15 results on '"Dias, Ana C."'
Search Results
2. Life cycle assessment of the production of Portland cement: a Southern Europe case study.
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Stafford, Fernanda N., Dias, Ana C., Arroja, Luís, Labrincha, João A., and Hotza, Dachamir
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PORTLAND cement industry , *PRODUCT life cycle assessment , *BUSINESS enterprises , *RAW materials , *ENERGY consumption - Abstract
The cement industry is one of the most important industries worldwide and the world cement production is projected to grow by 0.8–1.2% per year, reaching between 3700 and 4400 megatonnes in 2050. This industry is facing important challenges in the context of restrained use of raw materials and need of optimization of energy consumption. The aim of this paper is to assess environmental impacts of using wastes as a partial replacement of fossil fuels in a representative cement plant in Southern Europe. Refuse derived fuel and scrap tires were employed as alternative fuels. The environmental profile of the cement manufacturing was presented through a cradle-to-gate life cycle assessment approach based on primary data collected in the plant. The functional unit was 1 tonne of ordinary Portland cement and the impact assessment was based on indicators that comply with the ISO 14040. The impact categories studied were abiotic depletion, acidification, eutrophication, photochemical oxidant formation and global warming. The processes that most contributed to these impact categories were the atmospheric emissions in the kiln, the consumption of fossil fuels, and the consumption of electricity in the mills. [ABSTRACT FROM AUTHOR]
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- 2016
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3. Uncovering the potential of aqueous solutions of deep eutectic solvents on the extraction and purification of collagen type I from Atlantic codfish (Gadus morhua).
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Bisht, Meena, Martins, Margarida, Dias, Ana C. R. V., Ventura, Sónia P. M., and Coutinho, João A. P.
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ATLANTIC cod , *LACTIC acid , *SOLVENT extraction , *AQUEOUS solutions , *CODFISH , *COLLAGEN , *EUTECTICS - Abstract
Marine fish industries discard huge amounts of fish waste every year, which in turn impose problems of environmental pollution and loss of economic value. About 75% of the total weight of fish is discarded in the form of skins, bones, fins, heads, guts, and scales, which contain high levels of collagen type I. Generally, major sources for commercial collagens are the skin and bone of pigs and cows; however, these sources are chiefly associated with the risk of transference of zoonotic diseases or religious issues. Traditional protocols applied to the extraction of collagen are outdated, mainly with respect to present demands to develop more sustainable processes. This work explores the use of sustainable solvents, such as deep eutectic solvents (DES), to develop a more efficient, cost-effective and biocompatible process to extract collagen from waste from the fish industry waste. The extraction of collagen from the skin of Atlantic cod (Gadus morhua) using aqueous solutions of various eutectic solvents was studied, and after selection of the best solvent, an aqueous solution of urea (U) and lactic acid (LA) at a molar ratio of 1 : 2 (U : LA 1 : 2), the collagen extracted was properly characterized using SDS-PAGE, CD, FTIR, and XRD, and shown to be of type I. The results obtained here demonstrate an improvement in the yield and quality of the extracted collagen when eutectic mixtures were applied instead of acetic acid. After optimization of the process conditions, a maximum extraction yield of 6% was obtained for the aqueous solution of U : LA 1 : 2 at 0.75 M. The present work demonstrates the potential use of codfish skin waste and an aqueous solution of a DES to develop a more environmentally-friendly process to obtain high-quality collagen type I. It is an effort to convince industries to valorize their own residues under the guidelines of a circular economy. [ABSTRACT FROM AUTHOR]
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- 2021
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4. Carbon storage and CO2 dynamics from wood products harvested in Brazil during 1900–2016.
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Sanquetta, Carlos R., Tomé, Margarida, Dias, Ana C., Maas, Greyce C.B., Sanquetta, Felipe T.I., and Corte, Ana P.D.
