities Act of 1990 and federal funding concerning barrier-free playgrounds

The Americans with Disabilities Act (ADA) of 1990 was created to prohibit discrimination against disabled persons in our society. The goal of the ADA as a comprehensive civil rights law is to "ensure equal opportunity and complete participation, independent living and economic self-sufficiency" for disabled persons (U.S. Department of Justice, 2008). As part of Title II and III of the ADA, states and local governments are required to provide people with disabilities the same chance to engage in and benefit from all programs and services including recreational facilities and activities as every other citizen. Recreational facilities and related structures must comply with accessibility standards when creating new structures or renovating existing ones. Through a systematic literature review of articles accessed through online databases, articles relating to children with disabilities, their quality of life and their experience gained through play were reviewed, analyzed and synthesized. Additionally, the ADA's Final Rule regarding accessible playgrounds was evaluated through a descriptive analysis which yielded the following five components relating the importance of barrier-free playgrounds to children with disabilities: appropriate dimensions for children, integration of the play area, variety of activity and stimulation, availability of accessible play structures to communities, and financial feasibility. These components were used as evaluation criteria to investigate the degree to which the ADA's Final Rule document met these criteria. An evaluation of two federal funding sources, the Urban Parks and Recreation Renewal Program (UPARR) and the Land and Water Conservation Fund (LWCF), was also conducted which revealed three components relating the two programs' ability to support the realization of the ADA's Final Rule which included: current budget for the program, ability of local communities to attain funds, and level of ADA compliance required to receive funding. Majority of the evaluation of the Final Rule concluded it be adequate in development of barrier-free playgrounds although there are some portions of the guidelines that would benefit from further elucidation. Both funding programs were concluded to not adequately support the development of barrier-free playgrounds and therefore it was recommended that their funding be re-instated or increased as necessary. ^

The prevalence of periodontal disease in Mexican American vs European American elderly in three distinct socio-economic communities in San Antonio, Texas

Purpose of the Study: This study evaluated the prevalence of periodontal disease between Mexican American elderly and European American elderly residing in three socio-economically distinct neighborhoods in San Antonio, Texas. ^ Study Group: Subjects for the original protocol were participants of the Oral Health: San Antonio Longitudinal Study of Aging (OH: SALSA), which began with National Institutes of Health (NIH) funding in 1993 (M.J. Saunders, PI). The cohort in the study was the individuals who had been enrolled in Phases I and III of the San Antonio Heart Study (SAHS). This SAHS/SALSA sample is a community-based probability sample of Mexican American and European American residents from three socio-economically distinct San Antonio neighborhoods: low-income barrio, middle-income transitional, and upper-income suburban. The OH: SALSA cohort was established between July 1993 and May 1998 by sampling two subsets of the San Antonio Heart Study (SAHS) cohort. These subsets included the San Antonio Longitudinal Study of Aging (SALSA) cohort, comprised of the oldest members of the SAHS (age 65+ yrs. old), and a younger set of controls (age 35-64 yrs. old) sampled from the remainder of the SAHS cohort. ^ Methods: The study used simple descriptive statistics to describe the sociodemographic characteristics and periodontal disease indicators of the OH: SALSA participants. Means and standard deviations were used to summarize continuous measures. Proportions were used to summarize categorical measures. Simple m x n chi square statistics was used to compare ethnic differences. A multivariable ordered logit regression was used to estimate the prevalence of periodontal disease and test ethnic group and neighborhood differences in the prevalence of periodontal disease. A multivariable model adjustment for socio-economic status (income and education), gender, and age (treated as confounders) was applied. ^ Summary: In the unadjusted and adjusted model, Mexican American elderly demonstrated the greatest prevalence for periodontitis, p < 0.05. Mexican American elderly in barrio neighborhoods demonstrated the greatest prevalence for severe periodontitis, with unadjusted prevalence rates of 31.7%, 22.3%, and 22.4% for Mexican American elderly barrio, transitional, and suburban neighborhoods, respectively. Also, Mexican American elderly had adjusted prevalence rates of 29.4%, 23.7%, and 20.4% for barrio, transitional, and suburban neighborhoods, respectively. ^ Conclusion: This study indicates that the prevalence of periodontal disease is an important oral health issue among the Mexican American elderly. The results suggest that the socioeconomic status of the residential neighborhood increased the risk for severe periodontal disease among the Mexican American elderly when compared to European American elderly. A viable approach to recognizing oral health disparities in our growing population of Mexican American elderly is imperative for the provision of special care programs that will help increase the quality of care in this minority population.^

