Soil aquifer treatment was simulated in 1 m laboratory soil columns containing silica sand under saturated and unsaturated soil conditions to examine the effect of travel length through the unsaturated zone on the removal of wastewater organic matter, the effect of soil type on dissolved organic carbon removal and also the type of microorganisms involved in the removal process. Dissolved organic carbon removal and nitrification did enhance when the wastewater travelled a longer length through the unsaturated zone. A similar consortium of microorganisms was found to exist in both saturated and unsaturated columns. Microbial concentrations however were lowest in the soil column containing silt and clay in addition to silica sand. The presence of silt and clay was detrimental to DOC removal efficiency under saturated soil conditions due to their negative effect on the hydraulic performance of the soil column and microbial growth.

Diese Arbeit konzentriert sich auf die Anwendbarkeit eines hochmodernen 100 kHz BB-VSFG-Spektrometers, das kürzlich im SALSA Photonics Lab entwickelt wurde, für die Analyse der Grenzflächenstruktur von Alkylketten, des sie umgebenden Wassers und der Phosphatkopfgruppen von Phospholipidschichten. Zunächst wurden Phospholipid-Doppelschichten, die mehrere Komponenten enthalten, bei Laserwiederholraten von 5, 10, 50 und 100 kHz mit konstanter Pulsenergie untersucht. Die BB-VSFG-Spektren legen nahe, dass die Phospholipid-Doppelschichten während der Messungen ohne wärmeinduzierte Veränderungen stabil waren. Darüber hinaus bot die Erhöhung der Laserwiederholungsrate eine praktikable Möglichkeit, Spektren in kurzen Datenerfassungszeiten zu erhalten, ohne dass das Signal-Rausch-Verhältnis beeinträchtigt wurde. Die extrem kurze Aufnahmezeit von 500 ms, die hohe spektrale Auflösung und alle verwendeten Pulsparameter sorgen dafür, dass bei Messungen unter Umgebungsbedingungen keine thermisch bedingten Photoschäden auftreten. Es wurde eine systematische Untersuchung von ein- und zweikomponentigen Phospholipid-Monoschichten in Abhängigkeit von der Oberflächenspannung und dem Mischungsverhältnis für verschiedene Kombinationen an Polarisationen durchgeführt und die Abhängigkeit der Schwingungsspektren untersucht. Die Struktur von Alkylketten und umgebendem Wasser wurde anhand derselben Modellsystemen analysiert. Bislang nicht beobachtete Schwingungsbanden und Spektren von Monolagen mit geringer Oberflächenbedeckung wurden mit einem bisher nicht erreichten Signal-Rausch-Verhältnis gemessen und beschrieben. Die Struktur von Phospholipid-Monolagen mit identischen Kopfgruppen und unterschiedlichen Ketten wurde analysiert und verglichen. Die Spektren bestätigten die Anwesenheit von Wassermolekülen in der Nähe der Phosphat- und Cholingruppen der Phospholipid-Monolagen. / This thesis focuses on the applicability of a state-of-the-art 100 kHz BB-VSFG spectrometer recently developed at the SALSA Photonics Lab and on the analysis of the interfacial structure of alkyl chains, surrounding water, and the phosphate head groups of phospholipid layers. First, multi-component phospholipid bilayers were studied at laser repetition rates of 5, 10, 50, and 100 kHz at constant pulse energy. The spectra suggest that the phospholipid bilayers were stable during the measurements with no heat-induced distortions. Moreover, an increase in the laser repetition rate provided a feasible route to obtain spectra in short data acquisition times without compromising the signal-to-noise ratio. The extremely short acquisition time of 500 ms, the high spectral resolution, and all applied pulse parameters ensured no thermal induced photodamages occur during the measurements. A systematic study of one- and two-component phospholipid monolayers as a function of surface tension and mixture ratio at different polarization combinations was performed and the dependence of the vibrational spectra was explored. The structure of alkyl chains and surrounding water was analyzed using the same model systems. Vibrational modes that were previously unseen and spectra of monolayers at low surface coverage were reported for the first time with an unprecedented signal-to-noise ratio. The structure of phospholipid monolayers containing identical head groups and different chains was analyzed and compared. The order of the phospholipid molecules as a function of the composition of the monolayers was inferred from the spectral data. The influence of the hydration and/or changes in the orientation of the phosphate group was visible from the spectra as well.

