Distribution of indole-3-acetic acid in Petunia hybrida shoot tip cuttings and relationship between auxin transport, carbohydrate metabolism and adventitious root formation.

To determine the contribution of polar auxin transport (PAT) to auxin accumulation and to adventitious root (AR) formation in the stem base of Petunia hybrida shoot tip cuttings, the level of indole-3-acetic acid (IAA) was monitored in non-treated cuttings and cuttings treated with the auxin transport blocker naphthylphthalamic acid (NPA) and was complemented with precise anatomical studies. The temporal course of carbohydrates, amino acids and activities of controlling enzymes was also investigated. Analysis of initial spatial IAA distribution in the cuttings revealed that approximately 40 and 10% of the total IAA pool was present in the leaves and the stem base as rooting zone, respectively. A negative correlation existed between leaf size and IAA concentration. After excision of cuttings, IAA showed an early increase in the stem base with two peaks at 2 and 24h post excision and, thereafter, a decline to low levels. This was mirrored by the expression pattern of the auxin-responsive GH3 gene. NPA treatment completely suppressed the 24-h peak of IAA and severely inhibited root formation. It also reduced activities of cell wall and vacuolar invertases in the early phase of AR formation and inhibited the rise of activities of glucose-6-phosphate dehydrogenase and phosphofructokinase during later stages. We propose a model in which spontaneous AR formation in Petunia cuttings is dependent on PAT and on the resulting 24-h peak of IAA in the rooting zone, where it induces early cellular events and also stimulates sink establishment. Subsequent root development stimulates glycolysis and the pentosephosphate pathway

Estudio experimental sobre la reacción álcali-sílice en el hormigón producida por áridos de reacción lenta

