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71.
Enhanced extraction of proteins using cholinium‐based ionic liquids as phase‐forming components of aqueous biphasic systems
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Maria V. Quental Magda Caban Matheus M. Pereira Piotr Stepnowski João A. P. Coutinho Mara G. Freire 《Biotechnology journal》2015,10(9):1457-1466
Aqueous biphasic systems (ABS) composed of ionic liquids (ILs) are promising platforms for the extraction and purification of proteins. In this work, a series of alternative and biocompatible ABS composed of cholinium‐based ILs and polypropylene glycol were investigated. The respective ternary phase diagrams, tie‐lines, tie‐line lengths and critical points were determined at 25°C. The extraction performance of these systems for commercial bovine serum albumin (BSA) was then evaluated. The stability of BSA at the IL‐rich phase was ascertained by size exclusion high‐performance liquid chromatography and Fourier transform infrared spectroscopy. Appropriate ILs lead to the complete extraction of BSA for the IL‐rich phase, in a single step, while maintaining the protein's native conformation. Furthermore, to evaluate the performance of these systems when applied to real matrices, the extraction of BSA from bovine serum was additionally carried out, revealing that the complete extraction of BSA was maintained and achieved in a single step. The remarkable extraction efficiencies obtained are far superior to those observed with typical polymer‐based ABS. Therefore, the proposed ABS may be envisaged as a more effective and biocompatible approach for the separation and purification of other value‐added proteins. 相似文献
72.
Rodolpho Santos Telles Menezes Seán Gary Brady Ant?nio Freire Carvalho Marco Antonio Del Lama Marco Ant?nio Costa 《PloS one》2015,10(3)
The Neotropical Region harbors high biodiversity and many studies on mammals, reptiles, amphibians and avifauna have investigated the causes for this pattern. However, there is a paucity of such studies that focus on Neotropical insect groups. Synoeca de Saussure, 1852 is a Neotropical swarm-founding social wasp genus with five described species that is broadly and conspicuously distributed throughout the Neotropics. Here, we infer the phylogenetic relationships, diversification times, and historical biogeography of Synoeca species. We also investigate samples of the disjoint populations of S. septentrionalis that occur in both northwestern parts of South America through Central American and the Brazilian Atlantic rainforests. Our results showed that the interspecific relationships for Synoeca could be described as follows: (S. chalibea + S. virginea) + (S. cyanea + (S. septentrionalis/S. surinama)). Notably, samples of S. septentrionalis and S. surinama collected in the Atlantic Forest were interrelated and may be the result of incomplete lineage sorting and/or mitochondrial introgression among them. Our Bayesian divergence dating analysis revealed recent Plio-Pleistocene diversification in Synoeca. Moreover, our biogeographical analysis suggested an Amazonian origin of Synoeca, with three main dispersal events subsequently occurring during the Plio-Pleistocene. 相似文献
73.
Milton Maciel Jr. Fábia da Silva Pereira Cruz Marli Tenório Cordeiro Márcia Archer da Motta Klécia Marília Soares de Melo Cassemiro Rita de Cássia Carvalho Maia Regina Célia Bressan Queiroz de Figueiredo Ricardo Galler Marcos da Silva Freire Joseph Thomas August Ernesto T. A. Marques Rafael Dhalia 《PLoS neglected tropical diseases》2015,9(4)
Attenuated yellow fever (YF) virus 17D/17DD vaccines are the only available protection from YF infection, which remains a significant source of morbidity and mortality in the tropical areas of the world. The attenuated YF virus vaccine, which is used worldwide, generates both long-lasting neutralizing antibodies and strong T-cell responses. However, on rare occasions, this vaccine has toxic side effects that can be fatal. This study presents the design of two non-viral DNA-based antigen formulations and the characterization of their expression and immunological properties. The two antigen formulations consist of DNA encoding the full-length envelope protein (p/YFE) or the full-length envelope protein fused to the lysosomal-associated membrane protein signal, LAMP-1 (pL/YFE), aimed at diverting antigen processing/presentation through the major histocompatibility complex II precursor compartments. The immune responses triggered by these formulations were evaluated in H2b and H2d backgrounds, corresponding to the C57Bl/6 and BALB/c mice strains, respectively. Both DNA constructs were able to induce very strong T-cell responses of similar magnitude against almost all epitopes that are also generated by the YF 17DD vaccine. The pL/YFE formulation performed best overall. In addition to the T-cell response, it was also able to stimulate high titers of anti-YF neutralizing antibodies comparable to the levels elicited by the 17DD vaccine. More importantly, the pL/YFE vaccine conferred 100% protection against the YF virus in intracerebrally challenged mice. These results indicate that pL/YFE DNA is an excellent vaccine candidate and should be considered for further developmental studies. 相似文献
74.
