Recycling of cellulases in lignocellulosic hydrolysates using alkaline elution

The recovery of cellulases from lignin, lignocellulosic hydrolysates and cellulose by alkaline washes at pH 9 and 10 was examined. The effect of the pH on the structural stability of purified Cel7A was analyzed by circular dichroism. Purified Cel7A showed conformational changes at pH 9 and 10 that were reversible at pH 4.8. Temperature influenced the enzymatic hydrolysis of wheat straw and may be critical for the efficiency of cellulase recycling from wheat straw hydrolysates. Operation at moderate temperatures (37 °C) resulted in a rate of saccharification 19% higher than that obtained at 50° C, improving cellulase recycling by 49%. Over 60% of the enzyme activity on the synthetic substrate 4-methylumbelliferyl-β-d-cellobioside (MUC) may be recovered by using a simple alkaline wash. This is thus a promising strategy for enzyme recycling that is simple to implement at industrial scale, economical and effective.The authors acknowledge funding through FP7 KACELLE (Kalundborg Cellulosic Ethanol) project for supporting his work. We also thank Dr. Mai Ostergaard Haven for critical reviewing of the manuscript and Dr. Jane Lindedam for supplying the lignin

Evaluation of the biocompatibility of root canal sealers using subcutaneous implants

The purpose of this study was to evaluate in vivo the biocompatibility of Endométhasone, Pulp Canal Sealer EWT and AH-Plus root canal sealers after implantation in rat connective tissue. Twenty-four Wistar-Furth rats were used. Polyethylene tubes were filled with the sealers and implanted into specific dorsal subdermal tissue sites of the rats. Implants were removed after 3, 7 and 30 days, fixed and processed for glycol methacrylate-embedding technique to be examined microscopically. On the 3rd day, there was a mild inflammatory reaction to Pulp Canal Sealer EWT implants, but a severe response to the other sealers with presence of acute inflammatory cells. On the 7th day, tissue organization was more evident with attenuation of the inflammatory reaction, especially for the AH-Plus implants. On the 30th day, connective tissue with few inflammatory cells was observed in contact with all sealer implants. In this time interval, the tissue in contact with Pulp Canal Sealer EWT implants was more organized, while the tissue close to Endométhasone and AH-Plus implants showed a mild persistent inflammatory reaction and had similar results to each other. In conclusion, the sealers had a similar pattern of irritation, which was more severe in the beginning and milder with time, in such a way that all sealers showed a persistent mild reaction. Pulp Canal Sealer EWT yielded better tissue organization than Endométhasone and AH-Plus, which, in turn, showed similar results to each other

Quantitative And Qualitative Analysis Of Microorganisms In Root-filled Teeth With Persistent Infection: Monitoring Of The Endodontic Retreatment.

The aim of this study was to investigate in vivo microorganisms detected in root-filled teeth with post-treatment apical periodontitis and quantify colony-forming units (CFU) during endodontic retreatment. Fifteen root-filled teeth had their previous gutta-percha removed and were randomly instrumented before being divided into three groups and medicated with either [Ca(OH)2 + 2% CHX gel], [Ca(OH)2 + 0.9% NaCl] or 2% CHX gel. Samples were taken after removal of gutta-percha (S1), after chemomechanical preparation using 2% CHX gel (S2), and after inter-appointment dressing (S3) for 7 or 14 days later. Cultivable bacteria recovered from infected root canals at the three stages were counted and identified by means of culture and PCR assay (16S rDNA). Quantitative data were statistically analyzed by using Mann-Whitney test in which pairs of groups were compared (P < 0.05). CFU counts decreased significantly from S1 to S2 (P < 0.05). No significant difference was found between S2 and S3 (P = 0.3093) for all three experimental groups. Chemomechanical preparation and intra-canal dressing promoted significant median reductions of 99.61% and 99.57%, respectively, in the number of bacteria compared to S1 samples. A total of 110 cultivable isolates were recovered by culture technique from 32 different species and 7 different genera. Out of the 13 target species-specific primer of bacteria analyzed, 11 were detected during endodontic retreatment. The great majority of taxa found in post-treatment samples were Gram-positive bacteria, although Gram-negative bacteria were found by molecular methods. Moreover, our results showed that gutta-percha removal and chemomechanical preparation are effective for root canal disinfection, whereas additional intra-canal dressing did not improve disinfection.7302-

