Miniaturization of aqueous two-phase extraction for biological applications

Aqueous two-phase extraction (ATPE) is a biocompatible liquid-liquid (L-L) separation technique that has been under research for several decades towards the purification of biomolecules, ranging from small metabolites to large animal cells. More recently, with the emergence of rapid-prototyping techniques for fabrication of microfluidic structures with intricate designs, ATPE gained an expanded range of applications utilizing physical phenomena occurring exclusively at the microscale. Studies of ATPSs at nanoliter-scale are further extending the range of applications of these systems by taking advantage of rapid diffusion times, increased degree of control of individual liquid streams and droplets, continuous flow and the integration of multi-dimensional separation modes. Several examples of microfluidic ATPS platforms are described. The partition of molecules between two co-flowing liquid streams confined within a microchannel was successfully demonstrated by the on-line extraction of a fluorescein isothiocyanate (FITC) labeled immunoglobulin G (IgG) from a salt rich flow to a PEG rich flow. IgG diffusion to the PEG-rich phase was complete after 16 cm of channel using flow rates of 1 and 0.2 μL/min for the salt and PEGrich phases respectively. Besides proteins, ATPS have also been used to separate other more complex biomolecules in microfluidics such as virus-like particles. The potential of miniaturization as a high-throughput screening tool has also been explored. The developed setup allowed the screening of a wide range of concentrations inside the microchannel by varying the flow rates of the solutions while using sub-mL volumes for each ATPS-forming system. As a novel demonstration of the integrative potential of ATPE as a microfluidic sample preparation module, a microfluidic device comprising two modules was developed and used to perform a complex matrix clean-up in-line with an immunoassay. References: Silva, D. F., Azevedo, A. M., Fernandes, P., Chu, V. et al., Design of a microfluidic platform for monoclonal antibody extraction using an aqueous two-phase system. J. Chromatogr. A 2012, 1249, 1–7. Jacinto, M. J., Soares, R. R. G., Azevedo, A. M., Chu, V. et al., Optimization and miniaturization of aqueous two phase systems for the purification of recombinant human immunodeficiency virus-like particles from a CHO cell supernatant. Sep. Purif. Technol. 2015, 154, 27–35. Silva, D. F. C., Azevedo, A. M., Fernandes, P., Chu, V. et al., Determination of aqueous two phase system binodal curves using a microfluidic device. J. Chromatogr. A 2014, 1370, 115–120. Soares, R. R., Novo, P., Azevedo, A. M., Fernandes, P. et al., On-chip sample preparation and analyte quantification using a microfluidic aqueous two-phase extraction coupled with an immunoassay. Lab Chip 2014, 14, 4284–429

Exercício de intensidade moderada durante a gravidez é seguro para o feto? Ensaio clínico aberto

