Buried clay pot irrigation for efficient and controlled water delivery

This study presents water flow (WF) into soil from several pitchers buried in the soil up to their neck and filled with water,under natural atmospheric conditions for a period of two years. Variation in daily WF into soil indicated a direct correlation with moisture deficit (MD) in atmosphere. WF increases linearly with MD for non rainy days. WF without hydraulic head through all pots varied in the order air>soil>water. Base line flow in water with respect to air was < 5%. WF for pots with hydraulic head was also in the order air>soil>water, but with significant increase in WF. Hydraulic conductivity Ks was in the order air>soil>water.Ks in water was independent of MD, whereas for air and soil, Ks increased with MD. Thus total WF is partially under hydraulic head and partly due to pull effect through capillary pores on pot wall either due to MD in air or prevailing soil water tension in soil.

Assessing economically viable carbon reductions for the production of ammonia from biomass gasification

Greenhouse gas emissions from fertiliser production are set to increase before stabilising due to the increasing demand to secure sustainable food supplies for a growing global population. However, avoiding the impacts of climate change requires all sectors to decarbonise by a very high level within several decades. Economically viable carbon reductions of substituting natural gas reforming with biomass gasification for ammonia production are assessed using techno-economic and life cycle assessment. Greenhouse gas savings of 65% are achieved for the biomass gasification system and the internal rate of return is 9.8% at base-line biomass feedstock and ammonia prices. Uncertainties in the assumptions have been tested by performing sensitivity analysis, which show, for example with a ±50% change in feedstock price, the rate of return ranges between -0.1% and 18%. It would achieve its target rate of return of 20% at a carbon price of £32/t CO, making it cost competitive compared to using biomass for heat or electricity. However, the ability to remain competitive to investors will depend on the volatility of ammonia prices, whereby a significant decrease would require high carbon prices to compensate. Moreover, since no such project has been constructed previously, there is high technology risk associated with capital investment. With limited incentives for industrial intensive energy users to reduce their greenhouse gas emissions, a sensible policy mechanism could target the support of commercial demonstration plants to help ensure this risk barrier is resolved. © 2013 The Authors.

Assessing economically viable carbon reductions for the production of ammonia from biomass gasification

Greenhouse gas emissions from fertiliser production are set to increase before stabilising due to the increasing demand to secure sustainable food supplies for a growing global population. However, avoiding the impacts of climate change requires all sectors to decarbonise by a very high level within several decades. Economically viable carbon reductions of substituting natural gas reforming with biomass gasification for ammonia production are assessed using techno-economic and life cycle assessment. Greenhouse gas savings of 65% are achieved for the biomass gasification system and the internal rate of return is 9.8% at base-line biomass feedstock and ammonia prices. Uncertainties in the assumptions have been tested by performing sensitivity analysis, which show, for example with a ±50% change in feedstock price, the rate of return ranges between -0.1% and 18%. It would achieve its target rate of return of 20% at a carbon price of £32/t CO, making it cost competitive compared to using biomass for heat or electricity. However, the ability to remain competitive to investors will depend on the volatility of ammonia prices, whereby a significant decrease would require high carbon prices to compensate. Moreover, since no such project has been constructed previously, there is high technology risk associated with capital investment. With limited incentives for industrial intensive energy users to reduce their greenhouse gas emissions, a sensible policy mechanism could target the support of commercial demonstration plants to help ensure this risk barrier is resolved. © 2013 The Authors.