Ativação alcalina de cinzas volantes utilizando solução combinada de NaOH e Ca(OH)2

Este trabalho teve como objetivo avaliar o comportamento da resistência à compressão, ao longo do tempo, em amostras de argamassas à base de cinzas volantes álcali-ativadas. Como ativador alcalino foi utilizada uma solução combinada de NaOH e Ca(OH)2. A cura das amostras foi realizada de duas formas distintas. Na primeira, as amostras foram mantidas em estufa à 70°C/12 hs e à temperatura constante de 22°C em sala climatizada até a idade de ensaio. Apesar de serem obtidas resistências da ordem de 20 MPa nas primeiras 24 hs, houve decréscimo da resistência a partir da idade de 7 dias. Verificado esse comportamento, um segundo método de cura foi adotado. Para isso, o traço utilizado para o preparo de novas argamassas foi o mesmo adotado no método anterior. Porém, foram submetidas à temperatura constante de 70°C até a idade de ensaio. Nesse caso, as amostras apresentaram decréscimo mais acentuado da resistência em menor espaço de tempo, quando comparado às amostras curadas no primeiro método. Como estudos complementares foram realizadas análises microestruturais da CV antes e depois da ativação, utilizando o MEV/EDS.The main objective of this work was to evaluate the development of the compressive strength, along time, in alkali-activated fly ash mortars. As alkaline activator a combined solution of NaOH and Ca(OH)2 was used. The cure of the samples was carried through two distinct forms. In the first one, samples were kept in oven at 70°C during the first 12 hours and at constant room temperature of 22°C afterwards, until the age of testing. Compressive strength of 20 MPa magnitude were achieved during the first 24 hours, however it started to decrease after the age of 7 days. Considering this behaviour, second method of curing was adopted for the same mixing proportions. However, mortar samples were kept in oven at 70°C until the age of testing. In this case, compressive strength results decreased more significantly and at earlier ages, when compared with the results of samples cured using the first method. As complementary to understand the decreasing in compressive strength, microstructural analyses of the fly ash, before and after the activation, have been carried out, using the SEM/EDS