To determine the effect of treadmill walking on maternal heart rate (MHR) and cardiotocographic parameters (basal fetal heart rate [FHR], active fetal movements [AFM], number of accelerations and decelerations, and short-term variation [STV] and long-term variation [LTV] of fetal heart rate) in pregnant women at 36 weeks. Methods A nonrandomized, open clinical trial involving 88 healthy pregnant women submitted to moderate intensity walking and computed cardiotocography in 3 20-minute periods (resting, treadmill walking, and postexercise recovery). Results The mean FHR decreased during walking (resting: 137 bpm; treadmill: 98 bpm; recovery: 140 bpm; p <0.001), with bradycardia occurring in 56% of the fetuses in the first 10minutes of exercise, and in 47% after 20minutes. Bradycardia was not detected in the other phases. The mean STV and HV were 7.9, 17.0, and 8.0 milliseconds ( p <0.001) and 7.6, 10.8 and 7.6 bpm ( p =0.002) in the resting, walking and recovery phases, respectively. The mean number of fetal movements in 1 hour was 29.9, 22.2 and 45.5, respectively, in the 3 periods ( p <0.001). In overweight/obese women, the mean FHR was lower ( p =0.02). Following the logistic regression analysis, two variables remained significantly associated with bradycardia: maternal fitness in the 28 (th) week of pregnancy (protective effect) and maternal weight (increased risk). Conclusion In healthy fetuses, physical exercise proved to be safe, since, although FHR and AFM decreased during treadmill walking, an increase in SVT and LTV was observed419531538CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQFUNDAÇÃO DE APOIO À PESQUISA DO ESTADO DA PARAÍBA - FAPESQUniversal 480761/2008-6PPSUS2009Determinar o efeito da caminhada em esteira sobre a frequencia cardiaca materna (FCM) e parametros cardiotocograficos (batimentos cardiofetais basais [BCFs], movimentos ativos fetais [MAFs], numero de aceleracoes e desaceleracoes e variabilidade de curta [STV] e longa [LTV] duracAo da frequencia cardiaca fetal) em gestantes na 36 (a) semana. Metodos Foi realizado um ensaio clinico nAo randomizado e aberto com 88 gestantes saudaveis submetidas a caminhada de moderada intensidade na esteira e a cardiotocografia computadoriza em 3 momentos de 20 minutos (antes, durante e apos a caminhada). Resultados A media dos BCFs diminuiu durante a caminhada, retornando a niveis previos (antes: 137 bpm; durante: 98 bpm; apos: 140 bpm; p <0,001), com bradicardia ocorrendo em 56% dos fetos nos primeiros 10 minutos do exercicio, e em 47% apos 20 minutos. A bradicardia fetal nAo foi observada em outros momentos (antes ou depois). As medias da STV e da LTV foram 7,9, 17,0 e 8,0 milissegundos ( p <0,001) e 7,6, 10,7 e 7,6 bpm ( p =0,002) antes, durante e apos a caminhada, respectivamente. A media dos numeros dos MAFs em 1 hora foi 29,9, 22,2 e 45,5, respectivamente, nos tres momentos ( p <0,001). Nas mulheres com sobrepeso/obesidade, a media da FCM foi menor ( p =0,02). Apos a analise de regressAo logistica, duas variaveis permaneceram significativamente associadas a bradicardia: aptidAo maternal na 28 (a) semana de gravidez (efeito protetor) e peso materno (aumento do risco). ConclusAo Em fetos saudaveis, o exercicio fisico mostrou-se seguro, uma vez que, embora os BCFs e os MAFs diminuam durante a caminhada na esteira, foi observado um aumento da SVT e da LT

School environment associates with lung function and autonomic nervous system activity in children : a cross-sectional study

Children are in contact with local environments, which may affect respiratory symptoms and allergic sensitization. We aimed to assess the effect of the environment and the walkability surrounding schools on lung function, airway inflammation and autonomic nervous system activity. Data on 701 children from 20 primary schools were analysed. Lung function, airway inflammation and pH from exhaled breath condensate were measured. Pupillometry was performed to evaluate autonomic activity. Land use composition and walkability index were quantified within a 500 m buffer zone around schools. The proportion of effects explained by the school environment was measured by mixed-effect models. We found that green school areas tended to be associated with higher lung volumes (FVC, FEV1 and FEF25-75%) compared with built areas. FVC was significantly lower in-built than in green areas. After adjustment, the school environment explained 23%, 34% and 99.9% of the school effect on FVC, FEV1, and FEF25-75%, respectively. The walkability of school neighbourhoods was negatively associated with both pupil constriction amplitude and redilatation time, explaining -16% to 18% of parasympathetic and 8% to 29% of sympathetic activity. Our findings suggest that the environment surrounding schools has an effect on the lung function of its students. This effect may be partially mediated by the autonomic nervous system.Peer reviewe

Access to dental services and related factors: a home-based population study

Objetivou-se identificar fatores associados à falta de acesso aos serviços odontológicos. Estudo transversal, amostra complexa probabilística por conglomerados em dois estágios de 857 participantes, com mais de 18 anos em um município de grande porte populacional. Foram feitas análises múltiplas através da regressão logística e multivariada em árvores de decisão. Considerou-se como variável dependente o acesso aos serviços odontológicos. Identificou-se que 10,3% não obtiveram acesso. Nas análises múltipla e multivariada constataram-se associação com a idade, na regressão logística constatou-se maior chance de falta de acesso a cada ano de idade incrementado, entre aqueles com menor renda per capita e entre os que classificaram a aparência dos dentes e gengivas como “regular/ruim/péssima”. A falta de acesso aos serviços odontológicos é maior entre os mais vulneráveis socialmente. Há necessidade de incremento na alocação de recursos públicos que promovam educação em saúde e gere conhecimento sobre como acessar os serviços quando necessitar, tendo em foco os cuidados odontológicos como um direito humano e que possibilitem que não ocorra a falta de acesso à medida que os usuários envelhecem ou entre os com baixa renda e também os insatisfeitos com a aparência bucal24310211032The scope of this study was to identify factors associated with lack of access to dental services. It involved a cross-sectional study and a probabilistic complex sample by conglomerates in two stages with 857 participants over 18 years of age in a large city. Multiple analyses by means of logistic and multivariate regression in decision trees were made. The lack of access to dental services was considered a dependent variable. It was identified that 10.3% did not have access. In the multiple and multivariate analyses an association with age was verified and in the logistic regression a greater possibility of lack of access was found for each year of increased age, among those with the lowest per capita income and those who ranked appearance of teeth and gums as “fair/poor/very poor.” The lack of access to dental services was greater among the most socially vulnerable. There is a pressing need to increase the allocation of public resources to promote health education and provide knowledge about how to access services when they are needed, focusing on dental care as a human right and ensuring that lack of access does not occur as users get older or among those with low income and also those dissatisfied with their oral appearanc

