Ultra-high-resolution 3D imaging of atherosclerosis in mice with synchrotron differential phase contrast: a proof of concept study.

The goal of this study was to investigate the performance of 3D synchrotron differential phase contrast (DPC) imaging for the visualization of both macroscopic and microscopic aspects of atherosclerosis in the mouse vasculature ex vivo. The hearts and aortas of 2 atherosclerotic and 2 wild-type control mice were scanned with DPC imaging with an isotropic resolution of 15 μm. The coronary artery vessel walls were segmented in the DPC datasets to assess their thickness, and histological staining was performed at the level of atherosclerotic plaques. The DPC imaging allowed for the visualization of complex structures such as the coronary arteries and their branches, the thin fibrous cap of atherosclerotic plaques as well as the chordae tendineae. The coronary vessel wall thickness ranged from 37.4 ± 5.6 μm in proximal coronary arteries to 13.6 ± 3.3 μm in distal branches. No consistent differences in coronary vessel wall thickness were detected between the wild-type and atherosclerotic hearts in this proof-of-concept study, although the standard deviation in the atherosclerotic mice was higher in most segments, consistent with the observation of occasional focal vessel wall thickening. Overall, DPC imaging of the cardiovascular system of the mice allowed for a simultaneous detailed 3D morphological assessment of both large structures and microscopic details.

Soft tissue artifact distribution on lower limbs during treadmill gait: Influence of skin markers' location on cluster design.

Segment poses and joint kinematics estimated from skin markers are highly affected by soft tissue artifact (STA) and its rigid motion component (STARM). While four marker-clusters could decrease the STA non-rigid motion during gait activity, other data, such as marker location or STARM patterns, would be crucial to compensate for STA in clinical gait analysis. The present study proposed 1) to devise a comprehensive average map illustrating the spatial distribution of STA for the lower limb during treadmill gait and 2) to analyze STARM from four marker-clusters assigned to areas extracted from spatial distribution. All experiments were realized using a stereophotogrammetric system to track the skin markers and a bi-plane fluoroscopic system to track the knee prosthesis. Computation of the spatial distribution of STA was realized on 19 subjects using 80 markers apposed on the lower limb. Three different areas were extracted from the distribution map of the thigh. The marker displacement reached a maximum of 24.9mm and 15.3mm in the proximal areas of thigh and shank, respectively. STARM was larger on thigh than the shank with RMS error in cluster orientations between 1.2° and 8.1°. The translation RMS errors were also large (3.0mm to 16.2mm). No marker-cluster correctly compensated for STARM. However, the coefficient of multiple correlations exhibited excellent scores between skin and bone kinematics, as well as for STARM between subjects. These correlations highlight dependencies between STARM and the kinematic components. This study provides new insights for modeling STARM for gait activity.

A comprehensive examination of adolescent gambling

The purposes of this study were: a) to examine the prevalence and consequences associated with adolescent gambling, b) to examine the factors which influence adolescent gambling,. c) to detennine what factors discriminate among four groups of gamblers (no-risk/non-gamblers, low-risk gamblers, at-risk gamblers, and high-risk/problematic gamblers), and d) to examine the relation of gambling to nine other risk behaviours (i.e., alcohol use, smoking, marijuana use, hard drug use, sexual activity, minor delinquency, major delinquency, direct aggression, and indirect aggression). Adolescents (N = 3,767) from 25 secondary schools completed a twohour survey that assessed involvement in risk be~aviours as well as potential predictors from a wide range of contexts (school, neighbourhood, family, peer, and intrapersonal). The majority of adolescents reported gambling, although the frequency of gambling participation was low. The strongest predictors/discriminators of gambling involvement were gender, unstructured activities, structured activities, and risk attitudes/perceptions. In addition, the examination of the co-occurrence of gambling with other risk behaviours revealed that for high-risk/problem gamblers, the top three most frequent co-occurring high-risk behaviours were direct aggression, minor delinquency and alcohol. This study was the first to examine the continuum of gambling involvement (i.e., non-gambling to high risk/problematic gambling) using a comprehensive set ofpotential predictors with a large sample of secondary school students. The findings of this study support past research and theories (e.g., Theory of Triadic Influence) which suggest the importance ofproximal variables in predicting risk behaviors. The next step, however, will be to examine the direct and indirect 1 effects of the ultimate (e.g., temperament), distal (e.g., parental relationship), and proximal variables (e.g., risk attitudes/perceptions) on gambling involvement in a longitudinal study.

