Persistent organic pollutants (POPs) and adipose tissue hypoxia have been shown to independently affect adipocyte functions. The goals of this study were to (1) determine the effect of PCB-77, PCB-153, and DDE on the differentiation of human preadipocytes, and (2) investigate the cross-talk between PCB-77 and hypoxia in differentiated human adipocytes. First, human preadipocytes were exposed to PCB-77, PCB-153, or DDE during the entire 14-day differentiation period. We found no effect of low POP levels on lipid accumulation. Second, differentiated human adipocytes were exposed to a combination of PCB-77 and hypoxia. We demonstrated gene-specific cross-talk between PCB-77 and hypoxia, showing an additive effect of PCB-77 on VEGF, MCP-1, and adiponectin, as well as an inhibition of PCB-77-induced expression of CYP1A1 by hypoxia. This work has expanded our understanding of the role of POPs and hypoxia in differentiated human adipocytes.

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2005. / Härtill 4 uppsatser.

Ο ακριβής ρόλος των μεταγραφικών παραγόντων AP-1, NF-κB και PPARγ στην καρκινογένεση του παχέος εντέρου δεν είναι πλήρως εξακριβωμένος. Είναι γνωστό ότι ο υποκινητής του γονιδίου cox-2 περιλαμβάνει περιοχές πρόσδεσης των μεταγραφικών παραγόντων AP-1 και NF-κB. ΣΚΟΠΟΣ Με σκοπό την διερεύνηση του ρόλου των AP-1, NF-κB και PPARγ στην καρκινογένεση του παχέος εντέρου, εξετάστηκε η έκφραση των c-FOS, pc-JUN (φωσφορυλιωμένος-ενεργός c-JUN), pIκB-α (φωσφορυλιωμένος IκB-α – δείκτης ενεργοποίησης του μονοπατιού του ΝF-κB), CBP (κοινός μεταγραφικός συνενεργοποιητής των AP-1, NF-κB και PPARγ), EGFR, p53, PPARγ και COX-2, τόσο στον φυσιολογικό εντερικό βλεννογόνο όσο και σε αδενοκαρκινώματα παχέος εντέρου. ΥΛΙΚΟ-ΜΕΘΟΔΟΣ Εφαρμόστηκε ανοσοϊστοχημική μεθοδολογία σε τομές μονιμοποιημένες σε ουδέτερη φορμόλη και εγκλεισμένες σε παραφίνη από 60 ασθενείς με καρκίνο παχέος εντέρου. Για κάθε ασθενή δημιουργήθηκε ένα “μοριακό προφίλ”, τόσο για τα φυσιολογικά επιθηλιακά κύτταρα όσο και για τα καρκινικά. Οι στατιστικές σχέσεις μεταξύ των μεταγραφικών παραγόντων και των γονιδίων-στόχων μελετήθηκαν με την χρήση του Spearman’s rho συντελεστή συσχετίσεως, και έγινε περαιτέρω ανάλυση και επιβεβαίωση των αποτελεσμάτων με την βοήθεια μη-παραμετρικής ανάλυσης Kruskall-Wallis. ΑΠΟΤΕΛΕΣΜΑΤΑ Ο pΙκΒ-α βρέθηκε υπερεκφρασμένος στο 81.7% των αδενοκαρκινωμάτων σε σχέση με το γειτονικό φυσιολογικό επιθήλιο και η έκφρασή του συσχετιζόταν θετικά με την COX-2 (Spearman’s rho = 0.513, P < 0.001). H έκφραση του PPARγ βρέθηκε ελαττωμένη στο 77.3% των αδενοκαρκινωμάτων και παρουσίαζε αρνητική συσχέτιση με την έκφραση της COX-2 (Spearman’s rho = -0.412, P < 0.001) και του pIκB-α (Spearman’s rho = -0.444, P < 0.001). Ο CBP βρέθηκε υπερεκφρασμένος στο 50% των αδενοκαρκινωμάτων σε σχέση με το γειτονικό φυσιολογικό επιθήλιο και η έκφρασή του συσχετιζόταν θετικά με την COX-2 (Spearman’s rho = 0.461, P < 0.001). Οι EGFR και pc-JUN βρέθηκαν υπερεκφρασμένοι στο 41.7% και στο 43.3% των αδενοκαρκινωμάτων, αντίστοιχα, σε σχέση με το γειτονικό φυσιολογικό επιθήλιο και η έκφρασή τους συσχετιζόταν θετικά με την COX-2 (Spearman’s