No description available.

Phospholipids are dominant components of bacterial cell membranes, where they create double layers. Bacteria differ in their phospholipid composition determination of which can help in identification of important groups of microorganisms. Phospholipid composition of bacteria is influenced by many environmental factors, therefore its variation can be observed within one bacterial stem also. Because of its simplicity, thin layer chromatography is usually applied to identification and determination of bacterial phospholipids. Disadvantage of this method are the high demands of time, carefulness and skills of the analytical personnel. The increasing interest in the phospholipid double-layer promotes the detailed investigation of their fatty acid composition because the more detailed analyses allows for more information yield about bacteria. Gas chromatography hyphenated with mass spectrometry seems to be the best choice for these purposes. Fatty acid identity and total fatty acid content in phospholipid molecules could be determined by this method. Additionally, number, position and isomerism of double bonds and presence of other functional groups on hydrocarbon chain could be determined. Whereas a suitable and...

INTRODUÇÃO: Os transtornos mentais psicóticos são condições frequentes na população geral e estão associados a grande morbidade e disfuncionalidade. Apesar disso, as bases fisiopatológicas destes transtornos ainda estão em investigação. Estudos neuropatológicos post-mortem e de neuroimagem in vivo sugerem haver comprometimento da microestrutura de substância branca (SB) cerebral nestas condições clínicas, associado a alterações da conectividade cerebral. No entanto, nenhuma investigação prévia de neuroimagem avaliou especificamente se tais anormalidades microestruturais podem ser dependentes do estado clínico do paciente, i.e., se tais alterações podem variar de acordo com a fase da doença. Outra linha de investigação biológica em psicoses aponta para alterações na atividade da fosfolipase A2 (PLA2), uma enzima essencial a diversas funções na homeostase cerebral, incluindo manutenção de membrana celular, mielinização e atividade inflamatória. Estudos prévios sugerem haver relação entre alterações na atividade desta enzima e as fases da esquizofrenia. Entretanto, não há estudos translacionais que tenham avaliado como tais alterações se relacionam com anormalidades microestruturais de SB em pacientes psicóticos. OBJETIVOS: Investigar a hipótese de que alterações de microestrutura de SB presentes em pacientes em fase aguda do primeiro episódio psicótico (PEP) sejam potencialmente reversíveis após estabilização clínica; investigar também possíveis alterações estado-dependentes da atividade de PLA2 no PEP; e examinar interações entre manifestações clínicas, microestrutura de SB cerebral e atividade de PLA2 na fisiopatologia do PEP. METODOLOGIA: Pacientes em PEP não afetivo foram avaliados em dois períodos no tempo: durante a fase aguda da doença (T0); após remissão estável de sintomas (T1). Um grupo controle de voluntários saudáveis (CS) também foi avaliado longitudinalmente. Para investigar alterações de microestrutura de SB estado-dependentes, análises voxel-a-voxel de mapas cerebrais de índices de anisotropia (fractional anisotropy, FA) e difusividade (trace, TR) foram conduzidas, assim como o cálculo de correlações entre tais índices de DTI, variáveis clínicas e atividade de PLA2. A atividade dos três principais subgrupos de PLA2 em plaquetas foi estimada através de um método radioenzimático. RESULTADOS: 25 pacientes PEP e 51 CS foram avaliados em T0, com coleta de dados clínico-demográficos, ressonância magnética (RM) e amostra de sangue. Destes, 21 PEP e 36 CS realizaram a segunda aquisição de RM. No baseline (T0), os pacientes PEP apresentaram redução difusa de FA (p < 0,05, FDR), afetando principalmente SB fronto-límbica e fascículos associativos, projetivos e comissurais. As análises longitudinais demonstraram que a remissão clínica se associou a aumentos de FA em tratos de SB acometidos em T0 (p < 0,001, não corrigido), além de robustas correlações inversas entre aumentos de FA e redução sintomas ao longo do tempo (p < 0,05, FDR). As análises de PLA2 não demostraram efeitos estado-dependentes ou