Genetic transformation of sweet oranges with the D4E1 gene driven by the AtPP2 promoter

The objective of this work was to produce transgenic 'Pêra' and 'Valência' sweet orange plants using the D4E1 gene driven by the Arabidopsis thaliana phloem protein (AtPP2) promoter and to quantify transgene expression in different transformation events. Genetic transformation experiments were carried out with epicotyl segments co‑cultivated with Agrobacterium tumefaciens. Six plants from 'Pêra' sweet orange and seven plants from 'Valência' sweet orange were confirmed as different transgenic events by means of the polymerase chain reaction (PCR) and the Southern blot techniques. Transgene expression was quantified using real‑time quantitative PCR. D4E1 gene expression levels vary from 5 up to 50 times among different transformation events.

Beta-adrenergic blockade and intravenous nutrient-induced thermogenesis in lean and obese women.

Continuous respiratory exchange measurements were performed in nine obese and eight lean women for 1 h before, 3 h during, and 1 h after the intravenous administration of a nutrient mixture infused at twice the postabsorptive resting energy expenditure (REE). This experiment was conducted without or with beta-adrenergic blockade (iv propranolol). Propranolol administration did not change the postabsorptive REE [i.e., 1.03 +/- 0.07 before vs. 1.01 +/- 0.02 kcal/min after administration in lean women and 1.16 +/- 0.04 vs. 1.15 +/- 0.03 kcal/min (NS) in obese women]. The mean overall thermogenic response expressed as a percentage of the infused energy was similar in both groups and was not significantly blunted after propranolol infusion [6.9 +/- 0.4 vs. 5.9 +/- 0.6% in the lean women and 7.5 +/- 0.5 vs. 7.1 +/- 0.6% (NS) in the obese women]. During beta-adrenergic blockade the rate of lipid oxidation decreased in the lean group but was unchanged in the obese group and the glycemic response to nutrient administration was significantly higher in both groups than without propranolol. It is concluded that beta-adrenergic blockade has no effect on REE and on intravenous nutrient-induced thermogenesis in both lean and obese women.

Influence of rootstocks and pruning times on yield and on nutrient content and extraction in 'Niagara Rosada' grapevine

The objective of this work was to evaluate the influence of rootstocks and pruning times on yield and on nutrient content and extraction by pruned branches and harvested bunches of 'Niagara Rosada' grapevine in subtropical climate. The rootstocks 'IAC 766', 'IAC 572', 'IAC 313', 'IAC 571-6', and '106-8 Mgt' were evaluated. Treatments consisted of a combination between five rootstocks and three pruning times. At pruning, fresh and dry matter mass of branches were evaluated to estimate biomass accumulation. At harvest, yield was estimated by weighing of bunches per plant. Branches and bunches were sampled at pruning and at harvest, respectively, for nutrient content analysis. Nutrient content and dry matter mass of branches and bunches were used to estimate total nutrient extraction. 'Niagara Rosada' grapevine grafted onto the 'IAC 572' rootstock had the highest yield and dry matter mass of bunches, which were significantly different from the ones observed in 'Niagara Rosada'/'IAC 313'. 'Niagara Rosada' grafted onto the 'IAC 572' rootstock extracted the largest quantity of K, P, Mg, S, Cu, and Fe, differing from 'IAC 313' and 'IAC 766' in K and P extraction, and from '106-8 Mgt' in Mg and S extraction. Winter pruning results in higher yield, dry matter accumulation by branches, and total nutrient content and extraction.

On the transcriptional role of NLRC5 and the function of NLRC5-driven MHC class I expression in the immune system