rho = 0.444, P < 0.001 και Spearman’s rho = 0.431, P < 0.001, αντίστοιχα). Επιπλέον, η έκφραση του p53 εμφάνιζε θετική συσχέτιση με την έκφραση της COX-2 (Spearman’s rho = 0.435, P = 0.004). ΣΥΜΠΕΡΑΣΜΑΤΑ Η ανάλυση των παραπάνω αποτελεσμάτων οδηγεί στο συμπέρασμα ότι η επαγωγή του μονοπατιού EGFR–MAPK–AP-1 και του μονοπατιού του ΝF-κB, καθώς και η ελάττωση της έκφρασης του PPARγ, είναι σημαντικά γεγονότα κατά την διάρκεια της καρκινογένεσης του παχέος εντέρου. Οι σχέσεις μεταξύ των μεταγραφικών παραγόντων AP-1, NF-κB, PPARγ και της COX-2 οδήγησαν στην δημιουργία ενός μοριακού μοντέλου, το οποίο εξηγεί την υπερέκφραση της COX-2 στα αδενοκαρκινώματα του παχέος εντέρου. Η κλινική σημασία αυτού του μοντέλου για την χημειοπροφύλαξη και την θεραπεία του καρκίνου παχέος εντέρου συζητείται διεξοδικά. / The exact role of AP-1, NF-κB and PPARγ in the development of colon cancer remains to be elucidated. Cox-2 promoter contains transcriptional regulatory elements for various transcription factors including AP-1 and NF-κB. OBJECTIVE The present study evaluated the expression of c-FOS, pc-JUN (phosphorylated-active c-JUN), pIκB-α (phosphorylated IκB-α, a signaling intermediate of the NF-κB pathway), CBP (a known AP-1, NF-κB and PPARγ transcriptional coactivator), EGFR, p53, PPARγ and COX-2 in normal colonic epithelial cells and colon adenocarcinoma cells. METHOD Immunohistochemical methodology was performed on formalin-fixed, paraffin-embedded sections from 60 patients with colon adenocarcinomas. A “molecular profile” was created for each patient both for normal colonic epithelial cells and colon adenocarcinoma cells. Relationships between transcription factors and downstream molecular targets were evaluated by Spearman’s rho correlation coefficient and all results were further validated by nonparametric Kruskall-Wallis test. RESULTS pIκB-α was overexpressed in 81.7% of adenocarcinomas as compared to adjacent normal colon and correlated positively with COX-2 (Spearman’s rho = 0.513, P < 0.001). PPARγ was down-regulated in 77.3% of the adenocarcinomas and correlated inversely with COX-2 (Spearman’s rho = -0.412, P = 0.001) and pIκB-α (Spearman’s rho = -0.444, P < 0.001). CBP was induced in 50% of the cases and its expression correlated positively with COX-2 (Spearman’s rho = 0.461, P < 0.001). Induction of EGFR and pc-JUN was detected in 41.7% and 43.3% of the cases, respectively, and their expression correlated positively with COX-2 (Spearman’s rho = 0.444, P < 0.001 and Spearman’s rho = 0.431, P < 0.001, respectively). Moreover, p53 expression correlated positively with COX-2 (Spearman’s rho = 0.435, P = 0.004). CONCLUSIONS The results of this study indicate that activation of the EGFR–MAPK–AP-1 and NF-κB pathways as well as down-regulation of PPARγ expression are important events in colon carcinogenesis. The correlations between AP-1, NF-κB, PPARγ and COX-2 lead to the proposal of a dynamic molecular model that explains COX-2 induction in colon adenocarcinomas. The clinical implications of this model in chemoprevention and treatment of colorectal cancer are thoroughly discussed.