COFFEE: A HEALTH FUEL-BLOT POPULAR DRINKING

Now, the days begin with cups of coffee worldwide. Caffeine is the main component of coffee, which is vastly consumed as a psychoactive agent, and in varieties of dietary supplements. Day by day coffee and caffeinated-consumption areas are expanding. Only a single cup of coffee contains thousands of biochemical. Otherwise, during roasting, some of which turn to convert other chemicals moieties. Thus, the coffee is an interesting item to the drug scientists. Upon this jackpot, a number of researches have been done on coffee and its chemical components; in which many postulations are still in contentious and some are unclear to the coffee users. Upon going through the stand-point, this study has been snapshot to sketch a complete overview on coffee and its components. Our finding depicts constituents of coffee to have antioxidant, anti-inflammatory, anti-Alzheimer's disease, anti-Parkinson's disease, and cardioprotective activities. But the anti-cancerous effect of coffee components is not clear yet. In conclusion, coffee, and its constituents are in important in phytopharmacological research.Keywords: Coffee, Coffee components, Health-effect

miRNA-31 Improves Cognition and Abolishes Amyloid-beta Pathology by Targeting APP and BACE1 in an Animal Model of Alzheimer's Disease

Alzheimer's disease (AD) is the most common form of dementia worldwide, characterized by progressive memory impairment, behavioral changes, and, ultimately, loss of consciousness and death. Recently, microRNA (miRNA) dysfunction has been associated with increased production and impaired clearance of amyloid-β (Aβ) peptides, whose accumulation is one of the most well-known pathophysiological markers of this disease. In this study, we identified several miRNAs capable of targeting key proteins of the amyloidogenic pathway. The expression of one of these miRNAs, miR-31, previously found to be decreased in AD patients, was able to simultaneously reduce the levels of APP and Bace1 mRNA in the hippocampus of 17-month-old AD triple-transgenic (3xTg-AD) female mice, leading to a significant improvement of memory deficits and a reduction in anxiety and cognitive inflexibility. In addition, lentiviral-mediated miR-31 expression significantly ameliorated AD neuropathology in this model, drastically reducing Aβ deposition in both the hippocampus and subiculum. Furthermore, the increase of miR-31 levels was enough to reduce the accumulation of glutamate vesicles in the hippocampus to levels found in non-transgenic age-matched animals. Overall, our results suggest that miR-31-mediated modulation of APP and BACE1 can become a therapeutic option in the treatment of AD.This work was financed by the European Regional Development Fund (ERDF), through the Centro 2020 Regional Operational Programme under the project CENTRO-01-0145-FEDER-000008: BrainHealth 2020 and through the COMPETE 2020 - Operational Programme for Competitiveness and Internationalisation and Portuguese national funds via FCT - Fundação para a Ciência e a Tecnologia, under the project POCI- 01-0145-FEDER-007440 (reference UID/NEU/04539/2013). This work was also supported by the FCT Investigator Programme (IF/ 00694/2013 to J.P.), the Marie Curie Carrier Integration Grant (PCIG13-GA-2013-618525 to J.P.), HEALTHYAGING 2020 (CENTRO-01-0145-FEDER-000012 to A.T.B.-V.), and Bial Foundation Grant 264/16. A.T.B.-V., J.G., and A.L.C. are recipients of fellowships from the FCT (Grants PTDC/BIM-MEC/0651/2012, SFRH/ BPD/120611/2016, and SFRH/BPD/108312/2015)

Antioxidant, anti-acetylcholinesterase and cytotoxic activities of ethanol extracts of peel, pulp and seeds of exotic Brazilian fruits Antioxidant, anti-acetylcholinesterase and cytotoxic activities in fruits