Major histocompatibility complex (MHC) molecules are of crucial importance for the immune system to recognize and defend the body against external attacks. Foreign antigens are presented by specialized cells, called antigen presenting cells, to T lymphocytes in the context of MHC molecules, thereby inducing T cell activation. In addition, MHC molecules are essential for Natural Killer (NK) cell biology, playing a role in NK cell education and activation. Recently, the NOD-like receptor (NLR) family member NLRC5 (NLR caspase recruitment domain containing protein 5) was found to act as transcriptional regulator of MHC class I, in particular in T and NK cells. Its role in MHC class I expression is however minor in dendritic cells (DCs). This raised the question of whether inflammatory conditions, which augment the levels of NLRC5 in DCs, could increase its contribution to MHC class I expression. Our work shows that MHC class I transcript and intracellular levels depend on NLRC5, while its role in MHC class I surface expression is instead negligible. We describe however a general salvage mechanism that enables cells with low intracellular MHC class I levels to nevertheless maintain relatively high MHC class I on the cell surface. In addition, we lack a thorough understanding of NLRC5 target gene specificity and mechanism of action. Our work delineates the unique consensus sequence in MHC class I promoters required for NLRC5 recruitment and pinpoints conserved features conferring its specificity. Furthermore, through genome-wide analyses, we confirm that NLRC5 regulates classical MHC class I genes and identify novel target genes all encoding non-classical MHC class I molecules exerting an array of functions in immunity and tolerance. We finally asked why a dedicated factor co-regulates MHC class I expression specifically in T and NK lymphocytes. We show that deregulated NLRC5 expression affects the education of NK cells and alters the crosstalk between T and NK cells, leading to NK cell-mediated killing of T lymphocytes. Altogether this thesis work brings insights into molecular and physiological aspects of NLRC5 function, which might help understand certain aspects of immune responses and disorders. -- Les molécules du complexe majeur d'histocompatibilité (CMH) sont essentielles au système immunitaire pour l'initiation de la réponse immunitaire. En effet, l'activation des lymphocytes T nécessite la reconnaissance d'un antigène étranger présenté par les cellules présentatrices d'antigènes sur une molécule du CMH. Les molécules du CMH ont également un rôle fondamental pour la fonction des cellules Natural Killer (NK) puisqu'elles sont nécessaires à leur processus d'éducation et d'activation. Récemment, NLRC5 (NLR caspase recruitment domain containing protein 5), un membre de la famille des récepteurs de type NOD (NLRs), a été décrit comme un facteur de transactivation de l'expression des gènes du CMH de classe I. A l'état basai, cette fonction transcriptionnelle est essentielle dans les lymphocytes T et NK, alors que ce rôle reste mineur pour l'expression des molécules du CMH de classe I dans les cellules dendritiques (DCs). Dans des conditions inflammatoires, l'expression de NLRC5 augmente dans les DCs. Notre travail démontre que, dans ces conditions, les transcrits et les niveaux intracellulaires des molécules du CMH de classe I augmentent aussi d'une façon dépendante de NLRC5. A contrario, le rôle de NLRC5 sur les niveaux de molécules de surface reste minoritaire. Cette observation nous a conduits à l'identification d'un mécanisme général de compensation qui permet aux cellules de maintenir des niveaux relativement élevés de molécules de CMH de class I à leur surface malgré de faibles niveaux intracellulaires. De plus, il semblait nécessaire de s'orienter vers une approche plus globale afin de déterminer l'étendue de la fonction transcriptionnelle de NLRC5. Par une approche du génome entier, nous avons pu décrire une séquence consensus conservée présente dans les promoteurs des gènes du CMH de classe I, sur laquelle NLRC5 est spécifiquement recruté. Nous avons pu également identifier de nouveaux gènes cibles codant pour des molécules de CMH de classe I non classiques impliqués dans l'immunité et la tolérance. Finalement, nous nous sommes demandé quel est l'intérêt d'avoir un facteur transcriptionnel, en l'occurrence NLRC5, qui orchestre l'expression du CMH de classe I dans les lymphocytes T et NK. Nous montrons que la dérégulation de l'expression de NLRC5 affecte l'éducation des cellules NK et conduit à la mort cellulaire des lymphocytes T médiée par les cellules NK. Dans l'ensemble ce travail de thèse contribue à la caractérisation du rôle de NLRC5, tant au niveau moléculaire que physiologique, ce qui présente un intérêt dans le cadre de la compréhension de certains aspects physiopathologique de la réponse immunitaire.