La finalidad de esta investigación se enmarca dentro de los estudios sobre hormigones de presas llevados a cabo en el Laboratorio Central de Estructuras y Materiales del CEDEX. En España se han diagnosticado 18 obras afectadas tanto por la reacción álcali-sílice de tipo rápido como por la de tipo lento. Dos de de las obras, fabricadas con áridos graníticos, no presentan signos de deterioro pero en laboratorio se han hallado productos expansivos. En las 16 presas españolas restantes, el hormigón de 10 de ellas estaba fabricado con áridos graníticos, de las cuales, 7 están afectadas por la reacción de tipo lento. Sin embargo, en las clasificaciones internacionales de las rocas potencialmente reactivas se establece habitualmente que las rocas graníticas son inocuas o de reactividad muy baja. En los casos puntuales encontrados en la literatura, la reactividad de este tipo de áridos se encuentra frecuentemente asociada al cuarzo microcristalino que contienen y ocasionalmente se asocia esta reactividad también al cuarzo deformado y/o microfisurado. En la tesis doctoral de Víctor Daniel Lanza Fernández, también realizada en el CEDEX, se han tratado los áridos de reacción rápida, dando una descripción detallada de los componentes que intervienen en este tipo de reacción y presentando un ensayo eficaz para su detección en el laboratorio. La investigación desarrollada en la presente tesis doctoral se ha centrado en los áridos de reacción lenta, mucho menos estudiados. A partir del estudio bibliográfico realizado sobre este tipo de reacción se ha detectado ciertas lagunas, basadas principalmente en la falta de un método de detección en el laboratorio para los áridos de reacción lenta. No se ha encontrado un procedimiento en el estudio petrográfico que sea fiable y cuantificable para este tipo de áridos. El ensayo acelerado de barras de mortero, que para los áridos de reacción rápida supone un método rápido y fiable, falla en la detección de los áridos de reacción lenta al aplicar los límites normalizados. Si bien en publicaciones recientes se ha propuesto la posibilidad de ampliar el tiempo de tratamiento hasta los 90 días, la duración del ensayo es más larga de lo deseado. Para resolver estas lagunas, se han tomado áridos de obras reales afectadas por la reacción álcali-sílice (en algún caso también de las canteras de donde se extrajeron los áridos para la ejecución), con lo que la reactividad de estos áridos queda demostrada por su comportamiento en obra. Al objeto de aumentar la muestra de ensayo y utilizar también áridos inocuos, se han tomado muestras de canteras, tanto en uso como abandonadas. Sobre estos áridos se ha realizado una caracterización completa con los ensayos actualmente disponibles para el estudio de su reactividad con los álcalis del cemento: estudio petrográfico, ensayo acelerado de barras de mortero y Gel-Pat modificado. En la investigación se ha desarrollado una metodología de tinción de geles álcali-sílice para la cuantificación de los mismos en el interior de las barras de mortero. Se ha relacionado el volumen de gel generado en la reacción con la expansión producida, tanto para áridos lentos, como rápidos, estableciendo analogías y diferencias en el comportamiento de ambos. La cuantificación de este tipo de compuestos realizada en las barras de mortero, abre la posibilidad de estudiar la capacidad expansiva que presentan en testigos de hormigón de obras afectadas. Este dato podría ser una herramienta importante para el desarrollo de modelos matemáticos que puedan predecir el futuro comportamiento de las estructuras afectadas por la reacción álcali-sílice. La tinción ha sido asimismo utilizada para establecer un método de detección de áridos reactivos basado en el ensayo de Gel-Pat modificado. La metodología de detección propuesta para estos áridos es cuantitativa y proporciona los resultados en 14 días, suponiendo una ventaja importante sobre los métodos actuales, que permiten detectarlos en 90 días (de forma orientativa en 56 días). This research is part of the studies on durability of concrete dams carried out in the Laboratorio Central de Estructuras y Materiales of CEDEX during the last years. At the present time, 18 public works affected by alkali-silica reaction have been diagnosed in Spain. In 12 of them the concrete was mixed with granitic aggregates, 7 of which were damaged by the slow alkali-silica reaction, 3 by the rapid alkali-silica reaction and in the other 2 cases, although there was no visual evidence of the reaction in field, the laboratory tests showed the presence of expansive products inside the concrete. However, in the international classifications of potentially reactive aggregates granitic rocks are pointed out as innocuous or as low reactivity and usually their reactivity is attributed to the presence of microcrystalline quartz and occasionally with the strained and microcracked quartz. The rapid reactive aggregates were deeply studied in a previous research (Víctor Daniel Lanza’s Thesis) also carried out in CEDEX. In this research, a detailed description of the components involved in the rapid alkali-silica reaction was given. Also, a fast method to detect them in the laboratory was developed. The research of the present PhD Thesis is focused in the slow-reactive aggregates, much less studied. The state of the art has shown some gaps in the knowledge about this reaction, mainly the absence of a reliable method to detect slow-reactive aggregates. On one side, there is no systematic and quantitative petrographic test for these aggregates. On the other side, the accelerated mortar bar test has been proved to be effective to detect rapid reactive aggregates but the standard limits fail in the detection of the slowreactive aggregates. In recent investigations it has been proposed extending the test until 90 days, but this period is considered too long. In this research, aggregates taken from affected structures have been used in the test, so their reactivity has been shown in practice. With the aim of increasing the number of samples aggregates from quarries have been also used. A method to stain alkali-silica gels has been applied to mortar bars, thereby quantifying the volume of gel generated inside. This volume has been related to the expansion of the bars, both for rapid for slow reactive aggregates, establishing similarities and differences. The quantification of the gel volume, applied to concrete cores extracted from affected structures, could be an important tool in mathematical models to predict the future behaviour of the structures affected by the alkali-silica reaction. The staining method has been also used to develop a method to detect slow reactive aggregates, based on the optimised Gel-Pat test, obtaining results in 14 days. This represents a major improvement compared to the actual methods (accelerated mortar bar test) which give an indicative value of reactivity at 56 days, and a final result at 90 days.

