Reconstructing the Density of States by History-Dependent Metadynamics

We present a novel method for the calculation of the energy density of states D(E) for systems described by classical statistical mechanics. The method builds on an extension of a recently proposed strategy that allows the free energy profile of a canonical system to be recovered within a pre-assigned accuracy,[A. Laio and M. Parrinello, PNAS 2002]. The method allows a good control over the error on the recovered system entropy. This fact is exploited to obtain D(E) more efficiently by combining measurements at different temperatures. The accuracy and efficiency of the method are tested for the two-dimensional Ising model (up to size 50x50) by comparison with both exact results and previous studies. This method is a general one and should be applicable to more realistic model systems

Translation and cultural validation of the Lequesne's algofunctional questionnaire for osteoarthritis of knee and hip for portuguese language

OBJECTIVE: translation, validation and cultural adaptation of the algofunctional index of Lequesne for the Portuguese language. PATIENTS AND METHODS: the original questionnaire, published in English, was translated by three English professors, and after back translated by others three professors. Four physiotherapists had congregated and comparing the translations, they had elaborated the first version. This was applied to 11 patients with diagnosis of osteoarthritis (OA) of knees and/or hips, and this version was recognized fully by the patients, who had not reported understanding problems. This version, considered as definitive, was them applied to others 73 patients in the following form: a time for observer 1 (A1), in the same day for observer 2 (A2) and one week later for observer 1 or 2 (A3). Jointly it was collected data on age, sex and applied questionnaire Westerm Ontario and McMaster Universities WOMAC in its version validated for Portuguese language. RESULTS: for osteoarthritis of knees, the patients of the pre-test (10 of 11 evaluated) had average age of 63 years (DP=9.3) and 2 were men. Its average grade for the index of Lequesne was of 14.9 (DP=5.1). The patients, evaluated with the definitive version, in number of 42, had in average 67.5 years (DP=8.7), 5 were men. Its average for the index of Lequesne was for A1=11.9 (DP=5.0), for A2=12.1 (DP=6.4) and A3=11.3 (DP=7.9). The intraclass correlation between A1 and A2 was of 0.99 and between A1 and A3 was of 0,99. The Pearson coefficient between A1 and WOMAC pain was of 0.800, WOMAC stiffness was of 0.640 and WOMAC function was 0.828, all with statistical significance. For osteoarthritis of hips, the patients of the pre-test (3 of the 11 evaluated) had average age of 67 years (DP=9.18) and were all women. Its average grade for the index of Lequesne was of 11.2 (DP=5.86). The patients, evaluated with the definitive version, in number of 37, had in average 66.9 years (DP=9.01), 8 were men. Its average for the index of Lequesne was for A1=12.5 (DP=5.6) for A2=12,5 (DP=5.7) and A3=14.1 (DP=6.3). The intraclass correlation between A1 and A2 was of 0.99 and between A1 and A3 was of 0.98. The coefficient of Pearson between A1 and WOMAC pain was of 0.759, WOMAC rigidity was of 0.659 and WOMAC function was 0.851, all also with statistical significance. CONCLUSION: the version for the Portuguese language (Brazil) of the algofunctional index of Lequesne, for evaluation of osteoarthritis of knees and hips is validated for use in Brazilian population.OBJETIVO: tradução, validação e adaptação cultural do índice algofuncional de Lequesne para a língua portuguesa. PACIENTES E MÉTODOS: o questionário original, publicado em língua inglesa, foi traduzido por três professores de inglês, e após, retraduzidos por outros três professores. Quatro fisioterapeutas se reuniram e, comparando as traduções, elaboraram a primeira versão. Esta foi aplicada a 11 pacientes com diagnóstico de osteoartrite de joelhos e/ou quadris, e esta versão foi reconhecida plenamente pelos pacientes, que não relataram problemas de compreensão. Passou-se, então, a aplicar esta versão, considerada como definitiva, a outros 73 pacientes da seguinte forma: uma vez pelo observador 1 (A1), no mesmo dia pelo observador 2 (A2) e uma semana mais tarde pelo observador 1 ou 2 (A3). Conjuntamente, colheu-se dados sobre idade e sexo, e aplicou-se o questionário Westerm Ontário and McMaster Universities (WOMAC) em sua versão validada para língua portuguesa. RESULTADOS: para osteoartrite de joelhos, os pacientes do pré-teste (10 de 11 avaliados) tinham idade média de 63 anos (DP=9,3) e dois eram homens. Sua nota média para o índice de Lequesne foi de 14,9 (DP=5,1). Os pacientes, avaliados com a versão definitiva, em número de 42, tinham em média 67,5 anos (DP=8,7), cinco eram homens. Sua média para o índice de Lequesne foi para A1=11,9 (DP=5,0), para A2=12,1 (DP=6,4) e A3=11,3 (DP=7,9). A correlação intraclasse entre A1 e A2 foi de 0,99 e entre A1 e A3 foi de 0,99. O coeficiente de Pearson entre A1 e WOMAC dor foi de 0,800, WOMAC rigidez foi de 0,640 e WOMAC função foi 0,828, todas estatisticamente significantes. Para osteoartrite de quadris, os pacientes do pré-teste (3 dos 11 avaliados) tinham idade média de 67 anos (DP=9,18) e todos eram mulheres. Sua nota média para o índice de Lequesne foi de 11,2 (DP=5,86). Os pacientes, avaliados com a versão definitiva, em número de 37, tinham em média 66,9 anos (DP=9,01), 8 eram homens. Sua média para o índice de Lequesne foi para A1=12,5 (DP=5,6), para A2=12,5 (DP=5,7) e A3=14,1 (DP=6,3). A correlação intraclasse entre A1 e A2 foi de 0,99 e entre A1 e A3 foi de 0,98. O coeficiente de Pearson entre A1 e WOMAC dor foi de 0,759, WOMAC rigidez foi de 0,659 e WOMAC função foi 0,851, todas também estatisticamente significantes. CONCLUSÃO: a versão em língua portuguesa (Brasil) do índice algofuncional de Lequesne, para avaliação de osteoartrite de joelhos e quadris está validada para uso em população brasileira.Universidade Federal de São Paulo (UNIFESP)Centro Universitário das Faculdades Metropolitanas UnidasUNIFESP, EPMSciEL

