Dominant negative phenotype of Bacillus thuringiensis Cry1Ab, Cry11Aa and Cry4Ba mutants suggest hetero-oligomer formation among different Cry toxins.

Background - Bacillus thuringiensis Cry toxins are used worldwide in the control of different insect pests important in agriculture or in human health. The Cry proteins are pore-forming toxins that affect the midgut cell of target insects. It was shown that non-toxic Cry1Ab helix a-4 mutants had a dominant negative (DN) phenotype inhibiting the toxicity of wildtype Cry1Ab when used in equimolar or sub-stoichiometric ratios (1:1, 0.5:1, mutant:wt) indicating that oligomer formation is a key step in toxicity of Cry toxins. Methodology/Principal Findings - The DN Cry1Ab-D136N/T143D mutant that is able to block toxicity of Cry1Ab toxin, was used to analyze its capacity to block the activity against Manduca sexta larvae of other Cry1 toxins, such as Cry1Aa, Cry1Ac, Cry1Ca, Cry1Da, Cry1Ea and Cry1Fa. Cry1Ab-DN mutant inhibited toxicity of Cry1Aa, Cry1Ac and Cry1Fa. In addition, we isolated mutants in helix a-4 of Cry4Ba and Cry11Aa, and demonstrate that Cry4Ba-E159K and Cry11Aa-V142D are inactive and completely block the toxicity against Aedes aegypti of both wildtype toxins, when used at sub-stoichiometric ratios, confirming a DN phenotype. As controls we analyzed Cry1Ab-R99A or Cry11Aa-E97A mutants that are located in helix a-3 and are affected in toxin oligomerization. These mutants do not show a DN phenotype but were able to block toxicity when used in 10:1 or 100:1 ratios (mutant:wt) probably by competition of binding with toxin receptors. Conclusions/Significance - We show that DN phenotype can be observed among different Cry toxins suggesting that may interact in vivo forming hetero-oligomers. The DN phenotype cannot be observed in mutants affected in oligomerization, suggesting that this step is important to inhibit toxicity of other toxin

Politicas agrarias y estrategias campesinas en la Cuenca del Canete : anexos 8 a 13

Ce document réunit plusieurs diagnostics établis à partir d'observations de terrain sur la géographie physique, l'organisation sociale, les pratiques paysannes dans la vallée de Canete et la vallée Pampas afin de déterminer les facteurs limitants de la production et des pratiques culturales , notamment en matière d'irrigation

Examination of relaxin and its receptors expression in pig gametes and embryos

<p>Abstract</p> <p>Background</p> <p>Relaxin is a small peptide also known as pregnancy hormone in many mammals. It is synthesized by both male and female tissues, and its secretions are found in various body fluids such as plasma serum, ovarian follicular fluid, utero-oviduct secretions, and seminal plasma of many mammals, including pigs. However, the presence and effects of relaxin in porcine gametes and embryos are still not well-known. The purpose of this study was to assess the presence of relaxin and its receptors RXFP1 and RXFP2 in pig gametes and embryos.</p> <p>Methods</p> <p>Immature cumulus-oocyte complexes (COCs) were aspirated from sows' ovaries collected at the abattoir. After <it>in vitro</it>-maturation, COCs were <it>in vitro</it>-fertilized and cultured. For studies, immature and mature COCs were separately collected, and oocytes were freed from their surrounding cumulus cells. Denuded oocytes, cumulus cells, mature boar spermatozoa, zygotes, and embryos (cleaved and blastocysts) were harvested for temporal and spatial gene expression studies. Sections of ovary, granulosa and neonatal porcine uterine cells were also collected to use as controls.</p> <p>Results</p> <p>Using both semi-quantitative and quantitative PCRs, relaxin transcripts were not detected in all tested samples, while RXFP1 and RXFP2 mRNA were present. Both receptor gene products were found at higher levels in oocytes compared to cumulus cells, irrespective of the maturation time. Cleaved-embryos contained higher levels of RXFP2 mRNA, whereas, blastocysts were characterized by a higher RXFP1 mRNA content. Using western-immunoblotting or <it>in situ </it>immunofluorescence, relaxin and its receptor proteins were detected in all samples. Their fluorescence intensities were consistently more important in mature oocytes than immature ones. The RXFP1 and RXFP2 signal intensities were mostly located in the plasma membrane region, while the relaxin ones appeared homogeneously distributed within the oocytes and embryonic cells. Furthermore, spermatozoa displayed stronger RXFP2 signal than RXFP1 after western-immunoblotting.</p> <p>Conclusion</p> <p>All together, our findings suggest potential roles of relaxin and its receptors during oocyte maturation, early embryo development, and beyond.</p