Relevance of CD40/CD40L downstream pathways to Alzheimer's disease

The amyloid cascade hypothesis places amyloid-β at the origin of Alzheimer's disease (AD). Amyloid-β (Aβ) is the product of the sequential cleavage of the amyloid precursor protein (APP) by the enzymes β- and γ-secretases. An inflammatory component to AD has been suggested in association with CD40 (a member of the tumor necrosis factor receptor superfamily (TNFRS) and its cognate ligand CD40L. In this study, I hypothesized that the neutralization of pro-inflammatory cytokines produced downstream of CD40/CD40L interaction would reduce APP processing. I also hypothesized that blocking the binding of different adaptor proteins to CD40 by mutating its cytoplasmic tail would result in significant reduction of the APP metabolites: Aβ, sAPPβ, sAPPα, CTFβ and CTFα. ^ Treatment with CD40L of human embryonic kidney cells over-expressing both APP and CD40 (HEK/APPsw/CD40) significantly increased levels of the cytokine granulocyte macrophage colony stimulating factor (GM-CSF). Neutralizing antibodies against GM-CSF mitigated the CD40L-induced production of Aβ in these cells. Treatment of the HEK/APPsw/CD40 cells with recombinant GM-CSF significantly increased Aβ levels. GM-CSF receptor gene silencing with shRNA significantly reduced Aβ levels to below base line in non-stimulated HEK/APPsw/CD40 cells. Silencing of the GM-CSF receptor also decreased APP endocytosis (therefore reducing the availability of APP to be cleaved in the endosomes). ^ Using CD40 mutants, I show that CD40L can increase levels of Aβ(1-40), Aβ(1-42), sAPPβ, sAPPα and CTFβ independently of TRAF signaling. TRAFs had been shown to be necessary for most CD40/CD40L-dependent signaling. An increase in mature/immature APP ratio after CD40L treatment of CD40wt and CD40-mutant cells was observed, reflecting alterations in APP trafficking. CD4OL treatment of a neuroblastoma cell line over-expressing CTFβ suggested that CD40L affected γ-secretase activity. Inhibition of γ-secretase activity significantly reduced sAPPβ levels in the CD40L treated HEK/APPsw CD40wt and the CD40-mutant cells. The latter suggests CD40/CD40L interaction primarily acts on γ-secretase and affects β-secretase via a positive feedback mechanism. ^ Taken together, the results of this dissertation suggest that GM-CSF operates downstream of CD40/CD40L interaction and that GM-CSF modulates Aβ production by influencing APP trafficking. Moreover, the data presented suggest that CD40/CD40L interaction can modulate APP processing via a mechanism independent of TRAF signaling. ^

Seabed photographs taken along OFOS profiles during POLARSTERN cruise PS96 (ANT-XXXI/2 FROSN)

Within the context of the overall ecological working programme Dynamics of Antarctic Marine Shelf Ecosystems (DynAMo) of the PS96 (ANT-XXXI/2) cruise of RV "Polarstern" to the Weddell Sea (Dec 2015 to Feb 2016), seabed imaging surveys were carried out along drift profiles by means of the Ocean Floor Observation System (OFOS) of the Alfred Wegener Institute, Helmholtz Centre for Polar and Marine Research (AWI) Bremerhaven. The setup and mode of deployment of the OFOS was similar to that described by Bergmann and Klages (2012, doi:10.1016/j.marpolbul.2012.09.018). OFOS is a surface-powered gear equipped with two downward-looking cameras installed side-by-side: one high-resolution, wide-angle still camera (CANON® EOS 5D Mark III; lens: Canon EF 24 f/1.4L II, f stop: 13, exposure time: 1/125 sec; in-air view angles: 74° (horizontal), 53° (vertical), 84° (diagonal); image size: 5760 x 3840 px = 21 MPix; front of pressure resistant camera housing consisting of plexiglass dome port) and one high-definition color video camera (SONY® FCB-H11). The system was vertically lowered over the stern of the ship with a broadband fibre-optic cable, until it hovers approximately 1.5 m above the seabed. It was then towed after the slowly sailing ship at a speed of approximately 0.5 kn (0.25 m/s). The ship's Global Acoustic Positioning System (GAPS), combining Ultra Short Base Line (USBL), Inertial Navigation System (INS) and satellite-based Global Positioning System (GPS) technologies, was used to gain highly precise underwater position data of the OFOS. During the profile, OFOS was kept hanging at the preferred height above the seafloor by means of the live video feed and occasional minor cable-length adjustments with the winch to compensate small-scale bathymetric variations in seabed morphology. Information on water depth and height above the seafloor were continuously recorded by means of OFOS-mounted sensors (GAPS transponder, Tritech altimeter). Three lasers, which are placed beside the still camera, emit parallel beams and project red light points, arranged as an equilateral triangle with a side length of 50 cm, in each photo, thus providing a scale that can be used to calculate the seabed area depicted in each image and/or measure the size of organisms or seabed features visible in the image. In addition, the seabed area depicted was estimated using altimeter-derived height above seafloor and optical characteristics of the OFOS still camera. In automatic mode, a seabed photo, depicting an area of approximately 3.45 m**2 (= 2.3 m x 1.5 m; with variations depending on the actual height above ground), was taken every 30 seconds to obtain series of "TIMER" stills distributed at regular distances along the profiles that vary in length depending on duration of the cast. At a ship speed of 0.5 kn, the average distance between seabed images was approximately 5 m. Additional "HOTKEY" photos were taken from interesting objects (organisms, seabed features, such as putative iceberg scours) when they appeared in the live video feed (which was also recorded, in addition to the stills, for documentation and possible later analysis). If any image from this collection is used, please cite the reference as given above.

