La Cooperativa Arroceros Villa Elisa, un buen ejemplo de la tradición cooperativista de Entre Ríos (Argentina)

Así como la Historia reconoce a Entre Ríos como cuna de la colonización, es también en esta provincia donde desde el comienzo del siglo XX se desarrolla el cooperativismo agrario que presta importantes servicios al asociado en materia de abastecimiento, comercialización y transformación. Prestación que se traduce en un uso más racional de la tierra, un mayor volumen de negocios, el mejoramiento en la calidad del producto, la utilización eficiente del capital, el aumento de la demanda por la ampliación de los mercados y la introducción de servicios que cada agricultor por si sólo no podría alcanzar. El presente artículo se propone analizar la trayectoria de una cooperativa arrocera -fundada en la década de 1970- ubicada en la localidad entrerriana de Villa Elisa que en mayor o menor media ha ido cumpliendo con esas prestaciones y que hoy se posiciona como el tercer exportador nacional de arroz y el primero de gestión cooperativa. Más allá de las cuestiones económicas, se prestará especial atención a la articulación interinstitucional de la empresa y a los cambios introducidos a nivel organizacional

La Cooperativa Arroceros Villa Elisa, un buen ejemplo de la tradición cooperativista de Entre Ríos (Argentina)

Así como la Historia reconoce a Entre Ríos como cuna de la colonización, es también en esta provincia donde desde el comienzo del siglo XX se desarrolla el cooperativismo agrario que presta importantes servicios al asociado en materia de abastecimiento, comercialización y transformación. Prestación que se traduce en un uso más racional de la tierra, un mayor volumen de negocios, el mejoramiento en la calidad del producto, la utilización eficiente del capital, el aumento de la demanda por la ampliación de los mercados y la introducción de servicios que cada agricultor por si sólo no podría alcanzar. El presente artículo se propone analizar la trayectoria de una cooperativa arrocera -fundada en la década de 1970- ubicada en la localidad entrerriana de Villa Elisa que en mayor o menor media ha ido cumpliendo con esas prestaciones y que hoy se posiciona como el tercer exportador nacional de arroz y el primero de gestión cooperativa. Más allá de las cuestiones económicas, se prestará especial atención a la articulación interinstitucional de la empresa y a los cambios introducidos a nivel organizacional

Direct and indirect economic impacts of drought in the agri-food sector in the Ebro River basin (Spain).

The economic evaluation of drought impacts is essential in order to define efficient and sustainable management and mitigation strategies. The aim of this study is to evaluate the economic impacts of a drought event on the agricultural sector and measure how they are transmitted from primary production to industrial output and related employment. We fit econometric models to determine the magnitude of the economic loss attributable to water storage. The direct impacts of drought on agricultural productivity are measured through a direct attribution model. Indirect impacts on agricultural employment and the agri-food industry are evaluated through a nested indirect attribution model. The transmission of water scarcity effects from agricultural production to macroeconomic variables is measured through chained elasticities. The models allow for differentiating the impacts deriving from water scarcity from other sources of economic losses. Results show that the importance of drought impacts are less relevant at the macroeconomic level, but are more significant for those activities directly dependent on water abstractions and precipitation. From a management perspective, implications of these findings are important to develop effective mitigation strategies to reduce drought risk exposure.