Discovery of faint double-peak Halpha emission in the halo of low redshift galaxies

Aiming at the detection of cosmological gas being accreted onto galaxies of the local Universe, we examined the Halpha emission in the halo of 164 galaxies in the field of view of the Multi-Unit Spectroscopic Explorer Wide survey (\musew ) with observable Halpha (redshift < 0.42). An exhaustive screening of the corresponding Halpha images led us to select 118 reliable Halpha emitting gas clouds. The signals are faint, with a surface brightness of 10**(-17.3 pm 0.3) erg/s/cm2/arcsec2. Through statistical tests and other arguments, we ruled out that they are created by instrumental artifacts, telluric line residuals, or high redshift interlopers. Around 38% of the time, the Halpha line profile shows a double peak with the drop in intensity at the rest-frame of the central galaxy, and with a typical peak-to-peak separation of the order of pm 200 km/s. Most line emission clumps are spatially unresolved. The mass of emitting gas is estimated to be between one and 10**(-3) times the stellar mass of the central galaxy. The signals are not isotropically distributed; their azimuth tends to be aligned with the major axis of the corresponding galaxy. The distances to the central galaxies are not random either. The counts drop at a distance > 50 galaxy radii, which roughly corresponds to the virial radius of the central galaxy. We explore several physical scenarios to explain this Halpha emission, among which accretion disks around rogue intermediate mass black holes fit the observations best.Comment: pay attention to the last sentence of the abstract! Accepted for publication in Ap

A panchromatic spatially-resolved analysis of nearby galaxies -- I. Sub-kpc scale Main Sequence in grand-design spirals

We analyse the spatially resolved relation between stellar mass (M$_{\star}$) and star formation rate (SFR) in disk galaxies (i.e. the Main Sequence, MS). The studied sample includes eight nearby face-on grand-design spirals, e.g. the descendant of high-redshift, rotationally-supported star-forming galaxies. We exploit photometric information over 23 bands, from the UV to the far-IR, from the publicly available DustPedia database to build spatially resolved maps of stellar mass and star formation rates on sub-galactic scales of 0.5-1.5 kpc, by performing a spectral energy distribution fitting procedure that accounts for both the observed and the obscured star formation processes, over a wide range of internal galaxy environments (bulges, spiral arms, outskirts). With more than 30 thousands physical cells, we have derived a definition of the local spatially resolved MS per unit area for disks, $\log(\Sigma_{SFR})$=0.82log$(\Sigma_{*})$-8.69. This is consistent with the bulk of recent results based on optical IFU, using the H$\alpha$ line emission as a SFR tracer. Our work extends the analysis at lower sensitivities in both M$_{\star}$ and SFR surface densities, up to a factor $\sim$ 10. The self consistency of the MS relation over different spatial scales, from sub-galactic to galactic, as well as with a rescaled correlation obtained for high redshift galaxies, clearly proves its universality.Comment: 21 pages, 15 figures. Accepted for publication in MNRA

Gene therapy with Angiotensin-(1-9) preserves left ventricular systolic function after myocardial infarction

