Indiana University-Purdue University Indianapolis (IUPUI) / The passive properties of the venous wall are important in the development of venous pathology. Increase in venous pressure due to retrograde flow (reflux) and obstruction of venous flow by intrinsic and extrinsic means are the two possible mechanisms for venous hypertension. Reflux is the prevailing theory in the etiology of venous insufficiency. The objective of this thesis is to quantify the passive biomechanical response and structural remodeling of veins subjected to chronic venous reflux and hypertension. To investigate the effects of venous reflux on venous mechanics, the tricuspid valve was injured chronically in canines by disrupting the chordae tendineae. The conventional inflation-extension protocol in conjunction with intravascular ultrasound (IVUS) was utilized to investigate the passive biomechanical response of both control common iliac veins (from 9 dogs) and common iliac veins subjected to chronic venous reflux and hypertension (from 9 dogs). The change in thickness and constituent composition as a result of chronic venous reflux and hypertension was quantified using multiphoton microscopy (MPM) and histological evaluation. Biomechanical results indicate that the veins stiffened and became less compliant when exposed to eight weeks of chronic venous reflux and hypertension. The mechanical stiffening was found to be a result of a significant increase in wall thickness (p < 0.05) and a significant increase in the collagen to elastin ratio (p < 0.05). After eight weeks of chronic reflux, the circumferential Cauchy stress significantly reduced (p < 0.05) due to wall thickening, but was not restored to control levels. This provided a useful model for development and further analysis of chronic venous insufficiency and assessment of possible intervention strategies.

Einführung - Periphere Blutmonozyten spielen eine Rolle in der Pathogenese der Arteriosklerose. Bei spontan hypertensiven Ratten wurden im Vergleich zu normotonen Wistar-Kyoto-Ratten signifikant erhöhte Zahlen aktivierter Monozyten beobachtet [Liu 1996]. Wir untersuchten Monozyten von Patienten mit essentieller Hypertonie und von gesunden Probanden hinsichtlich möglicher Unterschiede in der Aktivierung anhand ihrer Adhäsion an von uns kultivierte (HAEC) und isolierte (HSVEC, HUVEC, HUAEC) humane Endothelzellen. Wir bestimmten die Adhäsionsmoleküle ICAM-1, VCAM-1 sowie E-Selektin und analysierten die Superoxidfreisetzung von humanem Endothel und Monozyten. Methoden - Humane periphere Blutmonozyten wurden mittels Dichtegradienten-zentrifugation und Plastikadhärenz isoliert und mit LPS, Angiotensin II (Ang II), und Ang II nach Vorinkubation mit dem AT1-Antagonisten Eprosartan stimuliert. Die Monozyten wurden auf einschichtigem Endothel ausgesät und die Adhäsion als Prozentsatz der initial gesäten Zellen erfasst. Die durch PMA induzierte Superoxidfreisetzung des Endothels oder der Monozyten wurde mittels Chemilumineszenz bestimmt. Ergebnisse - Monozyten von Patienten mit essentieller Hypertonie hefteten sich im Vergleich zu gesunden Probanden spontan und nach Stimulation mit Ang II signifikant verstärkt an HAEC und HUVEC-Monolayer an. Die Spiegel von ICAM-1 und VCAM-1 waren bei Patienten mit arterieller Hypertonie im Vergleich zu den gesunden Kontrollpersonen signifikant erhöht. Die Chemilumineszenzaktivität postkonfluenter Endothelzellen erhöhte sich nach Stimulation mit Ang II im Vergleich zur Messung ohne vorherige Stimulierung. Nach Stimulation mit PMA oder mit Ang II wurden bei Hypertonikern signifikant höhere Werte für die Chemilumineszenzaktivität der Monozyten gemessen als bei gesunden Kontrollpersonen. Schluss - Mit diesen Versuchen an humanen Monozyten und Endothelzellen wurde ein weiterer Beweis für die Aktivierung der Monozyten von Patienten mit essentieller Hypertonie erbracht. Meine Ergebnisse unterstützen die Sicht einer Monozytenbeteiligung an der Pathogenese atherosklerotischer Läsionen, die mit arterieller Hypertonie in Zusammenhang stehen. / Introduction - Peripheral blood monocytes are involved in the pathogenesis of atherosclerosis. Significantly elevated numbers of activated monocytes were observed in spontaneously hypertensive rats compared to those in normotensive Wistar-Kyoto rats [Liu 1996]. We isolated and cultivated human endothelial cells and examined monocytes from patients with arterial hypertension and healthy volunteers to identify possible differences in their adhesion behavior to human endothelial cells (HAEC, HSVEC, HUVEC, HUAEC). We determined the levels of ICAM-1, VCAM-1 and E-selectin, and we analyzed superoxide release by human endothelium and human monocytes. Methods - Peripheral blood monocytes were isolated by density gradient centrifugation and plastic adherence. Subsets of the samples were stimulated with LPS, Angiotensin II, Angiotensin II following preincubation with the AT1-antagonist eprosartan or left without a stimulant. After incubation, monocytes were seeded onto confluent monolayers of human aortic endothelial cells and the adhesion was determined as the percentage of the initially seeded cells. Oxygen species release induced by PMA was analyzed for endothelium and monocytes in suspension by chemiluminescence. Results - Peripheral blood monocytes of patients with essential hypertension performed a significantly increased spontaneous adhesion and adhesion following stimulation with Angiotensin II to HAEC- and HUVEC-monolayers. Levels of human soluble adhesion molecules ICAM-1 and VCAM-1 were significantly raised in hypertensive patients. Chemiluminescence activity of post confluent endothelial cells was increased after stimulation with Angiotensin II compared to the measurement before stimulation. Following stimulation with PMA or Angiotensin II, significantly higher chem-iluminescence levels were measured in hypertensive patients compared to healthy volunteers. Conclusion - These data indicate that monocytes of patients with essential hypertension may be preactivated. My results support the view of a monocyte involvement in the pathogenesis of atherosclerotic lesions that are related to arterial hypertension.