Patterns of faunal change at an upper Cambrian trilobite extinction event, Nolichucky Formation, Tennessee and Virginia

During the Upper Cambrian there were three mass extinctions, each of which eliminated at least half of the trilobite families living in North American shelf seas. The Nolichucky Formation preserves the record of one of these extinction events at the base of the Steptoean Stage. Sixty-six trilobite collections were made from five sections In Tennessee and Virginia. The lower Steptoean faunas are assigned to one low diversity, Aphelaspis-dominated biofacies, which can be recognized in several other parts of North America. In Tennessee, the underlying upper Marjuman strata contain two higher diversity biofacies, the Coosella-Glaphyraspis Biofacies and the Tricrepicephalus-Norwoodiid Biofacies. At least four different biofacies are present in other parts of North America: the Crepicephalus -Lonchocephalus Biofacies, the Kingstonia Biofacies, the Cedaria Biofacies, and the Uncaspis Biofacies. A new, species-based zonation for the Nolichucky Formation imcludes five zones, three of which are new. These zones are the Crepicephalus Zone, the Coosella perplexa Zone, the Aphelaspis buttsi Zone, the A. walcotti Zone and the A. tarda Zone. The Nolichucky Formation was deposited within a shallow shelf basin and consists largely of subtidal shales with stormgenerated carbonate interbeds. A relative deepening is recorded In the Nolichucky Formation near the extinction, and is indicated In some sections by the appearance of shale-rich, distal storm deposits above a carbonate-rich, more proximal storm deposit sequence. A comparable deepening-upward sequence occurs near the extinction in the Great Basin of southwestern United States and in central Texas, and this suggests a possible eustatic control. In other parts of North America, the extinction IS recorded In a variety of environmental settings that range from near-shore to slope. In shelf environments, there is a marked decrease in diversity, and a sharp reduction in biofacies differentiation. Although extinctions do take place in slope environments, there IS no net reduction in diversity because of the immigration of several new taxa.

