Effect of pH and temperature on the global compactness, structure, and activity of cellobiohydrolase Cel7A from Trichoderma harzianum

Due to its elevated cellulolytic activity, the filamentous fungus Trichoderma harzianum (T. harzianum) has considerable potential in biomass hydrolysis application. Cellulases from Trichoderma reesei have been widely used in studies of cellulose breakdown. However, cellulases from T. harzianum are less-studied enzymes that have not been characterized biophysically and biochemically as yet. Here, we examined the effects of pH and temperature on the secondary and tertiary structures, compactness, and enzymatic activity of cellobiohydrolase Cel7A from T. harzianum (Th Cel7A) using a number of biophysical and biochemical techniques. Our results show that pH and temperature perturbations affect Th Cel7A stability by two different mechanisms. Variations in pH modify protonation of the enzyme residues, directly affecting its activity, while leading to structural destabilization only at extreme pH limits. Temperature, on the other hand, has direct influence on mobility, fold, and compactness of the enzyme, causing unfolding of Th Cel7A just above the optimum temperature limit. Finally, we demonstrated that incubation with cellobiose, the product of the reaction and a competitive inhibitor, significantly increased the thermal stability of Th Cel7A. Our studies might provide insights into understanding, at a molecular level, the interplay between structure and activity of Th Cel7A at different pH and temperature conditions.

Linking origin of the electric field-assisted beta-PbF2 crystallization in lead oxyfluoroborate glasses below T (g) to simultaneous cathode/anode-compensated electrochemical reactions

Full validation of the electrochemical mechanisms so far postulated as driving force of electric field-assisted non-spontaneous crystallization development in given glasses has suffered experimental restrictions. In this work, we looked into origin of this phenomenon in lead oxyfluoroborate glasses, resulting in beta-PbF2 growth even below the corresponding glass transition temperatures, through achieving a systematic study of not only Pt,Ag/Glass/Ag,Pt- but also Pt,Ag/Glass/YSZ:PbF2/Ag,Pt-type cells, where YSZ:PbF2 represents a two-phase system (formed by Y2O3-doped ZrO2 and PbF2). It is demonstrated that crystallization induction in these glasses involves Pb2+ ions reduction at the cathode, the phenomenon being, however, confirmed only when the F- ions were simultaneously also able to reach the anode for oxidation, after assuring either a direct glass-anode contact or percolation pathways for free fluoride migration across the YSZ:PbF2 mixtures. A further support of this account is that the electrochemically induced beta-PbF2 phase crystallizes showing ramified-like microstructure morphology that arises, accordingly, from development of electroconvective diffusion processes under electric field action.

The effects of river discharge and seasonal winds on the shelf off southeastern South America

The Rio de la Plata waters form a low salinity tongue that affects the circulation, stratification and the distributions of nutrients and biological species over a wide extent of the adjacent continental shelf. The plume of coastal waters presents a seasonal meridional displacement reaching lower latitudes (28,S) during austral winter and 32 degrees S during summer. Historical data suggests that the wind causes the alongshore shift, with southwesterly (SW) winds forcing the plume to lower latitudes in winter while summer dominant northeasterly (NE) winds force its southward retreat. To establish the connection between wind and outflow variations on the distribution of the coastal waters, we conducted two quasi-synoptic surveys in the region of Plata influence on the continental shelf and slope of southeastern South America, between Mar del Plata, Argentina and the northern coast of Santa Catarina, Brazil. We observed that: (A) SW winds dominating in winter force the northward spreading of the plume to low latitudes even during low river discharge periods; (B) NE winds displace the plume southward and spread the low salinity waters offshore over the entire width of the continental shelf east of the Plata estuary. The southward retreat of the plume in summer leads to a volume decrease of low salinity waters over the shelf. This volume is compensated by an increase of Tropical waters, which dominate the northern shelf. The subsurface transition between Subantarctic and Subtropical Shelf Waters, the Subtropical Shelf Front, and the subsurface water mass distribution, however, present minor seasonal variations. Along shore winds also influence the dynamics and water mass variations along the continental shelf area. In areas under the influence of river discharge, Subtropical Shelf Waters are kept away from the coastal region. When low salinity waters retreat southward, NE winds induce a coastal upwelling system near Santa Marta Cape. In summer, solar radiation promotes the establishment of a strong thermocline that increases buoyancy and further enhances the offshore displacement of low salinity waters under the action of NE winds. (C) 2008 Elsevier Ltd. All rights reserved.