Dinoflagellates of ODP Leg 104 holes

Dinoflagellate cysts, pollen, and spores were studied from 78 samples of the Eocene to Miocene section of ODP Site 643 at the outer Wring Plateau. Dinoflagellate cysts ranging from less than 1,000 to rarely over 30,000 per gram of sediment in the Paleogene, and generally between 50,000 and 100,000 in the Miocene were present. The shift to conspicuously higher cyst frequencies takes place in the lowermost Miocene section and appears to reflect increased cyst recruitment rather than a change in sedimentation rate. Of the 179 dinoflagellate cyst forms whose ranges were recorded, 129 are known species. Fifteen assemblage zones have been recognized, although the upper Eocene is missing and no substantial lower Eocene was recorded at Site 643. Norwegian Sea and Rockall Plateau zonations were compared with this study. Detailed correlation with existing onshore section zonations was difficult because key zonal species are inadequately represented; however, the middle to upper Miocene zonation established for Denmark is applicable. Pollen and spores occur with relatively low frequencies, and palynodebris is generally absent, in contrast to the observations from DSDP Leg 38. Thirty-nine samples from Eocene to Miocene sediments at Site 642 were studied and correlated with Site 643. A lower Eocene cyst assemblage present in Hole 642D is older than the questionably lower Eocene assemblage from Site 643. Site 642 has a lower Eocene to lower Miocene hiatus.

La arquitectura residencial en El Real Sitio de El Pardo (1885 - 1965): Orígenes, tipos y configuración del trazado urbano