Petrography, geochemistry and structure of the Timmins area, Ontario

Regional structural analysis of the Timmins area indicates four major periods of tectonic deformation. The DI deformation is characterized by a series of isoclinal FI folds which are outlined in the study area by bedding, pillow tops and variolitic flows. The D2 deformation developed the Porcupine Syncline and refolded the Fl folds about a NE. axis. A pervasive S2 foliation developed during low grade (greenschist) regional metamorphism associated with the D2 deformation. The S2 foliation developed south of the Destor-Porcupine Break. The third phase of tectonic D3 deformation is recognized by the development of a S3 sub-horizontal crenulation cleavage which developed on the plane of the S2 foliation. No meso scopic folds are associated with this deformation. The 8 3 crenulation cleavage is observed south of the Destor-Porcupine Break. The D4 tectonic deformation is recorded as a subvertical S4 crenulation cleavage which developed on the plane of the S2 foliation and also offsets the S3 crenulation cleavage. Macroscopic F4 folds have refolded the F2 axial plane. No metamorphic recrystallization is associated with this deformation. The S4 crenulation cleavage is observed south of the Destor-Porcupine Break. Petrographic evidence indicates that the Timmins area has been subjected to pervasive regional low grade (greenschist) metamorphism which has recrystallized the original mineralogy. South of the study are~ the Donut Lake ultramafic lavas have been subjected to contact medium grade (amphibolite facies) metamorphism associated with the intrusion of the Peterlong Lake Complex. The Archean volcanic rocks of the Timmins area have been subdivided into komatiitic, tholeiitic and calcalkaline suites based on Zr, Ti0 2 and Ni. The three elements were used because of their r e lative immobility during subsequent metamorphic events. Geochemical observations in the Timmins area indicates that the composition of the Goose Lake and Donut Lake Formations are a series of peridotitic, pyroxenitic and basaltic komatiites. The Lower Schumacher Formation is a sequence of basaltic komatiites while the upper part of the Lower Schumacher Formation is an intercalated sequence of basaltic komatiites and low Ti0 2 tholeiites. The variolitic flows are felsic tholeiites in composition and geochemical evidenc e sugg ests that they developed as a n immiscible splitting of a tholeiitic magma. The Upper Schumacher Formation is a sequence of tholeiitic rocks dis p laying a mild iron enrichment. The Krist and Boomerang Formations are the felsic calc-alkaline rocks of the study area which are characteristically pyroclastic. The Redstone Fo rmation is dominantly a calc-alkali ne sequence of volcani c rocks whose minor mafic end me mbers exposed in 1t.he study hav e basaltic komatiitic compositions. Geochemical evidence sugges ts that the Keewatin-type se dimentary rocks have a composition similar to a quartz diorite or a granodiorite. Fi e l d obs ervations and petrographic evidence suggests that they were derived fr om a distal source and now repr esent i n part a turbidite sequence. The Timiskaming-type sedimentary rocks approach the c omp osi t ion of the felsic calc-alkaline rocks of the study area . The basal conglomerate in the study are a sugge s ts that th e uni t was derived fr om a proximal source. Petrographic and ge ochemical evidence suggests that the peridotitic and pyroxenitic komatiites originated as a 35-55% partial melt within the mantle, in excess of 100 Km. depth. The melt ros e as a diapir with the subsequent effusion of the ultramafic lavas, The basaltic komatiites and tholeiitic rocks originated in the mantle from lesser degrees of partial melting and fractionated in low pressure chambers. Geochemical evidence suggests a "genetic link" between the basaltic komatiites and tholeiites, The calc-alkaline rocks developed as a result of the increa.se In PO in the magma chamber. The felsic calcalkaline rocks are a late stage effusion possibly the last major volcanic eruptions in the area.

A sequential analysis of therapist scaffolding and child concept formation in narrative therapy

Narrative therapy is a postmodern therapy that takes the position that people create self-narratives to make sense of their experiences. To date, narrative therapy has compiled virtually no quantitative and very little qualitative research, leaving gaps in almost all areas of process and outcome. White (2006a), one of the therapy's founders, has recently utilized Vygotsky's (1934/1987) theories of the zone of proximal development (ZPD) and concept formation to describe the process of change in narrative therapy with children. In collaboration with the child client, the narrative therapist formalizes therapeutic concepts and submits them to increasing levels of generalization to create a ZPD. This study sought to determine whether the child's development proceeds through the stages of concept formation over the course of a session, and whether therapists' utterances scaffold this movement. A sequential analysis was used due to its unique ability to measure dynamic processes in social interactions. Stages of concept formation and scaffolding were coded over time. A hierarchical log-linear analysis was performed on the sequential data to develop a model of therapist scaffolding and child concept development. This was intended to determine what patterns occur and whether the stated intent of narrative therapy matches its actual process. In accordance with narrative therapy theory, the log-linear analysis produced a final model with interactions between therapist and child utterances, and between both therapist and child utterances and time. Specifically, the child and youth participants in therapy tended to respond to therapist scaffolding at the corresponding level of concept formation. Both children and youth and therapists also tended to move away from earlier and toward later stages of White's scaffolding conversations map as the therapy session advanced. These findings provide support for White's contention that narrative therapists promote child development by scaffolding child concept formation in therapy.