Persistent organic pollutants (POPs) and adipose tissue hypoxia have been shown to independently affect adipocyte functions. The goals of this study were to (1) determine the effect of PCB-77, PCB-153, and DDE on the differentiation of human preadipocytes, and (2) investigate the cross-talk between PCB-77 and hypoxia in differentiated human adipocytes. First, human preadipocytes were exposed to PCB-77, PCB-153, or DDE during the entire 14-day differentiation period. We found no effect of low POP levels on lipid accumulation. Second, differentiated human adipocytes were exposed to a combination of PCB-77 and hypoxia. We demonstrated gene-specific cross-talk between PCB-77 and hypoxia, showing an additive effect of PCB-77 on VEGF, MCP-1, and adiponectin, as well as an inhibition of PCB-77-induced expression of CYP1A1 by hypoxia. This work has expanded our understanding of the role of POPs and hypoxia in differentiated human adipocytes.

Les opiacés demeurent des molécules incontournables dans le traitement des douleurs moyennes à sévères. Si leur efficacité dans le traitement de la douleur aiguë est incontestable, leur utilisation chronique est responsable de nombreux effets indésirables comprenant une hypersensibilité à la douleur et une tolérance à leurs effets analgésiques. Une partie de ces effets secondaires résulteraient de l’activation de systèmes anti-opioïdes endogènes, comme les neuropeptides RF-amide, dont des études précédentes suggèrent une complémentarité de fonctionnement dans la modulation de la douleur. Le premier axe de travail de cette thèse fut de développer les outils moléculaires afin d’étudier la possibilité d’interactions fonctionnelles et d’hétérodimérisation de ces récepteurs, en particulier GPR103 et NPFF1R. Nous avons ainsi pu générer et caractériser des lignées cellulaires exprimant les différents récepteurs à peptide RF-amide avec un fluorophore fusionné à leur extrémité amino-terminale. En parallèle, nous avons pu développer au cours d’une collaboration fructueuse avec deux équipes de chimistes un ligand à dualité d’action, agoniste opioïdergique et antagoniste des récepteurs NPFF1R et NPFF2R. Chez la souris, nous avons montré que l’administration sous-cutanée de ce composé produit une analgésie longue durée, qui n’est pas atténuée par le développement de tolérance analgésique ou d’hyperalgésie après une semaine d’administration quotidienne. Le syndrome de sevrage, précipité par la naltrexone est plus faible après l’administration chronique de ce composé qu’avec l’agoniste opioïdergique de référence. De plus, grâce à ses caractéristiques d’agoniste biaisé sur le récepteur MOR, cette molécule induit une plus faible dépression respiratoire chez la souris. / Opioid analgesics continue to be the cornerstones for treating moderate to severe pain. However, upon chronic administration, their efficiency is limited because of prominent side effects, such as tolerance and dependence. One hypothesis for the occurrence of these side effects is that the chronic stimulation of the opioid system may trigger its endogenous counterparts, anti-opioid systems, producing hyperalgesia and analgesic tolerance. Previous data from our lab and others suggest that RF-amide peptide receptors can modulate pain signalling through cross-interactions. We developed cell lines expressing fluorescent RF-amide receptors for the study of functional crosstalk and heterodimerization between RF-amide peptide receptors, i.e. GPR103 and NPFF1R. Through a productive collaboration with two teams of chemists, we identified and characterized multitarget peptidomimetic compounds that combined G protein-biased agonism and NPFFR antagonism. In accordance with in vitro results, we observed that acute subcutaneous administration of this compound produced long-lasting antinociceptive effects with less respiratory depression in mice. No hypersensitivity nor analgesic tolerance developed after chronic administration. Altogether, this molecule showed potent antinociceptive effect with limited side effects upon acute and chronic administration.

Soft robots have the potential to transform the way robots interact with their environment. This is due to their low inertia and inherent ability to more safely interact with the world without damaging themselves or the people around them. However, existing sensing for soft robots has at least partially limited their ability to control interactions with their environment. Tactile sensors could enable soft robots to sense interaction, but most tactile sensors are made from rigid substrates and are not well suited to applications for soft robots that can deform. In addition, the benefit of being able to cheaply manufacture soft robots may be lost if the tactile sensors that cover them are expensive and their resolution does not scale well for manufacturability. Soft robots not only need to know their interaction forces due to contact with their environment, they also need to know where they are in Cartesian space. Because soft robots lack a rigid structure, traditional methods of joint estimation found in rigid robots cannot be employed on soft robotic platforms. This requires a different approach to soft robot pose estimation. This thesis will discuss both tactile force sensing and pose estimation methods for soft-robots. A method to make affordable, high-resolution, tactile sensor arrays (manufactured in rows and columns) that can be used for sensorizing soft robots and other soft bodies isReserved developed. However, the construction results in a sensor array that exhibits significant amounts of cross-talk when two taxels in the same row are compressed. Using the same fabric-based tactile sensor array construction design, two different methods for cross-talk compensation are presented. The first uses a mathematical model to calculate a change in resistance of each taxel directly. The second method introduces additional simple circuit components that enable us to isolate each taxel electrically and relate voltage to force directly. This thesis also discusses various approaches in soft robot pose estimation along with a method for characterizing sensors using machine learning. Particular emphasis is placed on the effectiveness of parameter-based learning versus parameter-free learning, in order to determine which method of machine learning is more appropriate and accurate for soft robot pose estimation. Various machine learning architectures, such as recursive neural networks and convolutional neural networks, are also tested to demonstrate the most effective architecture to use for characterizing soft-robot sensors.