UM ESTUDO PRELIMINAR DAS INFLUÊNCIAS DO MERCADO DE COMMODITIES NA VARIAÇÃO DO CUSTO DA ALIMENTAÇÃO DO TRABALHADOR: ANÁLISE DE RISCO DA SEGURANÇA ALIMENTAR E NUTRICIONAL

A Segurança Alimentar e Nutricional do trabalhador brasileiro é uma das formas de garantia de saúde e conseqüente aumento de produtividade refletido pelo compromisso do PAT Programa de Alimentação do Trabalhador que completou 30 anos em 2006. O objetivo do estudo é relacionar a variação de preços das commodities de alimentos presentes na refeição do trabalhador de acordo com o sistema normativo do PAT, identificando possíveis conseqüências negativas à Segurança Alimentar do Trabalhador no Brasil. A metodologia, por meio da análise de regressão múltipla, utiliza os dados de ICV Índice de Custo de Vida - de alimentação fora do domicílio como proxy a alimentação do trabalhador. Os resultados sugerem que os alimentos que apresentaram variação negativa com os tipos de refeições analisadas, como a carne suína e a soja, podem ser substituídos por outro valor nutricional semelhante como a carne bovina, o frango e o óleo de milho. Apenas a variação do preço do feijão foi seguida pelo aumento do preço do prato comercial, cuja alternativa é a mudança na maneira de oferecer a refeiç ão ao trabalhador, como os restaurantes por quilo. Diante do cenário exposto, há indícios de que a variação de preços dos alimentos no mercado de commodities teria pouca influência no preço final das refeições, mas a discussão em torno da futura escassez de oferta de alimentos devido ao aumento da população mundial e a mudança dos padrões alimentares com as sociedades urbanizadas insere o risco da SAN no contexto político e econômico da nutrição.(AU)

UM ESTUDO PRELIMINAR DAS INFLUÊNCIAS DO MERCADO DE COMMODITIES NA VARIAÇÃO DO CUSTO DA ALIMENTAÇÃO DO TRABALHADOR: ANÁLISE DE RISCO DA SEGURANÇA ALIMENTAR E NUTRICIONAL

A Segurança Alimentar e Nutricional do trabalhador brasileiro é uma das formas de garantia de saúde e conseqüente aumento de produtividade refletido pelo compromisso do PAT Programa de Alimentação do Trabalhador que completou 30 anos em 2006. O objetivo do estudo é relacionar a variação de preços das commodities de alimentos presentes na refeição do trabalhador de acordo com o sistema normativo do PAT, identificando possíveis conseqüências negativas à Segurança Alimentar do Trabalhador no Brasil. A metodologia, por meio da análise de regressão múltipla, utiliza os dados de ICV Índice de Custo de Vida - de alimentação fora do domicílio como proxy a alimentação do trabalhador. Os resultados sugerem que os alimentos que apresentaram variação negativa com os tipos de refeições analisadas, como a carne suína e a soja, podem ser substituídos por outro valor nutricional semelhante como a carne bovina, o frango e o óleo de milho. Apenas a variação do preço do feijão foi seguida pelo aumento do preço do prato comercial, cuja alternativa é a mudança na maneira de oferecer a refeiç ão ao trabalhador, como os restaurantes por quilo. Diante do cenário exposto, há indícios de que a variação de preços dos alimentos no mercado de commodities teria pouca influência no preço final das refeições, mas a discussão em torno da futura escassez de oferta de alimentos devido ao aumento da população mundial e a mudança dos padrões alimentares com as sociedades urbanizadas insere o risco da SAN no contexto político e econômico da nutrição.(AU)

Ephrin-dependent growth and pruning of hippocampal axons

Neuronal connections are arranged topographically such that the spatial organization of neurons is preserved by their termini in the targets. During the development of topographic projections, axons initially explore areas much wider than the final targets, and mistargeted axons are pruned later. The molecules regulating these processes are not known. We report here that the ligands of the Eph family tyrosine kinase receptors may regulate both the initial outgrowth and the subsequent pruning of axons. In the presence of ephrins, the outgrowth and branching of the receptor-positive hippocampal axons are enhanced. However, these axons are induced later to degenerate. These observations suggest that the ephrins and their receptors may regulate topographic map formation by stimulating axonal arborization and by pruning mistargeted axons.