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AFFORESTATION , *REFORESTATION , *WOOD products , *FOREST management , *LAND use - Abstract
How relevant is the role of CO2 from harvested wood products (HWPs) for the Brazilian GHG emission balance? The C stocks and CO2 emissions and removals from the HWPs in Brazil during 1900–2016 were evaluated using three methodological approaches: stock-change approach (SCA), production approach (PA), and stock-change of domestic origin (SCDOA). Data from the FAO's FAOSTAT system on production, import and export of semi-finished products (sawnwood, wood-based panels and paper/paperboard) were analyzed, considering distinct land-use activities: deforestation (D), forest management (FM) and afforestation/reforestation (A/R). Two scenarios to evaluate the effect of HWPs from D were evaluated. The C stock in HWPs in 2016 reached 252 Mt C, by SCA. Contributions to C storage historically were higher from sawnwood, but wood-based panels and paper/paperboard have recently outperformed it. A/R is the major activity responsible for C storage in HWPs. The CO2 removal by HWPs was 521 Mt during 1990–2016, which corresponds to 1.1% of the country's emissions in the same period. It was concluded that Brazil has become a major producer and wood consumer and that HWPs are allies for a positive C balance and fulfillment of the commitments assumed by the country regarding climate change. AFOLU Agriculture, forestry and other land uses A/R Afforestation/reforestation D Deforestation FAO United Nations Food and Agriculture Organization FM Forest management HWPs Harvested wood products IBGE Brazilian Institute of Geography and Statistics PA Production approach PEVS Plant extraction and forestry SCA Stock-change approach SCDOA Stock-change of domestic origin approach [ABSTRACT FROM AUTHOR]
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- 2019
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5. Environmental implications of the use of agglomerated cork as thermal insulation in buildings.
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Sierra-Pérez, Jorge, Boschmonart-Rives, Jesús, Dias, Ana C., and Gabarrell, Xavier
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THERMAL properties of buildings , *THERMAL insulation , *ENERGY industries , *ENERGY consumption of buildings , *RENEWABLE energy sources - Abstract
The market for insulation material is playing a crucial role in Europe's energy transformation, due to its influence on energy consumption in buildings. The introduction of renewable materials for thermal insulation is recent, and little is known so far about its environmental implications. This study analyses the environmental performance of a cork insulation board, made of agglomerated cork from forestry cork wastes, by means of cradle-to-gate Life Cycle Assessment methodology. The results indicate that the use of natural insulation materials does not necessarily imply a reduction of environmental impacts due to manufacturing processes with a low technological development. In this case, the most influential stage is the manufacturing stage, in which the board agglomeration and the cork trituration have the highest impacts. The most influential inputs are both the transport used during the life cycle and the large quantities of electricity and diesel in the manufacturing stage. Some strategies have been identified to reduce the environmental impact, such as promote the acquisition of local raw cork to reduce transportation from the manufacturer, improve the efficiency and productivity of manufacturing processes and improve the product design to help increase its market share. Moreover, the inclusion of biogenic carbon contained in forest-based building materials affects the Global Warming Potential results considerably. However, it is very important to consider how this biogenic carbon is calculated and how the product is managed after its lifetime. [ABSTRACT FROM AUTHOR]
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- 2016
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6. Diagnostic application of sensitive and specific phage-exposed epitopes for visceral leishmaniasis and human immunodeficiency virus coinfection.
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Ramos, Fernanda F., Tavares, Grasiele S. V., Ludolf, Fernanda, Machado, Amanda S., Santos, Thaís T. O., Gonçalves, Isabela A. P., Dias, Ana C. S., Alves, Patrícia T., Fraga, Vanessa G., Bandeira, Raquel S., Oliveira-da-Silva, João A., Reis, Thiago A. R., Lage, Daniela P., Martins, Vívian T., Freitas, Camila S., Chaves, Ana T., Guimarães, Nathalia S., Chávez-Fumagalli, Miguel A., Tupinambás, Unaí, and Rocha, Manoel O. C.
