Determination of Substrate Preferences for Desaturases and Elongases for Production of Docosahexaenoic Acid from Oleic Acid in Engineered Canola

Production of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in plant seed oils has been pursued to improve availability of these omega‐3 fatty acids that provide important human health benefits. Canola (Brassica napus), through the introduction of 10 enzymes, can convert oleic acid (OLA) into EPA and ultimately DHA through a pathway consisting of two elongation and five desaturation steps. Herein we present an assessment of the substrate specificity of the seven desaturases and three elongases that were introduced into canola by expressing individual proteins in yeast. In vivo feeding experiments were conducted with 14 potential fatty acid intermediates in an OLA to DHA pathway to determine the fatty acid substrate profiles for each enzyme. Membrane fractions were prepared from yeast expression strains and shown to contain active enzymes. The elongases, as expected, extended acyl‐CoA substrates in the presence of malonyl‐CoA. To distinguish between enzymes that desaturate CoA‐ and phosphatidylcholine‐linked fatty acid substrates, we developed a novel in vitro method. We show that a delta‐12 desaturase from Phytophthora sojae, an omega‐3 desaturase from Phytophthora infestans and a delta‐4 desaturase from Thraustochytrium sp., all prefer phosphatidylcholine‐linked acyl substrates with comparatively low use of acyl‐CoA substrates. To further validate our method, a delta‐9 desaturase from Saccharomyces cerevisiae was confirmed to use acyl‐CoA as substrate, but could not use phosphatidylcholine‐linked substrates. The results and the assay methods presented herein will be useful in efforts to improve modeling of fatty acid metabolism and production of EPA and DHA in plants.     

Hierarchical Functional Maps between Subdivision Surfaces

We propose a novel approach for computing correspondences between subdivision surfaces with different control polygons. Our main observation is that the multi‐resolution spectral basis functions that are open used for computing a functional correspondence can be compactly represented on subdivision surfaces, and therefore can be efficiently computed. Furthermore, the reconstruction of a pointwise map from a functional correspondence also greatly benefits from the subdivision structure. Leveraging these observations, we suggest a hierarchical pipeline for functional map inference, allowing us to compute correspondences between surfaces at fine subdivision levels, with hundreds of thousands of polygons, an order of magnitude faster than existing correspondence methods. We demonstrate the applicability of our results by transferring high‐resolution sculpting displacement maps and textures between subdivision models.     

An explicit structure‐preserving numerical scheme for EPDiff

We present a new structure‐preserving numerical scheme for solving the Euler‐Poincaré Differential (EPDiff) equation on arbitrary triangle meshes. Unlike existing techniques, our method solves the difficult non‐linear EPDiff equation by constructing energy preserving, yet fully explicit, update rules. Our approach uses standard differential operators on triangle meshes, allowing for a simple and efficient implementation. Key to the structure‐preserving features that our method exhibits is a novel numerical splitting scheme. Namely, we break the integration into three steps which rely on linear solves with a fixed sparse matrix that is independent of the simulation and thus can be pre‐factored. We test our method in the context of simulating concentrated reconnecting wavefronts on flat and curved domains. In particular, EPDiff is known to generate geometrical fronts which exhibit wave‐like behavior when they interact with each other. In addition, we also show that at a small additional cost, we can produce globally‐supported periodic waves by using our simulated fronts with wavefronts tracking techniques. We provide quantitative graphs showing that our method exactly preserves the energy in practice. In addition, we demonstrate various interesting results including annihilation and recreation of a circular front, a wave splitting and merging when hitting an obstacle and two separate fronts propagating and bending due to the curvature of the domain.     

Conformal Flattening by Curvature Prescription and Metric Scaling

We present an efficient method to conformally parameterize 3D mesh data sets to the plane. The idea behind our method is to concentrate all the 3D curvature at a small number of select mesh vertices, called cone singularities, and then cut the mesh through those singular vertices to obtain disk topology. The singular vertices are chosen automatically. As opposed to most previous methods, our flattening process involves only the solution of linear systems of Poisson equations, thus is very efficient. Our method is shown to be faster than existing methods, yet generates parameterizations having comparable quasi‐conformal distortion.     

Mechanical properties and linear infrared dichroism of thin films of polyurethane nanocomposites

Morphological, mechanical, and Fourier transform infrared dichroic investigations were performed on neat polyurethane (PU) polymer matrix and PU+CaCO₃ nanocomposite thin films to determine how the nanofiller influenced the mechanical properties. The measurements were performed on strips that were cut from the prepared films in parallel and perpendicular directions with respect to the direction of film preparation. Optical microscopy of PU and the PU+CaCO₃ nanocomposite revealed the strain‐induced transition from a continuous spherulitic morphology to a fiberlike structure. The stress–strain behavior of the neat PU and PU+CaCO₃ nanocomposite films showed significant differences at large strain regimes. The experimental results suggest that the mechanical properties were strongly related to the orientational behavior of the separated phases. The orientation of the hard and soft segments was analyzed by the orientation function calculated from the IR absorbances. A correlation between the orientations of segments, tensile properties, and hardness of the investigated polymer films was established. © 2008 Wiley Periodicals, Inc. J Appl Polym Sci, 2008     