The reproductive biology of the two-banded sea bream (Diplodus vulgaris) from the southwest coast of Portugal

The study of the reproduction of Diplodus vulgaris (Geoff.) as part of a base-line study of the fishery resources of the southwest coast of Portugal, was based on the analysis of the spawning season, gonad maturation, size at maturity, fecundity and hermaphroditism. The spawning season is relatively long, from December to March, with peaks in January and February. No significant differences were found either in the sex ratio (M:F = 1.01) over the year or by size. The size at first maturity (L-50) for all sexes and undetermined individuals combined was 18.33 cm total length (TL), with no significant differences between males and females. The estimated L-50 is considerably greater than the minimum legal size in Portugal of 15.0 cm. Mean absolute fecundity (F-a) was 131 127 oocytes, ranging from 31 523 to 250 608. The relationship between absolute fecundity and total length (TL) (F-a = 25 398 TL-484 426) and somatic weight (SW) (F-a = 878.8SW-71 416) was of the linear type. The mean number of oocytes per gram of somatic weight was 526, ranging from 194 to 887. The reproductive strategy of this species is characterized by a rudimentary hermaphroditism with possible protandry, as evidenced by the existence of individuals in sexual transition.

Controle on-line por atributos para o número de não-conformidades no item inspecionado com base em uma sequência de inspeção

This paper proposes a procedure to control on-line processes for attributes, using an Shewhart control chart with two control limits (warning limit and control limit) and will be based on a sequence of inspection (h). The inspection procedure is based on Taguchi et al. (1989), in which to inspect the item, if the number of non-conformities is higher than an upper control limit, the process needs to be stopped and some adjustment is required; and, if the last inspection h, from all items inspected present a number of non-conformities between the control limit and warning limit. The items inspected will suffer destructive inspection, being discarded after inspection. Properties of an ergodic Markov chain are used to get the expression of average cost per item and the aim was the determination of four optimized parameters: the sampling interval of the inspections (m); the constant W to draw the warning limit (W); the constant C to draw the control limit (C), where W £ C, and the length of sequence of inspections (h). Numerical examples illustrate the proposed procedure

Fontes de metionina em rações formuladas com base em aminoácidos totais ou digestíveis para frangas de reposição leves e semipesadas

Foram conduzidos dois experimentos com o objetivo de avaliar a utilização de duas fontes de metionina (em pó ou líquida) em rações formuladas com base em aminoácidos digestíveis ou totais para frangas de reposição leves e semipesadas, nas fases inicial (de 1 a 6 semanas de idade) e de crescimento (de 11 a 16 semanas de idade). Em cada experimento, foram utilizadas 384 aves distribuídas em delineamento inteiramente ao acaso, em esquema fatorial 2 × 2 × 2, composto de duas linhagens (Hy-Line W36 e Hy-Line Brown), duas fontes de metionina (HMTBA - 88% e DL-Metionina - 99%) e dois critérios de formulação da dieta (aminoácidos totais e digestíveis), totalizando oito tratamentos, cada um com seis repetições de oito aves. Foram avaliados os consumos de ração, proteína bruta e energia metabolizável, o peso e ganho de peso corporal e a conversão alimentar das aves. Não houve interação entre os fatores para nenhuma das variáveis estudadas. As aves da linhagem Brown apresentaram maior consumo de ração, peso corporal e ganho de peso. As fontes de metionina e os critérios de formulação de ração não influenciaram as características de desempenho. As aves das linhagens W36 e Brown da linhagem Hy-Line apresentam diferentes perfis de desenvolvimento corporal. A metionina em pó (DL-metionina - 99%) ou a líquida (HMTBA - 88%), assim como os critérios de formulação de ração (aminoácidos totais ou digestíveis) proporcionam desempenho semelhante em aves de reposição.