Que ce soit dans le domaine médical ou en contrôle non destructif, les systèmes d’imagerie ultrasonore sont devenus de plus en plus utilisés de nos jours. Leurs applications ne cessent de s’élargir et des performances toujours plus accrues sont vivement recherchées, afin d’améliorer la qualité des diagnostics réalisés. Nous sommes donc passés de l’utilisation de systèmes à base de transducteurs ultrasonores mono-élément à des systèmes utilisant des réseaux de transducteurs à une dimension (1D) et à deux dimensions (2D) composés d’éléments de plus en plus nombreux et petits. Néanmoins, un phénomène indésirable est fortement présent dans ces réseaux de transducteurs ultrasonores : il s’agit du couplage inter-éléments tendant à limiter leurs performances acoustiques et à modifier leur diagramme de rayonnement. Tout au long de ce travail de recherche, nous avons donc cherché à comprendre ce phénomène parasite et à apporter des solutions pour le réduire voire le supprimer. En se basant sur des modélisations éléments finis 2D et 3D et grâce à la fabrication de prototypes, nous avons d’une part, mis en évidence les différents types de couplages présents dans un réseau de transducteurs (acoustique, mécanique) et d’autre part, deux méthodes de correction basées l’une comme l’autre sur l’application de tensions convenables aux différents éléments du réseau ont été testées. La première méthode utilise les déplacements normaux moyens à la surface de chaque élément du réseau pour évaluer ces tensions, tandis que la deuxième fait appel aux courants motionnels parcourant chaque élément pour les déterminer. Les résultats numériques et expérimentaux concernant les déplacements et les diagrammes de rayonnement sont en bon accord. En outre, les deux méthodes s’avèrent particulièrement efficaces pour réduire le couplage inter-éléments. / Whether in medicine or in non-destructive testing, ultrasonic imaging systems have become increasingly used nowadays. Their applications continue to expand and good performances are needed to improve the quality of the diagnosis. Moreover, significant progress has been made since these systems were originally based on single element ultrasonic transducers and are now made of mono-dimensional (1D) and bi-dimensional (2D) elements arrays ever more numerous and smaller. However, an undesirable phenomenon is strongly present in the ultrasonic transducer arrays: it is the cross-talk, which limits their acoustic performances and modifies their radiation pattern. Throughout this research, we have attempted on one hand to understand this parasitic phenomenon and on the other hand to provide solutions in order to reduce it or even remove it. To highlight the cross-talk types (acoustic and mechanical) and to test the proposed correction methods, we developed two-dimensional (2D) and three-dimensional (3D) finite element modeling and fabricated some prototypes. Both correction methods rely on the application of suitable voltages to the array elements. The first method uses the average of the normal displacements at the surface of each element to evaluate the voltages, while the second one utilizes the motional currents through each element to determine them. The numerical and experimental results concerning the displacements and the radiation patterns are in good agreement. In addition to this, both methods have been efficiently performed to reduce the cross-talk.