Connexin Expression Is Altered in Liver Development of Yotari (dab1 -/-) Mice

Disabled-1 (Dab1) protein is an intracellular adaptor of reelin signaling required for prenatal neuronal migration, as well as postnatal neurotransmission, memory formation and synaptic plasticity. Yotari, an autosomal recessive mutant of the mouse Dab1 gene is recognizable by its premature death, unstable gait and tremor. Previous findings are mostly based on neuronal abnormalities caused by Dab1 deficiency, but the role of the reelin signaling pathway in nonneuronal tissues and organs has not been studied until recently. Hepatocytes, the most abundant cells in the liver, communicate via gap junctions (GJ) are composed of connexins. Cell communication disruption in yotari mice was examined by analyzing the expression of connexins (Cxs): Cx26, Cx32, Cx37, Cx40, Cx43 and Cx45 during liver development at 13.5 and 15.5 gestation days (E13.5 and E15.5). Analyses were performed using immunohistochemistry and fluorescent microscopy, followed by quantification of area percentage covered by positive signal. Data are expressed as a mean ± SD and analyzed by one-way ANOVA. All Cxs examined displayed a significant decrease in yotari compared to wild type (wt) individuals at E13.5. Looking at E15.5 we have similar results with exception of Cx37 showing negligible expression in wt. Channels formation triggered by pathological stimuli, as well as propensity to apoptosis, was studied by measuring the expression of Pannexin1 (Panx1) and Apoptosis-inducing factor (AIF) through developmental stages mentioned above. An increase in Panx1 expression of E15.5 yotari mice, as well as a strong jump of AIF in both phases suggesting that yotari mice are more prone to apoptosis. Our results emphasize the importance of gap junction intercellular communication (GJIC) during liver development and their possible involvement in liver pathology and diagnostics where they can serve as potential biomarkers and drug targets.     

Modeling the influence of vegetation and water pond on urban microclimate

The beneficial influence of trees and water ponds on summer comfort in urban spaces was studied experimentally in situ and in wind tunnels but the modeling needs further development to become effective in practical applications. This paper introduces a numerical approach based on coupling the CFD model of airflow, in which the influence of trees is considered as source terms, and the radiation exchange, completed with thermal conduction. The CFD, radiation and thermal conduction models use the same discretization grid at their common boundaries. The model was used to estimate the influence of trees and water ponds in a real town square. Comparison of results between two situations, with and without vegetation and water pond, indicate that surface temperatures are reduced in presence of trees and the comfort is improved.     

Q fever endocarditis in Romania: the first cases confirmed by direct sequencing

Infective endocarditis (IE) is a serious, life-threatening disease with highly variable clinical signs, making its diagnostic a real challenge. A diagnosis is readily made if blood cultures are positive, but in 2.5 to 31% of all infective endocarditis cases, routine blood cultures are negative. In such situations, alternative diagnostic approaches are necessary. Coxiella burnetii and Bartonella spp. are the etiological agents of blood culture-negative endocarditis (BCNE) most frequently identified by serology. The purpose of this study is to investigate the usefulness of molecular assays, as complementary methods to the conventional serologic methods for the rapid confirmatory diagnostic of Q fever endocarditis in patients with BCNE. Currently, detection of C. burnetii by culture or an antiphase I IgG antibody titers >800 represents a major Duke criterion for defining IE, while a titers of >800 for IgG antibodies to either B. henselae or B. quintana is used for the diagnosis of endocarditis due to Bartonella spp. We used indirect immunofluorescence assays for the detection of IgG titers for C. burnetii, B. henselae and B. quintana in 57 serum samples from patients with clinical suspicion of IE. Thirty three samples originated from BCNE patients, whereas 24 were tested before obtaining the blood cultures results, which finally were positive. The results of serologic testing showed that nine out of 33 BCNE cases exhibited antiphase I C. burnetii IgG antibody titer >800, whereas none has IgG for B. henselae or B. quintana. Subsequently, we used nested-PCR assay for the amplification of C. burnetii DNA in the nine positive serum samples, and we obtained positive PCR results for all analyzed cases. Afterwards we used the DNA sequencing of amplicons for the repetitive element associated to htpAB gene to confirm the results of nested-PCR. The results of sequencing allowed us to confirm that C. burnetii is the causative microorganism responsible for BCNE. In conclusion, the nested PCR amplification followed by direct sequencing is a reliable and accurate method when applied to serum samples, and it may be used as an additional test to the serological methods for the confirmatory diagnosis of BCNE cases determined by C. burnetii.     

Uric Acid and Oxidative Stress—Relationship with Cardiovascular,Metabolic, and Renal Impairment

Background: The connection between uric acid (UA) and renal impairment is well known due to the urate capacity to precipitate within the tubules or extra-renal system. Emerging studies allege a new hypothesis concerning UA and renal impairment involving a pro-inflammatory status, endothelial dysfunction, and excessive activation of renin–angiotensin–aldosterone system (RAAS). Additionally, hyperuricemia associated with oxidative stress is incriminated in DNA damage, oxidations, inflammatory cytokine production, and even cell apoptosis. There is also increasing evidence regarding the association of hyperuricemia with chronic kidney disease (CKD), cardiovascular disease, and metabolic syndrome or diabetes mellitus. Conclusions: Important aspects need to be clarified regarding hyperuricemia predisposition to oxidative stress and its effects in order to initiate the proper treatment to determine the optimal maintenance of UA level, improving patients’ long-term prognosis and their quality of life.     

Improving TiO2 activity in photo-production of hydrogen from sugar industry wastewaters

A series of TiO₂ photocatalysts dispersed on SiO₂, modified with metals (Mn, Cr, V) or non-metals (N or S) or covered with Au nanoparticles were prepared by incipient wetness impregnation, impregnation with urea or thiourea, and deposition-precipitation, respectively. The photocatalysts where characterized by chemical analysis, X-ray diffraction (XRD), diffuse reflectance UV-Vis spectroscopy (DR-UV-Vis) and Fourier transform infrared spectroscopy (FTIR). The photocatalytic activity was checked in the photogeneration of hydrogen from sugar solutions. The nature of the modifying species was found to influence the photoactivity and N and S led to more effective catalysts than metals. This behavior was attributed to the fact the N and S are less effective in forming recombination centers compared with the metals.