Coxiella burnetii Phagocytosis Is Regulated by GTPases of the Rho Family and the RhoA Effectors mDia1 and ROCK

The GTPases belonging to the Rho family control the actin cytoskeleton rearrangements needed for particle internalization during phagocytosis. ROCK and mDia1 are downstream effectors of RhoA, a GTPase involved in that process. Coxiella burnetii, the etiologic agent of Q fever, is internalized by the host´s cells in an actin-dependent manner. Nevertheless, the molecular mechanism involved in this process has been poorly characterized. This work analyzes the role of different GTPases of the Rho family and some downstream effectors in the internalization of C. burnetii by phagocytic and non-phagocytic cells. The internalization of C. burnetii into HeLa and RAW cells was significantly inhibited when the cells were treated with Clostridium difficile Toxin B which irreversibly inactivates members of the Rho family. In addition, the internalization was reduced in HeLa cells that overexpressed the dominant negative mutants of RhoA, Rac1 or Cdc42 or that were knocked down for the Rho GTPases. The pharmacological inhibition or the knocking down of ROCK diminished bacterium internalization. Moreover, C. burnetii was less efficiently internalized in HeLa cells overexpressing mDia1-N1, a dominant negative mutant of mDia1, while the overexpression of the constitutively active mutant mDia1-ΔN3 increased bacteria uptake. Interestingly, when HeLa and RAW cells were infected, RhoA, Rac1 and mDia1 were recruited to membrane cell fractions. Our results suggest that the GTPases of the Rho family play an important role in C. burnetii phagocytosis in both HeLa and RAW cells. Additionally, we present evidence that ROCK and mDia1, which are downstream effectors of RhoA, are involved in that processFil: Salinas Ojeda, Romina Paola. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Cienicas Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; ArgentinaFil: Ortiz Flores, Rodolfo Matias. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Cienicas Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; ArgentinaFil: Distel, Jesús Sebastián. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Cienicas Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; ArgentinaFil: Aguilera, Milton Osmar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Cienicas Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; ArgentinaFil: Colombo, Maria Isabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Cienicas Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; ArgentinaFil: Beron, Walter. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Cienicas Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentin

Getting Down to Specifics: Profiling Gene Expression and Protein-DNA Interactions in a Cell Type-Specific Manner.

The majority of multicellular organisms are comprised of an extraordinary range of cell types, with different properties and gene expression profiles. Understanding what makes each cell type unique, and how their individual characteristics are attributed, are key questions for both developmental and neurobiologists alike. The brain is an excellent example of the cellular diversity expressed in the majority of eukaryotes. The mouse brain comprises of approximately 75 million neurons varying in morphology, electrophysiology, and preferences for synaptic partners. A powerful process in beginning to pick apart the mechanisms that specify individual characteristics of the cell, as well as their fate, is to profile gene expression patterns, chromatin states, and transcriptional networks in a cell type-specific manner, i.e. only profiling the cells of interest in a particular tissue. Depending on the organism, the questions being investigated, and the material available, certain cell type-specific profiling methods are more suitable than others. This chapter reviews the approaches presently available for selecting and isolating specific cell types and evaluates their key features

Pictografías en Coctaca (Quebrada de Humahuaca, Jujuy, Argentina)

El sitio arqueológico de Coctaca (Quebrada de Humahuaca, Provincia de Jujuy, Argentina) es bien conocido en la literatura científica por la presencia en el mismo de extensos andenes para el cultivo. Como así de silos subterráneos y un disperso poblado que indican la dedicación totalmente agrícola de sus pobladores. Por los estilos cerámicos que se han hallado puede ubicárselo entre los años 1300-1400 de nuestra era. En este artículo se considera el reciente hallazgo en Coctaca de un paredón con pictografías (frescos rupestres ) que por sus motivos realizados en blanco, rojo y negro, permite deducir la intensa dedicación a la ganadería y conocimiento de un desarrollado arte textil. Lo último en razón de las guardas representadas

The challenge of rangeland degradation in a temperate semiarid region of Argentina: the Caldenal

The phytogeographical region known as the Caldenal comprises an area of approximately 10 million ha in the central temperate part of Argentina (35–40° S, 62–66° W). The original physiognomy of the vegetation consisted of a gramineous steppe with isolated shrubs and trees. Rangeland deterioration was initiated in the early-1900s with the introduction of livestock by the colonizing European ranchers. After a few decades of inappropriate use, the Caldenal proved to be a fragile environment. Major environmental impacts were: (a) The replacement of the most abundant palatable grasses Poa ligularis, Stipa clarazii, S. tenuis, Piptochaetium napostaense and Digitaria californica by low nutritive unpalatable grasses such as Stipa gynerioides, S. tenuissima, S. ambigua, S. bracychaeta and Elyonurus muticus; (b) The conversion of extensive areas dominated by grasses to a scrubland. There, frequent species are: Condalia microphylla, Lycium chilense, Prosopis alpataco, Larrea divaricata and Chuquiraga erinacea; (c) Depletion of plant cover resulting in incipient but increasing soil erosion in extensive areas. With the purpose of reversing this situation, the remit initiated 25 years ago was to develop management guidelines conducive to sustainable productivity while preserving the natural resources of the region. This work provides a comprehensive assessment of the impact of major stresses (drought, herbivory, fire) on grass and shrub responses, as well as more basic studies on the ecophysiology of these species. We have gained knowledge which is extremely useful to initiate rangeland utilization based on scientific information, facilitating increased sustainable rangeland productivity while preserving the natural resources.Fil: Fernandez, Osvaldo Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Centro de Recursos Naturales Renovables de la Zona Semiárida. Universidad Nacional del Sur. Centro de Recursos Naturales Renovables de la Zona Semiárida; ArgentinaFil: Gil, María Elena. Universidad Nacional del Sur. Departamento de Agronomía; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Distel, Roberto Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Centro de Recursos Naturales Renovables de la Zona Semiárida. Universidad Nacional del Sur. Centro de Recursos Naturales Renovables de la Zona Semiárida; Argentin