Sistema de coleta on-line do Censo Escolar da Educa????o B??sica (Educacenso)

O Educacenso ?? um sistema eletr??nico de coleta de informa????es educacionais composto por um aplicativo web, que permite a coleta, migra????o e altera????o de dados educacionais das escolas em todo o territ??rio nacional, e por um banco de dados relacional, que armazena de forma sistem??tica todas estas informa????es. O Educacenso representa uma inova????o, pois se trata de um banco de dados ??nico, de base nacional, alimentado por dados e informa????es que v??m diretamente das escolas, por meio da internet. Constitui-se no mais completo cadastro de escolas, alunos e docentes do pa??s. A constru????o de um banco de dados com informa????es individualizadas amplia as possibilidades de comunica????o com outras bases de dados do governo federal. O novo desenho metodol??gico e a utiliza????o de recursos de tecnologia da informa????o permitiram ampliar a precis??o e a fidedignidade dos dados do Censo Escolar, o que possibilita realiza????o de ajustes na pol??tica educacional e maior efetividade do gasto p??blico com a educa????o b??sica

Conhecendo o "valor do cliente" de um jornal on-line

Com o advento da Internet, uma questão parece peculiar ao marketing: conhecer o comportamento de um "novo" consumidor, o on-line. Uma proposta é o conhecimento do "valor do cliente", que é a percepção do consumidor sobre como atributos de produtos podem propiciar benefícios que o ajudem a atingir suas metas. A teoria de cadeias de meios-fim é a forma de se pensar sistematicamente nessa representação. O método mais amplamente utilizado para atingir as cadeias de meios-fim é o laddering. A pesquisa foi realizada com uma amostra de 78 usuários de um grande jornal on-line. Os principais "valores do cliente" dos usuários do jornal propiciaram conclusões importantes. Foram identificados leitores que se preocupam muito com seus objetivos pessoais, mas que também desejam interagir com os outros e que demonstraram interesse com questões sociais. Com base nisso, implicações gerenciais são discutidas e sugestões para futuras pesquisas são indicadas.

O conceito da qualidade nos produtos à base de Ginkgo biloba comercializados em Portugal

O extracto de Ginkgo biloba é o produto fitoterápico mais vendido na Europa. Em Portugal e muitos países, a maioria dos produtos à base de plantas são comercializados como suplementos alimentares, não estando garantidos, parâmetros de qualidade, segurança e eficácia. Realizouse um estudo, com recolha de informações, tendo por base uma amostra de 50 produtos à base de ginkgo. Da análise, verificou-se que 94% podiam ser encontrados à venda na internet, e desse total, 89% possuíam informação on-line quanto à composição. Apenas 40% referem a utilização do extracto padronizado de ginkgo e muitos recomendam doses superiores às referidas como terapêuticas.

In-line rheo-optical microstructural characterization of complex polymer systems

Tese de Doutoramento em Ciência e Engenharia de Polímeros e Compósitos.

Disease-causing mutations improving the branch site and polypyrimidine tract: pseudoexon activation of LINE-2 and antisense Alu lacking the poly(T)-tail.

Cryptic exons or pseudoexons are typically activated by point mutations that create GT or AG dinucleotides of new 5' or 3' splice sites in introns, often in repetitive elements. Here we describe two cases of tetrahydrobiopterin deficiency caused by mutations improving the branch point sequence and polypyrimidine tracts of repeat-containing pseudoexons in the PTS gene. In the first case, we demonstrate a novel pathway of antisense Alu exonization, resulting from an intronic deletion that removed the poly(T)-tail of antisense AluSq. The deletion brought a favorable branch point sequence within proximity of the pseudoexon 3' splice site and removed an upstream AG dinucleotide required for the 3' splice site repression on normal alleles. New Alu exons can thus arise in the absence of poly(T)-tails that facilitated inclusion of most transposed elements in mRNAs by serving as polypyrimidine tracts, highlighting extraordinary flexibility of Alu repeats in shaping intron-exon structure. In the other case, a PTS pseudoexon was activated by an A>T substitution 9 nt upstream of its 3' splice site in a LINE-2 sequence, providing the first example of a disease-causing exonization of the most ancient interspersed repeat. These observations expand the spectrum of mutational mechanisms that introduce repetitive sequences in mature transcripts and illustrate the importance of intronic mutations in alternative splicing and phenotypic variability of hereditary disorders.