correlações consistentes com os índices de DTI. CONCLUSÃO: Alterações da microestrutura de SB afetando tratos cerebrais essenciais para a integração de informação perceptual, cognição e emoções são detectáveis logo após o início do PEP e podem ser parcialmente revertidas em relação direta com a remissão de sintomas psicóticos agudos. Nossos achados reforçam a visão de que anormalidades de SB de tratos cerebrais são um componente neurobiológico central nos transtornos psicóticos agudos, e que a recuperação de tal patologia de SB pode levar à melhora clínica. Por outro lado, a atividade de PLA2 não parece ter associação direta com o estado de doença ou moderar as alterações microestruturais dinâmicas de SB aqui observadas. Estudos com amostras maiores e com um maior número de avaliações ao longo do tempo são necessários para confirmar e ampliar os resultados aqui apresentados / INTRODUCTION: Psychotic disorders are frequent conditions in the general population and are associated to severe morbidity and functional impairment. Notwithstanding, the pathophysiological basis of such disorders is still under investigation. Post-mortem neuropathologic investigations and in vivo neuroimaging studies have pointed to the occurrence of abnormalities in the microstructure of brain white matter (WM) in such clinical conditions, which are associated to changes in brain connectivity. However, no previous neuroimaging investigation has specifically examined whether such microstructural abnormalities would be state-dependent, i.e., whether such changes could relate to the illness phase. Another field of biological investigation in psychosis points to changes in the activity of phospholipase A2 enzyme (PLA2), which is essential to several functions implicated in brain homeostasis, such as the maintenance of cellular membrane, myelination and inflammatory activity. Previous studies suggest the existence of a relationship between changes on PLA2 activity and schizophrenia phase. Nonetheless, no translational study to date has examined the potential interplay between PLA2 activity and WM microstructural abnormalities in psychotic patients. OBJECTIVES: To investigate the hypothesis that WM microstructural changes observed in patients during the acute first-episode psychosis (FEP) are potentially reversible following clinical remission; to investigate possible state-dependent changes in PLA2 activity in FEP; and to examine interactions between clinical manifestations, brain WM microstructure and PLA2 activity in the pathophysiology of FEP. METODOLOGY: Patients with non-affective FEP were evaluated in two time points: during the acute phase (T0) and following sustained remission (T1). A control group of healthy volunteers (HC) was also longitudinally studied. In order to investigate state-dependent WM microstructure changes, voxelwise analyses of brain maps of anisotropy (fractional anisotropy, FA) and diffusivity (trace, TR) indexes were conducted, as well as correlations between such DTI metrics, clinical variables and PLA2 activity. The activity of the three main PLA2 subgroups was assessed in platelets using a radioenzymatic method. RESULTS: 25 FEP and 51 HC were evaluated at T0 (clinical and demographic data, MRI scanning, and blood collection). Out of these, 21 FEP and 36 HC also underwent a second MRI acquisition. At baseline (T0), FEP patients presented widespread reduction of FA (p < 0.05, FDR), affecting mainly fronto-limbic WM and associative, projective and commissural fasciculi. Longitudinal analyses showed that clinical remission was associated with FA increase in WM tracts that were affected at T0 (p < 0.001, uncorrected), besides robust inverse correlations between FA increase and symptoms reduction over time (p < 0.05, FDR). PLA2 analyses failed to show state-dependent effects or consistent correlations to DTI indexes. CONCLUSION: WM changes affecting brain tracts critical to the integration of perceptual information, cognition and emotions are detectable soon after the onset of FEP and may partially reverse in direct relation to the remission of acute psychotic symptoms. Our findings reinforce the view that WM abnormalities are a key neurobiological feature of acute psychotic disorders, and that recovery from such WM pathology can lead to amelioration of symptoms. In the other hand, it seems that PLA2 activity has no direct relationship to the disease state or modulatory effects on the dynamic WM changes observed herein. Studies with larger samples and with more time points evaluations are necessary to confirm and expand the findings reported herein