Physical volcanology, sedimentology, stratigraphy and petrochemistry of the Berry Creek metavolcanics: an Archean calc-alkaline complex, Lake of the Woods, Ontario

The steeply dipping, isoclinally folded early Precambrian (Archean) Berry Creek Metavolcanic Complex comprises primary to resedimented pyroclastic, epiclastic and autoclastic deposits. Tephra erupted from central volcanic edifices was dumped by mass flow mechanisms into peripheral volcanosedimentary depressions. Sedimentation has been essentially contemporaneous with eruption and transport of tephra. The monolithic to heterolithic tuffaceous horizons are interpreted as subaerial to subaqueous pumice and ash flows, secondary debris flows, lahars, slump deposits and turbidites. Monolithic debris flows, derived from crumble breccia and dcme talus, formed during downslope collapse and subsequent gravity flowage. Heterolithic tuff, lahars and lava flow morphologies suggest at least temporary emergence of the edifice. Local collapse may have accompanied pyroclastic volcanism. The tephra, produced by hydromagmatic to magmatic eruptions, were rapidly transported, by primary and secondary mechanisms, to a shallow littoral to deep water subaqueous fan developed upon the subjacent mafic metavolcanic platform. Deposition resulted from traction, traction carpet, and suspension sedimentation from laminar to turbulent flows. Facies mapping revealed proximal (channel to overbank) to distal facies epiclastics (greywackes, argillite) intercalated with proximal vent to medial fan facies crystal rich ash flows, debris flows, bedded tuff and shallow water to deep water lava flows. Framework and matrix support debris flows exhibit a variety of subaqueous sedimentary structures, e.g., coarse tail grading, double grading, inverse to normal grading, graded stratified pebbly horizons, erosional channels. Pelitic to psammitic AE turbidites also contain primary stru~tures, e.g., flames, load casts, dewatering pipes. Despite low to intermediate pressure greenschist to amphibolite grade metamorphism and variably penetrative deformation, relicts of pumice fragments and shards were recognized as recrystallized quartzofeldspathic pseudomorphs. The mafic to felsic metavolcanics and metasediments contain blasts of hornblende, actinolite, garnet, pistacitic epidote, staurolite, albitic plagioclase, and rarely andalusite and cordierite. The mafic metavolcanics (Adams River Bay, Black River, Kenu Lake, Lobstick Bay, Snake Bay) display _holeiitic trends with komatiitic affinities. Chemical variations are consistent with high level fractionation of olivine, plagioclase, amphibole, and later magnetite from a parental komatiite. The intermediate to felsic (64-74% Si02) metavolcanics generally exhibit calc-alkaline trends. The compositional discontinuity, defined by major and trace element diversity, can be explained by a mechanism involving two different magma sources. Application of fractionation series models are inconsistent with the observed data. The tholeiitic basalts and basaltic andesites are probably derived by low pressure fractionation of a depleted (high degree of partial melting) mantle source. The depleted (low Y, Zr) calc-alkaline metavolcanics may be produced by partial melting of a geochemically evolved source, e.g., tonalitetrondhjemite, garnet amphibolite or hydrous basalt.

Relation of temperament and authoritative parenting on subtypes of adolescent behaviour