Cinzas volantes álcali-ativadas com solução combinada de NaOH e Ca(OH)2

Este trabalho teve como objetivo avaliar o comportamento da resistência à compressão, ao longo do tempo, em amostras de argamassas à base de cinzas volantes álcali-ativadas. Como ativador alcalino foi utilizada uma solução combinada de NaOH e Ca(OH)2. A cura das amostras foi realizada de duas formas distintas. Na primeira, as amostras foram mantidas em estufa à 70°C/12 h e à temperatura constante de 22°C em sala climatizada até a idade de ensaio. Apesar de serem obtidas resistências da ordem de 20 MPa nas primeiras 24 h, houve decréscimo da resistência a partir da idade de 7 dias. Verificado esse comportamento, um segundo método de cura foi adotado. Para isso, o traço utilizado para o preparo de novas argamassas foi o mesmo adotado no método anterior. Porém, foram submetidas à temperatura constante de 70°C até a idade de ensaio. Nesse caso, as amostras apresentaram decréscimo mais acentuado da resistência em menor espaço de tempo, quando comparado às amostras curadas no primeiro método. Como estudos complementares foram realizadas análises microestruturais da CV antes e depois da álcali-ativação, utilizando o MEV/EDS. Nas imagens microestruturais foi possível identificar três morfologias distintas nas pastas álcali-ativadas: uma composta de regiões com aspecto denso; outra apresentando partículas de cinzas volantes parcialmente solubilizadas; e outra mostrando a formação de produtos em forma de cristais aciculares. Porém, apenas o uso do MEV/EDS não foi suficiente para que se pudesse entender o mecanismo de reação deletéria provocada entre os ativadores alcalinos e a cinza volante. Portanto, trabalhos futuros deverão contemplar o uso de ferramentas auxiliares às adotadas nesse trabalho com o objetivo de esclarecer tal mecanismo deletério.The main objective of this work was to evaluate over time the development of the compressive strength of alkali-activated fly ash mortars. A combined solution of NaOH and Ca(OH)2 was used as alkaline activator. The curing of the samples was carried out in two distinct ways. In the first one, samples were kept at 70°C during the first 12 hours after mixing and at 22°C afterwards, until the age of testing. Although compressive strengths of about 20 MPa were achieved during the first 24 hours, the strength started to decrease after 7 days. Considering this behaviour, a second method of curing was adopted. The mix proportions were the same as before, however the mortar samples were kept in an oven at 70°C until the age of testing. In this case, the compressive strength values decreased more significantly and in a shorter period of time, as compared to the results obtained for samples cured using the first method. Complementary studies were performed in order to better understand the observed reduction in compressive strength. Microstructural analyses of the fly ash, before and after the alkali-activation, have been carried out using Scanning Electron Microscopy (SEM) and Energy-Dispersive Spectroscopy (EDS). It was possible to identify three different morphologies in the alkali-activated pastes: one composed by regions with dense aspect; other showing partially solubilised particles of fly ash; and a third showing the formation of products with a needle-like shape. The information provided by the SEM/EDS experiments was not enough to enable the complete understanding of the mechanism of the deleterious reaction that took place among the alkaline activators and the fly ash. Therefore, future studies should address this question by using additional analysis tools in order to clarify this deleterious mechanism

Effect of Sheep Grazing, Stocking Rates and Dolomitic Limestone Application on the Floristic Composition of a Permanent Dryland Pasture, in the Montado Agroforestry System of Southern Portugal

The Montado is a complex agroforestry–pastoral ecosystem due to the interactions between soil–pasture–trees–animals and climate. The typical Montado soil has an acidic pH and manganese toxicity, which affect the pastureʹs productivity and pasture floristic composition (PFC). The PFC, on the other hand, can also be influenced by the type and intensity of grazing, which can lead to significant decreases in the amount of biomass produced and the biodiversity of species in the pasture. The objective of this study was to evaluate the effect of grazing type, by sheep, and different stocking rates on the PFC throughout the vegetative pasture cycle in areas with and without dolomitic limestone application. Thus, four treatments (P1UC to P4TC) were constituted: P1UC—without limestone application (U) and continuous grazing (CG); P2UD—U and deferred grazing (DG); P3TD—with the application of limestone (T) and DG; P4TC—T and CG. In DG plots, the placement and removal of the animals were carried out as a function of the average height of the pasture (placement—10 cm; removal—3 to 5 cm). The PFC was characterized in winter, at the peak of spring and in late spring. The PFC data were subjected to a multilevel pattern analysis (ISA). The combination of rainfall and temperature influenced the pasture growth rates and consequently the height of the pasture at different times of the year. Therefore, with the different growth rates of the pasture throughout the year, the sheep remain for different periods of time in the deferred grazing treatments. In the four treatments, 103 plant species were identified. The most representative botanical families in the four treatments were Asteraceae, Fabaceae and Poaceae. ISA identified 14 bioindicator species: eight for the winter period, three for the late spring vegetative period and three for the TC treatment

Corrole Isomers: Intrinsic Gas-phase Shapes Via Traveling Wave Ion Mobility Mass Spectrometry And Dissociation Chemistries Via Tandem Mass Spectrometry.