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VISCERAL leishmaniasis , *MIXED infections , *EPITOPES , *HIV , *LEISHMANIASIS , *ENZYME-linked immunosorbent assay , *SENSITIVITY & specificity (Statistics) - Abstract
The diagnosis of visceral leishmaniasis (VL) has improved with the search of novel antigens; however, their performance is limited when samples from VL/human immunodeficiency virus (HIV)-coinfected patients are tested. In this context, studies conducted to identify more suitable antigens to detect both VL and VL/HIC coinfection cases should be performed. In the current study, phage display was performed using serum samples from healthy subjects and VL, HIV-infected and VL/HIV-coinfected patients; aiming to identify novel phage-exposed epitopes to be evaluated with this diagnostic purpose. Nine non-repetitive and valid sequences were identified, synthetized and tested as peptides in enzyme-linked immunosorbent assay experiments. Results showed that three (Pep2, Pep3 and Pep4) peptides showed excellent performance to diagnose VL and VL/HIV coinfection, with 100% sensitivity and specificity values. The other peptides showed sensitivity varying from 50.9 to 80.0%, as well as specificity ranging from 60.0 to 95.6%. Pep2, Pep3 and Pep4 also showed a potential prognostic effect, since specific serological reactivity was significantly decreased after patient treatment. Bioinformatics assays indicated that Leishmania trypanothione reductase protein was predicted to contain these three conformational epitopes. In conclusion, data suggest that Pep2, Pep3 and Pep4 could be tested for the diagnosis of VL and VL/HIV coinfection. [ABSTRACT FROM AUTHOR]
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- 2021
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7. Integrative platform for the selective recovery of intracellular carotenoids and lipids from Rhodotorula glutinis CCT-2186 yeast using mixtures of bio-based solvents.
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Mussagy, Cassamo U., Santos-Ebinuma, Valéria C., Kurnia, Kiki A., Dias, Ana C. R. V., Carvalho, Pedro, Coutinho, João A. P., and Pereira, Jorge F. B.
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CAROTENOIDS , *ETHYL acetate , *RHODOTORULA , *LIPIDS , *MIXTURES , *SOLVENTS , *LIQUID-liquid extraction - Abstract
Natural bioactive compounds have been attracting growing interest from the industries as a "greener" alternative to synthetic raw materials/products. Rhodotorula glutinis yeast naturally synthesizes added value compounds such as lipids and carotenoids, commonly used for cosmetic, pharmaceutical, and food applications. R. glutinis constitutes a rigid cell-wall structure, requiring energy-saving and efficient cell disruption methods for a sustainable recovery of the intracellular compounds. A simple and ecofriendly technology using mixed bio-based solvents (biosolvents) was evaluated here as an alternative platform to permeabilize yeast cells and to improve the selective recovery of β-carotene, torularhodin, torulene and lipids. The extraction ability of pure and solvent mixtures (methanol, ethanol, ethyl acetate, isopropanol, cyclohexane and 2-methyl tetrahydrofuran) was initially screened, demonstrating the clear impact of using mixtures to improve the extraction yields (up to three-fold increase). After identifying ethyl acetate/ethanol/water as the solvent mixture with a greater capacity to extract carotenoids and lipids, the selective recovery of carotenoids and lipids was enhanced by optimizing the solvent mixture composition ratio. Envisioning the industrial application, an integrated biosolvent-based downstream platform was designed. Two different strategies were investigated to further isolate carotenoids and lipids from R. glutinis biomass and to recycle the ethyl acetate/ethanol/water mixture: (i) precipitation using cold acetone; (ii) sequential liquid–liquid extraction. The integrated process for each strategy was compared with a conventional extraction procedure in terms of recovery efficiencies and its environmental impact. Regardless of the strategy, it is shown that the mixture of ethyl acetate, ethanol and water (15/27/58% w/w) can be reused up to three consecutive extractive cycles, ensuring high extraction efficiency yields, while decreasing the process carbon footprint by about 75% compared to the conventional method. [ABSTRACT FROM AUTHOR]
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- 2020
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8. Continuous separation of cytochrome-c PEGylated conjugates by fast centrifugal partition chromatography.
