A combined approach for comparative exoproteome analysis of Corynebacterium pseudotuberculosis

Background: Bacterial exported proteins represent key components of the host-pathogen interplay. Hence, we sought to implement a combined approach for characterizing the entire exoproteome of the pathogenic bacterium Corynebacterium pseudotuberculosis, the etiological agent of caseous lymphadenitis (CLA) in sheep and goats. Results: An optimized protocol of three-phase partitioning (TPP) was used to obtain the C. pseudotuberculosis exoproteins, and a newly introduced method of data-independent MS acquisition (LC-MSE) was employed for protein identification and label-free quantification. Additionally, the recently developed tool SurfG+ was used for in silico prediction of sub-cellular localization of the identified proteins. In total, 93 different extracellular proteins of C. pseudotuberculosis were identified with high confidence by this strategy; 44 proteins were commonly identified in two different strains, isolated from distinct hosts, then composing a core C. pseudotuberculosis exoproteome. Analysis with the SurfG+ tool showed that more than 75% (70/93) of the identified proteins could be predicted as containing signals for active exportation. Moreover, evidence could be found for probable non-classical export of most of the remaining proteins. Conclusions: Comparative analyses of the exoproteomes of two C. pseudotuberculosis strains, in addition to comparison with other experimentally determined corynebacterial exoproteomes, were helpful to gain novel insights into the contribution of the exported proteins in the virulence of this bacterium. The results presented here compose the most comprehensive coverage of the exoproteome of a corynebacterial species so far

Alendronic acid as ionic liquid: New perspective on osteosarcoma

This work was supported by the Associate Laboratory for Green Chemistry LAQV which is financed by national funds from FCT/MCTES (UIDB/50006/2020). The authors also thank Fundacao para a Ciencia e Tecnologia for the projects PTDC/QUI-QOR/32406/2017, PEst-C/LA0006/2013 and RECI/BBBBQB/0230/2012), as well as one contract under Investigador FCT (L. C. Branco).Herein the quantitative synthesis of eight new mono-and dianionic Organic Salts and Ionic Liquids (OSILs) from alendronic acid (ALN) is reported by following two distinct sustainable and straightforward methodologies, according to the type of cation. The prepared ALN-OSILs were characterized by spectroscopic techniques and their solubility in water and biological fluids was determined. An evaluation of the toxicity towards human healthy cells and also human breast, lung and bone (osteosarcoma) cell lines was performed. Globally, it was observed that the monoanionic OSILs showed lower toxicity than the corresponding dianionic structures to all cell types. The highest cytotoxic effect was observed in OSILs containing a [C2OHMIM] cation, in particular [C2OHMIM][ALN]. The latter showed an improvement in IC50 values of ca. three orders of magnitude for the lung and bone cancer cell lines as well as fibroblasts in comparison with ALN. The development of OSILs with high cytotoxicity effect towards the tested cancer cell types, and containing an anti-resorbing molecule such as ALN may represent a promising strategy for the development of new pharmacological tools to be used in those pathological conditions.publishersversionpublishe

A Novel Approach for Bisphosphonates: Ionic Liquids and Organic Salts from Zoledronic Acid

Novel ionic liquids and organic salts based on mono- or dianionic zoledronate and protonated superbases, choline and n-alkylmethylimidazolium cations, were prepared and characterized by spectroscopic and thermal analyses. Most of the prepared salts display amorphous structures and very high solubility in water and saline solutions, especially the dianionic salts. Among the zoledronate-based ionic compounds, those containing choline [Ch] and methoxyethylmethylimidazolium [C3 OMIM] cations appear to have significant cytotoxicity against human osteosarcoma cells (MG63) and low toxicity toward healthy skin fibroblast cells. Because osteosarcoma is a bone pathology characterized by an increase in bone turnover rate, the results presented herein may be a promising starting point for the development of new ionic pharmaceutical drugs against osteosarcoma.info:eu-repo/semantics/publishedVersio

Branching annihilating random walks with parity conservation on a square lattice

Using Monte Carlo simulations we have studied the transition from an "active" steady state to an absorbing "inactive" state for two versions of the branching annihilating random walks with parity conservation on a square lattice. In the first model the randomly walking particles annihilate when they meet and the branching process creates two additional particles; in the second case we distinguish particles and antiparticles created and annihilated in pairs. Quite distinct critical behavior is found in the two cases, raising the question of what determines universality in this kind of systems.Comment: 4 pages, 4 EPS figures include

Phenotypic variance in fruits and seed germination of Campomanesia adamantium Camb.