INTRODUÇÃO: Os transtornos mentais psicóticos são condições frequentes na população geral e estão associados a grande morbidade e disfuncionalidade. Apesar disso, as bases fisiopatológicas destes transtornos ainda estão em investigação. Estudos neuropatológicos post-mortem e de neuroimagem in vivo sugerem haver comprometimento da microestrutura de substância branca (SB) cerebral nestas condições clínicas, associado a alterações da conectividade cerebral. No entanto, nenhuma investigação prévia de neuroimagem avaliou especificamente se tais anormalidades microestruturais podem ser dependentes do estado clínico do paciente, i.e., se tais alterações podem variar de acordo com a fase da doença. Outra linha de investigação biológica em psicoses aponta para alterações na atividade da fosfolipase A2 (PLA2), uma enzima essencial a diversas funções na homeostase cerebral, incluindo manutenção de membrana celular, mielinização e atividade inflamatória. Estudos prévios sugerem haver relação entre alterações na atividade desta enzima e as fases da esquizofrenia. Entretanto, não há estudos translacionais que tenham avaliado como tais alterações se relacionam com anormalidades microestruturais de SB em pacientes psicóticos. OBJETIVOS: Investigar a hipótese de que alterações de microestrutura de SB presentes em pacientes em fase aguda do primeiro episódio psicótico (PEP) sejam potencialmente reversíveis após estabilização clínica; investigar também possíveis alterações estado-dependentes da atividade de PLA2 no PEP; e examinar interações entre manifestações clínicas, microestrutura de SB cerebral e atividade de PLA2 na fisiopatologia do PEP. METODOLOGIA: Pacientes em PEP não afetivo foram avaliados em dois períodos no tempo: durante a fase aguda da doença (T0); após remissão estável de sintomas (T1). Um grupo controle de voluntários saudáveis (CS) também foi avaliado longitudinalmente. Para investigar alterações de microestrutura de SB estado-dependentes, análises voxel-a-voxel de mapas cerebrais de índices de anisotropia (fractional anisotropy, FA) e difusividade (trace, TR) foram conduzidas, assim como o cálculo de correlações entre tais índices de DTI, variáveis clínicas e atividade de PLA2. A atividade dos três principais subgrupos de PLA2 em plaquetas foi estimada através de um método radioenzimático. RESULTADOS: 25 pacientes PEP e 51 CS foram avaliados em T0, com coleta de dados clínico-demográficos, ressonância magnética (RM) e amostra de sangue. Destes, 21 PEP e 36 CS realizaram a segunda aquisição de RM. No baseline (T0), os pacientes PEP apresentaram redução difusa de FA (p < 0,05, FDR), afetando principalmente SB fronto-límbica e fascículos associativos, projetivos e comissurais. As análises longitudinais demonstraram que a remissão clínica se associou a aumentos de FA em tratos de SB acometidos em T0 (p < 0,001, não corrigido), além de robustas correlações inversas entre aumentos de FA e redução sintomas ao longo do tempo (p < 0,05, FDR). As análises de PLA2 não demostraram efeitos estado-dependentes ou correlações consistentes com os índices de DTI. CONCLUSÃO: Alterações da microestrutura de SB afetando tratos cerebrais essenciais para a integração de informação perceptual, cognição e emoções são detectáveis logo após o início do PEP e podem ser parcialmente revertidas em relação direta com a remissão de sintomas psicóticos agudos. Nossos achados reforçam a visão de que anormalidades de SB de tratos cerebrais são um componente neurobiológico central nos transtornos psicóticos agudos, e que a recuperação de tal patologia de SB pode levar à melhora clínica. Por outro lado, a atividade de PLA2 não parece ter associação direta com o estado de doença ou moderar as alterações microestruturais dinâmicas de SB aqui observadas. Estudos com amostras maiores e com um maior número de avaliações ao longo do tempo são necessários para confirmar e ampliar os resultados aqui apresentados / INTRODUCTION: Psychotic disorders are frequent conditions in the