Production and characterization of poly-(3-hydroxybutyrate) from recombinant Escherichia coli grown on cheap renewable carbon substrates

Although the biopolymer poly-(3-hydroxybutyrate), P[3HB], presents physicochemical properties that make it an alternative material to conventional plastics, its biotechnological production is quite expensive. As carbon substrates contribute greatly to P[3HB] production cost, the utilization of a cheaper carbon substrate and less demanding micro-organisms should decrease its cost. In the present study a 23 factorial experimental design was applied, aiming to evaluate the effects of using hydrolysed corn starch (HCS) and soybean oil (SBO) as carbon substrates, and cheese whey (CW) supplementation in the mineral medium (MM) on the responses, cell dried weigh (DCW), percentage P[3HB] and mass P[3HB] by recombinant Escherichia coli strains JM101 and DH10B, containing the P[3HB] synthase genes from Cupriavidus necator (ex-Ralstonia eutropha). The analysis of effects indicated that the substrates and the supplement and their interactions had positive effect on CDW. Statistically generated equations showed that, at the highest concentrations of HCS, SO and CW, theoretically it should be possible to produce about 2 g L(1) DCW, accumulating 50% P[3HB], in both strains. To complement this study, the strain that presented the best results was cultivated in MM added to HCS, SBO and CW ( in best composition observed) and complex medium (CM) to compare the obtained P[3HB] in terms of physicochemical parameters. The obtained results showed that the P[3HB] production in MM (1.29 g L(-1)) was approximately 20% lower than in CM (1.63 g L(-1)); however, this difference can be compensated by the lower cost of the MM achieved by the use of cheap renewable carbon sources. Moreover, using differential scanning calorimetry and thermogravimetry analyses, it was observed that the polymer produced in MM was the one which presented physicochemical properties (Tg and Tf) that were more similar to those found in the literature for P[3HB].

Stoichiometry, structure, and transport in the quasi-one-dimensional metal Li0.9Mo6O17

A correlation between lattice parameters, oxygen composition, and the thermoelectric and Hall coefficients is presented for single-crystal Li0.9Mo6O17, a quasi-one-dimensional (Q1D) metallic compound. The possibility that this compound is a compensated metal is discussed in light of a substantial variability observed in the literature for these transport coefficients.

On bifurcation of solutions of the Yamabe problem in product manifolds

We study local rigidity and multiplicity of constant scalar curvature metrics in arbitrary products of compact manifolds. Using (equivariant) bifurcation theory we determine the existence of infinitely many metrics that are accumulation points of pairwise non-homothetic solutions of the Yamabe problem. Using local rigidity and some compactness results for solutions of the Yamabe problem, we also exhibit new examples of conformal classes (with positive Yamabe constant) for which uniqueness holds. (C) 2011 Elsevier Masson SAS. All rights reserved.

Soil compressibility and least limiting water range of a constructed soil under cover crops after coal mining in Southern Brazil

The use of cover crops affects the support capacity of soil and least limiting water range to crop growth. The objective of this study was to quantify preconsolidation pressure (sigma(p)), compression index (CI) and least limiting water range (LLWR) of a reclaimed coal mining soil under different cover crops, in Candiota, RS, Brazil. In the experiment, with randomized blocks design and four replicates, the following cover crops (treatments) were evaluated: Hemarthria altissima (Poir.) Stapf & C.E. Hubbard, treatment 1 (T1), Paspalum notatum Flugge, treatment 4 (T4), Cynodon dactilon (L) Pers., treatment 5 (T5), control Brachiaria brizantha (Hochst.) Stapf, treatment 7 (T7) and without cover crop treatment 8 (reference treatment, T8). Soil compression and least limiting water range were evaluated with undisturbed samples at a depth of 0.00-0.05 m. In order to evaluate parameters of soil compressibility, the soil samples were saturated with water and subjected to -10 kPa matric potential and then submitted to a uniaxial compression test under the following pressures: 25, 50, 100, 200, 400, 800 and 1600 kPa. Cover crops decreased the preconsolidation pressure of constructed soils after coal mining and the greatest soil reclamation was obtained with the H. altissima cover crop, where the lowest degree of soil compactness and soil load capacity were observed. Soils cultivated under H. altissima or B. brizantha presented the highest least limiting water range and these two cover crops generated similar soil critical bulk density obtained by least limiting water range and soil load support capacity. (C) 2012 Elsevier B.V. All rights reserved.