Corrole and four of its isomers with subtle structural changes promoted by exchange of nitrogen and carbon atoms in the corrole ring have been studied by traveling wave ion mobility mass spectrometry and collision induced dissociation experiments. Significant differences in shapes and charge distributions for their protonated molecules were found to lead to contrasting gas phase mobilities, most particularly for corrorin, the most confused isomer. Accordingly, corrorin was predicted by B3LYP/6-31g(d,p) and collisional cross section calculations to display the most compact tri-dimensional structure, whereas NCC4 and corrole were found to be the most planar isomers. Better resolution between the corrole isomers was achieved using the more polarizable and massive CO(2) as the drift gas. Sequential losses of HF molecules were found to dominate the dissociation chemistry of the protonated molecules of these corrole isomers, but their unique structures caused contrasting labilities towards CID, whereas NCC4 showed a peculiar and structurally diagnostic loss of NH(3), allowing its prompt differentiation from the other isomers.108396-40

Enterotype May Drive the Dietary-Associated Cardiometabolic Risk Factors

Analyses of typical bacterial clusters in humans named enterotypes may facilitate understanding the host differences in the cardiometabolic profile. It stills unknown whether the three previously described enterotypes were present in populations living below the equator. We examined how the identification of enterotypes could be useful to explain the dietary associations with cardiometabolic risk factors in Brazilian subjects. In this cross-sectional study, a convenience sample of 268 adults (54.2% women) reported their dietary habits and had clinical and biological samples collected. In this study, we analyzed biochemical data and metagenomics of fecal microbiota (16SrRNA sequencing, V4 region). Continuous variables were compared using ANOVA, and categorical variables using chi-square test. Vsearch clustered the operational taxonomic units, and Silva Database provided the taxonomic signatures. Spearman coefficient was used to verify the correlation between bacteria abundances within each enterotype. One hundred subjects were classified as omnivore, 102 lacto-ovo-vegetarians, and 66 strict vegetarians. We found the same structure as the three previously described enterotypes: 111 participants were assigned to Bacteroides, 55 to Prevotella, and 102 to Ruminococcaceae enterotype. The Prevotella cluster contained higher amount of strict vegetarians individuals than the other enterotypes (40.0 vs. 20.7 and 20.6, p = 0.04). Subjects in this enterotype had a similar anthropometric profile but a lower mean LDL-c concentration than the Bacteroides enterotype (96 +/- 23 vs. 109 +/- 32 mg/dL, p = 0.04). We observed significant correlations between bacterial abundances and cardiometabolic risk factors, but coefficients differed depending on the enterotype. In Prevotella enterotype, Eubacterium ventriosum (r BMI = -0.33, p = 0.03, and r HDL-c = 0.33, p = 0.04), Akkermansia (r 2h glucose = -0.35, p = 0.02), Roseburia (r BMI = -0.36, p = 0.02 and r waist = -0.36, p = 0.02), and Faecalibacterium (r insulin = -0.35, p = 0.02) abundances were associated to better cardiometabolic profile. The three enterotypes previously described are present in Brazilians, supporting that those bacterial clusters are not population-specific. Diet-independent lower LDL-c levels in subjects from Prevotella than in other enterotypes suggest that a protective bacterial cluster in the former should be driving this association. Enterotypes seem to be useful to understand the impact of daily diet exposure on cardiometabolic risk factors. Prospective studies are needed to confirm their utility for predicting phenotypes in humans.FAPESPUniv Sao Paulo, Sch Publ Hlth, Dept Epidemiol, Sao Paulo, BrazilFundacao Oswaldo Cruz, Rene Rachou Res Ctr, Belo Horizonte, MG, BrazilUniv Fed Sao Paulo, Dept Prevent Med, Sao Paulo, BrazilUniv Sao Paulo, Sch Med, Heart Inst Incor, Lab Genet & Mol Cardiol, Sao Paulo, BrazilUniv Fed Sao Paulo, Dept Prevent Med, Sao Paulo, BrazilFAPESP: 2012/12626-9FAPESP: 2012/03880-9Web of Scienc