Η μικροχειρουργική τεχνική και η μεταφορά ελεύθερων αγγειούμενων κρημνών για την αποκατάσταση εκτεταμένων και σύνθετων ελλειμμάτων ογκολογικής ή τραυματικής αιτιολογίας της κεφαλής και του τραχήλου αποτελεί μέθοδο επιλογής. Με την τεχνική αυτή επιτυγχάνεται στον ίδιο χρόνο με την εκτομή, η λειτουργική και αισθητική αποκατάσταση με μειωμένο ποσοστό επιπλοκών και επίπτωση από την δότρια περιοχή. Στην παρούσα μελέτη καταγράφουμε και αναλύουμε την εμπειρία μας σε ασθενείς που αντιμετωπίστηκαν με μικροχειρουργική τεχνική και ελεύθερους αγγειούμενους κρημνούς. Την περίοδο 2003 έως 2010, σε 48 ασθενείς πραγματοποιήθηκαν 56 ελεύθεροι αγγειούμενοι κρημνοί. Οι 34 ασθενείς από τους 48 υπεβλήθηκαν συγχρόνως σε ογκολογική εκτομή λόγω Ca και σε αποκατάσταση, ενώ σε 14 από τους 48 ασθενείς αφορούσε έλλειμμα τραυματικής αιτιολογίας. Η τοπογραφία του ελλείμματος αφορούσε το τριχωτό της κεφαλής και του μετώπου σε 12 ασθενείς (25%), το μέσο τριτημόριο του προσώπου και τα παραρίνια σε 9 ασθενείς (18.7%), το κάτω τριτημόριο του προσώπου και τραχήλου σε 27 ασθενείς (56.25%). Σε 7 ασθενείς χρησιμοποιήθηκαν διπλοί ελεύθεροι αγγειούμενοι κρημνοί και σε 41 ασθενείς μονήρεις κρημνοί. Χρησιμοποιήθηκαν συνολικά 56 ελεύθεροι αγγειούμενοι κρημνοί. Σε 16 ασθενείς χρησιμοποιήθηκε φλεβικό μόσχευμα (33.3%). Οι κρημνοί επιλογής ήταν ο κερκιδικός κρημνός 28.5%, ο κρημνός της περόνης 17.8%, ο προσθιοπλάγιος κρημνός του μηρού 14.2%, ο μυοδερματικός κρημνός του ορθού κοιλιακού(VRAM) 12.5% και εγκάρσιος(ΤΡΑΜ) 5.3%, ο κρημνός του πλατέως ραχιαίου 8.9%, ο κρημνός του ισχνού προσαγωγού 7.1% και ο πρόσθιος οδοντωτός 1.7%. Οι μικροαγγειακές αναστομώσεις έγιναν στα αγγεία του τραχήλου σε ποσοστό 96.2%. Η επιβίωση των κρημνών ήταν σε ποσοστό 92.8% (52/56) και 4 κρημνοί απορρίφθηκαν (7.1%) λόγω θρόμβωσης. Ένας ασθενής απεβίωσε στην μετεγχειρητική περίοδο λόγω σοβαρών συστηματικών επιπλοκών. Ο σωστός σχεδιασμός, η επιλογή του κατάλληλου κρημνού, η εμπειρία στην μικροχειρουργική και η συνεργασία των εμπλεκόμενων ιατρικών ομάδων είναι αυτά που διασφαλίζουν το υψηλό ποσοστό επιτυχίας στην επανορθωτική μικροχειρουργική με πολύ καλό λειτουργικό και αισθητικό αποτέλεσμα. / Microsurgical free tissue transfer considered as the best choice for the reconstruction of head and neck extended and complex tissue defects due to tumor resection or trauma. A total of 48 patients underwent free tissue transfer between 2003-2010. There were 34 patients underwent one stage tumor resection and microsurgical free flap reconstruction and 16 patients for free flap reconstruction due to head and neck trauma. The defect in 12 patients 25% was on the scalp and forehead, the middle third in 9 patients 18.7% lower third in neck in 27 patients 56.25%. We used a combination of double free flaps for reconstruction in 7 cases and in 41 patients a single free flap. Vane grafts were used in 16 cases (33.3%). We used in total 56 free flaps with success rate 92.8% (52/56). Four flaps were lost due to anastomotic thromboses. Work horse flaps in our series include the radial forearm 28.5%, fibula 17.8%, ALT 14.2%, VRAM 12.5%, TRAM 5.3%, latissimus dorse 8.9%, gracilis 7.1% and serratus anterior 1.7%. The neck recipient vessels were used in 96.2%. One patient died in post surgical period after systemic complications. Preoperative surgical and reconstruction plan, flaps selection, high microsurgical experience and team collaboration are essential for the good functional and aesthetic results in microsurgery reconstruction of head and neck tissue defects.