Characterization of the effects of IL-17F on CHO cell line generation

CHO is the most commonly used mammalian host for the generation of cell lines allowing for the production of high quality therapeutic proteins. The generation of such cell lines is a lengthy and resource-intensive process requiring extensive screening in order to isolate candidates with optimal characteristics, such as growth, stability and productivity. For this reason, the biotechnology industry invests much effort in attempts to optimize CHO expression systems in order to streamline and shorten the cell line selection process. Based on preliminary observations of a facilitated selection of CHO-GS cell lines expressing members of the IL-17 cytokine family, this study investigates the use of IL-17F as a novel enhancing factor for CHO cell line generation. Using two different CHO expression systems (exploiting GS and DHFR-based selection), we demonstrated that IL-17F expression caused a significant increase in the occurrence of colonies during the selection process. All colonies selected produced substantial amounts of IL-17F, suggesting that benefits were conferred, during selection, to those cells expressing the cytokine. Furthermore, transgene expression levels were significantly increased when the selection pressure was raised to a level that would not normally be permissive for colony selection (i.e. 100 |o.M MSX for the CHO-GS expression system or 1000 nM MTX for the CHO-DHFR system). Finally, IL-17F expression was also found to enhance the rate of appearance of clones during single cell subcloning in the absence of selection pressure. Overall, these benefits have the potential to allow a substantial reduction in the length of cell line generation while significantly increasing cell line productivity. Nevertheless, we found that the high IL-17F expression levels required to convey enhancing effects was a limitation when attempting to co-express IL-17F and a recombinant soluble protein of therapeutic interest from independent CMV promoters within the same expression vector. In order to understand and overcome this limitation, studies were designed to characterize the IL-17F enhancing effect at the molecular and cellular level. Regular supplementation of recombinant biologically-active IL-17F into the culture medium during cell line selection was not able to reproduce the enhancing effects of endogenous IL-17F expression. In addition, increased IL-17F expression correlated with increased CHO-GS selection transgene expression at the single cell level. This data suggested a possible effect of IL-17F on viral promoter activity or transgene mRNA stability. It also provided direct evidence that the cells expressing the highest amounts of IL-17F obtained the most benefit. Overall data obtained from these study implied that IL-17F may act through an intracellular mechanism, possibly exerted during secretion. We therefore initiated experiments designed to determine the specific compartment(s) within which IL-17F triggers its effect. This work has identified IL-17F as a potentially powerful tool to optimize the CHO cell line generation process. The characterization of this enhancing effect at the molecular level has given us several insights into overcoming the current limitations, thus paving the way for the development of a viable technology that can be exploited within the biotechnology industry. - La CHO est la cellule hôte de mammifere la plus couramment utilisée dans la création de lignée cellulaire produisant des protéines thérapeutiques de haute qualité. La génération de ces lignées cellulaires est un processus long et exigeant l'utilisation de techniques de sélection robustes afin d'isoler des candidats possédants les caractéristiques optimales de croissance, de productivité et de stabilité d'expression. Les industries biopharmaceutiques ont investi beaucoup d'efforts afin d'optimiser les systèmes d'expression CHO dans le but raccourcir la longueur du procédé de sélection de lignées cellulaires et aussi d'en augmenter l'efficacité. A partir d'observations préliminaires obtenues lors de la génération de lignées cellulaires CHO- GS exprimant une cytokine appartenant à la famille des IL-17, nous avons réalisé une étude portant sur l'utilisation de l'IL-17F humaine (IL-17F) comme nouveau facteur d'optimisation pour la génération de lignées cellulaires CHO. Nous avons démontré, en utilisant les deux systèmes de sélection et d'expression CHO couramment utilisés (le premier exploitant la GS et l'autre basée sur la DHFR), que l'expression de l'IL-17F permet une augmentation significative de la fréquence d'apparition de colonies durant le processus de sélection de lignées cellulaires. Les différentes colonies sélectionnées expriment des quantités substantielles d'IL-17F, suggérant un effet bénéfique lors de la sélection qui serait exclusivement conféré aux cellules exprimant la cytokine. En outre, le niveau d'expression du transgene se trouve significativement augmenté lorsque la pression de sélection est portée à un niveau habituellement trop élevé pour permettre la sélection de colonies (soit 100 |JM MSX pour le système d'expression CHO-GS ou 1000 nM MTX pour le système CHO- DHFR). Enfin, l'expression d'IL-17F permet également d'améliorer la vitesse d'apparition de clones pendant une étape de sous-clonage en l'absence de pression de sélection. L'ensemble de ces effets bénéfiques permettent une réduction substantielle de la durée de génération de lignées cellulaires tout en augmentant considérablement la productivité des lignées obtenues. Néanmoins, nous avons constaté que la nécessité d'exprimer des niveaux élevés d'IL-17F afin obtenir l'ensemble de ses effets bénéfiques devient une contrainte lors de l'utilisation d'un vecteur d'expression composé de deux promoteurs CMV indépendants pour la co-expression de la cytokine et d'une protéine soluble présentant un intérêt thérapeutique. Afin de mieux comprendre et de surmonter cette limitation, plusieurs études ont été effectuées dans le but de mieux caractériser l'effet de IL-17F au niveau subcellulaire. L'apport régulier en IL-17F recombinante et biologiquement active dans le milieu de culture lors de la sélection de lignées cellulaires ne permet pas de reproduire les effets bénéfiques observés par l'expression endogène d'IL-17F. En outre, nous avons constaté que, lors de l'utilisation du système CHO- GS, l'augmentation d'expression de 1TL-17F est corrélée à un accroissement de l'expression du marqueur de sélection au niveau cellulaire. Ces résultats suggèrent un possible effet d'IL- 17F sur l'activité des promoteurs viraux et ainsi fournissent une preuve directe que les cellules exprimant de haut niveau d'IL-17F sont celles qui en profitent le plus. L'ensemble de ces observations mettrait en avant que l'effet d'IL-17F se ferait selon un mécanisme intracellulaire. Nous avons donc étudié le(s) compartiment(s) spécifique(s) dans lequel IL-17F pourrait exercer son effet. Ce travail a permis de définir IL-17F comme un puissant outil pour l'optimisation des procédés de génération de lignées cellulaires CHO. La caractérisation de cette amélioration de l'effet au niveau moléculaire nous a donné plusieurs indications sur la manière de dépasser les limitations actuelles, ouvrant ainsi la voie au développement d'une technologie viable qui peut être exploitée pars l'industrie biotechnologique.