Historia de la protección del patrimonio arquitectónico en España. 1933-1985

El trabajo de investigación que presentamos tiene como principal objetivo, la recopilación, el registro, el análisis y la reflexión sobre una época, tan trascendental como poco estudiada desde el ámbito arquitectónico, como es el período comprendido entre las dos normas de mayor relevancia en relación a la protección del Patrimonio Histórico Español del último siglo. Nos referimos a la Ley sobre Defensa, Conservación y Acrecentamiento del Patrimonio Histórico Nacional de 13 de mayo de 1933 y la Ley 13/1985, de 25 de junio, del Patrimonio Histórico Español. A través de la investigación realizada, se pretende aportar una visión integral de esta etapa, desde el enfoque arquitectónico, fundamentando la misma en el desarrollo de pautas metodológicas, abordadas desde la recopilación exhaustiva del material bibliográfico y documental para su posterior análisis. A partir de esta fase inicial, se han identificado los nexos comunes entre los estudios existentes sobre el patrimonio monumental español previos a la Guerra Civil y las investigaciones dedicadas a la historia de las últimas décadas del siglo XX. De esta forma, se ha procurado trazar un “puente” documental, con el que trasponer virtualmente el vacío bibliográfico existente. Históricamente, la protección del patrimonio histórico edificado y urbano, ha preocupado y ocupado a multitud de profesionales que, desde disciplinas dispares, han emprendido la tarea ímproba de comprender y explicar cuáles han sido los avatares, históricos y legales, que han marcado su evolución. Tal preocupación ha generado una bibliografía ingente y diversa, desde la protección formal y precisa, sobre uno u otro material, pasando por el marco historiográfico de las tendencias conservacionistas y las teorías decimonónicas, las filigranas formadas por las cuantiosas normas promulgadas desde la Novísima Recopilación, hasta la incidencia del planeamiento urbano en la tutela del patrimonio, incluidas la trama de competencias y yuxtaposiciones administrativas. Documentos de toda índole y profundidad científica, que como mosaicos hispanomusulmanes, dibujan el panorama patrimonial en el que la criba de material resulta una tarea, en ocasiones, inextricable. El título de este documento, en sí mismo, circunscribe la materia que ha sido el objeto de análisis durante el proceso de investigación, el Patrimonio Arquitectónico Monumental. El eje o núcleo basal de estudio se sitúa en los bienes inmuebles, los edificados, que, a su vez, ostentan la declaración de Bien de Interés Cultural, y que, por ende, pertenecen al Patrimonio Histórico Español. La metodología de trabajo se ha desarrollado de forma concéntrica, desde aspectos generales de la protección del patrimonio monumental, como el marco legal que antecede a la promulgación de la Ley de 1933, y el estado previo de los bienes susceptibles de ser preservados. Reconocemos en el ámbito legislativo, el fundamento orgánico que regula y dirige la tutela del patrimonio histórico español y la acción conservadora, y que delimita el ámbito a partir del cual se condiciona el devenir de los bienes culturales. Del esquema de situación surgido del análisis previo, se han detectado los factores claves en la transición hacia la Ley de Patrimonio Histórico Español; la evolución conceptual del “Patrimonio”, como apreciación genérica, y el testimonio de este progreso a través de los valores históricos, artísticos y culturales. El presente documento de investigación, consta de una primera fase, correspondiente al Capítulo 1, que se ha desarrollado a partir, principalmente, de la ordenación jurídica que rige el Patrimonio Histórico Español, a través de leyes, decretos, órdenes y disposiciones anexas, complementado con el material bibliográfico dedicado a la revisión histórica del proceso legal de la protección del patrimonio histórico-artístico. Si bien no ha sido nuestro propósito realizar un estudio pormenorizado del volumen jurídico e histórico que precede a la Ley de 1933, y que da inicio al período de estudio de la presente investigación, sí lo ha sido centrarnos en la elaboración de un extracto de aquellos elementos de la doctrina de mayor relevancia y repercusión en la protección del patrimonio histórico-artístico y/o monumental español. A lo largo de este estudio hemos comprobado lo que algunos juristas ya habían planteado, acerca de la profunda dispersión, ramificación, y diversificación de esfuerzos, tanto en la legislación específica como en la urbanística. Esta disgregación se ha extendido al ámbito de las medidas de reconocimiento caracterizado por la elaboración de múltiples catálogos e inventarios, con desigual transcendencia, alcance y utilidad. El resultado ha sido una división de esfuerzos, desdibujando el objetivo y convirtiendo