A Monte Carlo approach to the inverse problem of diffuse pollution risk in agricultural catchments

The hydrological and biogeochemical processes that operate in catchments influence the ecological quality of freshwater systems through delivery of fine sediment, nutrients and organic matter. Most models that seek to characterise the delivery of diffuse pollutants from land to water are reductionist. The multitude of processes that are parameterised in such models to ensure generic applicability make them complex and difficult to test on available data. Here, we outline an alternative - data-driven - inverse approach. We apply SCIMAP, a parsimonious risk based model that has an explicit treatment of hydrological connectivity. we take a Bayesian approach to the inverse problem of determining the risk that must be assigned to different land uses in a catchment in order to explain the spatial patterns of measured in-stream nutrient concentrations. We apply the model to identify the key sources of nitrogen (N) and phosphorus (P) diffuse pollution risk in eleven UK catchments covering a range of landscapes. The model results show that: 1) some land use generates a consistently high or low risk of diffuse nutrient pollution; but 2) the risks associated with different land uses vary both between catchments and between nutrients; and 3) that the dominant sources of P and N risk in the catchment are often a function of the spatial configuration of land uses. Taken on a case-by-case basis, this type of inverse approach may be used to help prioritise the focus of interventions to reduce diffuse pollution risk for freshwater ecosystems. (C) 2012 Elsevier B.V. All rights reserved.

Emergence of oriented circuits driven by synaptic pruning associated with spike-timing-dependent plasticity (STDP)

ABSTRACT: Massive synaptic pruning following over-growth is a general feature of mammalian brain maturation. Pruning starts near time of birth and is completed by time of sexual maturation. Trigger signals able to induce synaptic pruning could be related to dynamic functions that depend on the timing of action potentials. Spike-timing-dependent synaptic plasticity (STDP) is a change in the synaptic strength based on the ordering of pre- and postsynaptic spikes. The relation between synaptic efficacy and synaptic pruning suggests that the weak synapses may be modified and removed through competitive "learning" rules. This plasticity rule might produce the strengthening of the connections among neurons that belong to cell assemblies characterized by recurrent patterns of firing. Conversely, the connections that are not recurrently activated might decrease in efficiency and eventually be eliminated. The main goal of our study is to determine whether or not, and under which conditions, such cell assemblies may emerge out of a locally connected random network of integrate-and-fire units distributed on a 2D lattice receiving background noise and content-related input organized in both temporal and spatial dimensions. The originality of our study stands on the relatively large size of the network, 10,000 units, the duration of the experiment, 10E6 time units (one time unit corresponding to the duration of a spike), and the application of an original bio-inspired STDP modification rule compatible with hardware implementation. A first batch of experiments was performed to test that the randomly generated connectivity and the STDP-driven pruning did not show any spurious bias in absence of stimulation. Among other things, a scale factor was approximated to compensate for the network size on the ac¬tivity. Networks were then stimulated with the spatiotemporal patterns. The analysis of the connections remaining at the end of the simulations, as well as the analysis of the time series resulting from the interconnected units activity, suggest that feed-forward circuits emerge from the initially randomly connected networks by pruning. RESUME: L'élagage massif des synapses après une croissance excessive est une phase normale de la ma¬turation du cerveau des mammifères. L'élagage commence peu avant la naissance et est complété avant l'âge de la maturité sexuelle. Les facteurs déclenchants capables d'induire l'élagage des synapses pourraient être liés à des processus dynamiques qui dépendent de la temporalité rela¬tive des potentiels d'actions. La plasticité synaptique à modulation temporelle relative (STDP) correspond à un changement de la force synaptique basé sur l'ordre des décharges pré- et post- synaptiques. La relation entre l'efficacité synaptique et l'élagage des synapses suggère que les synapses les plus faibles pourraient être modifiées et retirées au moyen d'une règle "d'appren¬tissage" faisant intervenir une compétition. Cette règle de plasticité pourrait produire le ren¬forcement des connexions parmi les neurones qui appartiennent à une assemblée de cellules caractérisée par des motifs de décharge récurrents. A l'inverse, les connexions qui ne sont pas activées de façon récurrente pourraient voir leur efficacité diminuée et être finalement éliminées. Le but principal de notre travail est de déterminer s'il serait possible, et dans quelles conditions, que de telles assemblées de cellules émergent d'un réseau d'unités integrate-and¬-fire connectées aléatoirement et distribuées à la surface d'une grille bidimensionnelle recevant à la fois du bruit et des entrées organisées dans les dimensions temporelle et spatiale. L'originalité de notre étude tient dans la taille relativement grande du réseau, 10'000 unités, dans la durée des simulations, 1 million d'unités de temps (une unité de temps correspondant à une milliseconde), et dans l'utilisation d'une règle STDP originale compatible avec une implémentation matérielle. Une première série d'expériences a été effectuée pour tester que la connectivité produite aléatoirement et que l'élagage dirigé par STDP ne produisaient pas de biais en absence de stimu¬lation extérieure. Entre autres choses, un facteur d'échelle a pu être approximé pour compenser l'effet de la variation de la taille du réseau sur son activité. Les réseaux ont ensuite été stimulés avec des motifs spatiotemporels. L'analyse des connexions se maintenant à la fin des simulations, ainsi que l'analyse des séries temporelles résultantes de l'activité des neurones, suggèrent que des circuits feed-forward émergent par l'élagage des réseaux initialement connectés au hasard.