general population and are associated to severe morbidity and functional impairment. Notwithstanding, the pathophysiological basis of such disorders is still under investigation. Post-mortem neuropathologic investigations and in vivo neuroimaging studies have pointed to the occurrence of abnormalities in the microstructure of brain white matter (WM) in such clinical conditions, which are associated to changes in brain connectivity. However, no previous neuroimaging investigation has specifically examined whether such microstructural abnormalities would be state-dependent, i.e., whether such changes could relate to the illness phase. Another field of biological investigation in psychosis points to changes in the activity of phospholipase A2 enzyme (PLA2), which is essential to several functions implicated in brain homeostasis, such as the maintenance of cellular membrane, myelination and inflammatory activity. Previous studies suggest the existence of a relationship between changes on PLA2 activity and schizophrenia phase. Nonetheless, no translational study to date has examined the potential interplay between PLA2 activity and WM microstructural abnormalities in psychotic patients. OBJECTIVES: To investigate the hypothesis that WM microstructural changes observed in patients during the acute first-episode psychosis (FEP) are potentially reversible following clinical remission; to investigate possible state-dependent changes in PLA2 activity in FEP; and to examine interactions between clinical manifestations, brain WM microstructure and PLA2 activity in the pathophysiology of FEP. METODOLOGY: Patients with non-affective FEP were evaluated in two time points: during the acute phase (T0) and following sustained remission (T1). A control group of healthy volunteers (HC) was also longitudinally studied. In order to investigate state-dependent WM microstructure changes, voxelwise analyses of brain maps of anisotropy (fractional anisotropy, FA) and diffusivity (trace, TR) indexes were conducted, as well as correlations between such DTI metrics, clinical variables and PLA2 activity. The activity of the three main PLA2 subgroups was assessed in platelets using a radioenzymatic method. RESULTS: 25 FEP and 51 HC were evaluated at T0 (clinical and demographic data, MRI scanning, and blood collection). Out of these, 21 FEP and 36 HC also underwent a second MRI acquisition. At baseline (T0), FEP patients presented widespread reduction of FA (p < 0.05, FDR), affecting mainly fronto-limbic WM and associative, projective and commissural fasciculi. Longitudinal analyses showed that clinical remission was associated with FA increase in WM tracts that were affected at T0 (p < 0.001, uncorrected), besides robust inverse correlations between FA increase and symptoms reduction over time (p < 0.05, FDR). PLA2 analyses failed to show state-dependent effects or consistent correlations to DTI indexes. CONCLUSION: WM changes affecting brain tracts critical to the integration of perceptual information, cognition and emotions are detectable soon after the onset of FEP and may partially reverse in direct relation to the remission of acute psychotic symptoms. Our findings reinforce the view that WM abnormalities are a key neurobiological feature of acute psychotic disorders, and that recovery from such WM pathology can lead to amelioration of symptoms. In the other hand, it seems that PLA2 activity has no direct relationship to the disease state or modulatory effects on the dynamic WM changes observed herein. Studies with larger samples and with more time points evaluations are necessary to confirm and expand the findings reported herein