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Santos, João H. P. M., Ferreira, Ana M., Almeida, Mafalda R., Quinteiro, Paula S. G. N., Dias, Ana C. R. V., Coutinho, João A. P., Freire, Mara G., Rangel-Yagui, Carlota O., and Ventura, Sónia P. M.
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PARTITION chromatography , *CYTOCHROME c , *SUSTAINABLE design , *COUNTERCURRENT chromatography , *ECOLOGICAL impact , *CONTINUOUS processing - Abstract
Herein, the effective use of aqueous biphasic systems (ABS) in Fast Centrifugal Partition Chromatography (FCPC) for the purification of PEGylated cytochrome c conjugates is shown. High recoveries (between 88% and 100%) and purities (∼100%) were obtained. Both the unreacted cytochrome c and solvents may be recovered and reused, thus allowing the design of a sustainable process in a continuous regime for the isolation of bioconjugates. This process allowed the reduction of the complete E-factor and carbon footprint at circa 100% and 67%, respectively, reinforcing the important environmental contribution of recycling units. [ABSTRACT FROM AUTHOR]
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- 2019
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9. R-phycoerythrin extraction and purification from fresh Gracilaria sp. using thermo-responsive systems.
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Vicente, Filipa A., Cardoso, Inês S., Martins, Margarida, Gonçalves, Cátia V. M., Dias, Ana C. R. V., Domingues, Pedro, Coutinho, João A. P., and Ventura, Sónia P. M.
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SOLAR concentrators , *GRACILARIA , *CHEMICAL purification , *IONIC surfactants , *PROCESS optimization , *ECOLOGICAL impact - Abstract
R-phycoerythrin is a high added-value protein found in red macroalgae with several interesting properties. Despite the promising results found when R-phycoerythrin is used as an optically active center in luminescent solar concentrators (LSCs), it still has some problems that can be attributed to the low stability of the R-phycoerythrin in the presence of the specific contaminant proteins found in the crude extract. The development of downstream strategies able to reduce the use of environmentally hazardous solvents, while improving the purification without compromising the R-phycoerythrin structural integrity is still the biggest challenge to overcome. Aqueous micellar two-phase systems (AMTPS) appear as an appealing fractionation approach since they allow the processing of systems with larger water contents, while displaying great selectivity and biocompatibility with several biomolecules. Moroever, AMTPS that mix surfactants and surface-active ionic liquids are shown to significantly enhance protein purification. In this work, mixed AMTPS were applied to the R-phycoerythrin purification from red macroalgae. After the process optimization, this work proposes the application of two consecutive steps of purification as the final process to isolate R-phycoerythrin from the remaining proteins composing the crude extract, while maintaining the structural integrity of R-phycoerythrin, as requested to be used in the LSC. Besides a good performance, the two-step approach developed was also shown to have a lower environmental impact with a carbon footprint decrease of 16%, when compared with the conventional AMTPS. [ABSTRACT FROM AUTHOR]
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- 2019
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10. Ionic liquid-high performance extractive approach to recover carotenoids from Bactris gasipaes fruits.
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de Souza Mesquita, Leonardo M., Ventura, Sónia P. M., Braga, Anna R. C., Pisani, Luciana P., Dias, Ana C. R. V., and de Rosso, Veridiana V.