Tras la denominación de Real Sitio a mediados del siglo XVIII, bajo el reinado de Fernando VI, su sucesor Carlos III procedió a la incorporación a su Patrimonio de todos los Montes y Bosques de El Pardo. Comenzó entonces el proceso de planeamiento urbano y de construcción arquitectónica que finalizó en torno al año 1800. En lo sucesivo, no sólo se mantiene el curso de la conservación y consolidación de los edificios principales, sino que se realiza obra nueva de índole civil. Algunos edificios cambiaron de propiedad y de uso hasta que tras la Guerra Civil se procedió a la mayor transformación vivida por el Real Sitio. El intervalo que aquí se trata (1885 a 1965), no ha suscitado, en los estudios sobre El Pardo, atención suficiente al no acontecer obra nueva de carácter patrimonial ni ha sido objeto de análisis el trazado y la fisonomía del centro urbano residencial del pueblo que Carlos III configuró. Sin embargo se estima relevante analizar los cambios en la actividad residencial; en primer lugar porque coexiste con la arquitectura oficial y, por tanto, se entiende necesario un análisis global del conjunto y en segundo lugar porque facilita la comprensión sobre la imagen original de carácter histórico del conjunto de finales del siglo XVIII. Este marco temporal determina tres partes principales de estudio que estructuran la presente tesis, cuyas fechas establecen los intervalos históricos clave: Actuaciones sobre el núcleo urbano consolidado (1885-1931). Cese de la actividad constructiva (1931-1939). Propuestas regeneradoras y crecimiento acelerado (1939-1965). Dentro de ellos se establecen, a su vez, dos subcapítulos diferenciados con la finalidad de explicar los sucesos que pautaron los cambios trascendentales en la historia de El Pardo. En el estudio del estado de la cuestión se observa que en El Pardo, al igual que sucede en otros Reales Sitios, se investigan los edificios destacados como el Palacio, la Casita del Príncipe, la Casa de Oficios y la Casa de Infantes desde el punto de vista de su historia pero no desde la arquitectura ni de cómo esta afecta al desarrollo del trazado y por tanto al contexto urbano. Se manifiestan determinadas carencias de tratamiento gráfico que facilitarían la comprensión histórica mediante el análisis de la forma y cómo esta ha ido variando sustancialmente. El concepto de escala y orientación reordena el estudio, no sólo de estos edificios protagonistas sino de los que se entretejen a su alrededor y componen el conjunto histórico, lo cual aporta nuevas conclusiones al estado de la cuestión que aquí compete. El principal objetivo de la tesis es, por tanto, contribuir a la dimensión patrimonial mediante el estudio de la arquitectura residencial del pueblo de El Pardo y en cómo esta ha ido conformando y consolidando el entramado urbano original en torno a edificios de la realeza y corte. Analizar aquellos edificios que perduran, los que fueron reconstruidos, rehabilitados, y apuntar acontecimientos históricos que formularon la actual fisonomía. Sistematizar y reordenar sobre la traza actual los edificios que desaparecieron, nos da las pistas sobre las modificaciones en concepto de escala arquitectónica y urbana. El estudio de las fuentes y establecer una metodología de conexión de estas, ayuda a detectar dónde no se han dirigido aún los focos de interés así como las lagunas que han quedado por explorar con el fin de responder a nuevas hipótesis, conceder conclusiones y abrir otras líneas de investigación. Como conclusiones generales, la tesis aporta documentación nueva sobre el objeto de estudio, no solicitada, digitalizada o publicada con anterioridad. En ella se analizan los procesos de configuración, consolidación y transformación en el Real Sitio mediante la sistematización de estados comparativos. Con respecto al estudio de los diferentes contextos natural y urbano la tesis analiza cómo los accidentes naturales, el desarrollo de infraestructuras y el impulso de la agronomía afectaron a El Pardo a partir del siglo XIX, y estudia los procesos de configuración, consolidación y transformación en el Real Sitio mediante la sistematización de la documentación encontrada de manera gráfica y escrita. En relación al marco patrimonial arquitectónico, la tesis analiza los procesos edificatorios históricos. Se estudian, a su vez, cambios de ocupación o uso que derivaron en reformas, ampliaciones, obras de nueva planta e incluso en derribos, así como los proyectos no materializados o que se llevaron a cabo de manera parcial. Con respecto al análisis del momento histórico, la tesis analiza las posibles afectaciones, políticas, sociales y económicas en las etapas de Monarquía, Segunda República, Guerra Civil y Posguerra. Por último, la tesis abre cuatro vías de investigación (que ya se han tratado y avanzado en parte pero que escapan a los límites de este trabajo) que pueden plantear nuevas hipótesis, reportando así respuestas sobre objetos de estudio complementarios y paralelos al presente. Estas refieren a análisis más concretos sobre El Palacio Real de El Pardo y la Casa de Oficios, el Camino Real de Madrid a El Pardo desde la Puerta de Hierro, los cuarteles, puertas y portilleras del Monte de El Pardo y los proyectos desarrollados por el arquitecto Diego Méndez en los Reales Sitios para el Patrimonio Nacional. ABSTRACT Following the Royal Site denomination being granted in the mid-18th Century, during the reign of Ferdinand