Im Rahmen dieser Arbeit wurden fluoreszierende Sphingomyelin-Analoga zu Studium der sauren Sphingomyelinase (ASM) synthetisiert. Ausgehend von L-Serin wurde ein Sphingosin-Derivat mit natürlicher Stereochemie dargestellt. Anschließend wurde mittels Phosphorodichloridat-Chemie eine Aminoethylphosphat-Gruppe installiert. Zweifache Fluoreszenzmarkierung ergab Sonden mit der Fähigkeit zu Förster-Resonanzenergietransfer (FRET). Diese wurden als Substrate der ASM akzeptiert und erlaubten die Verfolgung der Enzymaktivität in vitro. Durch die Analyse der photophysikalischen Eigenschaften der Fluorophore wurde das allgemeine Konzept der Phasentrennungs-gestützten Signalverstärkung (PS) abgeleitet. Dieses Konzept wurde erfolgreich bestätigt durch die Synthese einer 30-mal leistungsfähigeren zweiten Generation der FRET-Sonde. Ein homogener Assay wurde entwickelt, der die Quantifizierung der ASM-Aktivität erlaubte. Unter Verwendung von gereinigter rekombinanter humaner ASM, HeLa-Zelllysaten oder Lysaten von murinen embryonalen Fibroblasten (MEFs) als Enzymquelle wurde ausschließlich unter den von der ASM bevorzugten Bedingungen eine vollständige und spezifische Hydrolyse der Sonde beobachtet. Des Weiteren erlaubte die Sonde die Detektion relativer Unterschiede der Aktivität der ASM in kultivierten MEFs mittels Fluoreszenzmikroskopie mit Zweiphotonenanregung (2PE). / Fluorescent sphingomyelin analogues have been synthesized to probe the acid sphingomyelinase (ASM). Starting from L-serine, a sphingosine with natural stereochemistry was synthesized. Subsequently, phosphorodichloridate chemistry was used to install an aminoethyl phosphate moiety. Dual fluorescent labeling afforded probes capable of Förster resonance energy transfer (FRET). They were recognized as substrates of ASM and allowed for monitoring of the enzyme’s activity in vitro. Through analysis of the fluorophores’ photophysical properties, the general concept of partition aided amplification of a FRET probe’s signal (PS) was developed. This concept was successfully confirmed by the synthesis of a second-generation probe with 30-fold improved response. A homogenous assay was developed, which allowed for a quantitation of ASM activity. Using either purified recombinant human ASM, or lysates of HeLa cells or mouse embryonic fibroblasts (MEFs) as an enzyme source, complete and specific cleavage was observed exclusively under conditions preferred by ASM. Furthermore, the probe enabled the detection of relative levels of ASM activity in cultivated MEFs using fluorescence microscopy with two-photon excitation (2PE).

The gastrointestinal tract (GIT) regulates nutrient uptake, secretes hormones and has a crucial gut flora and enteric nervous system. Of relevance for these functions are the G protein-coupled receptors (GPCRs) and the solute carriers (SLCs). The Adhesion GPCR subfamily is known to mediate neural development and immune system functioning, whereas SLCs transport e.g. amino acids, fatty acids (FAs) and drugs over membranes. We aimed to comprehensively characterize Adhesion GPCR and SLC gene expression along the rat GIT. Using qPCR we measured expression of 78 SLCs as well as all 30 Adhesion GPCRs in a twelve-segment GIT model. 21 of the Adhesion GPCRs had a widespread (≥5 segments) or ubiquitous (≥11 segments) expression. Restricted expression patterns were characteristic for most group VII members. Of the SLCs, we found the majority (56 %) of these transcripts to be expressed in all GIT segments. SLCs were predominantly found in the absorption-responsible gut regions. Both Adhesion GPCRs and SLCs were widely expressed in the rat GIT, suggesting important roles. The distribution of Adhesion GPCRs defines them as a potential pharmacological target. FAs constitute an important energy source and have been implicated in the worldwide obesity increase. FAs and their ratios – indices for activities of e.g. the desaturase enzymes SCD-1 (SCD-16, 16:1n-7/16:0), D6D (18:3n-6/18:2n-6) and D5D (20:4n-6/20:3n-6) – have been associated with e.g. overall mortality and BMI. We examined whether differences in FAs and their indices in five lipid fractions contributed to obesity susceptibility in rats fed a high fat diet (HFD), and the associations of desaturase indices between lipid fractions in animals on different diets. We found that on a HFD, obesity-prone (OP) rats had a higher SCD-16 index and a lower linoleic acid (LA) proportions in subcutaneous adipose tissue (SAT) than obesity-resistant rats. Desaturase indices were significantly correlated between many of the lipid fractions. The higher SCD-16 may indicate higher SCD-1 activity in SAT in OP rats, and combined with lower LA proportions may provide novel insights into HFD-induced obesity. The associations between desaturase indices show that plasma measurements can serve as proxies for some lipid fractions, but the correlations seem to be affected by diet and weight gain.