la acción del reconocimiento en múltiples empresas inconexas y de escasa trascendencia. Nuestra investigación avanza en el análisis de la protección del patrimonio, como concepto globalizador, con el desarrollo del Capítulo 2, en el que se incluye una serie de mecanismos directos e indirectos que, individualmente, suelen carecer de la fuerza efectiva que muchos de los monumentos o conjuntos monumentales requieren para sobrevivir al paso del tiempo y sus circunstancias. En primer lugar, en este segundo capítulo nos hemos centrado, específicamente, en el mecanismo regulado por la Ley del Patrimonio Histórico Español, y el régimen general de protección implementado a partir de su promulgación en 1985. En especial, consideraremos la declaración de Interés Cultural como grado máximo de protección y tutela de un bien, y su posterior inscripción en el Registro General correspondiente, dependiente del Ministerio de Educación, Cultura y Deporte. Este mecanismo representa el instrumento por antonomasia que condensa las facultades de tutela del Estado sobre un bien del que se considera poseedor y aglutinador de valores “culturales” —como cohesión de los valores históricos, artísticos, sociales, etc. — representativos de la idiosincrasia española, y sobre el cual no existen dudas sobre la necesidad de garantizar su permanencia a través de su conservación. En segunda instancia, hemos analizado el Planeamiento Urbanístico, como aglutinador de valores culturales contenidos en la ciudad y como contenedor de los efectos generados por el hombre a partir de su interacción con el medio en el que habita y se relaciona. En tercer término, hemos recopilado y estudiado la concepción de los catálogos, como noción genérica de protección. Desde hace siglos, este género ha estado definido como una herramienta capaz de intervenir en la protección del patrimonio histórico, aunque de una manera difícilmente cuantificable, mediante la identificación, enumeración y descripción de una tipología concreta de monumentos o grupos de ellos, contribuyendo al reconocimiento de los valores cualitativos contenidos en éstos. El tercer capítulo analiza el mecanismo directo de tutela que ejerce la Administración en el patrimonio monumental. La declaración de monumentalidad o de Bien de Interés Cultural y su inclusión en el Registro General de Protección. La protección teórica y la protección jurídica de un monumento, analizadas hasta el momento, resultan tan necesarias como pueriles si no van seguidas de su consumación. En el caso de este tipo de patrimonio monumental, toda acción que tenga como objeto resguardar los valores implícitos en un bien mueble o inmueble, y en su materia, implica el cumplimiento de la protección. Por último, el cuarto capítulo se convierte en el punto culminante, y por ende crucial, del proceso de protección del Patrimonio Cultural, el de la consumación de la intervención. La teoría, la crítica, la normativa y hasta las doctrinas más radicales en materia de protección del patrimonio cultural, carecen de sentido si no las suceden los hechos, la acción, en antítesis a la omisión o la desidia. De ello ha dado pruebas elocuentes la propia historia en multitud de ocasiones con la destrucción, por indolencia o desconocimiento, de importantes vestigios del patrimonio arquitectónico español. Por este motivo, y para ser consecuentes con nuestra tesis hemos recuperado, concentrado y analizado la documentación de obra de tres monumentos imprescindibles del patrimonio construido (la Catedral de Burgos, el Palacio-Castillo de la Aljafería en Zaragoza y la Muralla de Lugo). En ocasiones, al examinar retrospectivamente las intervenciones en monumentos de gran envergadura, física y cultural como catedrales o murallas, algunos investigadores han tenido la sospecha o prevención de que las actuaciones no han seguido un plan de actuación premeditado, sino que han sido el resultado de impulsos o arrebatos inconexos producto de la urgencia por remediar algún tipo de deterioro. En oposición a esto, y a través del estudio de las intervenciones llevadas a cabo en los tres monumentos mencionados, hemos podido corroborar que, a excepción de intervenciones de emergencia fruto de circunstancias puntuales, existe coherencia desde el proceso de análisis de situación de un bien a la designación de prioridades, que ha regido el proceso restaurador a lo largo de dos siglos. La evolución de las intervenciones realizadas en los monumentos analizados ha estado definida, además de por su complejidad, magnitud y singularidad constructiva, por el devenir de su estructura y su uso. En conclusión, la efectividad de la protección del patrimonio cultural español, radica en la concomitancia de múltiples aspectos, entre ellos: el cumplimiento acertado de las normas vigentes, específicas y accesorias; el conocimiento del