The last few decades have been a very exciting period in thedevelopment of micro-electronics and brought us to the brink ofimplementing entire systems on a single chip, on a hithertounimagined scale. However an unforeseen challenge has croppedup in the form of managing wires, which have become the mainbottleneck in performance, masking the blinding speed of activedevices. A major problem is that increasingly complicatedeffects need to be modelled, but the computational complexityof any proposed model needs to be low enough to allow manyiterations in a design cycle. This thesis addresses the issue of closed form modelling ofthe response of coupled interconnect systems. Following astrict mathematical approach, second order models for thetransfer functions of coupled RC trees based on the first andsecond moments of the impulse response are developed. The2-pole-1-zero transfer function that is the best possible fromthe available information is obtained for the signal path fromeach driver to the output in multiple aggressor systems. Thisallows the complete response to be estimated accurately bysumming up the individual waveforms. The model represents theminimum complexity for a 2-pole-1-zero estimate, for this classof circuits. Also proposed are new techniques for the optimisation ofwires in on-chip buses. Rather than minimising the delay overeach individual wire, the configuration that maximises thetotal bandwidth over a number of parallel wires isinvestigated. It is shown from simulations that there is aunique optimal solution which does not necessarily translate tothe maximum possible number of wires, and in fact deviatesconsiderably from it when the resources available for repeatersare limited. Analytic guidelines dependent only on processparameters are derived for optimal sizing of wires andrepeaters. Finally regular tiled architectures with a commoncommunication backplane are being proposed as being the mostefficient way to implement systems-on-chip in the deepsubmicron regime. This thesis also considers the feasibility ofimplementing a regular packet-switched network-on-chip in atypical future deep submicron technology. All major physicalissues and challenges are discussed for two differentarchitectures and important limitations are identified.

<p>The last few decades have been a very exciting period in thedevelopment of micro-electronics and brought us to the brink ofimplementing entire systems on a single chip, on a hithertounimagined scale. However an unforeseen challenge has croppedup in the form of managing wires, which have become the mainbottleneck in performance, masking the blinding speed of activedevices. A major problem is that increasingly complicatedeffects need to be modelled, but the computational complexityof any proposed model needs to be low enough to allow manyiterations in a design cycle.</p><p>This thesis addresses the issue of closed form modelling ofthe response of coupled interconnect systems. Following astrict mathematical approach, second order models for thetransfer functions of coupled RC trees based on the first andsecond moments of the impulse response are developed. The2-pole-1-zero transfer function that is the best possible fromthe available information is obtained for the signal path fromeach driver to the output in multiple aggressor systems. Thisallows the complete response to be estimated accurately bysumming up the individual waveforms. The model represents theminimum complexity for a 2-pole-1-zero estimate, for this classof circuits.</p><p>Also proposed are new techniques for the optimisation ofwires in on-chip buses. Rather than minimising the delay overeach individual wire, the configuration that maximises thetotal bandwidth over a number of parallel wires isinvestigated. It is shown from simulations that there is aunique optimal solution which does not necessarily translate tothe maximum possible number of wires, and in fact deviatesconsiderably from it when the resources available for repeatersare limited. Analytic guidelines dependent only on processparameters are derived for optimal sizing of wires andrepeaters.</p><p>Finally regular tiled architectures with a commoncommunication backplane are being proposed as being the mostefficient way to implement systems-on-chip in the deepsubmicron regime. This thesis also considers the feasibility ofimplementing a regular packet-switched network-on-chip in atypical future deep submicron technology. All major physicalissues and challenges are discussed for two differentarchitectures and important limitations are identified.</p>

This study investigated the role of opioid, adenosine, bradykinin, and calcitonin-gene related peptide (CGRP) receptors, and potential ‘cross-talk’ among suspected G-protein coupled receptors in a humoral model of remote ischemic preconditioning (rIPC) cardioprotection. Compared to Control dialysate (from non-preconditioned donor rabbit blood), rIPC dialysate (from remotely preconditioned blood) reduced cell death in rabbit cardiomyocytes following simulated ischemia and reperfusion. Non-selective, δ-, or κ-opioid receptor blockade and non-selective adenosine receptor blockade abolished rIPC dialysate protection; whereas, bradykinin B2 and CGRP receptor blockade had no effect. Non-selective adenosine receptor blockade fully and partially abolished protection by κ- and δ-opioid receptors, respectively. Multiple reaction monitoring mass spectrometry detected low levels of adenosine, and other preconditioning substances, in the dialysate. An increase in extracellular adenosine was not detected during opioid-induced preconditioning to explain this cross-talk. These results suggest that δ-opioid, κ-opioid, adenosine receptors, and opioid-adenosine cross-talk are involved in rIPC of freshly isolated cardiomyocytes.