Down-regulation of the CSLF6 gene results in decreased (1,3;1,4)-beta-D-glucan in endosperm of wheat

(1,3;1,4)-beta-d-Glucan (beta-glucan) accounts for 20% of the total cell walls in the starchy endosperm of wheat (Triticum aestivum) and is an important source of dietary fiber for human nutrition with potential health benefits. Bioinformatic and array analyses of gene expression profiles in developing caryopses identified the CELLULOSE SYNTHASE-LIKE F6 (CSLF6) gene as encoding a putative beta-glucan synthase. RNA interference constructs were therefore designed to down-regulate CSLF6 gene expression and expressed in transgenic wheat under the control of a starchy endosperm-specific HMW subunit gene promoter. Analysis of wholemeal flours using an enzyme-based kit and by high-performance anion-exchange chromatography after digestion with lichenase showed decreases in total beta-glucan of between 30% and 52% and between 36% and 53%, respectively, in five transgenic lines compared to three control lines. The content of water-extractable beta-glucan was also reduced by about 50% in the transgenic lines, and the M(r) distribution of the fraction was decreased from an average of 79 to 85 x 10(4) g/mol in the controls and 36 to 57 x 10(4) g/mol in the transgenics. Immunolocalization of beta-glucan in semithin sections of mature and developing grains confirmed that the impact of the transgene was confined to the starchy endosperm with little or no effect on the aleurone or outer layers of the grain. The results confirm that the CSLF6 gene of wheat encodes a beta-glucan synthase and indicate that transgenic manipulation can be used to enhance the health benefits of wheat products.

Guanine specific binding at a DNA junction formed by d[CG(5-BrU)ACG]2 with a topoisomerase poison in the presence of Co2+Ions†,‡

The structure of the duplex d[CG(5-BrU)ACG]2 bound to 9-bromophenazine-4-carboxamide has been solved through MAD phasing at 2.0 Å resolution. It shows an unexpected and previously unreported intercalation cavity stabilized by the drug and novel binding modes of Co2+ ions at certain guanine N7 sites. For the intercalation cavity the terminal cytosine is rotated to pair with the guanine of a symmetry-related duplex to create a pseudo-Holliday junction geometry, with two such cavities linked through the minor groove interactions of the N2/N3 guanine sites at an angle of 40°, creating a quadruplex-like structure. The mode of binding of the drug is shown to be disordered, with the major conformations showing the side chain bound to the N7 position of adjacent guanines. The other end of the duplex exhibits a terminal base fraying in the presence of Co2+ ions linking symmetry-related guanines, causing the helices to intertwine through the minor groove. The stabilization of the structure by the intercalating drug shows that this class of compound may bind to DNA junctions as well as duplex DNA or to strand-nicked DNA (‘hemi-intercalated'), as in the cleavable complex. This suggests a structural basis for the dual poisoning of topoisomerase I and II enzymes by this family of drugs.

7,8-dihydropyrido[2,3-d]pyrimidin-2-one; a bicyclic cytosine analogue capable of enhanced stabilisation of DNA duplexes

Incorporation of a bicyclic cytosine analogue, 3-beta-D-(2'-deoxyribofuranosyl)7,8- dihydropyrido[ 2,3-d] pyrimidine, into synthetic DNA duplexes results in a greatly enhanced thermal stability ( 3 - 4 degrees C per modification) compared to the corresponding unmodified duplex.

Comparison of D-region Doppler drift winds measured by the SuperDARN Finland HF radar over an annual cycle using the Kiruna VHF meteor radar

The SuperDARN chain of oblique HF radars has provided an opportunity to generate a unique climatology of horizontal winds near the mesopause at a number of high latitude locations, via the Doppler shifted echoes from sources of ionisation in the D-region. Ablating meteor trails form the bulk of these targets, but other phenomena also contribute to the observations. Due to the poor vertical resolution of the radars, care must be taken to reduce possible biases from sporadic-E layers and Polar Mesospheric Summer echoes that can affect the effective altitude of the geophysical parameters being observed. Second, there is strong theoretical and observational evidence to suggest that the radars are picking up echoes from the backward looking direction that will tend to reduce the measured wind strengths. The effect is strongly frequency dependent, resulting in a 20% reduction at 12 MHz and a 50% reduction at 10 MHz. A comparison of the climatologies observed by the Super-DARN Finland radar between September 1999 and September 2000 and that obtained from the adjacent VHF meteor radar located at Kiruna is also presented. The agreement between the two instruments was very good. Extending the analysis to the SuperDARN Iceland East radar indicated that the principles outlined above could be applied successfully to the rest of the SuperDARN network.