bien y de sus valores históricos, artísticos, y culturales; su catalogación o inclusión en los inventarios correspondientes; el compromiso de los agentes e instituciones de los cuales depende; la planificación de las tareas necesarias que garanticen tanto la salvaguarda estructural como la conservación de sus valores; y la incorporación de un plan de seguimiento que permita detectar eventuales peligros que atenten contra su conservación. Pero, la situación óptima estaría dada por un sistema en el que estos mecanismos —regulaciones específicas y urbanísticas, Declaraciones de Bien de Interés Cultural, Catálogos e Inventarios, etc. — funcionaran, de forma parcial o total, como una maquinaria, donde cada pieza operara con independencia relativa, pero en sintonía con los demás engranajes. Hasta el momento, la realidad dista mucho de esta situación, convirtiendo esta convivencia en una utopía. Tanto los legisladores, como las autoridades y los técnicos involucrados, deben tener presente que, de ellos, de los parámetros asignados por la legislación, de la implementación de los instrumentos estipulados por ésta y de las decisiones tomadas por cada uno de los poderes directivos de los órganos competentes, dependerá el alcance y efectividad de la protección, ya que en cada vertiente existe, en mayor o menor medida, un porcentaje de interpretación y subjetividad. ABSTRACT The research that we present has as the main objective to collect, record, analyzed and reflection on a time, that was little studied from the architectural field. It is the period between the two laws of most relevance to the protection of Spanish Historical Heritage of the last century. We refer to the Law on the Protection and Conservation of National Heritage of 1933 and Law 16/1985 of Spanish Historical Heritage. Through this research, it aims to provide a comprehensive view of the stage from the architectural approach, basing it on the development of methodological guidelines. The investigation was initiated by the bibliography and documentary for further analysis. After this initial phase, we have identified the common links between existing studies on the Spanish architectural heritage prior to the Civil War and dedicated research into the history of the late twentieth century. Thus, we have tried to draw a documental bridge, with which virtually transpose the gap that has existed. Historically, professionals from diverse disciplines have been worried and busy of the protection of the built and urban heritage. They have undertaken the daunting task of understanding and explaining the historical and legal difficulties, which have marked its evolution. This concern has generated an enormous and diverse literature, from formal and precise protection, in the framework of conservation historiographical trends and nineteenth-century theories. Also, they have studied the impact of urban planning in the protection of heritage, including the competences and administrative juxtapositions. They have generated a lot of documents of all kinds and scientific depth. The title of this document, in itself, circumscribes the matter that has been analyzed during this research process, the Monumental Architectural Heritage. The basal studio is located in the historical buildings, which, in turn, hold the declaration of cultural interest, and thus belong to the Spanish Historical Heritage. The work methodology was developed concentrically from general aspects of the protection of monuments, such as the legal framework that predates the enactment of the 1933 Act, and the previous state of the monuments that should be preserved. We recognize in the legislative sphere, the organic base that regulates and directs the tutelage of Spanish heritage and conservative action. The situation scheme emerged from the previous analysis, and we detected the key factors in the transition to the Spanish Historical Heritage Act; the conceptual evolution of the Heritage as a generic assessment, and witness this progress through historical, artistic and cultural values. This research paper consists of a first phase, corresponding to Chapter 1, which has developed from the legal regulation governing the Spanish Historical Heritage, through laws, decrees, orders and related provisions, supplemented the bibliography dedicated to the historical review of the legal process of protecting historical and artistic heritage. While it was not our intention to conduct a detailed study of the legal and historical volume preceding the 1933 Act, and that started the study period of this investigation, yes he has been focusing on the production of an extract from those elements of the doctrine with greater relevance and impact on the protection of Spanish art-historical and / or architectural heritage. Throughout our study we have seen what some jurists