Relationship between muscle strength and motor function in Duchenne muscular dystrophy

Measuring muscle strength and motor function is part of Duchenne muscular dystrophy (DMD) assessment. However, the relationship between these variables is controversial. Objective: To investigate the relationship between muscle strength and motor function and between these variables and age. Method: Muscle strength was measured by Medical Research Council (MRC) scale and motor function, by Motor Function Measure (MFM), in 40 non-ambulatory patients. Spearman tests investigated the relationships between muscle strength, motor function and age. Results: Total MRC and MFM scores were strongly related to each other (r = 0.94p 0.05). Strong and moderate relationships between partial muscle strength and motor function scores were found. Higher correlation coefficients were found between total scores and Dimensions 2 (axial/ proximal control) and 3 (distal control) of MFM. Conclusion: Muscle strength and motor function are strongly correlated and seem to decrease proportionally in DMD.Univ Sao Paulo, Fac Med, Dept Fisioterapia Fonoaudiol & Terapia Ocupac, Sao Paulo, SP, BrazilUniv Fed Sao Paulo, Fac Med, Dept Neurol, Sao Paulo, SP, BrazilUniv Fed Sao Paulo, Fac Med, Dept Neurol, Sao Paulo, SP, BrazilWeb of Scienc

Portable dehumidifiers as an original matrix for the study of inhalable nanoparticles in school

