Subtelomeric I-scei-mediated Double-strand Breaks Are Repaired By Homologous Recombination In Trypanosoma Cruzi

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Trypanosoma cruzi chromosome ends are enriched in surface protein genes and pseudogenes (e.g., trans-sialidases) surrounded by repetitive sequences. It has been proposed that the extensive sequence variability among members of these protein families could play a role in parasite infectivity and evasion of host immune response. In previous reports we showed evidence suggesting that sequences located in these regions are subjected to recombination. To support this hypothesis we introduced a double-strand break (DSB) at a specific target site in a T. cruzi subtelomeric region cloned into an artificial chromosome (pTAC). This construct was used to transfect T. cruzi epimastigotes expressing the I-SceI meganuclease. Examination of the repaired sequences showed that DNA repair occurred only through homologous recombination (HR) with endogenous subtelomeric sequences. Our findings suggest that DSBs in subtelomeric repetitive sequences followed by HR between them may contribute to increased variability in T. cruzi multigene families. © 2016 Chiurillo, Moraes Barros, Souza, Marini, Antonio, Cortez, Curto, Lorenzi, Schijman, Ramirez and da Silveira.7DEC11/51475-3, FAPESP, Fundação de Amparo à Pesquisa do Estado de São Paulo11/51693-0, FAPESP, Fundação de Amparo à Pesquisa do Estado de São Paulo306591/2015-4, CNPq, Conselho Nacional de Desenvolvimento Científico e TecnológicoFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Antihydrogen formation dynamics in a multipolar neutral anti-atom trap

Antihydrogen production in a neutral atom trap formed by an octupole-based magnetic field minimum is demonstrated using field-ionization of weakly bound anti-atoms. Using our unique annihilation imaging detector, we correlate antihydrogen detection by imaging and by field-ionization for the first time. We further establish how field-ionization causes radial redistribution of the antiprotons during antihydrogen formation and use this effect for the first simultaneous measurements of strongly and weakly bound antihydrogen atoms. Distinguishing between these provides critical information needed in the process of optimizing for trappable antihydrogen. These observations are of crucial importance to the ultimate goal of performing CPT tests involving antihydrogen, which likely depends upon trapping the anti-atom

Search For Trapped Antihydrogen

We present the results of an experiment to search for trapped antihydrogen atoms with the ALPHA antihydrogen trap at the CERN Antiproton Decelerator. Sensitive diagnostics of the temperatures, sizes, and densities of the trapped antiproton and positron plasmas have been developed, which in turn permitted development of techniques to precisely and reproducibly control the initial experimental parameters. The use of a position-sensitive annihilation vertex detector, together with the capability of controllably quenching the superconducting magnetic minimum trap, enabled us to carry out a high-sensitivity and low-background search for trapped synthesised antihydrogen atoms. We aim to identify the annihilations of antihydrogen atoms held for at least 130 ms in the trap before being released over ~30 ms. After a three-week experimental run in 2009 involving mixing of 10^7 antiprotons with 1.3 10^9 positrons to produce 6 10^5 antihydrogen atoms, we have identified six antiproton annihilation events that are consistent with the release of trapped antihydrogen. The cosmic ray background, estimated to contribute 0.14 counts, is incompatible with this observation at a significance of 5.6 sigma. Extensive simulations predict that an alternative source of annihilations, the escape of mirror-trapped antiprotons, is highly unlikely, though this possibility has not yet been ruled out experimentally.Comment: 12 pages, 7 figure

Jatropha curcasand Ricinus communisdisplay contrasting photosynthetic mechanisms in response to environmental conditions

