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1.
Salomé C. Vilchez Larrea Mariana Schlesinger María L. Kevorkian Mirtha M. Flawiá Guillermo D. Alonso Silvia H. Fernández Villamil 《PloS one》2013,8(6)
Trypanosoma cruzi, etiological agent of Chagas’ disease, has a complex life cycle which involves the invasion of mammalian host cells, differentiation and intracellular replication. Here we report the first insights into the biological role of a poly(ADP-ribose) glycohydrolase in a trypanosomatid (TcPARG). In silico analysis of the TcPARG gene pointed out the conservation of key residues involved in the catalytic process and, by Western blot, we demonstrated that it is expressed in a life stage-dependant manner. Indirect immunofluorescence assays and electron microscopy using an anti-TcPARG antibody showed that this enzyme is localized in the nucleus independently of the presence of DNA damage or cell cycle stage. The addition of poly(ADP-ribose) glycohydrolase inhibitors ADP-HPD (adenosine diphosphate (hydroxymethyl) pyrrolidinediol) or DEA (6,9-diamino-2-ethoxyacridine lactate monohydrate) to the culture media, both at a 1 µM concentration, reduced in vitro epimastigote growth by 35% and 37% respectively, when compared to control cultures. We also showed that ADP-HPD 1 µM can lead to an alteration in the progression of the cell cycle in hydroxyurea synchronized cultures of T. cruzi epimastigotes. Outstandingly, here we demonstrate that the lack of poly(ADP-ribose) glycohydrolase activity in Vero and A549 host cells, achieved by chemical inhibition or iRNA, produces the reduction of the percentage of infected cells as well as the number of amastigotes per cell and trypomastigotes released, leading to a nearly complete abrogation of the infection process. We conclude that both, T. cruzi and the host, poly(ADP-ribose) glycohydrolase activities are important players in the life cycle of Trypanosoma cruzi, emerging as a promising therapeutic target for the treatment of Chagas’ disease. 相似文献
2.
Luis Molina y Vedia Mónica Torruella Ricardo Attar Ernesto Podesta Juan A. Reig Mirtha M. Flawia Héctor N. Torres 《Biochemical and biophysical research communications》1983,113(3):778-783
A monoclonal antibody against soluble adenylate cyclase was obtained. The antibody inhibits cyclase activities from several lower eucaryotic organisms but not activities associated to testicular cytosol or turkey erythrocyte membranes. 相似文献
3.
Phillips OL Baker TR Arroyo L Higuchi N Killeen TJ Laurance WF Lewis SL Lloyd J Malhi Y Monteagudo A Neill DA Vargas PN Silva JN Terborgh J Martínez RV Alexiades M Almeida S Brown S Chave J Comiskey JA Czimczik CI Di Fiore A Erwin T Kuebler C Laurance SG Nascimento HE Olivier J Palacios W Patiño S Pitman NC Quesada CA Saldias M Lezama AT Vinceti B 《Philosophical transactions of the Royal Society of London. Series B, Biological sciences》2004,359(1443):381-407
Previous work has shown that tree turnover, tree biomass and large liana densities have increased in mature tropical forest plots in the late twentieth century. These results point to a concerted shift in forest ecological processes that may already be having significant impacts on terrestrial carbon stocks, fluxes and biodiversity. However, the findings have proved controversial, partly because a rather limited number of permanent plots have been monitored for rather short periods. The aim of this paper is to characterize regional-scale patterns of 'tree turnover' (the rate with which trees die and recruit into a population) by using improved datasets now available for Amazonia that span the past 25 years. Specifically, we assess whether concerted changes in turnover are occurring, and if so whether they are general throughout the Amazon or restricted to one region or environmental zone. In addition, we ask whether they are driven by changes in recruitment, mortality or both. We find that: (i) trees 10 cm or more in diameter recruit and die twice as fast on the richer soils of southern and western Amazonia than on the poorer soils of eastern and central Amazonia; (ii) turnover rates have increased throughout Amazonia over the past two decades; (iii) mortality and recruitment rates have both increased significantly in every region and environmental zone, with the exception of mortality in eastern Amazonia; (iv) recruitment rates have consistently exceeded mortality rates; (v) absolute increases in recruitment and mortality rates are greatest in western Amazonian sites; and (vi) mortality appears to be lagging recruitment at regional scales. These spatial patterns and temporal trends are not caused by obvious artefacts in the data or the analyses. The trends cannot be directly driven by a mortality driver (such as increased drought or fragmentation-related death) because the biomass in these forests has simultaneously increased. Our findings therefore indicate that long-acting and widespread environmental changes are stimulating the growth and productivity of Amazon forests. 相似文献
4.
Pirpignani ML Rivera E Hellman U Biscoglio de Jiménez Bonino M 《Archives of biochemistry and biophysics》2002,407(2):224-230
Vespid venoms contain Antigen 5, an important allergen whose primary structure and immunological behavior have been extensively studied from venoms of vespids of the Northern Hemisphere. We report herein structural and immunological aspects of Antigen 5 from Polybia scutellaris subspecies rioplatensis (vulgar name: camoati) found in South America. Mast cell degranulation, histamine release, and IgE induction experiments performed in mice allow us to suggest that P. scutellaris Antigen 5 is a variant with reduced IgE response and anaphylactic activity. Sequence data indicate that the protein has a 72.5-90.3% similarity to that of members of the vespid Antigen 5 family with an already known primary structure. Moreover, results suggest that the protein-a new member of an extracellular protein superfamily-could be a good candidate for immunotherapy related to vespid allergy. 相似文献
5.