Afin d’améliorer l’efficacité des traitements des pathologies articulaires, en tenant compte de leur complexité et de leur ampleur, des études récentes ont mis en évidence le rôle des assemblages lipidiques associés à la structure discontinue du fluide synovial dans le contrôle du fonctionnement tribologique articulaire. Ceci à conduit à la mise au point d’un modèle tribologique ex vivo (thèse AM Sfarghiu, 2006), proposant un « motif élémentaire » de la biolubrification articulaire, constitué de l’empilement d’interfaces phospholipidiques et de couches aqueuses. En utilisant ce modèle, l’objectif de ce travail a été d’étudier l’évolution des interfaces phospholipidiques du fluide synovial en présence de pathologies. Pour ce faire, une méthodologie nano-bio-tribologique alliant des analyses biochimiques, physicochimiques, nano-mécaniques et tribologiques a été utilisée. Les résultats de ces analyses montrent : l’influence de la faible rugosité des surfaces frottantes caractérisant les stades précoces des pathologies et celle des propriétés des interfaces phospholipidiques (liées à la variation de leur composition) sur la résistance mécanique, l’évolution au cours du frottement et la dégradation in situ des assemblages lipidiques des fluides synoviaux pathologiques. Le comportement des assemblages lipidiques est accentué par l’action des enzymes associées aux pathologies. Par conséquent, le fonctionnement articulaire dépend de la résistance mécanique des interfaces phospholipidiques et pour obtenir des coefficients de frottement très bas, l’accommodation de vitesse doit s’effectuer au niveau des couches d’hydratation qui entourent les ions présents dans la couche aqueuse. Ces résultats permettront de comprendre à court terme l’évolution des interfaces phospholipidiques dans les pathologies articulaires et, à plus long terme le bon enchaînement cause/conséquence responsable d’une pathologie articulaire afin de développer des traitements plus efficaces, ciblés et non prothétiques. / In order to improve the effectiveness of joint diseases’ treatments, given their complexity and magnitude, recent studies have highlighted the role of lipid assemblies associated with the discontinuous structure of the synovial fluid (SF) in the tribological performance of joint operation. Thus, an ex vivo tribological model (AM Sfarghiu, PhD thesis, 2006) providing a "basic pattern" for joint biolubrification was developed. It consists of the stack of phospholipidic interfaces and aqueous layers. Using this model, the objective of this work was to study the evolution of phospholipidic interfaces of SF within pathological state. Therefore, a nano-bio-tribological methodology combining biochemical, physicochemical, nano-mechanical and tribological analysis was used. The results of these analyses show: the influence of even small rubbing surfaces’ roughness characteristics of early stage illness and that of phospholipidic interfaces’ properties (related to their composition change) on the mechanical strength, changes in friction and in situ degradation of lipidic assemblies of pathological SF. The tribological operation is highlighted by enzymes’ associated with diseases. Thus, joint operation depends on the mechanical strength of phospholipidic interfaces and to obtain very low friction coefficients, velocity accommodation must be done at the level of hydration layers surrounding ions in the aqueous solution. These results would therefore allow better understanding of the evolution of phospholipidic interfaces in joint diseases and of the proper cause/consequence sequence responsible for a joint disease in order to develop more effective, targeted and non prosthetic treatments.