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FRUIT , *INDUSTRIAL capacity , *APPROPRIATE technology , *RAW materials , *ECOLOGICAL impact , *DATE palm - Abstract
Nowadays, one of the biggest challenges for society is the development of appropriate technologies to process the waste residue produced worldwide. In the food sector, the generated waste is estimated to be nearly billions of tons annually. Brazil is one of the most representative examples of the economic and industrial potential of underexplored residues and raw materials. The palm heart, scientifically known as Bactris gasipaes, mainly its fruits, is one of the many examples found in Brazilian flora. The fruits have significant amounts of carotenoids, namely, the all-trans-β-carotene, all-trans-lycopene and the rare all-trans-γ-carotene, which are considered as excellent raw materials of commercial interest. However, the main challenge that remains is their efficient recovery. This work proposes the development of a performant process of extraction mediated by the use of ionic liquid (IL)-based ethanolic solutions. Four ILs were examined, as well as the solid–liquid ratio R(S/L), number of extractions, the time of extraction, the co-solvent-ratio R(IL/E) and the homogenization method employed. After selecting the best solvent ([C4mim][BF4]) and process conditions (extraction yield of 172 ± 18 μgcarotenoids gdried biomass−1), the IL-ethanolic solution recyclability was tested by freezing/precipitating the IL (maximum of 94% of IL recovered), proving its success for at least 10 cycles while decreasing the process carbon footprint by 50% compared with the conventional method using acetone. [ABSTRACT FROM AUTHOR]
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- 2019
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11. Water footprint profile of crop-based vegetable oils and waste cooking oil: Comparing two water scarcity footprint methods.
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Caldeira, Carla, Quinteiro, Paula, Castanheira, Erica, Boulay, Anne-Marie, Dias, Ana C., Arroja, Luís, and Freire, Fausto
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VEGETABLE oils , *ECOLOGICAL impact , *WATER shortages , *BIODIESEL fuels , *WATER quality - Abstract
This paper compares the water footprint profiles of four feedstocks used for biodiesel production: palm, soya, rapeseed and waste cooking oil (WCO). The profiles include: (a) a water scarcity footprint related to freshwater consumption impacts and (b) a water quality degradation footprint related to freshwater degradation impacts. The water scarcity footprint was assessed using two impact assessment methods: one based on water stress indices (WSIs) and the other on the available water remaining (AWARE) indicator. The water degradation footprint was assessed considering the environmental mechanisms covered by the impact categories of eutrophication, aquatic acidification, human toxicity and freshwater ecotoxicity. The water scarcity profiles ranged from 0.002 to 2.11 world m 3 eq kg −1 oil (WSI method) and from 0.008 to 133.57 world m 3 eq kg −1 oil (AWARE method). Both methods showed that the cultivation stage assumes the primary role in the water scarcity footprint results and identified the same systems with higher water scarcity footprints. However, for the oil systems with closer results, the rank order given by each method is different due to the characterization factors of each method. Nevertheless, the results obtained with the AWARE method give more comprehensive water scarcity footprint results than those obtained when applying WSIs because AWARE considers the aquatic ecosystem water demand. The water degradation footprint of virgin oils is mainly caused by fertilizers and pesticides used in cultivation. WCO systems present lower impacts for all impact categories with the exception of human toxicity-cancer. The choice of locations with lower water scarcity to produce oil crops can be a determinant in the calculation of lower impacts. Moreover, optimizing fertilization schemes or choosing climatic conditions that require less fertilizers, pesticides and water consumption can reduce the impacts of the water footprint profile of vegetable oils. [ABSTRACT FROM AUTHOR]
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- 2018
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12. Separation of phenolic compounds by centrifugal partition chromatography.
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Santos, João H. P. M., Almeida, Mafalda R., Martins, Cláudia I. R., Dias, Ana C. R. V., Freire, Mara G., Coutinho, João A. P., and Ventura, Sónia P. M.