"Patron-driven acquisitions": la biblioteca propone y el lector dispone

Reseña de: Esposito, Joseph J.; Walker, Kizer; Ehling, Terry. PDA and the university press. A report prepared for The Andrew W. Mellon Foundation. [Baltimore, Md]: The Johns Hopkins University Press; [Ithaca, N.Y.]: Cornell University Library, Sept. 26th 2012. 65 p. Documento de trabajo disponible en línea en: . [Consulta: 10 de mayo de 2013].

Assessment on nutrient levels in the aerial biomass of irrigated guava in São Francisco Valley, Brazil

In perennial crops, fertilizers are applied to restore the amount of nutrient removed from the plant by harvesting. Nevertheless, the immobilized nutrient inside the whole plant should be considered. Based on it, the nutrient concentrations in the guava leaf and fruit and the nutrient contents removed by fruits and post-harvest pruning were evaluated in an irrigated orchard, in Petrolina, State of Pernambuco, Brazil. The greatest contents removed by fruits were N and K among the macronutrients, while Fe, Zn and Mn presented higher levels among the micronutrients. More than 60% of the total content of N, P, K, Mg, Fe, and B removed by pruning were immobilized in the leaf and non-marketable fruit.

Soil nutrient availability and its impact on fruit quality of Tahiti acid lime

The Tahiti acid lime in Brazil is mostly grown in the São Paulo State. The value of this crop production ranks among the ten most important fruits in the country. The Brazilian exports of Tahiti limes have increased in the last years with a corresponding increased demand for superior quality of fresh fruits, which is affected by mineral nutrients. Therefore, this study evaluated nutrient soil availability and its influence on nutritional status of trees based on the determination of leaf and fruit nutrient concentrations, fruit characteristics, and post harvest quality. Eleven commercial groves with trees older than 4-yr and differently managed were studied. Plots with six trees in each grove were sampled for soil (0-20 cm depth layer), leaf and fruit analyses with three replicates. Correlation coefficients were pair wised established for all variables. The results showed that N leaf concentration was well correlated with green color of fruit peel as measured by a color index (r = -0.71**), and which was optimum with Leaf-N around 22 g kg-1. Leaf-Ca was inversely correlated with fruit water loss after 14-day interval from harvest (r = -0.54*) demonstrating that Ca plays an important role in Tahiti fruit shelf-life. Data also suggested that increased fruit K concentration correlated with increased fruit water losses during storage (r >0.58*).

Diagnosis of the nutrient compositional space of fruit crops

Tissue analysis is a useful tool for the nutrient management of fruit orchards. The mineral composition of diagnostic tissues expressed as nutrient concentration on a dry weight basis has long been used to assess the status of 'pure' nutrients. When nutrients are mixed and interact in plant tissues, their proportions or concentrations change relatively to each other as a result of synergism, antagonism, or neutrality, hence producing resonance within the closed space of tissue composition. Ternary diagrams and nutrient ratios are early representations of interacting nutrients in the compositional space. Dual and multiple interactions were integrated by the Diagnosis and Recommendation Integrated System (DRIS) into nutrient indexes and by Compositional Nutrient Diagnosis into centered log ratios (CND-clr). DRIS has some computational flaws such as using a dry matter index that is not a part as well as nutrient products (e.g. NxCa) instead of ratios. DRIS and CND-clr integrate all possible nutrient interactions without defining an ad hoc interactive model. They diagnose D components while D-1 could be diagnosed in the D-compositional Hilbert space. The isometric log ratio (ilr) coordinates overcome these problems using orthonormal binary nutrient partitions instead of dual ratios. In this study, it is presented a nutrient interactive model as well as computation methods for DRIS and CND-clr and CND-ilr coordinates (CND-ilr) using leaf analytical data from an experimental apple orchard in Southwestern Quebec, Canada. It was computed the Aitchison and Mahalanobis distances across ilr coordinates as measures of nutrient imbalance. The effect of changing nutrient concentrations on ilr coordinates are simulated to identify the ones contributing the most to nutrient imbalance.