Des substances toxiques générées par les activités portuaires, urbaines et industrielles sont déversées à certains niveaux du milieu marin côtier marocain. Les mollusques peuvent concentrer les polluants et avoir des effets néfastes sur la santé humaine par l’intermédiaire de la chaîne alimentaire. Malgré le renforcement des mesures de sécurité alimentaire, l’implication de la pollution chimique des aliments dans les troubles métaboliques n’est pas à exclure. Pour prédire l’impact des polluants sur l’écosystème aquatique et sur la santé humaine, le développement d’outils de biosurveillance est nécessaire.Nous avons quantifié les métaux lourds (Cd, Cr et Pb) chez les moules (Mytilus galloprovincialis) issues de la côte atlantique marocaine (site industriel Jorf Lasfar (JL) et site touristique d’Oualidia (OL)) en raison de la proximité d’une plateforme d’extraction de phosphate puis caractérisé leurs profils lipidiques (acides gras, cholestérol, oxystérols, phytostérols et phospholipides). Les extraits lipidiques totaux de moules ont été testés in vivo sur des rats pour déterminer leurs effets sur les paramètres biochimiques plasmatiques et in vitro sur une lignée de cellules β pancréatiques murine (MIN-6) en conditions normo et hyperglycémique. Les effets des extraits de moules JL et OL ont été comparés par rapport à ceux de moules d’origine espagnole (ES) destinées à la consommation en France.Les métaux lourds dans les moules JL dépassent les normes internationales. Les concentrations métalliques dans tous les extraits lipidiques sont à l’état de trace. Les moules JL et OL sont moins riches en acides gras insaturés, plus riches en oxystérols et en phospholipides par rapport aux moules ES, suggérant un stress environnemental. Les extraits lipidiques des moules JL et OL administrés à des rats, ont provoqué une perturbation des paramètres plasmatiques, notamment des taux de glucose, créatinine, triglycérides et transaminases avec une augmentation de cholestérol-HDL. In vitro seuls les extraits lipidiques JL et OL induisent la mort des cellules MIN-6 par un processus non apoptotique. Ce processus est associé à une dépolarisation mitochondriale, une déstabilisation lysosomale et une augmentation de la perméabilité de la membrane cytoplasmique, paramètres mesurés par cytométrie en flux dans une démarche cytomique. Ils provoquent aussi une surproduction de H2O2, une augmentation d’activité catalase, une diminution du glutathion réduit, une peroxydation lipidique et une forte stimulation de la sécrétion d’insuline avec un effet plus marqués en présence des extraits lipidiques JL.Globalement, les lipides de moules JL induisent des effets néfastes in vivo et in vitro par rapport à ceux provenant de OL et ES. Une étude épidémiologique à large échelle dans le contexte des maladies métaboliques pourrait être pertinente chez les populations consommatrices de ces moules. / Toxic substances generated by various human activities are spilled on different area of the Moroccan coast. Shellfishes can concentrate pollutants and have some adverse effects on human health through the food chain. Despite the strengthening of food safety rules, the involvement of chemical pollution of food on metabolic disorders is not known. To predict the impact of pollutants on the aquatic ecosystem and human health, the development of appropriate biomonitoring tools is required.We quantified heavy metals (Cd, Cr and Pb) in mussels (Mytilus galloprovincialis) from two sites of Moroccan Atlantic coast (industrial site Jorf Lasfar (JL) and touristic site Oualidia (OL)) due to the proximity of a phosphate extraction platform, and further characterized their lipid profiles (fatty acids, cholesterol, oxysterols, phospholipids and phytosterols). Total lipid extracts of mussels were tested in vivo in rats to determine their effects on biochemical plasmatic parameters and in vitro on a β pancreatic murine cell line (MIN-6) in normo-and hyperglycemic conditions. The effects of JL and OL mussel extracts were compared to mussels from Spain (ES) used for human consumption in France. Heavy metals in JL mussels exceed international standard level. Metal concentrations in all lipid extracts are present in small quantity. JL and OL mussels are less enriched in unsaturated fatty acids, oxysterols and contain higher levels of phospholipids than ES mussels, suggesting an environmental stress. The lipid extracts of JL and OL mussels administered to rats induce a disruption of plasmatic parameters (glucose, creatinine, transaminases and triglycerides) with an increase of HDL-cholesterol. In vitro, only JL and OL lipid extracts induce MIN-6 cell death by a non-apoptotic process. This process is associated with mitochondrial depolarization, lysosomal destabilization and an increase of the cytoplasmic membrane permeability, parameters measured by flow cytométrie in a cytomic context. They also induce an overproduction of H2O2, an increase of catalase activity, a decrease of reduced glutathion, lipid peroxidation and a strong stimulation of insulin secretion with a more marked effect in presence of JL lipid extracts.Overall, JL mussel lipids induce various side effects in vivo and in vitro, which are more pronounced that those observed with OL and ES. A large-scale epidemiological study could be of interest to confirm the potential side effects of these mussels to favor metabolic disorders.