BACKGROUND: Angiotensin-(1-9) [Ang-(1-9)] is a novel peptide of the counter-regulatory axis of the renin angiotensin system previously demonstrated to have therapeutic potential in hypertensive cardiomyopathy when administered via osmotic minipump in mice. Here, we investigate whether gene transfer of Ang-(1-9) is cardioprotective in a murine model of myocardial infarction (MI). OBJECTIVES: To evaluate effects of Ang-(1-9) gene therapy on myocardial structural and functional remodeling post infarction. METHODS: C57BL/6 mice underwent permanent left anterior descending coronary artery ligation and cardiac function was assessed using echocardiography for 8 weeks followed by a terminal measurement of left ventricular (LV) pressure-volume loops. Ang-(1-9) was delivered by adeno-associated viral vector via single tail vein injection immediately following induction of MI. Direct effects of Ang-(1-9) on cardiomyocyte excitation–contraction coupling and cardiac contraction were evaluated in isolated mouse and human cardiomyocytes and in an ex vivo Langendorff perfused whole heart model. RESULTS: Gene delivery of Ang-(1-9) significantly reduced sudden cardiac death post-MI. Pressure–volume measurements revealed complete restoration of end systolic pressure, ejection fraction, end systolic volume and the end diastolic pressure–volume relationship by Ang-(1-9) treatment. Stroke volume and cardiac output were significantly increased versus sham. Histological analysis revealed only mild effects on cardiac hypertrophy and fibrosis, but a significant increase in scar thickness. Direct assessment of Ang-(1-9) on isolated cardiomyocytes demonstrated a positive inotropic effect via increasing calcium transient amplitude and increasing contractility. Ang-(1-9) increased contraction in the Langendorff model through a protein kinase A-dependent mechanism. CONCLUSIONS: Our novel findings show that Ang-(1-9) gene therapy preserves LV systolic function post-MI, restoring cardiac function. Furthermore, Ang-(1-9) has a direct effect on cardiomyocyte 3 calcium handling through a protein kinase A-dependent mechanism. These data highlight Ang-(1-9) gene therapy as a potential new strategy in the context of MI

Correlated optical, X-ray, and $-ray flaring activity seen with INTEGRAL during the 2015 outburst of V404 Cygni

Reproduced with permission from Astronomy & Astrophysics. © 2015 ESO.After 25 years of quiescence, the microquasar V404 Cyg entered a new period of activity in June 2015. This X-ray source is known to undergo extremely bright and variable outbursts seen at all wavelengths. It is therefore an object of prime interest to understand the accretion-ejection connections. These can, however, only be probed through simultaneous observations at several wavelengths. We made use of the INTEGRAL instruments to obtain long, almost uninterrupted observations from 2015 June 20th, 15:50 UTC to June 25th, 4:05 UTC, from the optical V-band, up to the soft γ-rays. V404 Cyg was extremely variable in all bands, with the detection of 18 flares with fluxes exceeding 6 Crab (20--40 keV) within 3 days. The flare recurrence can be as short as ∼ 20~min from peak to peak. A model-independent analysis shows that the >6 Crab flares have a hard spectrum. A simple 10--400 keV spectral analysis of the off-flare and flare periods shows that the variation in intensity is likely to be due to variations of a cut-off power law component only. The optical flares seem to be at least of two different types: one occurring in simultaneity with the X-ray flares, the other showing a delay greater than 10 min. The former could be associated with X-ray reprocessing by either an accretion disk or the companion star. We suggest that the latter are associated with plasma ejections that have also been seen in radio.Peer reviewe

MAXI J1659-152: the shortest orbital period black-hole binary

Following the detection of a bright new X-ray source, MAXI J1659-152, a series of observations was triggered with almost all currently flying high-energy missions. We report here on XMM-Newton, INTEGRAL and RXTE observations during the early phase of the X-ray outburst of this transient black-hole candidate. We confirm the dipping nature in the X-ray light curves. We find that the dips recur on a period of 2.4139+/-0.0005 hrs, and interpret this as the orbital period of the system. It is thus the shortest period black-hole X-ray binary known to date. Using the various observables, we derive the properties of the source. The inclination of the accretion disk with respect to the line of sight is estimated to be 60-75 degrees. The companion star to the black hole is possibly a M5 dwarf star, with a mass and radius of about 0.15 M_sun and 0.23 R_sun, respectively. The system is rather compact (orbital separation is about 1.35 R_sun) and is located at a distance of roughly 7 kpc. In quiescence, MAXI J1659-152 is expected to be optically faint, about 28 mag in the V-band.Comment: 5 pages, 4 figures, to be published in the proceedings of the 4th International MAXI Workshop `The First Year of MAXI: Monitoring variable X-ray sources', 2010 Nov 30 - Dec 2, Tokyo, Japa