A mutation in the transmembrane/luminal domain of the ryanodine receptor is associated with abnormal Ca2+ release channel function and severe central core disease

Central core disease is a rare, nonprogressive myopathy that is characterized by hypotonia and proximal muscle weakness. In a large Mexican kindred with an unusually severe and highly penetrant form of the disorder, DNA sequencing identified an I4898T mutation in the C-terminal transmembrane/luminal region of the RyR1 protein that constitutes the skeletal muscle ryanodine receptor. All previously reported RYR1 mutations are located either in the cytoplasmic N terminus or in a central cytoplasmic region of the 5,038-aa protein. The I4898T mutation was introduced into a rabbit RYR1 cDNA and expressed in HEK-293 cells. The response of the mutant RyR1 Ca2+ channel to the agonists halothane and caffeine in a Ca2+ photometry assay was completely abolished. Coexpression of normal and mutant RYR1 cDNAs in a 1:1 ratio, however, produced RyR1 channels with normal halothane and caffeine sensitivities, but maximal levels of Ca2+ release were reduced by 67%. [3H]Ryanodine binding indicated that the heterozygous channel is activated by Ca2+ concentrations 4-fold lower than normal. Single-cell analysis of cotransfected cells showed a significantly increased resting cytoplasmic Ca2+ level and a significantly reduced luminal Ca2+ level. These data are indicative of a leaky channel, possibly caused by a reduction in the Ca2+ concentration required for channel activation. Comparison with two other coexpressed mutant/normal channels suggests that the I4898T mutation produces one of the most abnormal RyR1 channels yet investigated, and this level of abnormality is reflected in the severe and penetrant phenotype of affected central core disease individuals.

A neuroendocrine role for chemerin in hypothalamic remodelling and photoperiodic control of energy balance

We thank Donna Wallace and the animal house staff for their help with the animal studies. We thank Pat Bain for help in preparing the figures. This work was supported by the Biotechnology and Biological Science Research Council (BBSRC) grant number BB/K001043/1 (G.H., A.W.R., P.N.S., P.J.Mc. and P.J.M.) and the Scottish Government (A.W.R., L.M.T., C.D.M. and P.J.M.).

Copy Number Variations in DISC1 and DISC1-Interacting Partners in Major Mental Illness

Acknowledgements We would like to thank all of the patients, relatives and control individuals who participated in the study. We are indebted to the late Prof. Walter Muir, Chair of Developmental Psychiatry and Honorary Consultant in Learning Disability Psychiatry, University of Edinburgh, who initiated these studies and whose work was dedicated to the welfare of the patients who generously participated. We are also grateful to Mrs. Pat Malloy for her assistance with DNA collection and MAQ assays screening of the Scottish samples. The Scottish sample collection was supported by a grant from the Chief Scientist Office (CSO), part of the Scottish Government Health and Social Care Directorates. This research was funded by grants from the CSO to B.S.P. (grant CZB/4/610), The Academy of Medical Sciences/Wellcome Trust to M.J. (grant R41455) and The RS Macdonald Charitable Trust (grant D21419 together with J.H.), the Swedish Research Council (grants 2003-5158 and 2006-4472), the Medical Faculty, Umeå University, and the County Councils of Västerbotten and Norrbotten, Sweden, as well as by grants from the Fund for Scientific Research Flanders (FWO-F), the Industrial Research Fund (IWT) and the Special Research Fund of the University of Antwerp, Belgium. M.J. is funded by a Wellcome Trust Clinical Research Fellowship for MB PhD graduates (R42811). We acknowledge the contribution of the personnel of the VIB Genetic Service Facility (http://www.vibgeneticservicefacility.be/) for the genetic analysis of the Swedish samples. Research nurses Gunnel Johansson, Lotta Kronberg, Tage Johansson and Lisbeth Bertilsson are thankfully acknowledged for their help and expertise. The Betula Study was funded by the Swedish Research Council (grants 345-2003-3883 and 315-2004-6977). We also acknowledge the contribution by the staff in the Betula project

A systematic review of the cost-effectiveness of non-surgical obesity interventions in men

Crown Copyright © 2015. Published by Elsevier Ltd. All rights reserved.