had already raised, about the scattering, branching and diversification of efforts, both in specific law and in urban law. This disaggregation has been extended to the field of recognition measures characterized by the development of multiple catalogs and inventories, with varying significance, scope and usefulness. The result has been a division of efforts, blurring the objective and turning the action of the recognition in multiple attempts little consequence. Our research advances in the analysis of heritage protection, as globalization concept in the Chapter 2, which includes a number of direct and indirect mechanisms that individually, often lack the effective force that many of monuments have required to survive the test of time and circumstances. First, in this second chapter we focused specifically on the mechanism regulated by the Spanish Historical Heritage Act, and the general protection regime implemented since its enactment in 1985 . In particular, we consider the declaration of cultural interest as maximum protection and protection of cultural assets, and their subsequent entry in the relevant General Register under the Ministry of Education, Culture and Sports . This mechanism is the instrument par excellence that condenses the powers of state care about a cultural asset, and which represents the cohesion of the historical, artistic, social values , etc. Secondly, we analyzed the Urban Planning, as a unifying cultural value in the city and as a container for the effects caused by man from its interaction with the environment in which he lives and relates. Thirdly, we have collected and studied the origin of catalogs, as generic notion of protection. For centuries, this genre has been defined as a tool to intervene in the protection of historical heritage, although difficult to quantify, through the identification, enumeration and description of a particular typology of monuments, and that contributing to the recognition of qualitative values contained therein. The third chapter analyzes the direct mechanism of protection performed by the Administration in the monuments with the statement of Cultural asset and inclusion in the General Protection Register. The theoretical and legal protection of a monument is as necessary as puerile if they are not followed by intervention. For this type of architectural heritage, any action which has the aim to safeguard the values implicit in the cultural asset involves protection compliance. Finally, the fourth chapter becomes the highlight, because it treated of the end process of the cultural heritage protection, the consummation of the intervention. The theory, the criticism, the rules and even the radical doctrines on the protection of cultural heritage, are meaningless if they do not take place the facts, the action, in antithesis to the omission. The history of the architectural heritage has given eloquent proof by itself. A lot of vestiges have been lost, in many times, for the destruction, through indolence or unknowledge. For this reason, and to be consistent with our thesis, we have collected and analyzed the projects documentation of three monuments (the Burgos Cathedral, the Aljafería Palace-Castle in Zaragoza and the Wall of Lugo). Sometimes, some researchers have suspected that there had not been planning. They suspect that the projects have been the result of different emergency situations. In opposition of this, we confirm that, except for emergency interventions result of specific circumstances, there have been a process of analysis to conclude in the priorities designation, which has guided the restoration process over two centuries. The complexity, magnitude and constructive uniqueness have defined the evolution of intervention. In conclusion, the effectiveness of the protection of Spanish cultural heritage lies in the conjunction of many aspects, including: the successful implementation of existing, specific and ancillary standards; the knowledge of good and its historical, artistic and cultural values; the cataloging and inclusion in the relevant inventories; and the commitment of the actors and institutions on which it depends. These planning tasks are necessary to ensure both structural safeguards as conservation values; and the introduction of a monitoring plan to detect possible dangers that threaten its conservation. But, the optimal situation would be given by a system in which these urban-regulations and specific mechanisms, would work together like a machine, where each piece operated with relative independence, but in tune with the other gears. So far, the reality is far from this situation, turning this coexistence in a utopia. Both legislators and officials and technicians involved must be aware that the effectiveness and scope of protection depends on your insight and commitment.