Over the years, researchers have investigated direct, conditional, and meditational pathways of adolescent aggression in relation to both temperament and parenting behaviours. However, no study to date has considered these relations with respect to a measure of aggression differentiated by form (e.g., overt, relational) and function (e.g., proactive, reactive). The present study examined the differential association of adolescent temperament and authoritative parenting on four subtypes of aggression. Participants included mothers, fathers, and one adolescent (between the ages of 10-19) from 663 families, recruited through random digit dialing. Parents reported on their child's temperament and occurrence of aggressive behaviours in addition to the perception of their own authoritative parenting. Adolescents reported on their own temperament and aggressive behaviours as well as on both their mother and father's authoritative parenting. Multiple regression analyses confirmed predictions that some aspects of temperament and authoritative parenting provide motivation towards the engagement of different aggressive behaviours. For example, higher negative affect was related to reactive types of aggression, whereas a strong desire for novel or risky behaviours related to proactive aggression. However, differences in effortful control altered the trajectory for both relationships. Higher levels of self-regulation reduced the impact of negative affect on reactive-overt aggression. Greater self-regulation also reduced the impact of surgency on proactive-overt aggression when age was a factor. Structural equation modeling was then used to assess the process through which adolescents become more or less susceptible to impulsive behaviours. Although the issue ofbi-directionality cannot be ruled out, temperament characteristics were the proximal correlate for aggression subtypes as opposed to authoritative parenting dimensions. Effortful control was found to partially mediate the relation between parental acceptancelinvolvement and reactive-relational and reactive-overt aggression, suggesting that higher levels of warmth and support as perceived by the child related to increased levels of self-regulation and emotional control, which in tum lead to less reactive-relational and less reactive-overt types of aggression in adolescents. On the other hand, negative affect partially mediated the relation between parental psychological autonomy granting and these two subtypes of aggression, supporting predictions that higher levels of autonomy granting (perceived independence) related to lower levels of frustration, which in tum lead to less reactive-relational and reactive-overt aggression in adolescents. Both findings provide less evidence for the evocative person-environment correlation and more support for temperament being an open system shaped by experience and authoritative parenting dimensions. As one of the first known studies examining the differential association of authoritative parenting and temperament on aggression subtypes, this study demonstrates the role parents can play in shaping and altering their children's temperament and the effects it can have on aggressive behaviour.

Sarcolemmal lipid analysis from mechanically skinned rat muscle fibres.

Membrane lipid composition, which includes phospholipid (PL) headgroup, and fatty acid (FA) saturation, has been shown to affect cellular function. The sarcolemma (SL) membrane is integral to skeletal muscle function and health. Previous studies assessing SL lipid composition are limited as they have 1) restricted analysis to a PL level and neglected FA composition and 2) relied on aggressive membrane isolation procedures resulting in t-tubule and sarcoplasmic reticulum contamination and unknown levels of nuclear and mitochondrial contamination. Thus, to overcome these limitations, this study assessed a method of individually skinned skeletal muscle fibres as an alternative to analyze complete sarcolemmal membrane lipid composition. The major findings of this study were 1) complete SL lipid composition can be obtained 2) the SL had higher sphingomyelin content than previous studies and 3) the SL membrane had minimal nuclear and mitochondrial contamination and was void of contamination from sarcoplasmic reticulum and t-tubules.

Spectral Reflectance Characteristics of Kimberlite Indicator Minerals and Other Selected Mineral Grains

High chromium content in kimberlite indicator minerals such as pyrope garnet and diopside is often correlated with the presence of diamonds. In this study, kimberlite indicator minerals were examined using visible light reflectance spectroscopy to determine if chromium content can be correlated with spectral absorption features. The depth of absorption features in the visible spectral region were correlated with the molecular percentage of chromium and other first series transition metal elements obtained by electron microprobe data. In the visible part of the spectrum, chromium is evident by 3 absorption features in the pyrope reflectance spectrum; one isolated and narrow feature at the wavelength 689 nm was used to correlate with the chromium mol %. The isolation of this feature in the pyrope spectra is advantageous since it is not directly affected by other proximal absorption bands that could be caused by other transition metals. Analysis of the feature indicates that as grain volume increases the depth of the absorption feature will also increase. Clustering grain volumes into fractions yields better correlation between absorption depth and mol % chromium. Other types of garnet (almandine, grossular, spessartine) and kimberlite indicator minerals (olivine, diopside, chromite, ilmenite) were analyzed to determine if other absorption features could be used to predict the proportion of specific transition metal elements. Diopside in particular illustrates the same isolated chromium absorption feature as pyrope and may indicate mol percent but needs further study with larger sample sets.