Pedro Paulo de Abreu Manso Barbara C. E. P. Dias de Oliveira Patrícia Carvalho de Sequeira Yuli Rodrigues Maia de Souza Jessica Maria dos Santos Ferro Igor José da Silva Luzia Fátima Gon?alves Caputo Priscila Tavares Guedes Alexandre Araujo Cunha dos Santos Marcos da Silva Freire Myrna Cristina Bonaldo Marcelo Pelajo-Machado 《PLoS neglected tropical diseases》2015,9(9)
The yellow fever (YF) 17D vaccine is one of the most effective human vaccines ever created. The YF vaccine has been produced since 1937 in embryonated chicken eggs inoculated with the YF 17D virus. Yet, little information is available about the infection mechanism of YF 17DD virus in this biological model. To better understand this mechanism, we infected embryos of Gallus gallus domesticus and analyzed their histopathology after 72 hours of YF infection. Some embryos showed few apoptotic bodies in infected tissues, suggesting mild focal infection processes. Confocal and super-resolution microscopic analysis allowed us to identify as targets of viral infection: skeletal muscle cells, cardiomyocytes, nervous system cells, renal tubular epithelium, lung parenchyma, and fibroblasts associated with connective tissue in the perichondrium and dermis. The virus replication was heaviest in muscle tissues. In all of these specimens, RT-PCR methods confirmed the presence of replicative intermediate and genomic YF RNA. This clearer characterization of cell targets in chicken embryos paves the way for future development of a new YF vaccine based on a new cell culture system. 相似文献
75.
Diogo G. Garcia Lidia M. F. Amorim Mauro V. de Castro Faria Aline S. Freire Ricardo E. Santelli Clóvis O. Da Fonseca Thereza Quirico-Santos Patricia Burth 《Molecular and cellular biochemistry》2010,342(1-2):29-34
Indoleamine 2,3-dioxygenase (IDO) is an enzyme that suppresses adaptive T-cell immunity by catabolizing tryptophan from the cellular microenvironment. Inhibition of IDO pathway might enhance the efficacy of immunotherapeutic strategies for cancer. We synthesized 1-alkyl-tryptophan targeted IDO inhibitors and compared their effects on IDO expression and activity in dendritic cells (DCs) with the common IDO inhibitor 1-methyl-dl-tryptophan (1-MT). The IDO gene expression was examined by RT-PCR and realtime PCR. The toxicity of these analogs on the proliferation of DCs was detected by MTT assay. All of these analogs inhibited IDO expression and activity induced by IFN-γ and showed no cytotoxicity to DCs at 100 μM. 1-MT intensively suppressed IDO1 expression and activity in DCs, and 1-propyl-tryptophan (1-PT) and 1-isopropyl-tryptophan (1-isoPT) moderately inhibited them. 1-Butyl-tryptophan (1-BT) and 1-ethyl-tryptophan (1-ET) mainly inhibited IDO2 expression. Our results suggest that those analogs differed in their inhibitory activity on IDO expression may give us a clue for developing active IDO inhibitors. 相似文献
76.
Bárbara Santos Pires da Silva Eloísa Bonfá Júlio César Bertacini de Moraes Carla Gonçalves Schain Saad Ana Cristina de Medeiros Ribeiro Célio Roberto Gonçalves Jozélio Freire de Carvalho 《Biologicals》2010,38(5):567-569
The objective of this study was to evaluate the influence of anti-tumor necrosis factor (anti-TNF) in juvenile idiopathic arthritis (JIA), ankylosing spondylitis (AS) or psoriatic arthritis (PsA). Sixty-two patients were investigated: 7 JIA; 37 AS; and 18 PsA. Caucasian race accounted for 79% and 29% were female. Mean age was 40.4 ± 12.6years. None of the patients had a history of diabetes, and none had used oral hypoglycemic agents or insulin. Treatment was with adalimumab, infliximab and etanercept. Glucose, inflammatory markers and prednisone dose were assessed at baseline, as well as after three and six months of treatment. The mean erythrocyte sedimentation rate was significantly lower at three months and six months than at baseline (13.7 ± 18.0 and 18 ± 22.5 vs. 27.9 ± 23.4 mm; p = 0.001). At baseline, three months and six months, we found the following: mean C-reactive protein levels were comparable (22.1 ± 22.7, 14.5 ± 30.7 and 16.0 ± 23.8 mg/L, respectively; p = 0.26); mean glucose levels remained unchanged (90.8 ± 22.2 mg/dl, 89.5 ± 14.6 mg/dl and 89.8 ± 13.6 mg/dl, respectively; p = 0.91); and mean prednisone doses were low and stable (3.9 ± 4.9 mg/day, 3.7 ± 4.8 mg/day and 2.6 ± 4.0 mg/day, respectively; p = 0.23). During the first six months of treatment, anti-TNF therapy does not seem to influence glucose metabolism in JIA, AS or PsA. 相似文献
77.
Prodocimo V Galvez F Freire CA Wood CM 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》2007,177(5):519-528
Na+ and Ca2+ regulation were compared in two euryhaline species, killifish (normally estuarine-resident) and rainbow trout (normally freshwater-resident)
during an incremental salinity increase. Whole-body unidirectional fluxes of Na+ and Ca2+, whole body Na+ and Ca2+, and plasma concentrations (trout only), were measured over 1-h periods throughout a total 6-h protocol of increasing salinity
meant to simulate a natural tidal flow. Killifish exhibited significant increases in both Na+ influx and efflux rates, with efflux slightly lagging behind efflux up to 60% SW, but net Na+ balance was restored by the time killifish reached 100% SW. Whole body Na+ did not change, in agreement with the capacity of this species to tolerate daily salinity fluctuations in its natural habitat.
