Impacte da nova regulamentação na certificação energética de edifícios

Dissertação de Mestrado, Engenharia Civil, Especialização em Construção, Instituto Superior de Engenharia, Universidade do Algarve, 2016

Mejora del comportamiento térmico de los morteros de cal aditivados y su empleo en la rehabilitación de inmuebles = Improvement of thermal behaviour of additived lime mortars and their use in rehabilitation of buildings

La envolvente de la edificación es la responsable de equilibrar el intercambio energético entre el interior y el exterior, por lo tanto cualquier actuación encaminada a la reducción del consumo energético ha de establecer, como uno de sus objetivos prioritarios, la mejora del comportamiento de la misma. Las edificaciones anteriores a 1940 constituyen la mayor parte de las existentes en áreas rurales y centros urbanos. En ellas, la repercusión de la fachada sobre las transmitancias globales pone de manifiesto la necesidad de intervención. Sin embargo, su elevada inercia térmica y los importantes saltos térmicos característicos de gran parte de España plantean la importancia de que aquélla se efectúe por el exterior. A tal respecto, la falta de disponibilidad de espesor suficiente para implantar sistemas tipo SATE deriva en que, frecuentemente, la única solución viable sea la de aislar por el interior perdiendo con ello la capacidad de acumulación térmica del muro y con el asociado riesgo de condensaciones. La amplia tradición en el empleo de revestimientos, especialmente en base de cal, permiten que éstos sean utilizados no sólo como elemento estético o de protección de la obra de fábrica antigua sino también para la mejora del comportamiento térmico del soporte, si se aprovecha el mecanismo de transmisión térmica por radiación. Éste es el objetivo de la presente Tesis Doctoral en la que se estudia la modificación de las propiedades radiantes de los morteros de revestimiento para la mejora de la eficiencia energética de las construcciones históricas, principalmente las constituidas por muros monolíticos, aunque podría ser de aplicación a otro tipo de construcciones compuestas por diversas capas. Como punto de partida, se estudió y revisó la documentación disponible sobre las investigaciones de las tres áreas científico-tecnológicas que convergen en la Tesis Doctoral: rehabilitación, material y comportamiento térmico, a partir de lo cual se comprobó la inexistencia de estudios similares al objeto de la presente Tesis Doctoral. Complementariamente, se analizaron los revestimientos en lo concerniente a los materiales que los constituyen, la composición de las mezclas y características de cada una de las capas así como al enfoque que, desde el punto de vista térmico, se estimaba más adecuado para la obtención de los objetivos propuestos. Basándonos en dichos análisis, se preseleccionaron ochenta materiales que fueron ensayados en términos de reflectancia y emisividad para elegir cuatro con los que se continuó la investigación. Éstos, junto con la cal elegida para la investigación y el árido marmóreo característico de la última capa de revestimiento, fueron caracterizados térmicamente, de forma pormenorizada, así como química y físicamente. Los fundamentos teóricos y los estudios preliminares desarrollados con distintos materiales, en estado fresco y endurecido, fueron empleados en la dosificación de componentes de las mezclas, en dos proporciones distintas, para el estudio del efecto del agregado. Éstas se ensayaron en estado fresco, para comprobar su adecuación de puesta en obra y prever su VI adherencia al soporte, así como en estado endurecido a 28 y 90 días de curado, para conocer las propiedades que permitieran prever su compatibilidad con aquél y estimar el alcance de la reducción de transferencias térmicas lograda. Además, se estudiaron las características generales de las mezclas que sirvieron para establecer correlaciones entre distintas propiedades y entender los resultados mecánicos, físicos (comportamiento frente al agua) y energéticos. Del estudio conjunto de las distintas propiedades analizadas se propusieron dos mezclas, una blanca y otra coloreada, cuyas características permiten garantizar la compatibilidad con la obra de fábrica antigua, desde el punto de vista físico y mecánico, y preservar la autenticidad de los revestimientos, en cuanto a la técnica de aplicación tradicional en sistemas multicapa. El comportamiento térmico de las mismas, sobre una obra de fábrica de 40 cm de espesor, se estimó, en estado estacionario y pseudo-transitorio, demostrándose reducciones del flujo térmico entre 16-48%, en condiciones de verano, y entre el 6-11%, en invierno, en función de la coloración y de la rugosidad de la superficie, en comparación con el empleo de la mezcla tradicional. Por lo que, se constata la viabilidad de los materiales compuestos propuestos y su adecuación al objetivo de la investigación. VII ABSTRACT The envelope is responsible for balancing the energy exchange between the inside and outside in buildings. For this reason, any action aimed at reducing energy consumption must establish, as one of its key priorities, its improvement. In rural areas and urban centers, most of the constructions were built before 1940. In them, the impact of the façade on the global transmittance highlights the need for intervention. However, its high thermal inertia and fluctuation of temperatures in the majority of Spain bring up that it should be placed outside the insulation. In this regard, the lack of availability of enough thickness to implement systems such as ETICS results in that often the only viable solution is to isolate the interior, losing thereby the wall’s heat storage capacity with the associated risk of condensation. The tradition in the use of renders, especially lime-based, allows us to use them not only as an aesthetic element or to protect the ancient masonry, but also for improved thermal performance of the support by taking advantage of the heat transfer mechanism by radiation. This is the aim of this Doctoral Thesis in which the modification of the radiative properties of lime mortars for renders to improve the energy efficiency of historic buildings, mainly composed of monolithic walls, is studied, although it could be applied to other structures composed of several layers. As a starting point, the available literature in the three scientific-technological areas that converge at the Doctoral Thesis: rehabilitation, material and thermal behaviour, was reviewed, and confirmed the absence of researches similar to this Doctoral Thesis. Additionally, the renders were studied with regard to the materials that constitute them, the composition of the mixtures and the characteristics of each layer, as well as to the approach which, from a thermal point of view, was deemed the most suitable for achieving the objectives sets. Based on thre aforementioned analysis, eighty materials tested in terms of reflectance and emissivity were pre-selected, to choose four with which the research was continued. Common marble sand, used in the last layer of the renders, together with the appointed materials and hydrated lime were characterized thermally, in detail, as well as chemically and physically. The theoretical bases and preliminary studies with different materials, in fresh and hardened state, were used in the dosage of the composition of the mixtures. In order to study their effect they were used in two different proportions, that is, ten mixtures in total. These were tested in their fresh state to evaluate their setting-up suitability and foresee their adhesion to the support, as well as in their hardened state, at 28 and 90 days of curing, to establish the properties which enabled us to anticipate their compatibility with the old masonry walls and estimate the scope of the reduction of heat transfers achieved. In addition, the general characteristics of the mixtures used to establish correlations and to understand the mechanical, physical and energy results were studied. Two mixtures, one white and one colored, were proposed as the result of the different properties analysed, whose characteristics allow the guarantee of mechanical and physical compatibility VIII with the old masonry and preserve the authenticity of the renders. The thermal behavior of both, applied on a masonry wall 40 cm thick, was estimated at a steady and pseudo-transient state, with heat flow reductions between 16-48% during summertime and 6-11% during wintertime, depending on the color and surface roughness, compared to the use of the traditional mixture. So, the viability of the proposed composite materials and their fitness to the aim of the research are established.

A descriptive analysis of the seasonal variation of physical oceanographic characteristics in the northern region of the Todos os Santos Bay (Bahia, Brazil)

The results obtained in the August and December 2003, August 2004 and January 2005 oceanographic campaigns in the northern region of the Todos os Santos Bay (lat. 12º44.5'S; long. 038º35.00'W) between the Madre de Deus and Maré islands are analyzed. Instruments of continuous and discrete samplings were used to measure hydrographic properties currents and tides. The water mass of the northern region of the bay is forced by semidiurnal and mesotides of form number 0.08 and the lunar component M2 height was estimated at 91cm. The time series of the surface currents indicated movements in the N/S direction, forced by the tide with maximum magnitudes of 0.73 m.s-1 on the December 2003 campaign. However, in August 2004 the currents were dominated by the wind stress forcing, with a maximum speed of 1.85 m.s-1 and SE direction. Near the bottom, the influence of the tide is not as evident, with a decrease in intensity due to internal and bottom friction, with a maximum velocity of 0.17 m.s-1. The thermal and haline structures were weakly horizontally, as well as vertically stratified, with extreme values varying in the intervals 23ºC (August, 2004) to 28ºC (December, 2003) and 31.0 psu (August, 2003) to 36.0 psu (December, 2003), respectively. Some conclusions may be drawn from these results: i) The signs of the dilution of the fresh water discharges of the Caípe, Mataripe and São Paulo rivers in the region under the influence of the RLAM were observed only during the winter periods, but in the summer the region was flooded by waters of oceanic origin and the salinities above 36.0 indicated TW mass intrusion; ii) The N-S circulation near the RLAM is strongly dominated by the tide, and the importance of the M2 component was unequivocal, however, the E-W component presented some tidal modulation away from abrupt bottom topographical changes, and iii) The residual series, calculated as the difference between the original and modeled, is about ¼ of the original and confirmed its semidiurnal character.

Secretory activity and endocrine regulation of male accessory glands in the blood-sucking bug Panstrongylus megistus (Hemiptera: Reduviidae)

The epithelial cells of Panstrongylus megistus male accessory glands (MAG) present ultrastructural characteristics of a secretory cell. Their secretory products are accumulated in the lumen of the four MAG lobes. During the first 8 days of adult life a strong secretion activity occurs, accumulating enough material to produce the first spermatophore. Cerebral neurosecretions as well as juvenile hormone are both involved in MAG secretory activity regulation. Juvenile hormone seems to be the responsible for the stimulation of most protein synthesis in male accessory glands. Cerebral neurosecretion seems to be necessary to stimulate juvenile hormone production and release by the corpus allatum. Furthermore, neurosecretion is required for some polypeptides synthesis by MAG. Although topic application of precocene II to adult males does not reproduce the same effects on MAG as does allatectomy, this compound causes strong reduction on male reproductive capacity.

Regulation of adaptive behaviour during fasting by hypothalamic Foxa2.

The lateral hypothalamic area is considered the classic 'feeding centre', regulating food intake, arousal and motivated behaviour through the actions of orexin and melanin-concentrating hormone (MCH). These neuropeptides are inhibited in response to feeding-related signals and are released during fasting. However, the molecular mechanisms that regulate and integrate these signals remain poorly understood. Here we show that the forkhead box transcription factor Foxa2, a downstream target of insulin signalling, regulates the expression of orexin and MCH. During fasting, Foxa2 binds to MCH and orexin promoters and stimulates their expression. In fed and in hyperinsulinemic obese mice, insulin signalling leads to nuclear exclusion of Foxa2 and reduced expression of MCH and orexin. Constitutive activation of Foxa2 in the brain (Nes-Cre/+;Foxa2T156A(flox/flox) genotype) results in increased neuronal MCH and orexin expression and increased food consumption, metabolism and insulin sensitivity. Spontaneous physical activity of these animals in the fed state is significantly increased and is similar to that in fasted mice. Conditional activation of Foxa2 through the T156A mutation expression in the brain of obese mice also resulted in improved glucose homeostasis, decreased fat and increased lean body mass. Our results demonstrate that Foxa2 can act as a metabolic sensor in neurons of the lateral hypothalamic area to integrate metabolic signals, adaptive behaviour and physiological responses.

Bcl9/Bcl9l are critical for Wnt-mediated regulation of stem cell traits in colon epithelium and adenocarcinomas.

Canonical Wnt signaling plays a critical role in stem cell maintenance in epithelial homeostasis and carcinogenesis. Here, we show that in the mouse this role is critically mediated by Bcl9/Bcl9l, the mammalian homologues of Legless, which in Drosophila is required for Armadillo/beta-catenin signaling. Conditional ablation of Bcl9/Bcl9l in the intestinal epithelium, where the essential role of Wnt signaling in epithelial homeostasis and stem cell maintenance is well documented, resulted in decreased expression of intestinal stem cell markers and impaired regeneration of ulcerated colon epithelium. Adenocarcinomas with aberrant Wnt signaling arose with similar incidence in wild-type and mutant mice. However, transcriptional profiles were vastly different: Whereas wild-type tumors displayed characteristics of epithelial-mesenchymal transition (EMT) and stem cell-like properties, these properties were largely abrogated in mutant tumors. These findings reveal an essential role for Bcl9/Bcl9l in regulating a subset of Wnt target genes involved in controlling EMT and stem cell-related features and suggest that targeting the Bcl9/Bcl9l arm of Wnt signaling in Wnt-activated cancers might attenuate these traits, which are associated with tumor invasion, metastasis, and resistance to therapy.

Gac/Rsm signal transduction pathway of gamma-proteobacteria: from RNA recognition to regulation of social behaviour.

In many gamma-proteobacteria, the conserved GacS/GacA (BarA/UvrY) two-component system positively controls the expression of one to five genes specifying small RNAs (sRNAs) that are characterized by repeated unpaired GGA motifs but otherwise appear to belong to several independent families. The GGA motifs are essential for binding small, dimeric RNA-binding proteins of a single conserved family designated RsmA (CsrA). These proteins, which also occur in bacterial species outside the gamma-proteobacteria, act as translational repressors of certain mRNAs when these contain an RsmA/CsrA binding site at or near the Shine-Dalgarno sequence plus additional binding sites located in the 5' untranslated leader mRNA. Recent structural data have established that the RsmA-like protein RsmE of Pseudomonas fluorescens makes specific contacts with an RNA consensus sequence 5'-(A)/(U)CANGGANG(U)/(A)-3' (where N is any nucleotide). Interaction with an RsmA/CsrA protein promotes the formation of a short stem supporting an ANGGAN loop. This conformation hinders access of 30S ribosomal subunits and hence translation initiation. The output of the Gac/Rsm cascade varies widely in different bacterial species and typically involves management of carbon storage and expression of virulence or biocontrol factors. Unidentified signal molecules co-ordinate the activity of the Gac/Rsm cascade in a cell population density-dependent manner.

Regulation of bistable ICEclc transfer in pseudomonas knackmussii B13

L'élément génétique intégratif et conjugatif auto-transférable de 103 kb qui se trouve dans le génome de Pseudomonas knackmussii B13 (ICEc/c) confère la capacité de dégrader le 3-chlorobenzoate et le 2-aminophénol. L'élément ICE c/c peut être transféré par conjugaison de la souche B13 à diverses bêta- et gamma- protéobactéries. Seule une sous-population de 3 à 5% des cellules transfère l'élément, les cellules dites "compétentes pour le transfert". L'acquisition de la compétence pour le transfert est vraisemblablement la conséquence d'une régulation bistable, conduisant une partie des cellules au transfert de l'élément ICE c/c tandis que, dans les autres, l'élément reste quiescent et ne se transfère pas. À ce jour, les mécanismes et les acteurs moléculaires qui régulent l'activation bistable de l'élément sont restés inconnus. Mon travail de doctorat visait à identifier les éléments bistables du régulon de la compétence pour le transfert et d'analyser les fondements moléculaires de la bistabilité de l'élément ICE c/c chez P. knackmussii. Le premier chapitre introduit le thème du transfert génétique horizontal avec un accent particulier sur les éléments intégratifs et conjugatifs (ICE) et ICEcIc. L'état actuel des connaissances sur l'organisation génétique, la régulation, l'intégration et le transfert de différents modèles de ICEs est exposé en détail. En outre, je m'étends sur les phénomènes d'hétérogénéité et de bistabilité phénotyplques, qu'on peut distinguer dans une population isogénique dans des conditions de culture homogènes, et qui sont susceptibles de jouer un rôle dans le transfert de l'élément ICE c/c, dans la mesure où il ne s'active et n'est transférable que dans une très petite sous-population de cellules. Dans le chapitre 2, je présente une analyse globale des régions promotrices minimales des gènes appartenant au régulon de la compétence pour le transfert de l'élément ICE c/c. Nous avons étudié les caractéristiques d'expression des promoteurs et, s'ils s'avéraient bistables, leur activation dans le temps par comparaison avec le mutant lntB13. Pour ce faire, nous avons utilisé des fusions de promoteurs avec des gènes rapporteurs et testé l'expression bistable chez P. knackmussii par microscopie à épifluorescence. Pour six promoteurs présentant une expression bistable, nous avons employé de la microscopie temporelle pour déterminer la chronologie de leur expression par rapport à Pint et PinR. Parmi eux, nous avons identifié deux gènes exprimés précocement et trois gènes exprimés tardivement dans le processus d'acquisition de la compétence de transfert. Dans le chapitre 3, j'expose une analyse d'expression génétique pour l'un des groupes de gènes dont la transcription est la plus élevée dans la région conservée de ICE c/c, les gènes orf81655-orf68241 contenus dans une région de 14 kb. Nous montrons d'abord que cet opéron fait partie du même régulon bistable que intB13 et inrR et analysons les caractéristiques génétiques qui conduisent à une transcription élevée. Nous étudions les fonctions biologiques de ce groupe de gènes par des délétlons ciblées et montrons que certaines d'entre elles empêchent le transfert de l'élément. Nous approfondissons la caractérlsatlon de I'orf8l655 en construisant une fusion transcrlptionnelle avec le gène codant pour la protéine fluorescente verte (egfp) (en utilisant le système minl-Tn5). L'expression de Vorf81655 dans des cellules individuelles est comparée au signal mesuré par hybridation in situ en fluorescence (FISH) sur le ARN messager du gène. En utilisant FISH, des délétlons du promoteur et de l'analyse directe de transcription, nous avons localisé la région promotrice du groupe de gènes. En outre, nous avons utilisé des mutations dirigées pour comprendre la bistabilité de cette région promotrice, caractérisée par une transcription très élevée et une traduction lente de l'ARN messager.  Dans le chapitre 4, nous nous efforçons de comprendre comment la bistabilité est générée au sein du régulon te de l'élément ICE c/c. Pour ce faire, nous avons tenté de reconstituer une expression bistable, dans un hôte qui ne présente pas de bistabilité naturellement, à partir d'éléments génétiques individuels. L'hôte choisi est Pseudomonas putida dans lequel nous avons introduit une copie unique de Pint, PinR ou PaipA fusionnés à la egfp, construits qui permettent d'observer l'apparition de bistabilité. Nous avons ensuite construit différents assemblages de composants génétiques de l'élément ICE c/c, en nous concentrant sur la région parA-inrR. En effet, nous avons pu démontrer qu'une expression bistable apparaît dans P. putida grâce à ces éléments en l'absence de l'élément ICE c/c complet. À noter que la plupart des construits génétiques activent PaipA ou P|,,R, mais qu'un seul recrée la bistabilité de Pint, ce qui suggère que la région parA-inrR permet à la fois d'engendrer la bistabilité et d'opérer la transition entre les promoteurs précoces et les promoteurs tardifs du régulon de la bistabilité. Dans le chapitre 5, nous concluons sur une discussion de la pertinence de nos résultats et sur de futures perspectives de recherche. -- The 103-kb self-transmissible integrative and conjugative element (ICE) of Pseudomonas knackmussii B13 (ICEc/c) confers the capacity to degrade 3- chlorobenzoate and 2-aminophenol. ICEc/c can be conjugated from strain B13 to a variety of Beta- and Gammaproteobacteria. Interestingly, ICE c/c transfer is observed in a subpopulatlon of cells (3-5%) only, the so-called 'transfer competent' cells. The formation of transfer competence (tc) is thought to be the consequence of a 'bistable' decision, which forces those cells to follow the developmental path which leads to ICEc/c transfer, whereas in others ICE c/c remains silent and does not transfer. So far, the mechanisms and molecular partners generating this bistable transfer activation in cells of P. knackmussii B13 remain mostly unidentified. This thesis aimed at understanding the extent of the tc bistability regulon and to dissect the molecular basis of bistabillty formation of ICEc/c in P. knackmussii. The first chapter is a general Introduction on horizontal gene transfer (HGT) with particular emphasis on ICEs and ICE c/c. The emphasis is made on the current knowledge about the HGT gene organization, regulation and specific integration and transfer aspects of the different ICEs models. Furthermore, I focus on the phenomena of phenotypic heterogeneity and bistability (the property of two distinguishable phenotypes existing within an isogenic population under homogeneous conditions), which may play a particular role in ICEc/c behaviour, since ICE activation and transfer only occurs in a very small subpopulation of cells. In Chapter Two, I focus on a global analysis of the different core promoters that might belong to the ICEc/c tc pathway regulon. We studied both expression patterns of ICEc/c promoters and, once being identified as "bistable", their temporal activation compared to that of intB13. In order to do this, we used promoter reporter fusions and tested blstability expression in P. knackmussii using epifluorescence microscopy. For the 6 promoters that showed bistable expression, we used time-lapse microscopy to study the timing of promoter expression in comparison to that of P,,,t or PlnR. We could establish two "early" and 3 "late" phase promoters in the process of transfer competence. In Chapter Three, I focused my attention on analysis of gene expression of one of the most highly transcribed gene clusters in the conserved core region of ICEc/c, a 14-kb gene cluster formed by the genes orf81655-orf68241. First we showed that this operon is part of the same bistability 'regulon' as intB13 and inrR, and analysed the genetic features that lead to high transcription. We studied the potential biological function of this cluster for ICE c/c by making specific gene deletions, showing that some interrupt ICEc/c transfer. We further analysed the orfdl655 promoter by constructing transcriptional egfp fusion reporter strains using the miniTn5 delivery system. Expression of the orf81655 promoter in single cells was compared to signals measured by Fluorescence In Situ Hybridization (FISH) on orfSl655 mRNA. We localized the promoter region of the gene cluster using FISH, promoter deletions, and by direct transcript analysis. We further used site-directed mutagenesis to understand the bistability character of the promoter region and the extremely high transcription but low translation from this mRNA. In Chapter Four, we set out to understand how bistability is generated in the tc pathway of ICEc/c. For this we tried rebuilding bistable expression from ICEc/c individual gene components in a host, which normally does not display bistability. As host we used P. putida without ICEc/c but with a single copy Pint-, PlnR- or PalpA- egfp fusion that enabled us to verify bistability formation. Subsequently, we built different assemblages of ICEc/c gene components, focusing on the parA-inrR region. Indeed, we found that bistable expression can be build from those components in P. putida without ICEc/c. Interestingly, most genetic constructs activated PaipA or PlnR, but only one resulted in bistable activation of PinT. This suggests that the parA-inrR region acts as a bistability "generator", but also as a bistability "relay" from early to late promoters in the tc pathway hierarchy. In the final fifth chapter, we conclude with a discussion of the relevance of the present thesis and the resulting perspectives for future studies.

A DESCRIPTIVE ANALYSIS OF THE SEASONAL VARIATION OF PHYSICAL OCEANOGRAPHIC CHARACTERISTICS IN THE NORTHERN REGION OF THE TODOS OS SANTOS BAY (BAHIA, BRAZIL)

The results obtained in the August and December 2003, August 2004 and January 2005 oceanographic campaigns in the northern region of the Todos os Santos Bay (lat. 12 degrees 44.5`S; long. 038 degrees 35.00`W) between the Madre de Deus and Mare islands are analyzed. Instruments of continuous and discrete samplings were used to measure hydrographic properties currents and tides. The water mass of the northern region of the bay is forced by semidiurnal and mesotides of form number 0.08 and the lunar component M(2) height was estimated at 91cm. The time series of the surface currents indicated movements in the N/S direction, forced by the tide with maximum magnitudes of 0.73 m.s(-1) on the December 2003 campaign. However, in August 2004 the currents were dominated by the wind stress forcing, with a maximum speed of 1.85 m.s(-1) and SE direction. Near the bottom, the influence of the tide is not as evident, with a decrease in intensity due to internal and bottom friction, with a maximum velocity of 0.17 m.s(-1). The thermal and haline structures were weakly horizontally, as well as vertically stratified, with extreme values varying in the intervals 23 degrees C (August, 2004) to 28 degrees C (December, 2003) and 31.0 psu (August, 2003) to 36.0 psu (December, 2003), respectively. Some conclusions may be drawn from these results: i) The signs of the dilution of the fresh water discharges of the Caipe, Mataripe and Sao Paulo rivers in the region under the influence of the RLAM were observed only during the winter periods, but in the summer the region was flooded by waters of oceanic origin and the salinities above 36.0 indicated TW mass intrusion; ii) The N-S circulation near the RLAM is strongly dominated by the tide, and the importance of the M(2) component was unequivocal, however, the E-W component presented some tidal modulation away from abrupt bottom topographical changes, and iii) The residual series, calculated as the difference between the original and modeled, is about 1/4 of the original and confirmed its semidiurnal character.

Use of behavioural patterns to analyze transport stress in piglets and young bulls. The influence of individual coping characteristics in stress response.

In modern farm systems the economic interests make reducing the risks related to transport practice an important goal. An increasing attention is directed to the welfare of animals in transit, also considering the new existing facilities. In recent years the results coming from the study of animal farm behaviour were used as tool to assess the welfare. In this thesis were analyzed behavioural patterns, jointly with blood variables, to evaluate the stress response of piglets and young bulls during transport. Since the animal behaviour could be different between individuals and these differences can affect animal responses to aversive situations, the individual behavioural characteristics were taken in account. Regarding young bulls, selected to genetic evaluation, the individual behaviour was investigated before, during and after transport, while for piglets was adopted a tested methodology classification and behavioural tests to observe their coping characteristics. The aim of this thesis was to analyse the behavioural and physiological response of young bulls and piglets to transport practice and to investigate if coping characteristics may affect how piglets cope with aversive situations. The thesis is composed by four experimental studies. The first one aims to identify the best existent methodology classification of piglets coping style between those that were credited in literature. The second one investigated the differences in response to novel situations of piglets with different coping styles. The last studies evaluated the stress response of piglets and young bulls to road transportation. The results obtained show that transport did not affect the behaviour and homeostasis of young animals which respond in a different way from adults. However the understanding of individual behavioural characteristic and the use of behavioural patterns, in addition to blood analyses, need to be more investigated in order to be useful tools to assess the animal response in aversive situation.

ROLE OF MIR-19A RELEASED BY INFLAMMATORY BREAST CANCER CELLS IN THE REGULATION OF DENDRITIC CELL FUNCTIONS: IN VITRO MODEL OF CROSSTALK IN THE TUMOR MICROENVIRONMENT OF INFLAMMATORY BREAST CANCER

Inflammatory breast cancer (IBC) is a rare but very aggressive form of locally advanced breast cancer (1-6% of total breast cancer patients in United States), with a 5-year overall survival rate of only 40.5%, compared with 85% of the non-IBC patients. So far, a unique molecular signature for IBC able to explain the dramatic differences in the tumor biology between IBC and non-IBC has not been identified. As immune cells in the tumor microenvironment plays an important role in regulating tumor progression, we hypothesized that tumor-associated dendritic cells (TADC) may be responsible for regulating the development of the aggressive characteristics of IBC. MiRNAs can be released into the extracellular space and mediate the intercellular communication by regulating target gene expression beyond their cells of origin. We hypothesized that miRNAs released by IBC cells can induce an increased activation status, secretion of pro-inflammatory cytokines and migration ability of TADC. In an in vitro model of IBC tumor microenvironment, we found that the co-cultured of the IBC cell line SUM-149 with immature dendritic cells (iDCSUM-149) induced a higher degree of activation and maturation of iDCSUM-149 upon stimulation with lipopolysaccharide (LPS) compared with iDCs co-cultured with the non-IBC cell line SUM-159 (iDCSUM-159), resulting in: increased expression of the costimulatory and activation markers; higher production of pro-inflammatory cytokines (TNF-a, IL-6); and 3) higher migratory ability. These differences were due to the exosome-mediated transfer of miR-19a and miR-146a from SUM-149 and SUM-159, respectively, to iDCs, causing the downregulation of the miR-19a target genes PTEN, SOCS-1 and the miR-146a target genes IRAK1, TRAF6. PTEN, SOCS-1 and IRAK1, TRAF6 are important negative and positive regulator of cytokine- and TLR-mediated activation/maturation signaling pathway in DCs. Increased levels of IL-6 induced the upregulation of miR-19a synthesis in SUM-149 cells that was associated with the induction of CD44+CD24-ALDH1+ cancer stem cells (CSCs) with epithelial-to-mesenchymal transition (EMT) characteristics. In conclusion, in IBC tumor microenvironment IL-6/miR-19a axis can represent a self-sustaining loop able to maintain a pro-inflammatory status of DCs, leading to the development of tumor cells with high metastatic potential (EMT CSCs) responsible of the poor prognosis in IBC patients.

The formation and impact of hazard control policy:a study of the regulation of white lead paint in Britain

Government regulation of industrial hazards is examined in the context of the economic and technical processes of industrial development. Technical problems and costs of control are considered as factors in both the formation and impact of regulation. This thesis focuses on an historical case-study of the regulation of the hazard to painting workers from the use of lead pigments in paint. A regulatory strategy based on the prohibition of lead paints gained initial acceptance within the British state in 1911, but was subsequently rejected in favour of a strategy that allowed continued use of lead paint subject to hygiene precautions. The development of paint technology and its determinants, including concern about health hazards, are analysed, focusing on the innovation and diffusion into the paint industry of the major white pigments: white lead (PbC03 .PB(OH)2)and its substitutes. The process of regulatory development is examined, and the protracted and polarised regulatory d~bate contrasted to the prevailing 'consensual' methods of workplace regulation. The rejection of prohibition is analysed in terms of the different political and technical resources of those groups in conflict over this policy. This highlights the problems of consensus formation around such a strategy, and demonstrates certain constraints on state regulatory activity, particularly regarding industrial development. Member-states of the International Labour Organisation agreed to introduce partial prohibition of lead paint in 1921. Whether this was implemented is related to the economic importance of lead and non-lead metal and pigment industries to a nation. An analysis is made of the control of lead poisoning. The rate of control is related to the economic and technological trajectory of the regulated industry. Technical and organisational characteristics are considered as well as regulatory factors which range from voluntary compliance and informal pressures to direct legal requirements. The implications of this case-study for the analysis of the development and impacts of regulation are assessed.

Melatonin reduces LH, 17 beta-estradiol and induces differential regulation of sex steroid receptors in reproductive tissues during rat ovulation

Background: Melatonin is associated with direct or indirect actions upon female reproductive function. However, its effects on sex hormones and steroid receptors during ovulation are not clearly defined. This study aimed to verify whether exposure to long-term melatonin is able to cause reproductive hormonal disturbances as well as their role on sex steroid receptors in the rat ovary, oviduct and uterus during ovulation. Methods: Twenty-four adult Wistar rats, 60 days old (+/-250 g) were randomly divided into two groups. Control group (Co): received 0.9% NaCl 0.3 mL + 95% ethanol 0.04 mL as vehicle; Melatonin-treated group (MEL): received vehicle + melatonin [ 100 mu g/100 g BW/day] both intraperitoneally during 60 days. All animals were euthanized by decapitation during the morning estrus at 4 a. m. Results: Melatonin significantly reduced the plasma levels of LH and 17 beta-estradiol, while urinary 6-sulfatoximelatonin (STM) was increased at the morning estrus. In addition, melatonin promoted differential regulation of the estrogen receptor (ER), progesterone receptor (PR), androgen receptor (AR) and melatonin receptor (MTR) along the reproductive tissues. In ovary, melatonin induced a down-regulation of ER-alpha and PRB levels. Conversely, it was observed that PRA and MT1R were up-regulated. In oviduct, AR and ER-alpha levels were down-regulated, in contrast to high expression of both PRA and PRB. Finally, the ER-beta and PRB levels were down-regulated in uterus tissue and only MT1R was up-regulated. Conclusions: We suggest that melatonin partially suppress the hypothalamus-pituitary-ovarian axis, in addition, it induces differential regulation of sex steroid receptors in the ovary, oviduct and uterus during ovulation.

Remotely sensed estimates of vegetation structural characteristics in restored wetlands, Southern California

Traditional field sampling approaches for ecological studies of restored habitat can only cover small areas in detail, con be time consuming, and are often invasive and destructive. Spatially extensive and non-invasive remotely sensed data can make field sampling more focused and efficient. The objective of this work was to investigate the feasibility and accuracy of hand-held and airborne remotely sensed data to estimate vegetation structural parameters for an indicator plant species in a restored wetland. High spatial resolution, digital, multispectral camera images were captured from an aircraft over Sweetwater Marsh (San Diego County, California) during each growing season between 1992-1996. Field data were collected concurrently, which included plant heights, proportional ground cover and canopy architecture type, and spectral radiometer measurements. Spartina foliosa (Pacific cordgrass) is the indicator species for the restoration monitoring. A conceptual model summarizing the controls on the spectral reflectance properties of Pacific cordgrass was established. Empirical models were developed relating the stem length, density, and canopy architecture of cordgrass to normalized-difference-vegetation-index values. The most promising results were obtained from empirical estimates of total ground cover using image data that had been stratified into high, middle, and low marsh zones. As part of on-going restoration monitoring activities, this model is being used to provide maps of estimated vegetation cover.

Complement-mediated regulation of Trichomonas vaginalis infection in mice

Trichomonas vaginalis is a flagellated protozoan which causes trichomoniasis, a sexually transmitted disease of the human genitourinary tract, The importance of the alternative complement pathway in host defence against T. vaginalis was investigated in vitro. Kinetic studies utilising immunofixation following electrophoresis showed that both a strongly and weakly virulent strain of T, vaginalis activated murine serum C3. In vivo studies with congenic-resistant, C5-deficient, B10.D2/OSn- and C5-sufficient, B10.D2/nSn mice showed that the presence of C5 is a significant factor in the innate host resistance to primary infection with a strongly virulent, but not a weakly virulent trichomonad strain.