The physiological consequences of ingesting a toxic plant (Diplotaxis tenuifolia) influence subsequent foraging decisions by sheep (Ovis aries)

Toxins and nutrients interact and define herbivores' experiences with toxic plants. However, there are still open questions about the mechanisms by which nutrient-toxin interactions affect experience and as a consequence foraging decisions by consumers. This study provides a deeper insight into such mechanisms by using supplemental nutrients, a toxic plant typically avoided by herbivores (wild rocket; Diplotaxis tenuifolia), and a small ruminant (sheep; Ovis aries) as models. Thirty-six sheep were randomly assigned to four treatments (n = 9) where animals consumed: wild rocket (“DT”), wild rocket followed by a protein supplement (“DT + P”), wild rocket followed by a protein supplement + a mineral supplement containing iodine and copper (“DT + P + M”), or alfalfa pellets in amounts that paired the ingestion of wild rocket by DT (“CTRL”). Towards the end of the phase of exposure (day 35), DT showed the lowest intake of wild rocket, as well as reduced levels of plasma thyroid hormones (T3 and T4), alanine aminotransferase, and a trend towards reduced hemoglobin relative to DT + P and DT + P + M. Total concentration of serum proteins and albumins were greater in sheep fed the protein supplements, which have probably elicited a protective effect on toxin ingestion. Foraging behavior was then evaluated in an experimental arena where animals could select among randomly distributed buckets containing a fixed amount of wild rocket or variable amounts of barley grain (a preferred food). Regardless of barley grain availability, DT showed lower intake and lower times spent eating wild rocket than DT + P and DT + P + M. Unexpectedly, CTRL (without previous experience with wild rocket) ingested amounts of wild rocket comparable to those observed by DT + P and DT + P + M. A negative feeding experience with wild rocket is needed for animals to display the typical pattern of aversion commonly observed in grazing conditions.Fil: Catanese, Francisco Hernan. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Centro de Recursos Naturales Renovables de la Zona Semiarida. Universidad Nacional del Sur. Centro de Recursos Naturales Renovables de la Zona Semiarida; ArgentinaFil: Fernandez, Paola Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Centro de Recursos Naturales Renovables de la Zona Semiarida. Universidad Nacional del Sur. Centro de Recursos Naturales Renovables de la Zona Semiarida; ArgentinaFil: Villalba, Juan José. University of Utah; Estados UnidosFil: Distel, Roberto Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Centro de Recursos Naturales Renovables de la Zona Semiarida. Universidad Nacional del Sur. Centro de Recursos Naturales Renovables de la Zona Semiarida; Argentin

How the foraging decisions of a small ruminant are influenced by past feeding experiences with low-quality food

Feeding experiences with low-quality foods can be improved when these foods are ingested in close temporal association with foods of higher nutritional quality. However, preference for low-quality foods in nature seems to be rather insensitive to past positive experiences and more related to their intrinsic nutritional value. An explanation for this observation is still lacking, mainly because little is known about how herbivores use information about low-quality foods during foraging. Our objective was to provide original information about this issue using a small ruminant (sheep; Ovis aries) as animal model. We manipulated the sheep’s experience with a low-quality food (wheat straw) using a conditioning procedure (“oral-delay conditioning procedure”), and then we evaluated the use of this information in a simulated foraging scenario provided with wheat straw and a variable amount of a high-quality food in spatially separated feeding stations. Inclusion of wheat straw into the diet was strongly dependent on the availability of the high-quality food. We observed a threshold level in the availability of the high-quality food, which defined a zone of drastic change in the likelihood of inclusion of the wheat straw into the diet (i.e., acceptance or rejection of wheat straw). This threshold level did not change for sheep with (CS+) or without (CS-) a previous positive experience with wheat straw. However, once foraging conditions stimulated all sheep to start including the wheat straw into the diet (i.e., below the threshold level), the intake of this food was greater by CS+ sheep. This increased intake was not explained by a higher motivation to eat the wheat straw but to a greater amount of time spent foraging this food and less time spent searching for the preferred higher-quality alternative. We discuss these results based on optimal foraging models and learning models of diet selection.Fil: Catanese, Francisco Hernan. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Centro de Recursos Naturales Renovables de la Zona Semiárida. Universidad Nacional del Sur. Centro de Recursos Naturales Renovables de la Zona Semiárida; ArgentinaFil: Distel, Roberto Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Centro de Recursos Naturales Renovables de la Zona Semiárida. Universidad Nacional del Sur. Centro de Recursos Naturales Renovables de la Zona Semiárida; ArgentinaFil: Fernandez, Paola Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Centro de Recursos Naturales Renovables de la Zona Semiárida. Universidad Nacional del Sur. Centro de Recursos Naturales Renovables de la Zona Semiárida; ArgentinaFil: Villalba, Juan Jose. University of Utah; Estados Unido