Higher plants display different adaptive strategies in photosynthesis to cope with abiotic stress. In this study, photosynthetic mechanisms and water relationships displayed byJatropha curcasL. (physic nuts) andRicinus communisL. (castor bean), in response to variations in environmental conditions, were assessed.R. communis showed higher CO2 assimilation, stomatal and mesophyll conductance thanJ. curcas as light intensity and intercellular CO2 pressure increased. On the other hand,R. communis was less effective in stomatal control in response to adverse environmental factors such as high temperature, water deficit and vapor pressure deficit, indicating lower water use efficiency. Conversely,J. curcas exhibited higher photosynthetic efficiency (gas exchange and photochemistry) and water use efficiency under these adverse environmental conditions.R. communisdisplayed higher potential photosynthesis, but exhibited a lowerin vivo Rubisco carboxylation rate (Vcmax) and maximum electron transport rate (Jmax). During the course of a typical day, in a semiarid environment, with high irradiation, high temperature and high vapor pressure deficit, but exposed to well-watered conditions, the two studied species presented similar photosynthesis. Losing potential photosynthesis, but maintaining favorable water status and increasing non-photochemical quenching to avoid photoinhibition, are important acclimation mechanisms developed byJ. curcas to cope with dry and hot conditions. We suggest thatJ. curcas is more tolerant to hot and dry environments thanR. communis but the latter species displays higher photosynthetic efficiency under well-watered and non-stressful conditions

Salt stress induced damages on the photosynthesis of physic nut young plants

Salinity is a major limiting factor to crop productivity in the world especially in semiarid regions. The aim of this study was to evaluate the photosynthetic resistance of Jatropha curcas (L.) young plants subjected to salt stress. The experiment was carried out in a completely randomized design with treatments in a 2 x 3 factorial: two NaCl levels (0 and 100 mmol L-1) and three harvest times: 7 and 14 days of salt exposure and three days of recovery. Leaf Na+ and Cl-concentrations and the K+/Na+ ratios, after seven days of salt exposure, did not reach ionic toxic levels, suggesting that the NaCl-induced osmotic effects prevailed over the ionic ones. Under this condition, the salt stress caused reduction in leaf gas exchange parameters, such as CO2 fixation, stomatal conductance and transpiration. In contrast, salt stress did not change the photochemical efficiency of photossystem II. Conversely, after 14 days of treatment, Na+ and Clions reached very high concentrations, up to toxic levels in leaves. Under such conditions, both leaf gas exchange and photochemistry suffered strong impairment probably caused by ionic toxicity. The recovery treatment for 3d did not significantly decrease the leaf salt concentrations and no improvement was observed in the photosynthetic performance. Physic nut young plants are sensitive to high NaCl-salinity conditions, with high leaf Na+ and Cl- concentrations, low K+/Na+ ratio and great photosynthetic damages due to stomatal and biochemical limitations.A salinidade é um dos principais fatores que limitam a produtividade das culturas no mundo principalmente em regiões semiáridas. Avaliou-se a resistência da fotossíntese de plantas jovens de pinhãomanso (Jatropha curcas L.) submetidas ao estresse salino. O experimento foi realizado em delineamento inteiramente casualizado com tratamentos em fatorial 2 x 3: duas concentrações de NaCl (0 e 100 mmol L-1) e três tempos de avaliação (7 e 14 dias de exposição e três dias de recuperação). As concentrações de Na+ e Cl- e a relação K+/Na+ nas folhas, após sete dias de exposição ao sal, não indicaram níveis tóxicos, sugerindo os efeitos osmóticos induzidos pelo NaCl prevaleceram sobre as causas iônicas. Sob essas condições, o estresse salino causou redução nos parâmetros de trocas gasosas, como fixação de CO2, condutância estomática e transpiração, mas ao contrário, não alterou a eficiência fotoquímica do fotossistema II. Após 14 dias de tratamento, os íons salinos atingiram concentrações muito elevadas nas folhas, provavelmente atingindo níveis tóxicos. Em tais condições, as trocas gasosas e a atividade fotoquímica sofreram forte redução causada pelo estresse iônico. O tratamento de recuperação não induziu queda intensa nas concentrações dos íons salinos nas folhas e nenhuma melhoria foi observada no desempenho fotossintético. Plantas jovens de pinhão manso são sensíveis a condições de salinidade elevada por NaCl, mostrando altas concentrações de Na+ e Cl-, baixa razão K+/Na+ e danos fotossintéticos intensos causados tanto por limitações estomáticas como por limitações bioquímicas