Good air quality is documented as a significant factor of social justice. The human health hazards associated with air pollution are not distributed equally across cities; the most vulnerable people are more exposed to ambient air as they commute to work and wait for buses or trains at the stations. Aerosols play important roles in atmosphere quality and the climate; their oxidation at the nanoscale level may possibly increase the reactivity and toxicity of atmospheric particulates. Indoor school environments are characterized by high concentrations of different airborne particulate and gaseous pollutants. The documentation of nanoparticles (NPs), ultra-fine particles (UFPs), and micron-size particle species present in indoor primary schools are an important aspect in the recognition of their influence in respirational difficulties and decreased cognitive progress in children. This work utilizes the study of condensed water, sampled with portable dehumidifiers (PD), to describe NPs and UFPs in the vapor stage of enclosed zones. The acquired extracts were analyzed by advanced electron microscopy techniques. A total of 392 NPs and 251 UFPs were examined in a set of 22 samples acquired in moderately limited or inadequately ventilated indoor areas from several schools. Noting that NPs-related disorders happen at particular places of respirational structure, identification of site-specific NPs accumulation should be anticipated in direction to better verify the corresponding human health outcomes resulting from respirable NPs

Brewers' spent grain as substrates for production of cellulolytic and hemicellulolytic enzymes by different Aspergillus species

Brewers' spent grain (BSG) is the major by-product of the brewing industry, representing around 85% of the total by-products generated. It is a lignocellulosic material containing about 38% cellulose, 29% hemicellulose, chiefly arabinoxylans, and 13% lignin. The production of cellulolytic and hemicellulolytic enzymes using this material as substrate represent an eco-friendly strategy for the lignocellulosic biomass hydrolysis, generating fermentable sugars that can be converted into high- added value products, such as bioethanol, lactic acid, xylitol and others. Thus, this work aimed to evaluate the potential of cellulolytic and hemicellulolytic enzymes production by some Aspergillus species cultivated in BSG. Fungi were grown in minimum media, pH 6.5, with 1% BSG and inoculum was done with 107spores/mL, cultivated at 30°C, 120 rpm, for 5 days. Every 24 hours 2 mL of the samples were collected. The enzymatic activity was performed after the incubation of the crude extract with 1% Linear arabinan, Xylan from beechwood, Xyloglucan, Locust bean gum and CMC, at 50°C for 60 minutes and the reducing sugars were determined using dinitrosalicylic acid (DNS). Synthetic substrates (2 mM of PNP--L-arabinofuranoside, PNP--D-xylopyranoside, PNP--Dglucopyranoside and PNP--D-cellobioside) were also used at the same conditions. The extract from A. niveus showed the best arabinanase (0.284 U/mL) and -glucosidase (0.126 U/mL) activities after 48 and 96 hours of cultivation, respectively. On the other hand, the extract from A. brasiliensis presented the best activities of -L-arabinofuranosidase (0.129 U/mL), -xylosidase (0.265 U/mL) and xylanase (2.15 U/mL) when cultivated for 48 hours. After 72 hours, this fungus also showed the best activities for xyloglucanase (1.06 U/mL), mannanase (0.617 U/mL) and endoglucanase (0.254 U/mL). The extract produced by A. flavus presented the best cellobiohydrolase activity with 0.113 U/mL after 120 hours of cultivation. It is important to mention that A. awamori, A. clavatus and A. terreus also showed good levels of different enzymes produced but they were not the best producers. These data suggest the great potential of different cellulolytic and hemicellulolytic enzymes production using BSG as substrate, which represents an eco-friendly destination for the residues and can generate high-added value products with great biotechnological application.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Institutos Nacionais de Ciência e Tecnologia (INCT)Fundação para a Ciência e a Tecnologia (FCT)info:eu-repo/semantics/publishedVersio

Production of an enzymatic cocktail by Aspergillus awamori grown on corn straw with stirred tank bioreactor