Causal relationship between obesity and vitamin D status: bi-directional mendelian randomization analysis of multiple cohorts

BACKGROUND: Obesity is associated with vitamin D deficiency, and both are areas of active public health concern. We explored the causality and direction of the relationship between body mass index (BMI) and 25-hydroxyvitamin D [25(OH)D] using genetic markers as instrumental variables (IVs) in bi-directional Mendelian randomization (MR) analysis. METHODS AND FINDINGS: We used information from 21 adult cohorts (up to 42,024 participants) with 12 BMI-related SNPs (combined in an allelic score) to produce an instrument for BMI and four SNPs associated with 25(OH)D (combined in two allelic scores, separately for genes encoding its synthesis or metabolism) as an instrument for vitamin D. Regression estimates for the IVs (allele scores) were generated within-study and pooled by meta-analysis to generate summary effects. Associations between vitamin D scores and BMI were confirmed in the Genetic Investigation of Anthropometric Traits (GIANT) consortium (n = 123,864). Each 1 kg/m(2) higher BMI was associated with 1.15% lower 25(OH)D (p = 6.52×10⁻²⁷). The BMI allele score was associated both with BMI (p = 6.30×10⁻⁶²) and 25(OH)D (-0.06% [95% CI -0.10 to -0.02], p = 0.004) in the cohorts that underwent meta-analysis. The two vitamin D allele scores were strongly associated with 25(OH)D (p≤8.07×10⁻⁵⁷ for both scores) but not with BMI (synthesis score, p = 0.88; metabolism score, p = 0.08) in the meta-analysis. A 10% higher genetically instrumented BMI was associated with 4.2% lower 25(OH)D concentrations (IV ratio: -4.2 [95% CI -7.1 to -1.3], p = 0.005). No association was seen for genetically instrumented 25(OH)D with BMI, a finding that was confirmed using data from the GIANT consortium (p≥0.57 for both vitamin D scores). CONCLUSIONS: On the basis of a bi-directional genetic approach that limits confounding, our study suggests that a higher BMI leads to lower 25(OH)D, while any effects of lower 25(OH)D increasing BMI are likely to be small. Population level interventions to reduce BMI are expected to decrease the prevalence of vitamin D deficiency.

Interaction between allelic variations in vitamin D receptor and retinoid X receptor genes on metabolic traits

BACKGROUND: Low vitamin D status has been shown to be a risk factor for several metabolic traits such as obesity, diabetes and cardiovascular disease. The biological actions of 1, 25-dihydroxyvitamin D, are mediated through the vitamin D receptor (VDR), which heterodimerizes with retinoid X receptor, gamma (RXRG). Hence, we examined the potential interactions between the tagging polymorphisms in the VDR (22 tag SNPs) and RXRG (23 tag SNPs) genes on metabolic outcomes such as body mass index, waist circumference, waist-hip ratio (WHR), high- and low-density lipoprotein (LDL) cholesterols, serum triglycerides, systolic and diastolic blood pressures and glycated haemoglobin in the 1958 British Birth Cohort (1958BC, up to n = 5,231). We used Multifactor- dimensionality reduction (MDR) program as a non-parametric test to examine for potential interactions between the VDR and RXRG gene polymorphisms in the 1958BC. We used the data from Northern Finland Birth Cohort 1966 (NFBC66, up to n = 5,316) and Twins UK (up to n = 3,943) to replicate our initial findings from 1958BC. RESULTS: After Bonferroni correction, the joint-likelihood ratio test suggested interactions on serum triglycerides (4 SNP - SNP pairs), LDL cholesterol (2 SNP - SNP pairs) and WHR (1 SNP - SNP pair) in the 1958BC. MDR permutation model testing analysis showed one two-way and one three-way interaction to be statistically significant on serum triglycerides in the 1958BC. In meta-analysis of results from two replication cohorts (NFBC66 and Twins UK, total n = 8,183), none of the interactions remained after correction for multiple testing (Pinteraction >0.17). CONCLUSIONS: Our results did not provide strong evidence for interactions between allelic variations in VDR and RXRG genes on metabolic outcomes; however, further replication studies on large samples are needed to confirm our findings.