Ethanol extracts of powdered genipap (Genipa americana L), umbu (Spondia tuberosa A.) and siriguela (Spondia purpurea L) prepared from separate pulp, seeds and peel were investigated for their (i) antioxidant capacity, which was evaluated by various known methods; (ii) acetylcholinesterase (AChE) inhibitory activity; and (iii) cytotoxic effect on corneal epithelial cells of sheep. the highest values of total phenolic content were obtained with peel and seed extracts. Siriguela and umbu (seeds and peel) extracts displayed the highest antioxidant activities. Lipid peroxidation assays using mimetic biomembranes and mouse liver homogenates indicated that genipap pulp is a promising antioxidant. the investigation of phenols and organic acid contents revealed the presence of quercetin, citric and quinic acids, chlorogenic acid derivatives, among others, in several extracts, with the highest amount found in siriguela seeds. Genipap pulp and siriguela seed ethanol extracts presented an AChE inhibition zone similar to that of the positive control, carbachol. AChE inhibition assay with chlorogenic acid, one of the main constituents of siriguela seeds, revealed that this acid showed activity similar to that of the control physostigmine. These data suggest that these extracts are potentially important antioxidant supplements for the everyday human diet, pharmaceutical and cosmetic industries. (C) 2012 Elsevier B.V. All rights reserved.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)INCT-RedoxomaINCT-BioanaliticaUniv Fed Alagoas, Inst Quim & Biotecnol, BR-57072970 Maceio, AL, BrazilUniv Pernambuco, BR-56300000 Petrolina, PE, BrazilUniv Fed Alagoas, Rede Nordeste Biotecnol RENORBIO, BR-57072970 Maceio, AL, BrazilUniv Fed Alagoas, Inst Ciencias Biol & Saude, BR-57072970 Maceio, AL, BrazilUniv São Paulo, Inst Quim, Dept Bioquim, BR-05508000 São Paulo, BrazilUniversidade Federal de São Paulo, Inst Ciencias Ambientais Quim & Farmaceut, BR-09972270 Diadema, SP, BrazilUniv Estadual Campinas, Inst Biol, Dept Biol Vegetal, Campinas, SP, BrazilUniv Fed Ceara, Dept Quim Organ & Inorgan, BR-60021970 Fortaleza, CE, BrazilEmbrapa Tabuleiros Costeiros, Tabuleiro Martins, BR-57061970 Maceio, AL, BrazilUniversidade Federal de São Paulo, Inst Ciencias Ambientais Quim & Farmaceut, BR-09972270 Diadema, SP, BrazilFAPESP: 2008/58035-6Web of Scienc

Contribution of wildlife governmental centers to \ud conservation and biological study of sloths Bradypus \ud variegatus

Our study evaluated the anthropogenic threats to the common sloth in the Atlantic forest (AF) by analyzing data collected in wildlife governmental centers. Interesting, new biological data could also be reported. The main risks identified were the road network and falls, followed by domestic dogs and electric shocks. Tourism and preliminary evidences of locally distinct biological features were suggested as potential menaces to conservation of the AF common sloth populations. The birth season within the southeast appears to be the rainy season, but that is not the case in the northeast. Finally, the weight of adult individuals was reported not to vary between sexes. Data were confirmed by the highest number of individuals ever analyzed. Thus, although neotropical wildlife governmental centers usually cope with the lack of human resources, this study highlights that their records can be successfully used to add important biological information to the conservation and study of neotropical wildlife.The authors would like to thank to Camila L. Clozato, all the CETAS and DEPAVE-SP staff, and their collaborators, namely the Fire and Police Departments, health and environmental institutions, and citizens, who have been collecting animals and data across the years. We also acknowledge two anonymous reviewers for their helpful comments on an early version of this manuscript. S.M. Silva was supported by a Foundation for Science and Technology PhD grant (SFRH/ BD/40638/2007). N. Moraes-Barros was supported by CAPES. J. S. Morgante had a research grant from FAPESP (08/52207-0)

Adaptación cultural de la versión brasileña del Godin-Shephard Leisure-Time Physical Activity Questionnaire