In contrast, rainbow trout experienced a dramatic increase in Na+ influx (50-fold relative to FW values), but not Na+ efflux between 40 and 60% SW, resulting in a large net loading of Na+ at higher salinities (60–100% SW), and increases in plasma Na+ and whole body Na+ at 100% SW. Killifish were in negative Ca2+ balance at all salinities, whereas trout were in positive Ca2+ balance throughout. Ca2+ influx rate increased two- to threefold in killifish at 80 and 100% SW, but there were no concomitant changes in Ca2+ efflux. Ca2+ flux rates were affected to a larger degree in trout, with twofold increases in Ca2+ influx at 40% SW and sevenfold increases at 100% SW. Again, there was no change in Ca2+ efflux with salinity, so plasma Ca2+ concentration increased in 100% SW. As the killifish is regularly submitted to increased salinity in its natural environment,
it is able to rapidly activate changes in unidirectional fluxes in order to ensure ionic homeostasis, in contrast to the trout. 相似文献
78.
Ana Carolina Luchiari Cristiane Regina do Amaral Duarte Fúlvio Aurélio de Morais Freire Kari Nissinen 《Journal of Ethology》2007,25(2):169-175
We studied the colour preference of isolated Nile tilapia (Oreochromis niloticus) and whether previous residence or body size can affect environmental colour choice. In the first phase, a cylindrical tank
was divided into five differently coloured compartments (yellow, blue, green, white and red), a single fish was introduced
into the tank and the frequency at which this fish visited each compartment was recorded over a 2-day study period. An increasingly
larger fish (approx +2 cm in length each time) was then added into the tank on each of days 3, 5 and 7 (=four fish in the
tank by day 7), and the frequency at which each fish visited the different compartments of the tank was observed twice a day
to obtain visit frequency data on the differently sized fishes. This experiment was replicated six times. In the first phase,
the solitary fish established residence inside the yellow compartment on the first and second days. Following the
introduction of a larger fish, the smaller fish was displaced from the occupied compartment. Nile tilapia possibly shows this
preference for yellow as a function of its visual spectral sensitivity and/or the spectral characteristics of its natural
environment. Moreover, body size is an important factor in determining hierarchical dominance and territorial defence, and
dominant fish chose the preferred environmental colour compartment as their territory. 相似文献
79.
Corte AC Svoboda WK Navarro IT Freire RL Malanski LS Shiozawa MM Ludwig G Aguiar LM Passos FC Maron A Camargo ZP Itano EN Ono MA 《Mycopathologia》2007,164(5):225-228
The aim of this study was to evaluate the seroprevalence of Paracoccidioides brasiliensis infection in wild New World monkeys (Cebus sp. and Alouatta caraya). A total of 93 animals (Cebus sp., n = 68 and Alouatta caraya, n = 25) were captured in the Paraná River basin, Paraná State, Brazil and the serum samples were analyzed by ELISA and immunodiffusion
using P. brasiliensis gp43 and exoantigen as antigens, respectively. The seropositivity observed by ELISA was 44.1% and 60% for Cebus sp. and A. caraya, respectively, while by immunodiffusion test Cebus sp. showed positivity of 2.9% only. No significant difference was observed in relation to age and sex. This is the first
report of paracoccidioidomycosis in wild capuchin monkeys and in wild-black and golden-howler monkeys. The high positivity
to P. brasiliensis infection in both species evaluated in our study and the positivity by immunodiffusion test in Cebus sp. suggest that natural disease may be occurring in wild monkeys living in paracoccidioidomycosis endemic areas. 相似文献
80.
Enzymatic pre-hydrolysis applied to the anaerobic treatment of effluents from poultry slaughterhouses 总被引:1,自引:0,他引:1
Alessandra Bormann Garcia Vallado Denise Maria Guimares Freire Magali Christe Cammarota 《International biodeterioration & biodegradation》2007,60(4):219-225
A pool of hydrolases with 21.4 U g−1 lipase activity was produced through solid-state fermentation of the fungus Penicillium restrictum in waste from the Orbignya oleifera (babassu) oil processing industry. Enzymatic hydrolysis and anaerobic biodegradability tests were conducted on poultry slaughterhouse effluents with varying oil and grease contents (150–1200 mg l−1) and solid enzymatic pool concentrations (0.1–1.0% w/v). Enhanced anaerobic treatment efficiency relative to raw effluent was achieved when a 0.1% concentration of enzymatic pool was used in the pre-hydrolysis stage with 1200 mg oil and grease l−1 (chemical oxygen demand (COD) removal efficiency of 85% vs. 53% and biogas production of 175 ml vs. 37 ml after 4 d). 相似文献