Pereira CA Alonso GD Ivaldi S Bouvier LA Torres HN Flawiá MM 《The Journal of eukaryotic microbiology》2003,50(2):132-134
Arginine kinase catalyzes the transphosphorylation between phosphoarginine and ADP. Phosphoarginine is involved in temporal ATP buffering and inorganic phosphate regulation. Trypanosoma cruzi arginine kinase phosphorylates only L-arginine (specific activity 398.9 x mUE-min(-1) x mg(-1)), and is inhibited by the arginine analogs, agmatine, canavanine, nitroarginine, and homoarginine. Canavanine and homoarginine also produce a significant inhibition of the epimastigote culture growth (79.7% and 55.8%, respectively). Inhibition constants were calculated for canavanine and homoarginine (7.55 and 6.02 mM, respectively). In addition, two novel guanidino kinase activities were detected in the epimastigote soluble extract. The development of the arginine kinase inhibitors of T. cruzi could be an important feature because the phosphagens biosynthetic pathway in trypanosomatids is different from the one in their mammalian hosts. 相似文献
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7.
Costa H Distéfano AJ Marino-Buslje C Hidalgo A Berenguer J Biscoglio de Jiménez Bonino M Ferrarotti SA 《Applied microbiology and biotechnology》2012,94(1):123-130
Cyclodextrin glycosyltransferases (CGTases) are important enzymes in biotechnology because of their ability to produce cyclodextrin
(CD) mixtures from starch whose relative composition depends on enzyme source. A multiple alignment of 46 CGTases and Shannon
entropy analysis allowed us to find differences and similarities that could be related to product specificity. Interestingly,
position 179 has Gly in all the CGTases except in that from Bacillus circulans DF 9R which possesses Gln. The absence of a side chain at that position has been considered as a strong requirement for substrate
binding and cyclization process. Therefore, we constructed two mutants of this enzyme, Q179L and Q179G. The activity and kinetic
parameters of Q179G remained unchanged while the Q179L mutant showed a different CDs ratio, a lower catalytic efficiency,
and a decreased ability to convert starch into CDs. We show that position 179 is involved in CGTase product specificity and
must be occupied by Gly—without a side chain—or by amino acid residues able to interact with the substrate through hydrogen
bonds in a way that the cyclization process occurs efficiently. These findings are also explained on the basis of a structural
model. 相似文献
8.
Stings by insects from the Hymenoptera order are known to cause life-threatening allergic reactions and impair life quality. Despite the effectiveness of conventional vespid venom immunotherapy, more standardized and safer allergy vaccines are required and recombinant hypoallergenic variants are important clinical tools. Antigen 5 is a major allergen of vespid venoms and it was previously reported that Antigen 5 from Polybia scutellaris (Poly s 5) could be a hypoallergenic variant. In this work we assess the immunological behavior and allergenic activity of Poly s 5 in order to explore its suitability for specific immunotherapy. With this aim, recombinant Poly s 5 was expressed in Pichia pastoris and the presence of cross-reactive epitopes with Pol a 5, a known allergenic Antigen 5, was investigated both at the IgG and IgE levels, by ELISA assays and a basophil-mediator release assay respectively. A molecular model was also built to better understand the relationship between immunological and structural aspects. In mice, Poly s 5 induced IgG antibodies which cross-reacted with Pol a 5. However, Poly s 5 induced only minimal amounts of IgE and was a poor inducer of basophil-mediator release, even when the cells were sensitized with Pol a 5-specific IgE. Moreover, Poly s 5-specific serum showed a specific protective activity and was able to inhibit the Pol a 5-induced basophil degranulation. Structural analysis from the molecular model revealed that a few amino acid substitutions in the N-terminal region of Poly s 5 should lead to an alteration of the surface topography and electrostatic potential of the epitopes which could be responsible for its hypoallergenic behavior. These findings, taken as a whole, show that Poly s 5 is likely a naturally occurring hypoallergenic Antigen 5 variant. 相似文献
9.
Mariano A. Ferrari Mirtha N. Lewis † Miguel A. Pascual † Claudio Campagna ‡ 《Marine Mammal Science》2009,25(3):681-692
Southern elephant seals breed at Península Valdés (PV, Argentina) along 200 km of coastline. Annual pup counts at peak breeding season for the entire colony increased from 12,113 in 1995 to 14,350 in 2006. Two demographic subunits were identified in the North and South of PV with different trends in births numbers, sex ratios and harem sizes. Birth numbers increased in the South, but decreased sharply in the North. To explain the trends in the colony and subunits, a population model was proposed that integrates social structure (harem size and sex ratio) in a fertility function that quantifies the effects of the social structure on the number of births. We found that a better fit to census data results from our model compared to a linear one (χ1 2 = 4.027, P = 0.045) . The model was then used to test alternative hypotheses about the role of recruitment and migration on the dynamic of the two subunits. Results indicated the relevance of considering social structure in population models of gregarious and polygynous species, and is an additional tool for comparative studies between populations of elephant seals where long term census are available. 相似文献
10.
Martín Castellanos-Joya Guadalupe Delgado-Sánchez Leticia Ferreyra-Reyes Pablo Cruz-Hervert Elizabeth Ferreira-Guerrero Gabriela Ortiz-Solís Mirtha Irene Jiménez Leslie Lorena Salazar Rogelio Montero-Campos Norma Mongua-Rodríguez Renata Baez-Salda?a Miriam Bobadilla-del-Valle Jesús Felipe González-Roldán Alfredo Ponce-de-León José Sifuentes-Osornio Lourdes García-García 《PloS one》2014,9(9)