OBJETIVO: Realizar adaptação cultural para versão brasileira do questionário de atividade física no tempo de lazer e avaliar a validade de conteúdo, praticabilidade, aceitabilidade e confiabilidade. MÉTODOS: Foram realizadas as etapas de tradução, síntese, retrotradução, avaliação por comitê de especialistas e pré-teste, seguidos pela avaliação da praticabilidade, aceitabilidade e confiabilidade (teste-reteste). Os juízes avaliaram as equivalências semântico-idiomática, conceitual, cultural e metabólica. A versão adaptada foi submetida ao pré-teste (n = 20) e teste-reteste (n = 80) em indivíduos saudáveis e pacientes com doenças cardiovasculares, em Limeira, SP, entre 2010 e 2011. A proporção de concordância do comitê de juízes foi quantificada por meio do Índice de Validade de Conteúdo. A confiabilidade foi avaliada segundo critério de estabilidade, com intervalo de 15 dias entre as aplicações, a praticabilidade pelo tempo gasto na entrevista e a aceitabilidade pelo percentual de itens não respondidos e proporção de pacientes que responderam a todos os itens. RESULTADOS: A versão traduzida do questionário apresentou equivalências semântico-idiomática, conceitual, cultural e metabólica adequadas, com substituição de algumas atividades físicas mais adequadas para a população brasileira. A análise da praticabilidade evidenciou curto tempo de aplicação do instrumento (média de 3,0 min). Quanto à aceitabilidade, todos os pacientes responderam a 100% dos itens. A análise do teste-reteste sugeriu estabilidade temporal do instrumento (Índice de Correlação Intraclasse = 0,84). CONCLUSÕES: A versão brasileira do questionário apresentou propriedades de medida satisfatórias. Recomenda-se sua aplicação a populações diversas em estudos futuros, a fim de disponibilizar propriedades de medida robustas.OBJETIVO: Realizar adaptación cultural para la versión brasileña del cuestionario de actividad física en el tiempo de descanso y evaluar la validez del contenido, factibilidad, aceptabilidad y confiabilidad. MÉTODOS: Se realizaron las etapas de traducción, síntesis, retro-traducción, evaluación por comité de especialistas y pre-prueba, seguidos por la evaluación de la factibilidad, aceptabilidad y confiabilidad (prueba- re-prueba). Los árbitros evaluaron las equivalencias semántico-idiomática, conceptual, cultural y metabólica. La versión adaptada fue sometida a la pre-prueba (n=20) y prueba-re-prueba (n=80) en individuos saludables y pacientes con enfermedades cardiovasculares, en Limeira, SP, Brasil, entre 2010 y 2011. La proporción de concordancia del comité de árbitros fue cuantificada por medio de índice de validez de contenido. La confiabilidad fue evaluada según criterio de estabilidad, con intervalo de 15 días entre las aplicaciones, la factibilidad por el tiempo de gasto en la entrevista y la aceptabilidad por el porcentaje de ítems no respondidos y proporción de pacientes que respondieron a todos los ítems. RESULTADOS: Las versión traducida del cuestionario presentó equivalencias semántico-idiomática, conceptual, cultural y metabólica adecuadas, con sustitución de algunas actividades físicas más adecuadas para la población brasileña. El análisis de la factibilidad presentó corto tiempo de aplicación del instrumento (promedio de 3,0 min). Con relación a la aceptabilidad, todos los pacientes respondieron a 100% de los itens. El análisis de la prueba-re-prueba sugirió estabilidad temporal del instrumento (índice de correlación intraclase = 0,84). CONCLUSIONES: La versión brasileña del cuestionario presentó propiedades de medida satisfactorias. Se recomienda su aplicación a poblaciones diversas en estudios futuros, a fin de dejar disponibles las propiedades de medida robustas.OBJECTIVE: To conduct the cultural adaptation of the Brazilian version of the Godin-Shephard Leisure-Time Physical Activity Questionnaire (GSLTPAQ) and to assess its content validity, practicability, acceptability and reliability. METHODS: The stages of translation, synthesis, back translation, expert committee review and pre-test were carried out, followed by the evaluation of the practicability, acceptability and reliability (test-retest). The judges assessed its semantic, idiomatic, conceptual, cultural and metabolic equivalences. The adapted version was submitted to the pre-test (n = 20), and test-retest (n = 80), in healthy individuals and in those suffering from cardiovascular disease in Limeira, SP, Southeastern Brazil, between 2010 and 2011. The proportion of agreement of the committee of judges was assessed using the Content Validity Index. Reliability was assessed by the criterion of stability, with 15 days between applications. Practicability was evaluated by the time spent interviewing and acceptability was estimated as the percentage of unanswered items and the proportion of patients who responded to all items. RESULTS: The translated version of the questionnaire showed evidence of appropriate semantic-idiomatic, conceptual, cultural and metabolic equivalence, with substitutions of several physical activities more appropriate to the Brazilian population. The practicability analysis showed short time needed for the application of the instrument (mean 3.0 minutes). As for acceptability, all patients answered 100% of the items. The test-retest analysis suggested that stability was good (Intraclass Correlation Coefficient value of 0.84). CONCLUSIONS: The Brazilian version of the questionnaire showed satisfactory measures of the qualities in question. Its application to diverse populations in future studies is recommended in order to provide robust measures of these qualities