VI, his successor Charles III proceeded to include all the Forests and Woodlands of El Pardo in his heritage. That then gave rise to the process of town planning and architectural construction that was completed around 1800. Thereafter, not only the process of conservation and consolidation of the main buildings has been maintained, but new civil engineering works have also been carried out. Some buildings changed ownership and use until, after the Civil War, the greatest transformation experienced by the Royal Site was undertaken. The time frame this paper concerns (1885 to 1965), has not attracted sufficient attention in studies of El Pardo due to there having been no new works with heritage status, nor has there been an analysis of the layout and external appearance of the residential centre in the town once conceived by Charles III. However, it is considered relevant to analyse the changes in residential activity, firstly, because it coexists with the official architecture and, thus, it is considered necessary to perform a global analysis of the complex and, secondly, because it facilitates a historical understanding of the original appearance of the complex at the end of the 18th Century. This time framework defines three main parts of the study that provide the structure of this thesis, the dates of which establish the key historical time frames: Actions in the consolidated town centre (1885-1931). Cessation of construction works (1931-1939). Proposals of regeneration and accelerated growth (1939-1965). Two distinct sub-chapters are also established within these, in order to explain the events that marked the transcendental changes in the history of El Pardo. When studying the subject matter, it is noted that in El Pardo, as is the case in other Royal Sites, outstanding buildings such as the Palace, the Prince's Cottage, the Trades House and the Infantes House are usually researched strictly from the point of view of their history, but not from an architectural perspective, nor analysing how that affects the development of the site layout and thus the urban area. Specific shortcomings are evident in the graphic treatment that would have otherwise facilitated a historical understanding through the analysis of the shape and the way it has gradually undergone substantial variation. The concept of scale and orientation reorganises the study, not only of these key buildings, but also of those that are woven around them and make up the historic complex, allowing entirely new conclusions concerning the subject matter analysed herein. Therefore, the main purpose of this thesis is to outline our heritage through the study of the residential architecture of the town of El Pardo and the analysis of the way the original town has been built up and consolidated around the buildings erected by royalty and the court; to analyse the buildings that still remain, those that were rebuilt, refurbished, and to note historic events that shaped its current appearance. To this end, a systematic classification and reorganisation on the current urban layout of the buildings that have disappeared will give us the key to understand changes in the concept of architectural and urban scale. Studying the sources and establishing a methodology to connect them will help us detect those areas where the focus of interest has not concentrated yet, and will also reveal the gaps that remain unexplored, in order to respond to new hypotheses, reach new conclusions and open up new lines of research. As general conclusions, this thesis provides new documentation on the subject matter that had not been requested, digitized or published before. There we find an analysis of the processes of configuration, consolidation and transformation of the Royal Site through a systematic classification of comparative states. With regard to the study of the multiple natural and urban environments, this thesis analyses the way natural features, development of infrastructures and agricultural driving forces affected El Pardo as of the 19th Century, and it studies the processes of configuration, consolidation and transformation of the Royal Site by systematically classifying the documentation found in graphic and written documents. In relation to the architectural heritage framework, this thesis analyses historical building processes. Likewise, a study is also performed on the changes in land occupation or use that led to reforms, extensions, new buildings and even to demolitions, as well as on unrealized projects, or even on those that were partially implemented. As for the analysis of the historical time period, this thesis assesses the potential political, social and economic effects of the Monarchy, Second Republic, Civil War and Post-War Periods. Finally, this thesis opens up four lines of investigation (that have already been discussed and partially advanced, but which fall beyond the scope of this work) that could pose new hypotheses, thus giving answer to other subject matters parallel and complementary to the one assessed herein. These refer to more specific analyses of El Palacio Real de El Pardo (Royal Palace of El Pardo) and the Casa de Oficios (Trades House), the Royal Highway from Madrid to El Pardo from Puerta de Hierro, the barracks, gates and entrances to estates in the Woodlands of El Pardo and the projects developed on the Royal Sites by the architect Diego Méndez for the National Heritage.

