Different populations of RNA polymerase II in living mammalian cells

RNA polymerase II is responsible for transcription of most eukaryotic genes, but, despite exhaustive analysis, little is known about how it transcribes natural templates in vivo. We studied polymerase dynamics in living Chinese hamster ovary cells using an established line that expresses the largest (catalytic) subunit of the polymerase (RPB1) tagged with the green fluorescent protein (GFP). Genetic complementation has shown this tagged polymerase to be fully functional. Fluorescence loss in photobleaching (FLIP) reveals the existence of at least three kinetic populations of tagged polymerase: a large rapidly-exchanging population, a small fraction resistant to 5,6-dichloro-1-β-D-ribofuranosylbenzimidazole (DRB) but sensitive to a different inhibitor of transcription (i.e. heat shock), and a third fraction sensitive to both inhibitors. Quantitative immunoblotting shows the largest fraction to be the inactive hypophosphorylated form of the polymerase (i.e. IIA). Results are consistent with the second (DRB-insensitive but heat-shock-sensitive) fraction being bound but not engaged, while the third (sensitive to both DRB and heat shock) is the elongating hyperphosphorylated form (i.e. IIO)

Connecting Variability in Global Transcription Rate to Mitochondrial Variability

The authors demonstrate a connection between variability in the rate of transcription and differences in cellular mitochondrial content

Heteropolycompounds as catalysts for biomass product transformations

[EN] In the present review we show a variety of biomass product transformations through catalysis by both bulk and supported heteropolycompounds. The biomass sources considered include carbohydrates, oils and fats, and terpenes as main starting material groups. The products obtained and their applications are presented.We thank CONICET (PIP 003), Agencia Nacional de Promocion Cientifica y Tecnologica (Argentina) (PICT 0406), and Universidad Nacional de La Plata for financial support. GPR and HJT are members of CONICET. MJC and SI thank to Spanish Government-MINECO through Consolider Ingenio 2010-Multicat project for financial support.Sanchez, LM.; Thomas, HJ.; Climent Olmedo, MJ.; Romanelli, GP.; Iborra Chornet, S. (2016). Heteropolycompounds as catalysts for biomass product transformations. Catalysis Reviews: Science and Engineering. 58(4):497-586. doi:10.1080/01614940.2016.1248721S49758658

Spectral characterization of laser-accelerated protons with CR-39 nuclear track detector

CR-39 nuclear track material is frequently used for the detection of protons accelerated in laser-plasma interactions. The measurement of track densities allows for determination of particle angular distributions, and information on the kinetic energy can be obtained by the use of passive absorbers. We present a precise method of measuring spectral distributions of laser-accelerated protons in a single etching and analysis process. We make use of a one-to-one relation between proton energy and track size and present a precise calibration based on monoenergetic particle beams. While this relation is limited to proton energies below 1 MeV, we show that the range of spectral measurements can be significantly extended by simultaneous use of absorbers of suitable thicknesses. Examples from laser-plasma interactions are presented, and quantitative results on proton energies and particle numbers are compared to those obtained from a time-of-flight detector. The spectrum end points of continuous energy distributions have been determined with both detector types and coincide within 50-100 keV

Compuestos y reacciones quimicas naturales como motivadores del aprendizaje

Uno de los componentes importantes en el proceso de aprendizaje, se relaciona con la fuerte motivación intrínseca que el estudiante muestra hacia los contenidos que debe incorporar. Mas allá de los intereses propios de los alumnos es posible aportar, desde los materiales didácticos con los cuales se trabaja en el aula, información, imágenes, e inquietudes que, al relacionarse con temas aplicados de su entorno natural conduzcan a un aumento en la atención, como modo de mejorar la comprensión, retención y posterior uso activo de los conocimientos adquiridos (Teoría Uno, Perkins, 1995). En esta propuesta el aprendizaje de la escritura de fórmulas y ecuaciones químicas, con todos sus componentes asociados se presenta como material didáctico que describe compuestos y reacciones químicas observables en el entorno natural circundante del alumno