Isolation of recombinant antibody fragments (scFv) by phage display technology for detection of almond allergens in food products

Tree nut allergies are considered an important health issue in developed countries. To comply with the regulations on food labeling, reliable allergen detection methods are required. In this work we isolated almond-specific recombinant antibody fragments (scFv) from a commercial phage display library bypassing the use of live animals, hence being consistent with the latest policies on animal welfare. To this end an iterative selection procedure employing the Tomlinson I phage display library and a crude almond protein extract was carried out. Two different almond-specific scFv (named PD1F6 and PD2C9) were isolated after two rounds of biopanning, and an indirect phage ELISA was implemented to detect the presence of almond protein in foodstuffs. The isolated scFvs demonstrated to be highly specific and allowed detection of 40 ng mL?1 and 100 ng mL?1 of raw and roasted almond protein, respectively. The practical detection limit of the assay in almond spiked food products was 0.1 mg g?1 (110e120 ppm). The developed indirect phage ELISA was validated by analysis of 92 commercial food products, showing good correlation with the results obtained by a previously developed real-time PCR method for the detection of almond in foodstuffs. The selected phage clones can be affinity maturated to improve their sensitivity and genetically engineered to be employed in different assay formats.

p53-independent apoptosis induced by paclitaxel through an indirect mechanism

The efficacy of chemotherapeutic agents may be determined by a number of different factors, including the genotype of the tumor cell. The p53 tumor suppressor gene frequently is mutated in human tumors, and this may contribute to chemotherapeutic resistance. We tested the requirement for wild-type p53 in the response of tumor cells to treatment with paclitaxel (trade name Taxol), an antineoplastic agent that stabilizes cellular microtubules. Although paclitaxel is broadly effective against human tumor xenografts in mice, including some known to carry p53 mutations, we found that p53-containing mouse tumor cells were significantly more sensitive to direct treatment with this drug than were p53-deficient tumor cells. In an attempt to reconcile this apparent discrepancy, we examined the requirement for p53 in the cytotoxic effects of tumor necrosis factor α (TNF-α), a cytokine released from murine macrophages upon paclitaxel treatment. Conditioned medium from paclitaxel-treated macrophages was capable of inducing p53-independent apoptosis when applied to transformed mouse embryonic fibroblasts and was inhibitable by antibodies against TNF-α. Furthermore, in response to direct treatment with TNF-α, both wild-type and p53-deficient tumor cells underwent apoptosis to similar extents and with similar kinetics. Our results suggest that the efficacy of paclitaxel in vivo may be due not only to its microtubule-stabilizing activity, but its ability to activate local release of an apoptosis-inducing cytokine.

Expression of the Hox gene complex in the indirect development of a sea urchin

Hox complex genes control spatial patterning mechanisms in the development of arthropod and vertebrate body plans. Hox genes are all expressed during embryogenesis in these groups, which are all directly developing organisms in that embryogenesis leads at once to formation of major elements of the respective adult body plans. In the maximally indirect development of a large variety of invertebrates, the process of embryogenesis leads only to a free-living, bilaterally organized feeding larva. Maximal indirect development is exemplified in sea urchins. The 5-fold radially symmetric adult body plan of the sea urchin is generated long after embryogenesis is complete, by a separate process occurring within imaginal tissues set aside in the larva. The single Hox gene complex of Strongylocentrotus purpuratus contains 10 genes, and expression of eight of these genes was measured by quantitative methods during both embryonic and larval developmental stages and also in adult tissues. Only two of these genes are used significantly during the entire process of embryogenesis per se, although all are copiously expressed during the stages when the adult body plan is forming in the imaginal rudiment. They are also all expressed in various combinations in adult tissues. Thus, development of a microscopic, free-living organism of bilaterian grade, the larva, does not appear to require expression of the Hox gene cluster as such, whereas development of the adult body plan does. These observations reflect on mechanisms by which bilaterian metazoans might have arisen in Precambrian evolution.

Differential Epitope Tagging of Actin in Transformed Drosophila Produces Distinct Effects on Myofibril Assembly and Function of the Indirect Flight Muscle

We have tested the impact of tags on the structure and function of indirect flight muscle (IFM)-specific Act88F actin by transforming mutant Drosophila melanogaster, which do not express endogenous actin in their IFMs, with tagged Act88F constructs. Epitope tagging is often the method of choice to monitor the fate of a protein when a specific antibody is not available. Studies addressing the functional significance of the closely related actin isoforms rely almost exclusively on tagged exogenous actin, because only few antibodies exist that can discriminate between isoforms. Thereby it is widely presumed that the tag does not significantly interfere with protein function. However, in most studies the tagged actin is expressed in a background of endogenous actin and, as a rule, represents only a minor fraction of the total actin. The Act88F gene encodes the only Drosophila actin isoform exclusively expressed in the highly ordered IFM. Null mutations in this gene do not affect viability, but phenotypic effects in transformants can be directly attributed to the transgene. Transgenic flies that express Act88F with either a 6x histidine tag or an 11-residue peptide derived from vesicular stomatitis virus G protein at the C terminus were flightless. Overall, the ultrastructure of the IFM resembled that of the Act88F null mutant, and only low amounts of C-terminally tagged actins were found. In contrast, expression of N-terminally tagged Act88F at amounts comparable with that of wild-type flies yielded fairly normal-looking myofibrils and partially reconstituted flight ability in the transformants. Our findings suggest that the N terminus of actin is less sensitive to modifications than the C terminus, because it can be tagged and still polymerize into functional thin filaments.