Cellular Mechanisms of Resveratrol's Interaction with Mitochondrial Reactive Oxygen Species Metabolism

Resveratrol, a polyphenol found naturally in red wines, has attracted great interest in both the scientific community and the general public for its reported ability to protect against many of the diseases facing Western society today. While the purported health effects of resveratrol are well characterized, details of the cellular mechanisms that give rise to these observations are unclear. Here, the mitochondrial antioxidant enzyme Mn superoxide dismutase (MnSOD) was identified as a proximal target of resveratrol in vitro and in vivo. MnSOD protein and activity levels increase significantly in cultured cells treated with resveratrol, and in the brain tissue of mice given resveratrol in a high fat diet. Preventing the increase in MnSOD levels eliminates two of resveratrol’s more interesting effects in the context of human health: inhibition of proliferative cell growth and cytoprotection. Thus, the induction of MnSOD is a critical step in the molecular mechanism of resveratrol. Mitochondrial morphology is a malleable property that is capable of impeding cell cycle progression and conferring resistance against stress induced cell death. Using confocal microscopy and a novel ‘cell free’ fusion assay it was determined that concurrent with changes in MnSOD protein levels, resveratrol treatment leads to a more fused mitochondrial reticulum. This observation may be important to resveratrol’s ability to slow proliferative cell growth and confer cytoprotection. Resveratrol's biological activities, including the ability to increase MnSOD levels, are strikingly similar to what is observed with estrogen treatment. Resveratrol fails to increase MnSOD levels, slow proliferative cell growth and confer cytoprotection in the presence of an estrogen receptor antagonist. Resveratrol's effects can be replicated with the specific estrogen receptor beta agonist diarylpropionitrile, and are absent in myoblasts lacking estrogen receptor beta. Four compounds that are structurally similar to resveratrol and seven phytoestrogens predicted to bind to estrogen receptor beta were screened for their effects on MnSOD, proliferative growth rates and stress resistance in cultured mammalian cells. Several of these compounds were able to mimic the effects of resveratrol on MnSOD levels, proliferative cell growth and stress resistance in vitro. Thus, I hypothesize that resveratrol interacts with estrogen receptor beta to induce the upregulation of MnSOD, which in turn affects cell cycle progression and stress resistance. These results have important implications for the understanding of RES’s biological activities and potential applications to human health.

Association between disruption of CD4 receptor dimerization and increased human immunodeficiency virus type 1 entry

Affiliation: Département de microbiologie et immunologie, Faculté de médecine, Université de Montréal & Institut de Recherches Cliniques de Montréal

Régulation de la proprotéine convertase subtilisine / kexine type 9 (PCSK9) dans les cellules intestinales Caco-2/15

La proprotéine convertase subtilisine/kexine type 9 (PCSK9) favorise la dégradation post-transcriptionnelle du récepteur des lipoprotéines de faible densité (LDLr) dans les hépatocytes et augmente le LDL-cholestérol dans le plasma. Cependant, il n’est pas clair si la PCSK9 joue un rôle dans l’intestin. Dans cette étude, nous caractérisons les variations de la PCSK9 et du LDLr dans les cellules Caco-2/15 différentiées en fonction d’une variété d’effecteurs potentiels. Le cholestérol (100 µM) lié à l’albumine ou présenté en micelles a réduit de façon significative l’expression génique (30%, p<0,05) et l’expression protéique (50%, p<0,05) de la PCSK9. Étonnamment, une diminution similaire dans le LDLr protéique a été enregistrée (45%, p<0,05). Les cellules traitées avec le 25-hydroxycholestérol (50 µM) présentent également des réductions significatives dans l’ARNm (37%, p<0,01) et la protéine (75%, p<0,001) de la PCSK9. Une baisse des expressions génique (30%, p<0,05) et protéique (57%, p<0,01) a également été constatée dans le LDLr. Des diminutions ont aussi été observées pour la HMG CoA réductase et la protéine liant l’élément de réponse aux stérols SREBP-2. Il a été démontré que le SREBP-2 peut activer transcriptionnellement la PCSK9 par le biais de la liaison de SREBP-2 à son élément de réponse aux stérols situé dans la région proximale du promoteur de la PCSK9. Inversement, la déplétion du contenu cellulaire en cholestérol par l’hydroxypropyl-β-cyclodextrine a augmenté l’expression génique de la PCSK9 (20%, p<0,05) et son contenu protéique (540%, p<0,001), en parallèle avec les niveaux protéiques de SREBP-2. L’ajout des acides biliaires taurocholate et déoxycholate dans le milieu apical des cellules intestinales Caco-2/15 a provoqué une baisse d’expression génique (30%, p<0,01) et une hausse d’expression protéique (43%, p<0,01) de la PCSK9 respectivement, probablement via la modulation du FXR (farnesoid X receptor). Ces données combinées semblent donc indiquer que la PCSK9 fonctionne comme un senseur de stérols dans le petit intestin.