Compuestos y reacciones quimicas naturales como motivadores del aprendizaje

Uno de los componentes importantes en el proceso de aprendizaje, se relaciona con la fuerte motivación intrínseca que el estudiante muestra hacia los contenidos que debe incorporar. Mas allá de los intereses propios de los alumnos es posible aportar, desde los materiales didácticos con los cuales se trabaja en el aula, información, imágenes, e inquietudes que, al relacionarse con temas aplicados de su entorno natural conduzcan a un aumento en la atención, como modo de mejorar la comprensión, retención y posterior uso activo de los conocimientos adquiridos (Teoría Uno, Perkins, 1995). En esta propuesta el aprendizaje de la escritura de fórmulas y ecuaciones químicas, con todos sus componentes asociados se presenta como material didáctico que describe compuestos y reacciones químicas observables en el entorno natural circundante del alumno

Calibration and performance tests of detectors for laser-accelerated protons

We present the calibration and performance tests carried out with two detectors for intense proton pulses accelerated by lasers. Most of the procedures were realized with proton beams of 0.46-5.60 MeV from a tandem accelerator. One approach made use of radiochromic films, for which we calibrated the relation between optical density and energy deposition over more than three orders of magnitude. The validity of these results and of our analysis algorithms has been confirmed by controlled irradiation of film stacks and reconstruction of the total beam charge for strongly non-uniform beam profiles. For the spectral analysis of protons from repeated laser shots, we have designed an online monitor based on a plastic scintillator. The resulting signal from a photomultiplier directly measured on a fast oscilloscope is especially useful for time-of-flight applications. Variable optical filters allow for suppression of saturation and an extension of the dynamic range. With pulsed proton beams we have tested the detector response to a wide range of beam intensities from single particles to 3 ×105 protons per 100 ns time interval.Project funded by the Spanish Ministry of Economy and Competitiveness and co-funded with FEDER's funds within the INNPACTO 2011 program under Grant No. IPT-2011-0862-900000. This work was supported by the Spanish Plan Nacional de Investigacion Cientifica, Desarrollo e Innovacion Tecnologica (I+D+i) under Grant No. TEC 2013-48036-C3-1-R and the Valencian Local Government under Grants PROMETEOII/2013/010 and ISIC 2011/013. The work of A. J. Gonzalez is financed by CSIC with a JAE-Doc contract under Junta de Ampliacion de Estudios program, cofinanced by the European Social Fund.Peer Reviewe

Coadministration of the Three Antigenic Leishmania infantum Poly (A) Binding Proteins as a DNA Vaccine Induces Protection against Leishmania major Infection in BALB/c Mice

Highly conserved intracellular proteins from Leishmania have been described as antigens in natural and experimental infected mammals. The present study aimed to evaluate the antigenicity and prophylactic properties of the Leishmania infantum Poly (A) binding proteins (LiPABPs). Three different members of the LiPABP family have been described. Recombinant tools based on these proteins were constructed: recombinant proteins and DNA vaccines. The three recombinant proteins were employed for coating ELISA plates. Sera from human and canine patients of visceral leishmaniasis and human patients of mucosal leishmaniasis recognized the three LiPABPs. In addition, the protective efficacy of a DNA vaccine based on the combination of the three Leishmania PABPs has been tested in a model of progressive murine leishmaniasis: BALB/c mice infected with Leishmania major. The induction of a Th1-like response against the LiPABP family by genetic vaccination was able to down-regulate the IL-10 predominant responses elicited by parasite LiPABPs after infection in this murine model. This modulation resulted in a partial protection against L. major infection. LiPABP vaccinated mice showed a reduction on the pathology that was accompanied by a decrease in parasite burdens, in antibody titers against Leishmania antigens and in the IL-4 and IL-10 parasite-specific mediated responses in comparison to control mice groups immunized with saline or with the non-recombinant plasmid. The results presented here demonstrate for the first time the prophylactic properties of a new family of Leishmania antigenic intracellular proteins, the LiPABPs. The redirection of the immune response elicited against the LiPABP family (from IL-10 towards IFN-γ mediated responses) by genetic vaccination was able to induce a partial protection against the development of the disease in a highly susceptible murine model of leishmaniasisThe study was supported in Spain by grants from Ministerio de Ciencia e Innovación FIS PI11/00095 and FISPI14/00366 from the Instituto de Salud Carlos III within the Network of TropicalDiseases Research (VI P I+D+I 2008-2011, ISCIII -Subdirección General de Redes y Centros de Investigación Cooperativa (RD12/0018/0009)). This work was also supported in Brazil by a grant from CNPq (Ciencia sem Fronteiras-PVE 300174/2014-4). A CBMSO institutional grant from Fundación Ramón Areces is also acknowledged. EAFC is a grant recipient of CNPq. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscrip