Processos celulares que governam as NADPH oxidases vasculares em condições patológicas não estão claros ainda. Como os processos redox são parte intrínseca da resposta da célula ao estresse, temos investigado se o estresse oxidativo pode convergir com outros tipos de estresse via Nox(es). No presente estudo, focamos na inibição do proteasoma como uma condição relevante de estresse. A incubação de células musculares lisas com concentrações não apoptóticas de inibidores do proteasoma, MG132 e lactacistina, promoveu aumento na produção basal de superóxido e na atividade da NADPH oxidase, diminuição da atividade da SOD e da razão GSH/GSSG. Por outro lado, a inibição do proteasoma diminui a atividade da Nox após estímulo com Angiotensina II ou Tunicamicina, conhecido estressor do retículo endoplasmático. Em condições basais, MG132 induz a expressão de mRNA da Nox1, entretanto o aumento de Nox1 induzido por Angiotensina II foi diminuído na presença de MG132. O mesmo efeito ocorre com a indução de Nox4 pela Tunicamicina, que nesse caso foi drasticamente reduzida na presença de MG132. Além disso, tanto Angiotensina II quanto Tunicamicina induziram a atividade lítica do proteasoma 20S. A seguir, investigamos as conseqüências fisiológicas do MG132 na sinalização do estresse do RE, uma conhecida resposta mediada por Nox4. Células vasculares incubadas com MG132 induzem a expressão de marcadores do estresse do RE, GRP78 e XBP1, e também os marcadores mais tardios ATF4 e o próapoptótico CHOP/GADD153. Resultados similares ocorrem também com a Tunicamicina. Entretanto, a co-incubação de Tunicamicina e MG132 diminui e a sinalização do estresse do RE. AKT e p38 MAPK foram ativados por MG132, possivelmente como resposta ao estresse induzido pela inibição do proteasoma. Assim, a inibição do proteasoma bloqueia a NADPH oxidase, com aumento da atividade basal e expressão da Nox1 versus forte inibição da ativação e expressão da Nox4 frente ao estímulo. A inibição da Nox4 associa-se e pode contribuir para a inibição pelo MG132 da sinalização do estresse do RE. Portanto, o proteasoma parece exercer papel na integração de estresses celulares envolvendo a NADPH oxidase. A inibição do proteasoma pode ter papel na terapia de doenças associadas a estresse do RE. / Cellular processes governing vascular Nox family NADPH oxidases in disease conditions are unclear. Since redox processes are intrinsic to cell stress response, we asked whether oxidative stress merges with other types of stress via Nox(es). We focused on proteasome inhibition as a relevant stress condition. Vascular smooth muscle cells (VSMC) incubation with non-apoptotic concentrations of proteasome inhibitors MG132 or lactacystin promoted increased baseline superoxide generation (HPLC/DHE products) and NADPH oxidase activity, decreased SOD activity and GSH/GSSG ratio. Conversely, proteasome inhibitors decreased by Nox response to Angiotensin II (AngII) and abrogated Nox response to endoplasmic reticulum (ER) stressor tunicamycin. With MG132, basal Nox1 mRNA levels were increased, while Nox1 response to AngII was blunted. Moreover, MG132 abolished Nox4 mRNA levels TN-induced. Both AngII and TN (at 2 and 4 hs) promoted increased 20S proteasome lytic activity. We next assessed physiological consequences of MG132 in ER stress signaling, a known Nox4- mediated response. VSMC incubation with MG132 alone enhanced expression of the ER stress markers Grp78 and XBP1 and late markers such as ATF4 and proapoptotic CHOP/GADD153. Similar results occurred with the known ER stressor TN. However, co-incubation of TN and MG132 decreased Grp78, Grp94 and CHOP/GADD153, indicating that proteasome inhibition interrupts ER stress. AKT and p38 are activated by MG132 as response to stress and recover to survival. Thus, proteasome inhibition disrupts NADPH oxidase, with increased baseline activity and Nox1 expression vs. strong inhibition of stimulated Nox1 and Nox4 activation/expression. The later effect may underlie MG132-mediated inhibition of ER stress signaling. (Support: FAPESP, CNPq Milênio Redoxoma)

L’hypertension artérielle pulmonaire (HTAP) est une maladie mortelle caractérisée par un important remodelage vasculaire, principalement dû à l’hyperprolifération et à la résistance à l’apoptose des cellules du muscle lisse de l’artère pulmonaire (CML-AP). Récemment il a été montré que les CML-AP présentent un remodelage du métabolisme énergétique, avec une régulation négative de l’oxidation phosphorylante associée à une activation de la voie glycolytique, qui semble contribuer à leur phénotype particulier. La désacétylase sirtuine1 (SIRT1) est un important modulateur du métabolisme énergétique, notamment via son activation de peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC-1α), régulateur clé de la biogenèse mitochondriale. Dans cette étude, nous montrons pour la première fois que la prolifération des CML-AP de rat et humaines est caractérisée par une réduction de l’activité de SIRT1, et est augmentée suite à l’inhibition pharmacologique ou la sous-expression spécifique de SIRT1. De plus, suite à hypoxie chronique, des souris génétiquement déficientes en SIRT1 présentent un remodelage vasculaire plus important que celui observé chez les souris contrôles, ce qui est associé à une augmentation accentuée de l’hypertrophie et de la pression systolique du ventricule droit. Au contraire, l’activation pharmacologique de SIRT1 inhibe fortement la prolifération des CML-AP, et est associée à l’activation de la biogenèse mitochondriale. L’ensemble de ces résultats suggère que l'inactivation de SIRT1 joue un rôle causal dans l’hyperprolifération des CML-AP et cette enzyme pourrait être une nouvelle cible thérapeutique prometteuse pour le traitement de l’HTAP. / Pulmonary arterial hypertension (PAH) is a lethal disease characterized by an intensive vascular remodelling, mainly due to hyper-proliferation and resistance to apoptosis of pulmonary artery smooth muscle cells (PASMCs). Recently it has been found that PASMCs, similarly to cancer cells, demonstrate a shift in energy metabolism from oxidative phosphorylation towards glycolysis thus contributing to their particular phenotype. The deacetylase sirtuin1 (SIRT1) is an important modulator of energy metabolism, particularly via its activation of peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC-1α), the master regulator of mitochondrial biogenesis. Here we show for the first time that rat and human PASMC proliferation is characterised by a diminution of SIRT1 activity, and is potentiated by SIRT1 pharmacological inhibition or specific downregulation. Moreover, after chronic hypoxia exposure, SIRT1 KO mice display a more intense vascular remodelling compared to their control littermates and this is associated with an exacerbated increase in right ventricle systolic pressure and hypertrophy. Conversely, pharmacological SIRT1 activation strongly inhibits PASMC proliferation, and is associated with the activation of mitochondrial biogenesis. In general, the data obtained show that SIRT1 inactivation plays a causative role in PASMC proliferation and this enzyme could be a promising therapeutic target for PAH treatment.

Les deux nucléotides cycliques, AMPc et GMPc, sont des seconds messagers importants qui régulent une grande variété de fonctions cellulaires, en particulier la fonction contractile cardiovasculaire, la croissance des cardiomyocytaires et la prolifération des cellules musculaires lisses vasculaires. Les phosphodiestérases (PDE) dégradent les nucléotides cycliques et exercent un contrôle local de leur concentration intracellulaire. Une altération de la voie de signalisation des nucléotides cycliques est impliquée dans plusieurs situations pathologiques telles que l’hypertension artérielle systémique ou pulmonaire, l’athérosclérose et l'hypertrophie cardiaque. Ainsi, les PDE constituent de puissantes cibles thérapeutiques pour restaurer un contrôle correct des nucléotides cycliques. Onze familles de PDEs sont actuellement décrites, les PDE1-6 étant les plus étudiées et les PDE 7-11 représentant de nouvelles familles.L'objectif de cette thèse était d'étudier le rôle respectif de 4 familles de PDEs, la PDE1, famille stimulée par le complexe Ca2+/calmoduline, les PDE5 et PDE9 spécifiques du GMPc, et la PDE8 spécifique de l'AMPc, dans le contrôle des concentrations intracellulaires d'AMPc ([AMPc]i) et de GMPc ([GMPc]i) dans les cellules musculaires lisses aortiques de rat (CMLARs) et les myocytes cardiaques de rat en utilisant une approche pharmacologique facilitée par le développement de nouveaux inhibiteurs sélectifs de PDEs. Les activités d'hydrolyse d’AMPc et de GMPc ont été mesurées par dosage enzymatique, tandis que les [AMPc]i et [GMPc]i ont été suivies sur cellules isolées, in situ, en temps réel, grâce à l'utilisation de l'imagerie FRET (Fluorescence Resonance Energy Transfer). Dans les CMLARs en culture, une activité d'hydrolyse des nucléotides cycliques via les PDE1, PDE5 et PDE9 a été observée. Nous avons montré un rôle fonctionnel de la PDE1 non stimulée dans le contrôle de l’augmentation de la [GMPc]i induite par le peptide natriurétique de type C (CNP). Il est intéressant de noter que, lors de l’élévation de la concentration intracellulaire en Ca2+, la PDE1 exerce également un contrôle de la réponse GMPci induite par le monoxyde d’azote (NO) et de la réponse AMPc médiée par la stimulation des récepteurs β-adrénergiques (β-AR). La PDE5 exerce un rôle majeur dans la réponse GMPc provoquée par l'activation de la guanylyl cyclase (GC) soluble par le NO ou des GC membranaires par les peptides natriurétiques, CNP et ANP. En revanche, la PDE9 ne régule que la réponse GMPc induite par le NO dans les RASMC cultivées. Aucune activité ou fonction hydrolytique de l'AMPc n'a été révélée avec l'inhibiteur de la PDE8 dans les CMLARs ou les cardiomyocytes de rat. Dans ces cellules cardiaques, l'activité d'hydrolyse médiée par la PDE1 n'a été détectée que sur la réponse GMPc et uniquement en présence de Ca2 +/Calmoduline. L'inhibiteur de la PDE1 n'a que légèrement affecté la réponse AMPc médiée par les récepteurs β-AR, par augmentation du pic du signal FRET.En conclusion, notre travail démontre que dans les cellules musculaires lisses vasculaires, les PDE1, PDE5 et PDE9 exercent une régulation spécifique et locale des [AMPc]i et [GMPc]i, renforçant le rôle clé des PDEs dans la compartimentation subcellulaire de la signalisation des nucléotides cycliques. / The two cyclic nucleotides cAMP and cGMP are important second messengers that regulate a large variety of cellular functions, in particular cardiovascular contractile function, cardiomyocyte cell growth and vascular smooth muscle cell proliferation. Phosphodiesterases (PDEs) degrade cyclic nucleotides, and exert a fine local control of their intracellular concentration. Alteration of cyclic nucleotides signaling pathway is involved in several pathological situations such as systemic and pulmonary arterial hypertensions, atherosclerotic lesions and cardiac hypertrophy. Thus, PDEs constitute potent therapeutic targets to restore a right cyclic nucleotide function. Eleven families of PDEs are now described, PDE1-6 being the most studied and PDE 7-11 representing the new families.The aim of the present thesis was to investigate the respective role of 4 PDE families, the Ca2+/calmodulin-stimulated PDE1, the cGMP-specific PDE5 and PDE9, and the cAMP-specific PDE8, in controlling intracellular cAMP ([cAMP]i) and intracellular cGMP ([cGMP]i) concentrations in both rat aortic smooth muscle cells (RASMCs) and cardiac myocytes by using a pharmacological approach taken advantage of the development of new selective PDE inhibitors. Cyclic AMP- and cGMP-hydrolyzing activities were measured by enzymatic assay on cell lysate, whereas real-time [cAMP]i and [cGMP]i were followed in situ in isolated cells using Fluorescence Resonance Energy Transfer (FRET) imaging. In cultured RASMCs, PDE1, PDE5 and PDE9 hydrolyzing activities were observed. We showed a functional role of basal PDE1 in controlling [cGMP]i increased by the C-type Natriuretic Peptide (CNP). Interestingly, upon high intracellular Ca2+ concentration, PDE1 also regulated the Nitric Oxide (NO)-mediated [cGMP]i response and the β-adrenoceptor (β-AR)-mediated [cAMP]i response. PDE5 exerted a major role in degrading [cGMP]i produced by the activation of either the soluble guanylyl cyclase (GC) elicited by NO or the particulate GCs by the natriuretic peptides, CNP and ANP. By contrast, PDE9 only regulated NO-induced [cGMP]i increase in cultured RASMCs. No cAMP-hydrolyzing activity or function was revealed with the PDE8 inhibitor in RASMCs or cardiac myocytes. In rat cardiomyocytes, PDE1-mediated hydrolyzing activity was only detected on cGMP in the presence of Ca2+/calmodulin. Unexpectedly, PDE1 inhibition slightly affected the β-AR-mediated [cAMP]i response by increasing the peak of FRET signal.In conclusion, our work underscores the distinct role of PDE1, PDE5, and PDE9 in locally regulating the [cAMP]i and [cGMP]i, in vascular smooth muscle cells, strengthening the concept of PDEs as key actors of cyclic nucleotide subcellular compartmentation.

The objective of the thesis is to create software, which implements industrial PID controller enabling smooth switch-over and automatic tuning of parameters. The next objective is to create visualization for this controller. Whole controller have to be implemented into PLC Power Panel (B&R company). At the beginning of the thesis there is theoretical description of all implemented algorithms (discrete control algorithms derived from PID, smooth switch-over, antiwindup, identification algorithms). In the following there is designed program structure, which is state machine-shaped. All algorithms are implemented in ANSI C at first as s-function for Matlab/Simulink program, which enables to simulate and verify the controller. Implementation of controller into PLC Power Panel through the use of Automation Studio program from B&R company is described in the next part. Visualization is created in the same program. Simulations and verifications on mathematical and physical model demonstrates functionality of implemented algorithms, but also points out some problems associated with the use of identification algorithms in real world (noise, quantization in A/D and D/A converter).

L’appareil digestif est un organe vital qui assure la digestion des aliments, l’absorption des nutriments et l’élimination des déchets. Une des propriétés essentielles du tube digestif est la motricité digestive qui est définie comme l’ensemble des contractions nécessaires au transit du bol alimentaire depuis la bouche jusqu’à l’anus. Les acteurs de la motricité digestive sont le système nerveux entérique, les cellules interstitielles de Cajal, et les cellules musculaires lisses. Les cellules musculaires lisses et les cellules interstitielles de Cajal ont pour origine un progéniteur mésenchymateux commun. Les cellules dérivées du mésenchyme présentent une certaine plasticité et sont capables de transiter d’un état différencié contractile et fonctionnel à un état prolifératif et immature. Toutefois, un déséquilibre de cette balance au profit de l’état d’immaturité est à l’origine de désordres de motricité digestive. Les travaux de recherches développés par l’équipe ont pour objectifs d’étudier les mécanismes qui gouvernent la différenciation des progéniteurs mésenchymateux digestifs afin d’étudier ces mécanismes en conditions pathologiques. Dans cet optique, l’équipe a identifié le gène LIX1 (LImb eXpression 1) comme le premier marqueur moléculaire de l’immaturité du muscle lisse digestif et a mis en évidence son rôle dans le contrôle de la différenciation des progéniteurs mésenchymateux au travers de la régulation de l’oncogène YAP1 (McKey et al, 2016). Dans ce contexte, le travail de recherche que j’ai réalisé s’est principalement concentré sur l’étude de LIX1 et de ses protéines partenaires dans le contrôle de la différenciation des cellules musculaires lisses gastriques et leur plasticité en conditions pathologiques.Dans un premier temps, j’ai étudié la fonction de LIX1 dans un cancer mésenchymateux du tube digestif, les GISTs (GastroIntestinal Stromal Tumor). J’ai mis en évidence le rôle et la fonction de LIX1 dans l’agressivité et dans l’immaturité des GISTs. Dans un deuxième temps, j’ai participé à la caractérisation moléculaire de cellules dérivées de patients POIC (Pseudo Obstruction Intestinale Chronique) pour lesquelles nous avons mis en évidence un défaut de différenciation associé à une expression anormale de PDGFR-A. Dans un troisième temps, j’ai développé un modèle de cellules musculaires lisses gastriques humaines dont la différenciation est maîtrisable pour étudier le métabolisme au cours de la différenciation. L’ensemble des travaux montre que LIX1 et sa mécanistique participent à la plasticité des SMCs. / The digestive tract is a vital organ ensuring food digestion, nutrient absorption and waste excretion. One of the main properties of digestive tract is the motricity which is defined as the set of contractions that allows the transition of the food from the mouth to the anus. Cells involved in the regulation of digestive plasticity are the enteric nervous cells, the interstitial cells of Cajal and the smooth muscle cells. The interstitial cells of Cajal and smooth muscle cells derived from a common mesenchymal progenitor. Mesenchyme-derived cells have the unique capacity to switch from the contractile and functional state to an immaturity state. This plasticity is responsible for motricity disorders. Our work aims to identify the mechanisms involved in the differentiation of the mesenchymal progenitors and to study those mechanisms in pathological conditions. The team previously identified the LIX1 gene (LImb eXpression 1) as the first molecular marker of the digestive smooth muscle immaturity and demonstrated its role on the differentiation of mesenchymal progenitors through the control of YAP1 (McKey et al., 2016). In this context, during my thesis, I focused on LIX1 and the mitochondrial remodeling as a putative regulatory mechanism of mesenchymal-derived cells differentiation. First, I investigated and demonstrated the role and function of LIX1 in the aggressiveness and the immaturity of the GastroIntestinal Stromal Tumor (GIST) cells. In parallel, I participated to the characterization of cells derived from CPIO (Chronic Pseudo Intestinal Obstruction) patients. Finally, I developed a new model of human gastric smooth muscle cells to evaluate the metabolism during the SMC differentiation. Altogether, we showed that LIX1 and its downstream pathways control SMC plasticity.

Mechanically Stabilized Earth (MSE) walls retain soil on steep, unstable slopes with crest loads. Over the last decade, they are becoming quite popular due to their low cost-to-benefit ratio, design flexibility, and ease of construction. Like any civil infrastructure, MSE walls need to be continuously monitored according to transportation asset management criteria during and after the construction stage to ensure that their expected serviceability measures are met and to detect design and/or construction issues, which could lead to structural failure. Current approaches for monitoring MSE walls are mostly qualitative (e.g., visual inspection or examination). Besides being time consuming, visual inspection might have inconsistencies due to human subjectivity. Other monitoring approaches are based on using total station, geotechnical field instrumentations, and/or Static Terrestrial Laser Scanning (TLS). These instruments are capable of providing highly accurate, reliable performance measures. However, the underlying data acquisition and processing strategies are time-consuming and are not scalable. This research focuses on a comprehensive strategy using a Mobile LiDAR Mapping System (MLS) for the acquisition and processing of point clouds covering the MSE wall. The strategy produces standard serviceability measures, as defined by the American Association of State Highway and Transportation Officials (AASHTO) – e.g., longitudinal and transversal angular distortions. It also delivers a set of recently developed measures (e.g., out-of-plane offsets and 3D position/orientation deviations for individual panels constituting the MSE wall). Moreover, it is also capable of handling MSE walls with smooth or textured panels with the latter being the focus of this research due to its more challenging nature. For this study, an ultra-high-accuracy wheel-based MLS has been developed to efficiently acquire reliable data conducive to the development of the standard and new serviceability measures. To illustrate the feasibility of the proposed acquisition/processing strategy, two case studies in this research have been conducted with the first one focusing on the comparative performance of static and mobile LiDAR in terms of the agreement of the derived serviceability measures. The second case study aims at illustrating the feasibility of the proposed strategy in handling large textured MSE walls. Results from both case studies confirm the potential of using MLS for efficient, economic, and reliable monitoring of MSE walls.

Abnormal vascular smooth muscle cell (VSMC) proliferation is thought to play an important role in the pathogenesis of atherosclerosis. Adipocytes produce several paracrine bioactive substances that can affect VSMC growth and migration. Our study focused on the ability of epicardial adipocytes to produce bioactive substances together with studying of direct effect of these substances on the VSMC proliferation rate. The gene expression of human cytokines (IL-6, IL-8, IL-18, RANTES and MCP-1) and adipokines (leptin and adiponectin) was measured in primary cell lines of epicardial and visceral adipocytes, both in undifferentiated and mature statuses. Moreover, adipokine production (IL-6, IL-8, MCP-1, VEGF and adiponectin) in conditioned media obtained from above mentioned primary cell cultures of adipocytes was measured by a Luminex assay. The VSMC proliferation rate was measured after co-culturing with CM obtained from primary cell cultures of adipocytes. The epicardial preadipocytes showed an increased expression of IL-8 (3,25-fold, p<0,05) compared with visceral preadipocytes. The expression of the adiponectin in epicardial preadipocytes was markedly decreased in comparison of the expression in visceral preadipocytes (p< 0,0001). Moreover, the gene expression was dependent on the differentiation...

<div>Many practical applications benefit from the reconstruction of a smooth multivariate function from discrete data for purposes such as reducing file size or improving analytic and visualization performance. Among the different reconstruction methods, tensor product B-spline has a number of advantageous properties over alternative data representation. However, the problem of constructing a best-fit B-spline approximation effectively contains many roadblocks. Within the many free parameters in the B-spline model, the choice of the knot vectors, which defines the separation of each piecewise polynomial patch in a B-spline construction, has a major influence on the resulting reconstruction quality. Yet existing knot placement methods are still ineffective, computationally expensive, or impose limitations on the dataset format or the B-spline order. Moving beyond the 1D cases (curves) and onto higher dimensional datasets (surfaces, volumes, hypervolumes) introduces additional computational challenges as well. Further complications also arise in the case of undersampled data points where the approximation problem can become ill-posed and existing regularization proves unsatisfactory.</div><div><br></div><div>This dissertation is concerned with improving the efficiency and accuracy of the construction of a B-spline approximation on discrete data. Specifically, we present a novel B-splines knot placement approach for accurate reconstruction of discretely sampled data, first in 1D, then extended to higher dimensions for both structured and unstructured formats. Our knot placement methods take into account the feature or complexity of the input data by estimating its high-order derivatives such that the resulting approximation is highly accurate with a low number of control points. We demonstrate our method on various 1D to 3D structured and unstructured datasets, including synthetic, simulation, and captured data. We compare our method with state-of-the-art knot placement methods and show that our approach achieves higher accuracy while requiring fewer B-spline control points. We discuss a regression approach to the selection of the number of knots for multivariate data given a target error threshold. In the case of the reconstruction of irregularly sampled data, where the linear system often becomes ill-posed, we propose a locally varying regularization scheme to address cases for which a straightforward regularization fails to produce a satisfactory reconstruction.</div>

In Caught Between Jazz and Pop, I challenge the prevalent marginalization and malignment of smooth jazz in the standard jazz narrative. Furthermore, I question the assumption that smooth jazz is an unfortunate and unwelcomed evolutionary outcome of the jazz-fusion era. Instead, I argue that smooth jazz is a long-lived musical style that merits multi-disciplinary analyses of its origins, critical dialogues, performance practice, and reception. Chapter 1 begins with an examination of current misconceptions about the origins of smooth jazz. In many jazz histories, the origins of smooth jazz are defined as a product of the jazz-fusion era. I suggest that smooth jazz is a distinct jazz style that is not a direct outgrowth of any mainstream jazz style, but a hybrid of various popular and jazz styles. Chapters 2 through 4 contain eight case studies examining the performers of crossover jazz and smooth jazz. These performers have conceived and maintained distinct communicative connections between themselves and their audiences. In the following chapter, the unfair treatment of popular jazz styles is examined. Many early and influential jazz critics sought to elevate jazz to the status of art music by discrediting popular jazz styles. These critics used specific criteria and emphasized notions of anti-commerciality to support their theoretical positions. In Chapter 6, the studio recordings and live performances of smooth jazz are discussed. Critics frequently complain that most smooth jazz recordings feature glossy packaging and pristine studio editing, resulting in a too-perfect product. Although this aesthetic is the result of a unique series of interactions, recordings do not represent the complete musical nature of smooth jazz. Live performances contain important, but typically neglected aspects of smooth jazz. Live performances enable performers to extend solos, interact, and communicate directly to the audience. While recordings are a useful source for musical analysis, smooth jazz, like other styles of jazz, is an improvisatory music that utilizes multiple sites of production and cannot be accurately judged on recordings alone.

The dissertation focuses on two general categories of defining space: on space expression according to the expressive approach to aesthetics and, secondly, on space representation based on the mimetic aesthetics. The exploration of the surface structure of space employs the philosophical categories of smooth and striated space, formulated by Deleuze and Guattari in their Treatise on Nomadology, while the depths of the inner spaces, including the spaces of the human mind, are treated within the framework of Gaston Bachelard's phenomenology, stressing the importance of the symbolic meanings hidden in the unconscious. The primary texts in which the concept of space is explored range from the Brontë sisters' novels (Wuthering Heights, Jane Eyre) to Thomas Hardy's Wessex novels, and further on to the last novel of George Eliot, Daniel Deronda. Attention is given to the role of natural elements constituting space, with emphasis on the element of water.