Assistance de l’enseignement de la conception architecturale par la modélisation de savoir-faire des référents

Dans une époque de changements des moyens de représentation et communication en architecture, cette recherche porte sur l’enseignement de la conception architecturale et plus spécifiquement sur l’apport que l’informatique pourrait avoir dans ce processus. En nous basant sur une méthodologie qualitative, exploratoire et participative, nous y procédons par enchainement de questions, celle de départ étant la suivante: Comment l’enseignement de la conception architecturale pourrait tirer avantage des moyens numériques? Notre objectif est de proposer des méthodes et des outils d’apprentissage aux étudiants en architecture pour enrichir leurs démarches de conception grâce à l’ordinateur. Après une revue de la littérature dans le domaine, et un approfondissement de l’étude sur le rôle des référents architecturaux et sur la conception intégrée, nous avons procédé à une observation exploratoire du travail des étudiants en atelier d’architecture. Ces premières étapes de la recherche ont permis de dégager des discordances entre les positions théoriques et la pratique en l’atelier, pour concrétiser ultérieurement la question de recherche. Dans le but de discerner des méthodes efficaces et innovatrices pour répondre aux discordances identifiées, nous avons engagé une étude de la littérature sur les théories cognitives par rapport aux connaissances, l’apprentissage et la conception. Certaines stratégies ont pu être définies, notamment la nécessité de représentation multimodale des référents architecturaux, l’importance de représenter le processus et non seulement le résultat, ainsi que l’avantage d’inciter les étudiants à travailler dans leur ‘zone proximale’ de développement. Suite à ces recherches, une méthode d’enseignement complémentaire a été définie. Elle propose aux étudiants des explorations de l’objet en conception basées sur la manipulation des savoir-faire architecturaux. Cette méthode a été opérationnalisée d’un point de vue pédagogique ainsi que didactique et mise à l’épreuve auprès des étudiants en atelier. Un prototype de librairie de référents architecturaux interactifs (LibReArchI) a été créé dans ce but. Elle a été conçue en tant qu’environnement de conception et espace de partage de savoir-faire entre étudiants et enseignants. Les principaux résultats de cette recherche démontrent le rôle positif de la méthode proposée pour le transfert des savoir-faire architecturaux lors de l’apprentissage en atelier. Son potentiel d’assister la conception intégrée et de stimuler l’émergence d’idées a été constaté. Au niveau théorique, un modèle d’un cycle du processus de design avec le numérique a été esquissé. En conclusion, des avenues de développements futurs de cette recherche sont proposées.