Factors affecting berry composition of Tempranillo grapevines before the arrest of phloem transport.

It is already known that berry ripening is determined by the leaf area/fruit ratio, as well as temperature and leaf physiology. The aim of this work was to assess the influence of these parameters on Tempranillo cultivar throughout stage III of berry development.

Caracterización funcional de los genes sgb (suppressors of agb1-2 susceptibility to pathogens): Nuevos reguladores de la respuesta de inmunidad innata de Arabidopsis thaliana

Un porcentaje importante de las pérdidas de la producción agrícola se deben a las enfermedades que causan en los cultivos los hongos necrótrofos y vasculares. Para mejorar la productividad agrícola es necesario tener un conocimiento detallado de las bases genéticas y moleculares que regulan la resistencia de las plantas a este tipo de patógenos. En Arabidopsis thaliana la resistencia frente a patógenos necrótrofos, como el hongo Plectosphaerella cucumerina BMM (PcBMM), es genéticamente compleja y depende de la activación coordinada de distintas rutas de señalización, como las reguladas por las hormonas ácido salicílico (SA), ácido jasmónico (JA), etileno (ET) y ácido abscísico (ABA), así como de la síntesis de compuestos antimicrobianos derivados del Triptófano y de la integridad de la pared celular (Llorente et al., 2005, Hernández-Blanco et al., 2007; Delgado-Cerezo et al., 2012). Uno de los componentes claves en la regulación de la resistencia de las plantas a patógenos (incluidos hongos necrótrofos y biótrofos) es la proteína G heterotrimérica, un complejo proteico formado por tres subunidades (Gα, Gβ y Gγ), que también regula distintos procesos del desarrollo vegetal. En Arabidopsis hay un gen que codifica para la subunidad α (GPA1), otro para la β (AGB1), y tres genes para la subunidad γ (AGG1, AGG2 y AGG3). El complejo GPA1-AGB1-AGG (1-3) se activa y disocia tras la percepción de una señal específica, actuando el dímero AGB1-AGG1/2 como un monómero funcional que regula las respuestas de defensa (Delgado-Cerezo et al., 2012). Estudios transcriptómicos y análisis bioquímicos de la pared celular en los que se comparaban los mutantes agb1-2 y agg1 agg2, y plantas silvestres (Col-0) revelaron que la resistencia mediada por Gβ-Gγ1/2 no es dependiente de rutas de defensa previamente caracterizadas, y sugieren que la proteína G podría modular la composición/estructura (integridad) de la pared celular (Delgado-Cerezo et al., 2012). Recientemente, se ha demostrado que AGB1 es un componente fundamental de la respuesta inmune mediada por Pathogen- Associated Molecular Patterns (PTI), ya que los mutantes agb1-2 son incapaces de activar tras el tratamiento con PAMPs respuestas de inmunidad, como la producción de especies reactivas de oxígeno (ROS; Liu et al., 2013). Dada la importancia de la proteína G heterotrimérica en la regulación de la respuestas de defensa (incluida la PTI) realizamos un escrutinio de mutantes supresores de la susceptibilidad de agb1-2 al hongo necrótrofo, PcBMM, para identificar componentes adicionales de las rutas de señalización reguladas por AGB1. En este escrutinio se aislaron cuatro mutantes sgb (suppressors of agb1-2 susceptibility to pathogens), dos de los cuales, sgb10 y sgb11, se han caracterizado en la presente Tesis Doctoral. El mutante sgb10 es un segundo alelo nulo del gen MKP1 (At3g55270) que codifica la MAP quinasa-fosfatasa 1 (Bartels et al., 2009). Este mutante presenta lesiones espontáneas en plantas adultas y una activación constitutiva de las principales rutas de defensa (SA, JA y ET, y de metabolitos secundarios, como la camalexina), que explicaría su elevada resistencia a PcBMM y Pseudomonas syringae. Estudios epistáticos sugieren que la resistencia mediada por SGB10 no es dependiente, si no complementaria a la regulada por AGB1. El mutante sgb10 es capaz de restablecer en agb1-2 la producción de ROS y otras respuestas PTI (fosforilación de las MAPK6/3/4/11) tras el tratamiento con PAMPs tan diversos como flg22, elf18 y quitina, lo que demuestra el papel relevante de SGB10/MKP1 y de AGB1 en PTI. El mutante sgb11 se caracteriza por presentar un fenotipo similar a los mutantes irregular xylem (e.g. irx1) afectado en pared celular secundaria: irregularidades en las células xilemáticas, reducción en el tamaño de la roseta y altura de planta, y hojas con un mayor contenido de clorofila. La resistencia de sgb11 a PcBMM es independiente de agb1-2, ya que la susceptibilidad del doble mutante sgb11 agb1-2 es intermedia entre la de agb1-2 y sgb11. El mutante sgb11 no revierte la deficiente PTI de agb1-2 tras el tratamiento con flg22, lo que indica que está alterado en una ruta distinta de la regulada por SGB10. sgb11 presenta una sobreactivación de la ruta del ácido abscísico (ABA), lo