Focal adhesion kinase governs cardiac concentric hypertrophic growth by activating the AKT and mTOR pathways

The heart responds to sustained overload by hypertrophic growth in which the myocytes distinctly thicken or elongate on increases in systolic or diastolic stress. Though potentially adaptive, hypertrophy itself may predispose to cardiac dysfunction in pathological settings. The mechanisms underlying the diverse morphology and outcomes of hypertrophy are uncertain. Here we used a focal adhesion kinase (FAK) cardiac-specific transgenic mice model (FAK-Tg) to explore the function of this non-receptor tyrosine kinase on the regulation of myocyte growth. FAK-Tg mice displayed a phenocopy of concentric cardiac hypertrophy, reflecting the relative thickening of the individual myocytes. Moreover, FAK-Tg mice showed structural, functional and molecular features of a compensated hypertrophic growth, and preserved responses to chronic pressure overload. Mechanistically, FAK overexpression resulted in enhanced myocardial FAK activity, which was proven by treatment with a selective FAK inhibitor to be required for the cardiac hypertrophy in this model. Our results indicate that upregulation of FAK does not affect the activity of Src/ERK1/2 pathway, but stimulated signaling by a cascade that encompasses PI3K, AKT, mTOR, S6K and rpS6. Moreover, inhibition of the mTOR complex by rapamycin extinguished the cardiac hypertrophy of the transgenic FAK mice. These findings uncover a unique role for FAK in regulating the signaling mechanisms that governs the selective myocyte growth in width, likely controlling the activity of PI3K/AKT/mTOR pathway, and suggest that FAK activation could be important for the adaptive response to increases in cardiac afterload. This article is part of a Special Issue entitled "Local Signaling in Myocytes". (C) 2011 Elsevier Ltd. All rights reserved.

Numerical study of Mach number and thermal effects on sound radiation by a mixing layer

Mach number and thermal effects on the mechanisms of sound generation and propagation are investigated in spatially evolving two-dimensional isothermal and non-isothermal mixing layers at Mach number ranging from 0.2 to 0.4 and Reynolds number of 400. A characteristic-based formulation is used to solve by direct numerical simulation the compressible Navier-Stokes equations using high-order schemes. The radiated sound is directly computed in a domain that includes both the near-field aerodynamic source region and the far-field sound propagation. In the isothermal mixing layer, Mach number effects may be identified in the acoustic field through an increase of the directivity associated with the non-compactness of the acoustic sources. Baroclinic instability effects may be recognized in the non-isothermal mixing layer, as the presence of counter-rotating vorticity layers, the resulting acoustic sources being found less efficient. An analysis based on the acoustic analogy shows that the directivity increase with the Mach number can be associated with the emergence of density fluctuations of weak amplitude but very efficient in terms of noise generation at shallow angle. This influence, combined with convection and refraction effects, is found to shape the acoustic wavefront pattern depending on the Mach number.

A NOTE ON THE UNIQUENESS OF SOLUTIONS FOR THE YAMABE PROBLEM

Using recent results on the compactness of the space of solutions of the Yamabe problem, we show that in conformal classes of metrics near the class of a nondegenerate solution which is unique (up to scaling) the Yamabe problem has a unique solution as well. This provides examples of a local extension, in the space of conformal classes, of a well-known uniqueness criterion due to Obata.

Odd homoclinic orbits for a second order Hamiltonian system

The aim of this paper is to find an odd homoclinic orbit for a class of reversible Hamiltonian systems. The proof is variational and it employs a version of the concentration compactness principle of P. L. Lions in a lemma due to Struwe.

Differences in early biomechanical properties after myopic and hyperopic lasik using mechanical microkeratome Sub-Bowman's Keratomileusis (SBK)

Purpose: To evaluate biomechanical changes measured with the ORA (Ocular Response Analyzer (R); Reichert Ophthalmic Instruments, Buffalo, New York, USA) after Lasik with the Moria One Use Plus and to compare the biomechanics changes after myopic and hyperopic ablations. Methods: Fourteeneyes for hyperopia (H) and 19 eyes for myopia (M) were evaluated with the ORA preoperatively and 1 month after Lasik with thin flap (100 microns) using SBK-OUP (Sub-Bowman Keratomileusis-One Use Plus, Moria (R)). CH (Corneal Hysteresis), CRF (Corneal Resistance Factor), IOPg (gold-standard, Goldmann correlated Intraocular pressure), IOPcc (Corneal compensated Intraocular pressure) and more 38 variables derived from the corneal biomechanical response signal of the ORA were analyzed. The Wilcoxon test was used to assess differences between the variables before and after surgery for each group and the differences between the pre and postoperative (1 month) myopic eyes were compared with those obtained in hyperopic eyes, using the Mann-Whitney test. Results: There was a significant difference before and after Lasik in myopic and hyperopic eyes in IOPg (Wilcoxon, p<0.05), but not in IOPcc. Only myopic eyes showed a significant difference in CH and CRF measurements before and after LASIK, as well as 9 other biomechanical parameters (aspect1, h1, dive1, path1, p1area1, W11, H11, and w2 path11; Wilcoxon, p<0, 05), 8 of these being related to the first sign of flattening. Five parameters related to the sign of the second applanation showed significant variation only in the eyes before and after hyperopic Lasik (aspect2, h2, dive2, mslew2 and H21; Wilcoxon, p<0,05). There was a difference in both myopic and hyperopic on three parameters related to the applanation signal areas (p1area, and p2area p2area1; Wilcoxon, p<0.05). Differences in IOPg and p1area, before and after surgery were significantly higher in myopic eyes than in hyperopic eyes (Mann-Whitey, p<0.05). Conclusion: There are several significant differences in biomechanical parameters after Lasik with Moria OUP_SBK. Overall, the impact of myopic LASIK on corneal biomechanics is higher than of hyperopic Lasik. The parameters derived from the first sign of the ORA are more affected in myopic LASIK, whereas parameters derived from the second applanation are more affected in hyperopic LASIK.