The Role of Glucosidase I (Cwh41p) in the Biosynthesis of Cell Wall β-1,6-Glucan Is Indirect

CWH41, a gene involved in the assembly of cell wall β-1,6-glucan, has recently been shown to be the structural gene for Saccharomyces cerevisiae glucosidase I that is responsible for initiating the trimming of terminal α-1,2-glucose residue in the N-glycan processing pathway. To distinguish between a direct or indirect role of Cwh41p in the biosynthesis of β-1,6-glucan, we constructed a double mutant, alg5Δ (lacking dolichol-P-glucose synthase) cwh41Δ, and found that it has the same phenotype as the alg5Δ single mutant. It contains wild-type levels of cell wall β-1,6-glucan, shows moderate underglycosylation of N-linked glycoproteins, and grows at concentrations of Calcofluor White (which interferes with cell wall assembly) that are lethal to cwh41Δ single mutant. The strong genetic interactions of CWH41 with KRE6 and KRE1, two other genes involved in the β-1,6-glucan biosynthetic pathway, disappear in the absence of dolichol-P-glucose synthase (alg5Δ). The triple mutant alg5Δcwh41Δkre6Δ is viable, whereas the double mutant cwh41Δkre6Δ in the same genetic background is not. The severe slow growth phenotype and 75% reduction in cell wall β-1,6-glucan, characteristic of the cwh41Δkre1Δ double mutant, are not observed in the triple mutant alg5Δcwh41Δkre1Δ. Kre6p, a putative Golgi glucan synthase, is unstable in cwh41Δ strains, and its overexpression renders these cells Calcofluor White resistant. These results demonstrate that the role of glucosidase I (Cwh41p) in the biosynthesis of cell wall β-1,6-glucan is indirect and that dolichol-P-glucose is not an intermediate in this pathway.

Climate and infectious disease: Use of remote sensing for detection of Vibrio cholerae by indirect measurement

It has long been known that cholera outbreaks can be initiated when Vibrio cholerae, the bacterium that causes cholera, is present in drinking water in sufficient numbers to constitute an infective dose, if ingested by humans. Outbreaks associated with drinking or bathing in unpurified river or brackish water may directly or indirectly depend on such conditions as water temperature, nutrient concentration, and plankton production that may be favorable for growth and reproduction of the bacterium. Although these environmental parameters have routinely been measured by using water samples collected aboard research ships, the available data sets are sparse and infrequent. Furthermore, shipboard data acquisition is both expensive and time-consuming. Interpolation to regional scales can also be problematic. Although the bacterium, V. cholerae, cannot be sensed directly, remotely sensed data can be used to infer its presence. In the study reported here, satellite data were used to monitor the timing and spread of cholera. Public domain remote sensing data for the Bay of Bengal were compared directly with cholera case data collected in Bangladesh from 1992–1995. The remote sensing data included sea surface temperature and sea surface height. It was discovered that sea surface temperature shows an annual cycle similar to the cholera case data. Sea surface height may be an indicator of incursion of plankton-laden water inland, e.g., tidal rivers, because it was also found to be correlated with cholera outbreaks. The extensive studies accomplished during the past 25 years, confirming the hypothesis that V. cholerae is autochthonous to the aquatic environment and is a commensal of zooplankton, i.e., copepods, when combined with the findings of the satellite data analyses, provide strong evidence that cholera epidemics are climate-linked.

Indirect recognition of allopeptides promotes the development of cardiac allograft vasculopathy