Characterization of the murine high Km glucose transporter GLUT2 gene and its transcriptional regulation by glucose in a differentiated insulin-secreting cell line.

In pancreatic beta-cells, the high Km glucose transporter GLUT2 catalyzes the first step in glucose-induced insulin secretion by glucose uptake. Expression of the transporter has been reported to be modulated by glucose either at the protein or mRNA levels. In this study we used the differentiated insulinoma cell line INS-1 which expresses high levels of GLUT2 and show that the expression of GLUT2 is regulated by glucose at the transcriptional level. By run-on transcription assays we showed that glucose induced GLUT2 gene transcription 3-4-fold in INS-1 cells which was paralleled by a 1.7-2.3-fold increase in cytoplasmic GLUT2 mRNA levels. To determine whether glucose regulatory sequences were present in the promoter region of GLUT2, we cloned and characterized a 1.4-kilobase region of mouse genomic DNA located 5' of the translation initiation site. By RNase protection assays and primer extension, we determined that multiple transcription initiation sites were present at positions -55, -64, and -115 from the first coding ATG and which were identified in liver, intestine, kidney, and beta-cells mRNAs. Plasmids were constructed with the mouse promoter region linked to the reporter gene chloramphenicol acetyltransferase (CAT), and transiently and stably transfected in the INS-1 cells. Glucose induced a concentration-dependent increase in CAT activity which reached a maximum of 3.6-fold at 20 mM glucose. Similar CAT constructs made of the human GLUT2 promoter region and the CAT gene displayed the same glucose-dependent increase in transcriptional activity when transfected into INS-1 cells. Comparison of the mouse and human promoter regions revealed sequence identity restricted to a few stretches of sequences which suggests that the glucose responsive element(s) may be conserved in these common sequences.