Association of vitamin D status with arterial blood pressure and hypertension risk: a mendelian randomisation study

BACKGROUND: Low plasma 25-hydroxyvitamin D (25[OH]D) concentration is associated with high arterial blood pressure and hypertension risk, but whether this association is causal is unknown. We used a mendelian randomisation approach to test whether 25(OH)D concentration is causally associated with blood pressure and hypertension risk. METHODS: In this mendelian randomisation study, we generated an allele score (25[OH]D synthesis score) based on variants of genes that affect 25(OH)D synthesis or substrate availability (CYP2R1 and DHCR7), which we used as a proxy for 25(OH)D concentration. We meta-analysed data for up to 108 173 individuals from 35 studies in the D-CarDia collaboration to investigate associations between the allele score and blood pressure measurements. We complemented these analyses with previously published summary statistics from the International Consortium on Blood Pressure (ICBP), the Cohorts for Heart and Aging Research in Genomic Epidemiology (CHARGE) consortium, and the Global Blood Pressure Genetics (Global BPGen) consortium. FINDINGS: In phenotypic analyses (up to n=49 363), increased 25(OH)D concentration was associated with decreased systolic blood pressure (β per 10% increase, -0·12 mm Hg, 95% CI -0·20 to -0·04; p=0·003) and reduced odds of hypertension (odds ratio [OR] 0·98, 95% CI 0·97-0·99; p=0·0003), but not with decreased diastolic blood pressure (β per 10% increase, -0·02 mm Hg, -0·08 to 0·03; p=0·37). In meta-analyses in which we combined data from D-CarDia and the ICBP (n=146 581, after exclusion of overlapping studies), each 25(OH)D-increasing allele of the synthesis score was associated with a change of -0·10 mm Hg in systolic blood pressure (-0·21 to -0·0001; p=0·0498) and a change of -0·08 mm Hg in diastolic blood pressure (-0·15 to -0·02; p=0·01). When D-CarDia and consortia data for hypertension were meta-analysed together (n=142 255), the synthesis score was associated with a reduced odds of hypertension (OR per allele, 0·98, 0·96-0·99; p=0·001). In instrumental variable analysis, each 10% increase in genetically instrumented 25(OH)D concentration was associated with a change of -0·29 mm Hg in diastolic blood pressure (-0·52 to -0·07; p=0·01), a change of -0·37 mm Hg in systolic blood pressure (-0·73 to 0·003; p=0·052), and an 8·1% decreased odds of hypertension (OR 0·92, 0·87-0·97; p=0·002). INTERPRETATION: Increased plasma concentrations of 25(OH)D might reduce the risk of hypertension. This finding warrants further investigation in an independent, similarly powered study.

Application of dried blood spots to determine vitamin D status in a large nutritional study with unsupervised sampling: the Food4Me project

An efficient and robust method to measure vitamin D (25-hydroxy vitamin D3 (25(OH)D3) and 25-hydroxy vitamin D2 in dried blood spots (DBS) has been developed and applied in the pan-European multi-centre, internet-based, personalised nutrition intervention study Food4Me. The method includes calibration with blood containing endogenous 25(OH)D3, spotted as DBS and corrected for haematocrit content. The methodology was validated following international standards. The performance characteristics did not reach those of the current gold standard liquid chromatography-MS/MS in plasma for all parameters, but were found to be very suitable for status-level determination under field conditions. DBS sample quality was very high, and 3778 measurements of 25(OH)D3 were obtained from 1465 participants. The study centre and the season within the study centre were very good predictors of 25(OH)D3 levels (P<0·001 for each case). Seasonal effects were modelled by fitting a sine function with a minimum 25(OH)D3 level on 20 January and a maximum on 21 July. The seasonal amplitude varied from centre to centre. The largest difference between winter and summer levels was found in Germany and the smallest in Poland. The model was cross-validated to determine the consistency of the predictions and the performance of the DBS method. The Pearson's correlation between the measured values and the predicted values was r 0·65, and the sd of their differences was 21·2 nmol/l. This includes the analytical variation and the biological variation within subjects. Overall, DBS obtained by unsupervised sampling of the participants at home was a viable methodology for obtaining vitamin D status information in a large nutritional study.

Na verdade a senhora Blum gostaria de conhecer o leiteiro: procedimentos poéticos em Peter Bichsel

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

2-D ZO CRS stack by considering an acquisition line with smooth topography

Os dados sísmicos terrestres são afetados pela existência de irregularidades na superfície de medição, e.g. a topografia. Neste sentido, para obter uma imagem sísmica de alta resolução, faz-se necessário corrigir estas irregularidades usando técnicas de processamento sísmico, e.g. correições estáticas residuais e de campo. O método de empilhamento Superfície de Reflexão Comum, CRS ("Common-Reflection-Surface", em inglês) é uma nova técnica de processamento para simular seções sísmicas com afastamento-nulo, ZO ("Zero-Offset", em inglês) a partir de dados sísmicos de cobertura múltipla. Este método baseia-se na aproximação hiperbólica de tempos de trânsito paraxiais de segunda ordem referido ao raio (central) normal. O operador de empilhamento CRS para uma superfície de medição planar depende de três parâmetros, denominados o ângulo de emergência do raio normal, a curvatura da onda Ponto de Incid��ncia Normal, NIP ("Normal Incidence Point", em inglês) e a curvatura da onda Normal, N. Neste artigo o método de empilhamento CRS ZO 2-D é modificado com a finalidade de considerar uma superfície de medição com topografia suave também dependente desses parâmetros. Com este novo formalismo CRS, obtemos uma seção sísmica ZO de alta resolução, sem aplicar as correições estáticas, onde em cada ponto desta seção são estimados os três parâmetros relevantes do processo de empilhamento CRS.