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PHENOLS , *SEPARATION (Technology) , *PARTITION chromatography - Abstract
Phenolic compounds are ubiquitous biomolecules exhibiting a wide range of physiological properties, with application in the pharmaceutical and nutraceutical fields. In this work, aqueous biphasic systems (ABS) formed by polyethylene glycol and sodium polyacrylate, and inorganic salts or ionic liquids as electrolytes, were applied for the purification of caffeic, ferulic and protocatechuic acids (CA, FA, and PA, respectively), vanillin (VN) and syringaldehyde (SA), followed by the use of centrifugal partition chromatography (CPC) to reinforce the fractionation process scale-up. In single-step experiments in ABS, high selectivities and adequate partition coefficients (KCA = 2.78 ± 0.20; KPA = 0.44 ± 0.04; KFA = 0.23 ± 0.01; KVN = 1.12 ± 0.05 and KSA = 1.23 ± 0.02) were achieved using ABS formed by sodium chloride as the electrolyte. This system was further applied in CPC, allowing an efficient separation of the five phenolic compounds after the optimization of the equipment operational conditions, while demonstrating the potential of polymer-based ABS to be used in liquid–liquid chromatography. Finally, the recovery of the phenolic compounds (between 65 and 87%) with high purity from the ABS phases was demonstrated, allowing the reuse of the ABS phase-forming components, which was proved to be of low environmental impact. In fact, in a scenario where the polymeric phases are reused, the carbon footprint is decreased to 36%, as the consumption of new chemicals and water reduces considerably. [ABSTRACT FROM AUTHOR]
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- 2018
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13. Association between polymorphism of tumor necrosis factor alpha (tnfα) in the region -308 g/a with tinnitus in the elderly with a history of occupational noise exposure.
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Marchiori, Luciana, Dias, Ana, Gonçalvez, Alyne, Poly-Frederico, Regina, Doi, Marcelo, Marchiori, Luciana L M, Dias, Ana C M, Gonçalvez, Alyne S, Poly-Frederico, Regina C, and Doi, Marcelo Y
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TINNITUS , *CYTOKINES , *NOISE-induced deafness , *TUMOR necrosis factors , *GENETIC polymorphisms , *AUDIOMETRY , *DISEASE susceptibility , *INTERVIEWING , *NOISE , *POLYMERASE chain reaction , *OCCUPATIONAL hazards , *ENVIRONMENTAL exposure , *CROSS-sectional method , *OLD age , *DISEASE risk factors - Abstract
Context: Tinnitus is a common disorder that occurs frequently across all strata of population and has an important health concern and is often associated with different forms of the hearing loss of varying severity.Aims: To investigate the association between the polymorphism of tumor necrosis factor alpha (TNFα) in the region -308 G/A with the susceptibility to tinnitus in individuals with the history of exposure to occupational noise.Settings and Design: This was a cross-sectional study with a sample of 179 independent elderly people above 60 years of age.Materials and Methods: Information on exposure to occupational noise was obtained by interviews. Audiological evaluation was performed using pure tone audiometry and genotyped through polymerase chain reaction by restriction fragment length polymorphism.Statistical Analysis Used: Data were analyzed using the chi-square test and the odds ratio (OR), with the significance level set at 5%.Results: Among elderly with tinnitus (43.01%), 33.76% had a history of exposure to occupational noise. A statistically significant association was found between genotype frequencies of the TNFα gene in the -308 G/A region and the complaint of tinnitus (P = 0.04 and χ2 = 4.19). The elderly with the G allele were less likely to have tinnitus due to occupational noise exposure when compared to those carrying the A allele (OR = 2.74; 95% CI: 1.56-4.81; P < 0.0005).Conclusion: This study suggests an association between the TNFα with susceptibility to tinnitus in individuals with a history of exposure to occupational noise. [ABSTRACT FROM AUTHOR]- Published
- 2018
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14. High-through identification of T cell-specific phage-exposed mimotopes using PBMCs from tegumentary leishmaniasis patients and their use as vaccine candidates against Leishmania amazonensis infection.