que podría explicar su resistencia a PcBMM. La mutación sgb11 ha sido cartografiada en el cromosoma III de Arabidopsis entre los marcadores AthFUS6 (81,64cM) y nga6 (86,41cM) en un intervalo de aproximadamente 200 kb, que comprende genes, entre los que no se encuentra ninguno previamente descrito como IRX. El aislamiento y caracterización de SGB11 apoya la relevancia de la proteína G heterotrimérica en la regulación de la interconexión entre integridad de la pared celular e inmunidad. ABSTRACT A significant percentage of agricultural losses are due to diseases caused by necrotrophic and vascular fungi. To enhance crop yields is necessary to have a detailed knowledge of the genetic and molecular bases regulating plant resistance to these pathogens. Arabidopsis thaliana resistance to necrotrophic pathogens, such as Plectosphaerella cucumerina BMM (PcBMM) fungus, is genetically complex and depends on the coordinated activation of various signaling pathways. These include those regulated by salicylic acid (SA), jasmonic acid (JA), ethylene (ET) and abscisic acid (ABA) hormones and the synthesis of tryptophan-derived antimicrobial compounds and cell wall integrity (Llorente et al., 2005, Hernández-Blanco et al., 2007; Delgado-Cerezo et al., 2012). One key component in the regulation of plant resistance to pathogens (including biotrophic and necrotrophic fungi) is the heterotrimeric G-protein. This protein complex is formed by three subunits (Gα, Gβ and Gγ), which also regulates various plant developmental processes. In Arabidopsis only one gene encodes for subunits α (GPA1) and β (AGB1), and three genes for subunit γ (AGG1, AGG2 y AGG3). The complex GPA1- AGB1-AGG(1-3) is activated and dissociates after perception of an specific signal, AGB1- AGG1/2 acts as a functional monomer regulating defense responses (Delgado-Cerezo et al., 2012). Comparative transcriptomic studies and biochemical analyses of the cell wall of agb1-2 and agg1agg2 mutant and wild plants (Col-0), showed that Gβ-Gγ1/2-mediated resistance is not dependent on previously characterized defense pathways. In addition, it suggests that G protein may modulate the composition/structure (integrity) of the plant cell wall (Delgado-Cerezo et al., 2012). Recently, it has been shown that AGB1 is a critical component of the immune response mediated by Pathogen-Associated Molecular Patterns (PTI), as agb1-2 mutants are unable to activate immune responses such as oxygen reactive species (ROS) production after PAMPs treatment (Liu et al., 2013). Considering the importance of the heterotrimeric G protein in regulation of defense responses (including PTI), we performed a screening for suppressors of agb1-2 susceptibility to the necrotrophic fungus PcBMM. This would allow the identification of additional components of the signaling pathways regulated by AGB1. In this search four sgb mutants (suppressors of agb1-2 susceptibility to pathogens) were isolated, two of which, sgb10 and sgb11, have been characterized in this PhD thesis. sgb10 mutant is a second null allele of MKP1 gene (At3g55270), which encodes the MAP kinase-phosphatase 1 (Bartels et al., 2009). This mutant exhibits spontaneous lesions in adult plants and a constitutive activation of the main defense pathways (SA, JA and ET, and secondary metabolites, such as camalexin), which explains its high resistance to Pseudomonas syringae and PcBMM. Epistatic studies suggest that SGB10- mediated resistance is not dependent, but complementary to the regulated by AGB1. The sgb10 mutant is able to restore agb1-2 ROS production and other PTI responses (MAPK6/3/4/11 phosphorylation) upon treatment with PAMPs as diverse as, flg22, elf18 and chitin, demonstrating the relevant role of SGB10/MKP1 and AGB1 in PTI. sgb11 mutant is characterized by showing a similar phenotype to irregular xylem mutants (e.g. irx1), affected in secondary cell wall: irregular xylems cells, rosette size reduction and plant height, and higher chlorophyll content on leaves. The resistance of sgb11 to PcBMM is independent of agb1-2, as susceptibility of the double mutant agb1-2sgb11 is intermediate between agb1-2 and sgb11. The sgb11 mutant does not revert the deficient PTI response in agb1-2 after flg22 treatment, indicating that is altered in a pathway different to the one regulated by SGB10. sgb11 presents an over-activation of the abscisic acid pathway (ABA), which could explain its resistance to PcBMM. The sgb11 mutation has been mapped on chromosome III of Arabidopsis, between AthFUS6 (81.64 cM) and nga6 (86.41 cM) markers, in 200 kb interval, which does not include previously known IRX genes. The isolation and characterization of SGB11 supports the importance of heterotrimeric G protein in the regulation of the interconnection between the cell wall integrity and immunity.