Variance Effective Population Size for Dioecious Species

The concept of effective population size (N(e)) is an important measure of representativeness in many areas. In this research, we consider the statistical properties of the number of contributed gametes under practical situations by adapting Crow and Denninston's (1988) N(e) formulas for dioecious species. Three sampling procedures were considered. In all circumstances, results show that as the offspring sex ratio (r) deviates from 0.5, N(e) values become smaller, and the efficiency of gametic control for increasing N(e) is reduced. For finite populations, where all individuals are potentially functional parents, the reduction in N(e) due to an unequal sex ratio can be compensated for through female gametic control when 0.28 <= r <= 0.72. This outcome is important when r is unknown. When only a fraction of the individuals in a population is taken for reproduction, N(e) is meaningful only if the size of the reference population is clearly defined. Gametic control is a compensating factor in accession regeneration when the viability of the accession is around 70 or 75%. For germ-plasm collection, when parents are a very small fraction of the population, maximum N(e) will be approximately 47 and 57% of the total number of offspring sampled, with female gametic control, r varying between 0.3 and 0.5, and being constant over generations.

The effect of ploidy on the fatty acid profile during the reproductive cycle of female rainbow trout (Oncorhynchus mykiss)

Rainbow trout Oncorhynchus mykiss triploids are regularly produced in fish farms to improve growth because the triploid females do not develop ovaries during the reproductive cycle. In this study, the tissue fatty acid allocations in triploid and diploid females were compared during the reproductive cycle to determine whether the ploidy influences the fatty acid profile of fish produced in aquaculture. The ovaries, liver, and white muscle fatty acid contents of diploid and triploid females were analyzed during the reproductive cycle. Diploid females tend to accumulate more polyunsaturated fatty acids than triploids during some phases of the reproductive cycle, and this profile was compensated by an increase in saturated and monounsaturated fatty acids in triploid females. Arachidonic acid (C20:4n6) was the main n6 polyunsaturated fatty acid in the ovaries of diploid females during the most advanced phases of the reproductive cycle, and docosahexaenoic acid (C22:6n3) was the main n3 polyunsaturated fatty acid. In triploid females, the percentage of both of these polyunsaturated fatty acids was lower than in diploid females during the most advanced phases of the reproductive cycle. In general, the lack of ovary development altered the hepatic synthesis of some fatty acids, mainly monounsaturated and polyunsaturated fatty acids, decreasing the content of the main fatty acids in the white muscle and, consequently, the mobilization of these specific fatty acids to the ovaries.

The fatty acid profiles and energetic substrates of two Nile tilapia (Oreochromis niloticus, Linnaeus) strains, Red-Stirling and Chitralada, and their hybrid

Protein and lipid content as well as the fatty acid (FA) composition of storage tissues were analysed in two varieties of Oreochromis niloticus (Red-Stirling and Chitralada) and their hybrid. The animals were maintained in cages for 11 months. The samples were taken when the animals weighed 10, 50, 100, 250 and 500 g. The results showed that changes in the metabolic processes occur during an increase in body mass in both varieties of tilapia and also their hybrid, but that these differences are not found in animals collected at the commercial weight. The protein content of the fillet and liver decreased with growth and the same protein content associated with growth was found for fillet lipid content. The genetic variety did not influence the FA profile of the fillet, but different genotypes had different hepatic FA compositions. Even with the same lipid content, the hepatocytes of Chitralada accumulated higher levels of polyunsaturated fatty acids (PUFA) n6 in triglycerides and increased C22:6n3 in the hepatocyte membranes. The higher n6PUFA content was compensated by a lower fraction of saturated FA in the hepatocyte triglycerides. The skin of Chitralada also had higher n6PUFA and C22:6n3 contents, suggesting a higher ability to deposit PUFA in the skin due to alterations in the liver synthetic pathway.