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Carvalho, Gerusa B., Costa, Lourena E., Lage, Daniela P., Ramos, Fernanda F., Santos, Thaís T. O., Ribeiro, Patrícia A. F., Dias, Daniel S., Salles, Beatriz C. S., Lima, Mariana P., Carvalho, Lívia M., Dias, Ana C. S., Alves, Patrícia T., Franklin, Michelle L., Silva, Renata A. M., Duarte, Mariana C., Menezes-Souza, Daniel, Roatt, Bruno M., Chávez-Fumagalli, Miguel A., Goulart, Luiz Ricardo, and Teixeira, Antonio L.
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T cells , *LEISHMANIASIS , *IMMUNE response , *TUMOR necrosis factors , *IMMUNOGLOBULIN G , *VACCINES - Abstract
In the current study, phage-exposed mimotopes as targets against tegumentary leishmaniasis (TL) were selected by means of bio-panning cycles employing sera of TL patients and healthy subjects, besides the immune stimulation of peripheral blood mononuclear cells (PBMCs) collected from untreated and treated TL patients and healthy subjects. The clones were evaluated regarding their specific interferon- γ (IFN- γ) and interleukin-4 (IL-4) production in the in vitro cultures, and selectivity and specificity values were calculated, and those presenting the best results were selected for the in vivo experiments. Two clones, namely A4 and A8, were identified and used in immunization protocols from BALB/c mice to protect against Leishmania amazonensis infection. Results showed a polarized Th1 response generated after vaccination, being based on significantly higher levels of IFN- γ , IL-2, IL-12, tumour necrosis factor- α (TNF- α) and granulocyte-macrophage colony-stimulating factor (GM-CSF); which were associated with lower production of specific IL-4, IL-10 and immunoglobulin G1 (IgG1) antibodies. Vaccinated mice presented significant reductions in the parasite load in the infected tissue and distinct organs, when compared with controls. In conclusion, we presented a strategy to identify new mimotopes able to induce Th1 response in PBMCs from TL patients and healthy subjects, and that were successfully used to protect against L. amazonensis infection. [ABSTRACT FROM AUTHOR]
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- 2019
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15. Leishmania infantum β-Tubulin Identified by Reverse Engineering Technology through Phage Display Applied as Theranostic Marker for Human Visceral Leishmaniasis.
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Costa, Lourena E., Alves, Patrícia T., Carneiro, Ana Paula, Dias, Ana C. S., Fujimura, Patrícia T., Araujo, Galber R., Tavares, Grasiele S. V., Ramos, Fernanda F., Duarte, Mariana C., Menezes-Souza, Daniel, Briza, Peter, Briza, Fátima F., Coelho, Eduardo A. F., and Goulart, Luiz Ricardo
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LEISHMANIA infantum , *LEISHMANIASIS , *IMMUNOPRECIPITATION , *IMMUNOBLOTTING , *SERODIAGNOSIS - Abstract
Two Leishmania infantum mimotopes (B10 and C01) identified by phage display showed to be antigenic and immunogenic for visceral (VL) and tegumentary (TL) leishmaniasis; however, their biological targets in the parasites have not been identified. The aim of the present study was to investigate the native antigens expressing both mimotopes, and to use them in distinct immunological assays. For this, a subtractive phage display technology was used, where a combinatorial library of single-chain variable fragments (scFv) was employed and the most reactive monoclonal antibodies for each target were captured, being the target antigens identified by mass spectrometry. Results in immunoblotting and immunoprecipitation assays showed that both monoclonal scFvs antibodies identified the β-tubulin protein as the target antigen in L. infantum. To validate these findings, the recombinant protein was cloned, purified and tested for the serodiagnosis of human leishmaniasis, and its immunogenicity was evaluated in PBMC derived from healthy subjects and treated or untreated VL patients. Results showed high diagnostic efficacy, as well as the development of a specific Th1 immune response in the cell cultures, since higher IFN-γ and lower IL-10 production was found. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
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