Empilhamento pelo método superfície de reflexão comum 2-D com topografia e introdução ao caso 3-D

O método de empilhamento sísmico CRS simula seções sísmicas ZO a partir de dados de cobertura múltipla, independente do macro-modelo de velocidades. Para meios 2-D, a função tempo de trânsito de empilhamento depende de três parâmetros, a saber: do ângulo de emergência do raio de reflexão normal (em relação à normal da superfície) e das curvaturas das frentes de onda relacionadas às ondas hipotéticas, denominadas NIP e Normal. O empilhamento CRS consiste na soma das amplitudes dos traços sísmicos em dados de múltipla cobertura, ao longo da superfície definida pela função tempo de trânsito do empilhamento CRS, que melhor se ajusta aos dados. O resultado do empilhamento CRS é assinalado a pontos de uma malha pré-definida na seção ZO. Como resultado tem-se a simulação de uma seção sísmica ZO. Isto significa que para cada ponto da seção ZO deve-se estimar o trio de parâmetros ótimos que produz a máxima coerência entre os eventos de reflexão sísmica. Nesta Tese apresenta-se fórmulas para o método CRS 2-D e para a velocidade NMO, que consideram a topografia da superfície de medição. O algoritmo é baseado na estratégia de otimização dos parâmetros de fórmula CRS através de um processo em três etapas: 1) Busca dos parâmetros, o ângulo de emergência e a curvatura da onda NIP, aplicando uma otimização global, 2) busca de um parâmetro, a curvatura da onda N, aplicando uma otimização global, e 3) busca de três parâmetros aplicando uma otimização local para refinar os parâmetros estimados nas etapas anteriores. Na primeira e segunda etapas é usado o algoritmo Simulated Annealing (SA) e na terceira etapa é usado o algoritmo Variable Metric (VM). Para o caso de uma superfície de medição com variações topográficas suaves, foi considerada a curvatura desta superfície no algoritmo do método de empilhamento CRS 2-D, com aplicação a dados sintéticos. O resultado foi uma seção ZO simulada, de alta qualidade ao ser comparada com a seção ZO obtida por modelamento direto, com uma alta razão sinal-ruído, além da estimativa do trio de parâmetros da função tempo de trânsito. Foi realizada uma nálise de sensibilidade para a nova função de tempo de trânsito CRS em relação à curvatura da superfície de medição. Os resultados demonstraram que a função tempo de trânsito CRS é mais sensível nos pontos-médios afastados do ponto central e para grandes afastamentos. As expressões da velocidade NMO apresentadas foram aplicadas para estimar as velocidades e as profundidades dos refletores para um modelo 2-D com topografia suave. Para a inversão destas velocidades e profundidades dos refletores, foi considerado o algoritmo de inversão tipo Dix. A velocidade NMO para uma superfície de medição curva, permite estimar muito melhor estas velocidades e profundidades dos refletores, que as velocidades NMO referidas as superfícies planas. Também apresenta-se uma abordagem do empilhamento CRS no caso 3-D. neste caso a função tempo de trânsito depende de oito parâmetros. São abordadas cinco estratégias de busca destes parâmetros. A combinação de duas destas estratégias (estratégias das três aproximações dos tempos de trânsito e a estratégia das configurações e curvaturas arbitrárias) foi aplicada exitosamente no empilhamento CRS 3-D de dados sintéticos e reais.

Bryozoan biodiversity in Saint Peter and Saint Paul Archipelago, Brazil

Among the marine invertebrate groups recorded from oceanic islands, bryozoans stand out because they can live and reproduce in suboptimal habitats, which may enhance their dispersal capabilities. This study aimed to update the checklist of bryozoans known from the Saint Peter and Saint Paul Archipelago (ASPSP) and discusses their distribution. During the five expeditions conducted between 2007 and 2009, 22 species were found, of which 16 were new occurrences for the archipelago. The bryozoans were collected from different biotic (algae and invertebrates) and abiotic (rocks, rubble and wrecks) substrata. The bryozoan community in ASPSP includes: eight new and probably endemic species, five species that belong to widespread species complexes, three species known only from the Brazilian coast, two species reported from the Western Atlantic and one species recorded from oceanic islets in the Atlantic. Additionally, three species are widespread in tropical to subtropical waters. Margaretta buski can be highlighted as the most conspicuous and abundant species between 1045 m deep and acts as an "ecosystem engineer".