A synthetic cryIC gene, encoding a Bacillus thuringiensis δ-endotoxin, confers Spodoptera resistance in alfalfa and tobacco

Spodoptera species, representing widespread polyphagous insect pests, are resistant to Bacillus thuringiensis δ-endotoxins used thus far as insecticides in transgenic plants. Here we describe the chemical synthesis of a cryIC gene by a novel template directed ligation–PCR method. This simple and economical method to construct large synthetic genes can be used when routine resynthesis of genes is required. Chemically phosphorylated adjacent oligonucleotides of the gene to be synthesized are assembled and ligated on a single-stranded, partially homologous template derived from a wild-type gene (cryIC in our case) by a thermostable Pfu DNA ligase using repeated cycles of melting, annealing, and ligation. The resulting synthetic DNA strands are selectively amplified by PCR with short specific flanking primers that are complementary only to the new synthetic DNA. Optimized expression of the synthetic cryIC gene in alfalfa and tobacco results in the production of 0.01–0.2% of total soluble proteins as CryIC toxin and provides protection against the Egyptian cotton leafworm (Spodoptera littoralis) and the beet armyworm (Spodoptera exigua). To facilitate selection and breeding of Spodoptera-resistant plants, the cryIC gene was linked to a pat gene, conferring resistance to the herbicide BASTA.

Cyclin E, a redundant cyclin in breast cancer

Cyclin E is an important regulator of cell cycle progression that together with cyclin-dependent kinase (cdk) 2 is crucial for the G1/S transition during the mammalian cell cycle. Previously, we showed that severe overexpression of cyclin E protein in tumor cells and tissues results in the appearance of lower molecular weight isoforms of cyclin E, which together with cdk2 can form a kinase complex active throughout the cell cycle. In this study, we report that one of the substrates of this constitutively active cyclin E/cdk2 complex is retinoblastoma susceptibility gene product (pRb) in populations of breast cancer cells and tissues that also overexpress p16. In these tumor cells and tissues, we show that the expression of p16 and pRb is not mutually exclusive. Overexpression of p16 in these cells results in sequestering of cdk4 and cdk6, rendering cyclin D1/cdk complexes inactive. However, pRb appears to be phosphorylated throughout the cell cycle following an initial lag, revealing a time course similar to phosphorylation of glutathione S-transferase retinoblastoma by cyclin E immunoprecipitates prepared from these synchronized cells. Hence, cyclin E kinase complexes can function redundantly and replace the loss of cyclin D-dependent kinase complexes that functionally inactivate pRb. In addition, the constitutively overexpressed cyclin E is also the predominant cyclin found in p107/E2F complexes throughout the tumor, but not the normal, cell cycle. These observations suggest that overexpression of cyclin E in tumor cells, which also overexpress p16, can bypass the cyclin D/cdk4-cdk6/p16/pRb feedback loop, providing yet another mechanism by which tumors can gain a growth advantage.

Gonadotropin-releasing hormone analogue conjugates with strong selective antitumor activity

Conjugation of gonadotropin-releasing hormone (GnRH) analogues GnRH-III, MI-1544, and MI-1892 through lysyl side chains and a tetrapeptide spacer, Gly-Phe-Leu-Gly (X) to a copolymer, poly(N-vinylpyrrolidone-co-maleic acid) (P) caused increased antiproliferative activity toward MCF-7 and MDA-MB-231 breast, PC3 and LNCaP prostate, and Ishikawa endometrial cancer cell lines in culture and against tumor development by xenografts of the breast cancer cells in immunodeficient mice. MCF-7 cells treated with P-X-1544 and P-X-1892 displayed characteristic signs of apoptosis, including vacuoles in the cytoplasm, rounding up, apoptotic bodies, bleb formation, and DNA fragmentation. Conjugates, but not free peptides, inhibited cdc25 phosphatase and caused accumulation of Ishikawa and PC3 cells in the G2/M phase of the cell cycle after 24 h at lower doses and in the G1 and G2 phases after 48 h. Since P-X-peptides appear to be internalized, the increased cytotoxicity of the conjugates is attributed to protection of peptides from proteolysis, enhanced interaction of the peptides with the GnRH receptors, and/or internalization of P-X-peptide receptor complexes so that P can exert toxic effects inside, possibly by inhibiting enzymes involved in the cell cycle. The additional specificity of P-X-peptides compared with free peptides for direct antiproliferative effects on the cancer cells but not for interactions in the pituitary indicates the therapeutic potential of the conjugates.