Graft loss from chronic rejection has become the major obstacle to the long-term success of whole organ transplantation. In cardiac allografts, chronic rejection is manifested as a diffuse and accelerated form of arteriosclerosis, termed cardiac allograft vasculopathy. It has been suggested that T-cell recognition of processed alloantigens (allopeptides) presented by recipient antigen-presenting cells through the indirect pathway of allorecognition plays a critical role in the development and progression of chronic rejection. However, definitive preclinical evidence to support this hypothesis is lacking. To examine the role of indirect allorecognition in a clinically relevant large animal model of cardiac allograft vasculopathy, we immunized MHC inbred miniature swine with synthetic polymorphic peptides spanning the α1 domain of an allogeneic donor-derived swine leukocyte antigen class I gene. Pigs immunized with swine leukocyte antigen class I allopeptides showed in vitro proliferative responses and in vivo delayed-type hypersensitivity responses to the allogeneic peptides. Donor MHC class I disparate hearts transplanted into peptide-immunized cyclosporine-treated pigs not only rejected faster than unimmunized cyclosporine-treated controls (mean survival time = 5.5 +/−1.7 vs. 54.7 +/−3.8 days, P < 0.001), but they also developed obstructive fibroproliferative coronary artery lesions much earlier than unimmunized controls (<9 vs. >30 days). These results definitively link indirect allorecognition and cardiac allograft vasculopathy.

The contribution of trait-mediated indirect effects to the net effects of a predator

Many prey modify traits in response to predation risk and this modification of traits can influence the prey's resource acquisition rate. A predator thus can have a “nonlethal” impact on prey that can lead to indirect effects on other community members. Such indirect interactions are termed trait-mediated indirect interactions because they arise from a predator's influence on prey traits, rather than prey density. Because such nonlethal predator effects are immediate, can influence the entire prey population, and can occur over the entire prey lifetime, we argue that nonlethal predator effects are likely to contribute strongly to the net indirect effects of predators (i.e., nonlethal effects may be comparable in magnitude to those resulting from killing prey). This prediction was supported by an experiment in which the indirect effects of a larval dragonfly (Anax sp.) predator on large bullfrog tadpoles (Rana catesbeiana), through nonlethal effects on competing small bullfrog tadpoles, were large relative to indirect effects caused by density reduction of the small tadpoles (the lethal effect). Treatments in which lethal and nonlethal effects of Anax were manipulated independently indicated that this result was robust for a large range of different combinations of lethal and nonlethal effects. Because many, if not most, prey modify traits in response to predators, our results suggest that the magnitude of interaction coefficients between two species may often be dynamically related to changes in other community members, and that many indirect effects previously attributed to the lethal effects of predators may instead be due to shifts in traits of surviving prey.

IA-2, a transmembrane protein of the protein tyrosine phosphatase family, is a major autoantigen in insulin-dependent diabetes mellitus.

IA-2 is a 105,847 Da transmembrane protein that belongs to the protein tyrosine phosphatase family. Immunoperoxidase staining with antibody raised against IA-2 showed that this protein is expressed in human pancreatic islet cells. In this study, we expressed the full-length cDNA clone of IA-2 in a rabbit reticulocyte transcription/translation system and used the recombinant radiolabeled IA-2 protein to detect autoantibodies by immunoprecipitation. Coded sera (100) were tested: 50 from patients with newly diagnosed insulin-dependent diabetes mellitus (IDDM) and 50 from age-matched normal controls. Sixty-six percent of the sera from patients, but none of the sera from controls, reacted with IA-2. The same diabetic sera tested for autoantibodies to islet cells (ICA) by indirect immunofluorescence and glutamic acid decarboxylase (GAD65Ab) by depletion ELISA showed 68% and 52% positivity, respectively. Up to 86% of the IDDM patients had autoantibodies to IA-2 and/or GAD65. Moreover, greater than 90% (14 of 15) of the ICA-positive but GAD65Ab-negative sera had autoantibodies to IA-2. Absorption experiments showed that the immunofluorescence reactivity of ICA-positive sera was greatly reduced by prior incubation with recombinant IA-2 or GAD65 when the respective antibody was present. A little over one-half (9 of 16) of the IDDM sera that were negative for ICA were found to be positive for autoantibodies to IA-2 and/or GAD65, arguing that the immunofluorescence test for ICA is less sensitive than the recombinant tests for autoantibodies to IA-2 and GAD65. It is concluded that IA-2 is a major islet cell autoantigen in IDDM, and, together with GAD65, is responsible for much of the reactivity of ICA with pancreatic islets. Tests for the detection of autoantibodies to recombinant IA-2 and GAD65 may eventually replace ICA immunofluorescence for IDDM population screening.