Increased agroindustrial activity has led to the accumulation of large amounts of lignocellulosic material (LCM). LCM is nature's most abundant source of renewable carbon, representing a valuable industrial substrate with potential for many applications. Thus, the objective of this work was to screen for different holocellulases and to analyze the production of an Aspergillus awamori enzyme cocktail grown in corn straw (CS) using a stirred tank bioreactor. Screening was performed with A. clavatus, A. flavus, A. terreus, A. niveus, A. awamori and A. brasiliensis cultivated in minimal medium (MM), with 1% CS at 30 ºC or 37 ºC (for A. niveus), 120 rpm, for 5 days. Xylanase (XYN) and endoglucanase (EG) activities were evaluated by formation of reducing sugars using dinitrosalicylic acid (DNS). Cellobiohydrolase (CBH), -glucosidase (BGL) and -xylosidase (BXL) were determined by cleavage of PNP--D-cellobioside, PNP--D-glucopyranoside and PNP--D- xylopyranoside, respectively. After screening, a pre-inoculum was prepared with the best enzyme producer using a 500 mL MM flask and incubated at 30 °C, 120 rpm for 48 hours. The increase of enzyme production was performed in a Benchtop BioFlo 310 bioreactor, with 4.5 L of MM and 1% of CS, and was then inoculated the best enzyme producer. Cultivation was performed at 30 °C, pH 6.5, 275 rpm, air flow 2 v.v.m., for 5 days. During the screening, all fungi presented EG, CBH, BGL, XYN and BXL activities. However, A. awamori was chosen to continue the experiments because of its BXL activity which was 12.6 times higher than that produced by A. niveus. At the scale-up stage, XYN production (47.80 U/ mL) increased 4.1-fold compared to flask activity (11.52 U/mL). BXL also showed 1.6-times higher activity, as well as EG, CBH and BGL, which improved 2.3, 3.3 and 1.2 times their activities, respectively. It was concluded that the staggering of cocktail production improved the enzymatic activities and that corn straw is an excellent source of induction. Furthermore, this cocktail has the potential to be applied in the hydrolysis of different LCM due to the range of holocellulases present.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Institutos Nacionais de Ciência e Tecnologia (INCT)Fundação para a Ciência e a Tecnologia (FCT)info:eu-repo/semantics/publishedVersio

Immobilization and application of the recombinant xylanase GH10 of Malbranchea pulchella in the production of xylooligosaccharides from hydrothermal liquor of the eucalyptus (Eucalyptus grandis) wood chips

Xylooligosaccharides (XOS) are widely used in the food industry as prebiotic components. XOS with high purity are required for practical prebiotic function and other biological benefits, such as antioxidant and inflammatory properties. In this work, we immobilized the recombinant endo-1,4-β-xylanase of Malbranchea pulchella (MpXyn10) in various chemical supports and evaluated its potential to produce xylooligosaccharides (XOS) from hydrothermal liquor of eucalyptus wood chips. Values >90% of immobilization yields were achieved from amino-activated supports for 120 min. The highest recovery values were found on Purolite (142%) and MANAE-MpXyn10 (137%) derivatives, which maintained more than 90% residual activity for 24 h at 70 °C, while the free-MpXyn10 maintained only 11%. In addition, active MpXyn10 derivatives were stable in the range of pH 4.0–6.0 and the presence of the furfural and HMF compounds. MpXyn10 derivatives were tested to produce XOS from xylan of various sources. Maximum values were observed for birchwood xylan at 8.6 mg mL−1 and wheat arabinoxylan at 8.9 mg mL−1, using Purolite-MpXyn10. Its derivative was also successfully applied in the hydrolysis of soluble xylan present in hydrothermal liquor, with 0.9 mg mL−1 of XOS after 3 h at 50 °C. This derivative maintained more than 80% XOS yield after six cycles of the assay. The results obtained provide a basis for the application of immobilized MpXyn10 to produce XOS with high purity and other high-value-added products in the lignocellulosic biorefinery field.The authors gratefully acknowledge FAPESP (São Paulo Research Foundation, grants No: 2018/07522-6) and FCT (POCI-01-0145-FEDER-032206)—transnational cooperation project EcoTech, and National Institute of Science and Technology of Bioethanol, INCT, CNPq 465319/2014-9/FAPESP n ◦ 2014/50884- 5) for financial support. Research scholarships were granted to RCA, DA, and JCSS by FAPESP (Grant No: 2020/00081-4, 2020/15510-8, and 2019/21989-7, respectively), to CCVD and VEP by CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, Finance Code 001).info:eu-repo/semantics/publishedVersio