Les transporteurs sélectifs de cholestérol SR-BI, SR-BII, CD36 et ABCA1 contribuent au maintien de l’homéostasie du cholestérol intratesticulaire

Le testicule assure la production des spermatozoïdes et la sécrétion de la testostérone. Chaque fonction est assumée par un compartiment cellulaire distinct: l’épithélium séminifère et le tissu interstitiel. Le cholestérol, présent dans les deux compartiments, est un composé indispensable aux membranes cellulaires et un précurseur essentiel de la testostérone. Dans le compartiment interstitiel, environ 40 % du cholestérol utilisé pour la production hormonale est importé du sang à partir des lipoprotéines HDL et/ou LDL. Dans l’épithélium séminifère, la cellule de Sertoli assure le contrôle et le maintien de la spermatogenèse. Elle a la capacité de synthétiser du cholestérol à partir de l’acétate in vitro, néanmoins, il n’y a pas d’évidence qu’elle le fait in vivo. De plus il existe, au niveau des tubules séminifères, une barrière hémato-testiculaire qui empêche le libre passage de plusieurs composés sanguins, y compris le cholestérol. Nous avons testé l’hypothèse qu’il existe des moyens d’importation du cholestérol sanguin, mais aussi l’exportation du cholestérol intra-tissulaire, qui contourneraient cette barrière et qui contribueraient au maintien du taux intratubulaire du cholestérol compatible avec le bon déroulement de la spermatogenèse. Nous avons comparé les taux de variation de l’expression de l’ARNm et de la protéine des transporteurs sélectifs de cholestérol SR-BI, SR-BII, CD36 et ABCA1 aux taux de variation du cholestérol libre et estérifié au cours de la spermatogenèse chez les souris normales durant le développement postnatal. Afin de mieux apprécier le niveau d’implication de chacun de ces récepteurs, nous avons examiné comment la suppression du gène d’une enzyme comme la lypase hormono-sensible (HSL) ou de celui d’un transporteur de cholestérol comme SR-BI, CD36 ou NPC1 était compensée et comment cette suppression affectait le taux de cholestérol libre et estérifié dans chacun des deux compartiments cellulaires du testicule. Nous avons dans un premier temps mis au point une nouvelle technique d’isolation des testicules en fraction enrichie en tissu interstitiel (ITf) et en tubules séminifères (STf) qui a l’avantage de mieux préserver l’intégrité des formes phosphorylées et glycosylées des protéines comparée aux techniques préexistantes. Les résultats de nos analyses ont montré que l’expression de SR-BI et CD36 étaient maximales dans les ITf au moment où les souris ont complété leur maturité sexuelle et où le niveau de synthèse de la testostérone était maximal. Dans les tubules séminifères, l’expression maximale de SR-BI et le taux le plus élevé de cholestérol estérifié étaient mesurés de façon concomitante à 35 jours après la naissance, au moment où la première vague de l’activité spermatogénétique était complétée. L’expression de l’ABCA1 était maximale au moment où le taux de cholestérol était élevé et minimale au moment où le taux de cholestérol était le plus bas, alors que le niveau d’expression de CD36 était maximal chez l’adulte au moment où le taux de spermiation était le plus élevé. L’expression de SR-BII variait peu dans les deux compartiments cellulaires durant le développement. La suppression génétique de la HSL et de NPC1, qui cause une infertilité chez les souris mâles, était accompagnée d’une accumulation de cholestérol libre et estérifié dans les tubules séminifères. Par contre, la suppression génétique de SR-BI et CD36, qui ne causent pas d’infertilité chez les souris mâles était sans impact significatif sur le taux de cholestérol intratubulaire. Nous avons montré que l’invalidation génétique d’un transporteur sélectif ou d’une enzyme du métabolisme du cholestérol était accompagnée d’un ensemble de mécanismes de compensation visant à maintenir le taux de cholestérol libre aux niveaux semblables à ceux mesurés dans les fractions tissulaires de souris normales. Ensemble, nos résultats ont montré que l’expression des transporteurs sélectifs de cholestérol SR-BI, SR-BII, CD36 et ABCA1 variait en fonction de la spermatogenèse et du taux intratesticulaire du cholestérol suggérant leur contribution au maintien de l’homéostasie du cholestérol intratesticulaire.