Influence of rootstock on nutrient content in grape petioles

Genetic diversity of rootstocks can affect nutrient uptake and the nutrient status of grapevines. The rootstock influence on nutrient content in grape petioles was evaluated on three rootstocks competition experiments carried out at Vale do Rio do Peixe region, in the state of Santa Catarina, Brazil, with the cultivars Niagara Rosada, Concord, and Isabella, grafted on different rootstocks. Two soil liming depths were also evaluated in the Isabella experiment. The greatest rootstock effect was observed on K and Mg content and K/Mg ratio in the petioles. The Vitis vinifera x V. rotundifolia hybrid rootstocks VR 043-43 and VR 044-4 provided the highest K/Mg values and self rooted Isabella the lowest K/Mg ratio. The other tested rootstocks resulted in intermediate values. There was also significant effect on P content, but only in Niagara Rosada and Concord experiments. The depth of soil liming did not significantly affect K and Mg content in the Isabella experiment. The results indicate that rootstock must be considered for nutritional status evaluation and fertilizer recommendation regarding to K and Mg.

Spontaneous Emergence of Chirality in the Limited Enantioselectivity Model: Autocatalytic Cycle Driven by an External Reagent

The emergence of chirality in enantioselective autocatalysis for compounds unable to transform according to the Frank-like reaction network is discussed with respect to the controversial limited enantioselectivity (LES) model composed of coupled enantioselective and non-enantioselective autocatalyses. The LES model cannot lead to spontaneous mirror symmetry breaking (SMSB) either in closed systems with a homogeneous temperature distribution or in closed systems with a stationary non-uniform temperature distribution. However, simulations of chemical kinetics in a two-compartment model demonstrate that SMSB may occur if both autocatalytic reactions are spatially separated at different temperatures in different compartments but coupled under the action of a continuous internal flow. In such conditions, the system can evolve, for certain reaction and system parameters, toward a chiral stationary state; that is, the system is able to reach a bifurcation point leading to SMSB. Numerical simulations in which reasonable chemical parameters have been used suggest that an adequate scenario for such a SMSB would be that of abyssal hydrothermal vents, by virtue of the typical temperature gradients found there and the role of inorganic solids mediating chemical reactions in an enzyme-like role.

Assessment of critical competences required in managing end-user driven businesses in a global forest products compa

Työn pääasiallisena tavoitteena on selvittää ydinosaamista, jota tarvitaan kansainvälisen metsäteollisuusyrityksen loppukäyttäjävetoisessa liiketoiminnassa. Lisäksi tavoitteena on laatia kompetenssimalli, jota voidaan käyttää mm. rekrytoinnissa, koulutuksen ja työnkierron suunnittelussa. Tarkoituksena on tarkastella sekä tällä hetkellä että tulevaisuudessa tarvittavia ydinosaamisia. Työ keskittyy myyntihenkilöiden kompetenssien kuvaamiseen. Kirjallisuuden ja asiantuntijoiden haastattelujen avulla on selvitetty. kompetensseihin liittyviä asioita ja käsitteitä, kuten organisaation oppiminen, innovaatiot sekä asiakaslähtöisyys. Tämän jälkeen on kartoitettu case-yrityksen kartonkiyksikön myyntihenkilökunnan kompetensseja. Kompetenssikartoitus on tehty teemahaastattelujen avulla sekä lisäksi on käytetty työn aikana laadittua kompetenssimatriisia. Saatuja tuloksia on verrattu yhden asiakkaan näkemyksiin. Tulosten mukaan kolme tärkeintä tämän hetken kompetenssia ovat: holistinen näkemys asioista, suhteiden rakentaminen ja kielitaito. Tulevaisuuden tärkeimpiä kompetensseja puolestaan ovat: asiakaslähtöisyys, sähköinen kaupankäynti ja suhteiden rakentaminen. Lopuksi on käsitelty yleisiä metsäteollisuuden tulevaisuuden haasteita sekä annettu joitain kehitysideoita kompetenssien kehittämistä varten.