O objetivo deste trabalho foi avaliar a diversidade fenotípica da Campomanesia adamantium, com base nas características morfológicas dos frutos e emergência das sementes. Foram avaliadas dez populações em área de Cerrado Sentido Restrito, em Dourados-MS. A marcação das coordenadas de cada população foi obtida em função das diferenças fenotípicas apresentadas entre elas, distribuídas numa área de 32 ha. A coleta dos frutos foi realizada na segunda quinzena de dezembro de 2009, época de frutificação da espécie. Para o estudo da biometria, utilizou-se o delineamento experimental inteiramente casualizado, com dez tratamentos, correspondentes ás populações e dez repetições de dez frutos, correspondentes ás unidades experimentais. O estudo do comportamento germinativo das sementes foi desenvolvido em condições de casa de vegetação utilizando o delineamento blocos casualizados, com quatro repetições de dezesseis sementes, como unidade experimental, para cada população. No estudo biométrico dos frutos e no comportamento germinativo das sementes, observou-se a existência de grupos estatisticamente diferentes para cada característica avaliada. As populações de C. adamantium, em área nativa, apresentaram frutos com alta variabilidade fenotípica. Na casa de vegetação, as plântulas obtidas por propagação de sementes apresentaram uma menor proporção de diversidade fenotípica. Considerando-se as características dos frutos e propagativas, as populações 4, 9 e 10 destacam-se se como de maior potencial para estudos genéticos e de melhoramento.The aim of this work was to evaluate the phenotypic diversity of Campomanesia adamantium based on the morphological characteristics and seed emergence in ten populations from Cerrado (sensu stricto) area, in Dourados-MS. The establishment of coordinates of every population was obtained as a function of phenotypic difference showed among them, distributed in an area of 32 ha. Collect of fruits were done in the second period of fifteen days of December, 2009, fruitful period of the specie. For biometry study, a complete randomized experimental design was used, with ten treatments, which corresponded to populations and ten replications of ten fruits, which corresponded to experimental units. Germinative behavior study of seeds was carried out in green house conditions and use randomized block design, with four replications of sixteen seeds, as experimental unit, for every population. In biometry study of fruits and in germinative behavior of seeds, it was observed the presence of groups statistically different for every evaluated characteristic. Populations of C. adamantium, in native area, showed fruits with high phenotypic variability. In green house, seedlings by seed propagation showed a small proportion of phenotypic diversity. Considering characteristics of fruits and propagatives, populations 4, 9 and 10 are outstanding as the highest potential for genetic studies and for improvement.Facultad de Ciencias Agrarias y Forestale

Phenotypic variance in fruits and seed germination of Campomanesia adamantium Camb.

O objetivo deste trabalho foi avaliar a diversidade fenotípica da Campomanesia adamantium, com base nas características morfológicas dos frutos e emergência das sementes. Foram avaliadas dez populações em área de Cerrado Sentido Restrito, em Dourados-MS. A marcação das coordenadas de cada população foi obtida em função das diferenças fenotípicas apresentadas entre elas, distribuídas numa área de 32 ha. A coleta dos frutos foi realizada na segunda quinzena de dezembro de 2009, época de frutificação da espécie. Para o estudo da biometria, utilizou-se o delineamento experimental inteiramente casualizado, com dez tratamentos, correspondentes ás populações e dez repetições de dez frutos, correspondentes ás unidades experimentais. O estudo do comportamento germinativo das sementes foi desenvolvido em condições de casa de vegetação utilizando o delineamento blocos casualizados, com quatro repetições de dezesseis sementes, como unidade experimental, para cada população. No estudo biométrico dos frutos e no comportamento germinativo das sementes, observou-se a existência de grupos estatisticamente diferentes para cada característica avaliada. As populações de C. adamantium, em área nativa, apresentaram frutos com alta variabilidade fenotípica. Na casa de vegetação, as plântulas obtidas por propagação de sementes apresentaram uma menor proporção de diversidade fenotípica. Considerando-se as características dos frutos e propagativas, as populações 4, 9 e 10 destacam-se se como de maior potencial para estudos genéticos e de melhoramento.The aim of this work was to evaluate the phenotypic diversity of Campomanesia adamantium based on the morphological characteristics and seed emergence in ten populations from Cerrado (sensu stricto) area, in Dourados-MS. The establishment of coordinates of every population was obtained as a function of phenotypic difference showed among them, distributed in an area of 32 ha. Collect of fruits were done in the second period of fifteen days of December, 2009, fruitful period of the specie. For biometry study, a complete randomized experimental design was used, with ten treatments, which corresponded to populations and ten replications of ten fruits, which corresponded to experimental units. Germinative behavior study of seeds was carried out in green house conditions and use randomized block design, with four replications of sixteen seeds, as experimental unit, for every population. In biometry study of fruits and in germinative behavior of seeds, it was observed the presence of groups statistically different for every evaluated characteristic. Populations of C. adamantium, in native area, showed fruits with high phenotypic variability. In green house, seedlings by seed propagation showed a small proportion of phenotypic diversity. Considering characteristics of fruits and propagatives, populations 4, 9 and 10 are outstanding as the highest potential for genetic studies and for improvement.Facultad de Ciencias Agrarias y Forestale

Schistosome and liver fluke derived catechol-estrogens and helminth associated cancers.

Infection with helminth parasites remains a persistent public health problem in developing countries. Three of these pathogens, the liver flukes Clonorchis sinensis, Opisthorchis viverrini and the blood fluke Schistosoma haematobium, are of particular concern due to their classification as Group 1 carcinogens: infection with these worms is carcinogenic. Using liquid chromatography-mass spectrometry (LC-MS/MS) approaches, we identified steroid hormone like (e.g., oxysterol-like, catechol estrogen quinone-like, etc.) metabolites and related DNA-adducts, apparently of parasite origin, in developmental stages including eggs of S. haematobium, in urine of people with urogenital schistosomiasis, and in the adult stage of O. viverrini. Since these kinds of sterol derivatives are metabolized to active quinones that can modify DNA, which in other contexts can lead to breast and other cancers, helminth parasite associated sterols might induce tumor-like phenotypes in the target cells susceptible to helminth parasite associated cancers, i.e., urothelial cells of the bladder in the case of urogenital schistosomiasis and the bile duct epithelia or cholangiocytes, in the case of O. viverrini and C. sinensis. Indeed we postulate that helminth induced cancers originate from parasite estrogen-host epithelial/urothelial cell chromosomal DNA adducts, and here we review recent findings that support this conjecture