Les gisements à Sn-W de type veine et greisen sont des systèmes magmatiques-hydrothermaux dont l’exploitation fournit une part importante de la production mondiale de tungstène et qui représentent également une source importante d’étain. La formation de ces gisements résulte d’un continuum de processus magmatiques et hydrothermaux et implique un transport efficace et la focalisation des fluides minéralisateurs. Cette étude vise àaméliorer la compréhension des processus hydrodynamiques et géologiques impliqués lors du transport et du dépôt de métaux conduisant à la formation de ces gisements. Nous avons réalisé une étude pluridisciplinaire combinant (i) travail de terrain (étude géologique et structurale), (ii) reconstruction des paléo-circulations hydrothermales via l’analyse texturale des bandes de croissance des tourmalines, (iii) détermination expérimentale des changements de perméabilité induits par la greisenisation et (iv) modélisation numérique des écoulements péri-granitiques prenant en compte l’évolution de la perméabilité dynamique lors des interactions fluide-roche. Cette méthodologie a été appliquée au cas du gisement W-Sn-(Cu) de Panasqueira, qui constitue un site de référence pour étudier les processus magmatiques e thydrothermaux conduisant à la formation de gisements à Sn-W de classe mondiale. Les résultats obtenus démontrent que l’expulsion des fluides magmatiques minéralisés a déclenché la greisenisation des parties apicales (coupoles etapex) de l’intrusion granitique, entraînant la création de porosité (~ 8,5%) qui améliore significativement la perméabilité(de 10-20 à 10-17 m²) au sein du greisen massif composant le toit de l’intrusion. Le développement de ce niveau perméable constitue un drain important favorisant l'expulsion et la focalisation des fluides magmatiques minéralisateurs exsolvés lors de la cristallisation du granite sous-jacent. Cette focalisation des décharges hydrothermales (i) améliore significativement le transport des métaux, et (ii) favorise l'établissement de conditions de pression de fluide élevées qui couplées aux contraintes régionales compressives causent l'ouverture des veines minéralisées au toit de l’intrusion.Cette étude souligne l’importance des rétrocontrôles entre perméabilité dynamique et altération hydrothermale. Ces derniers constituent des mécanismes majeurs permettant d’améliorer significativement la circulation des fluides minéralisateurs et donc la formation de gisements hydrothermaux de grandes tailles / The vein and greisen Sn-W deposits are magmatic-hydrothermal systems that provide an important part of theworld W production and represent an important source of Sn. The formation of these deposits involves continuum ofmagmatic-hydrothermal processes and implies the transfer and the focusing of a large amount of mineralizing fluids. Thisstudy aims to improve understanding of hydrodynamic and geological processes involved during the transport and thedeposition of metals leading to the formation of these deposits. We have performed a complete study combining (i) fieldworks (geological and structural studies), (ii) fluid flow reconstruction via the textural analysis of tourmaline growth bands,(iii) experimental determination of permeability changes during greisenization, and (iv) numerical modeling of peri-graniticfluid flow accounting for magmatic fluid production and dynamic permeability related to fluid-rock interactions. Thismethodology was applied in the case of the world-class W-Sn-(Cu) Panasqueira deposit, which represents a referencesite to study magmatic-hydrothermal processes leading to the formation of large vein and greisen deposit. Our resultsdemonstrate that the releasing and the expulsion of ore-bearing magmatic fluids triggered greisenization of the apicalpart of granite intrusion, which caused generation of porosity (~8.5%) and therefore a significant increase of permeability(from 10-20 to 10-17 m²) in massive greisen composing the granite’s roof. The development of this permeable pathwayconstitutes an important drain promoting the expulsion and the focusing of magmatic fluids produced during thecrystallization of the underlying granite. This enhancement of magmatic fluids expulsion (i) promotes significantly fluidflux and transfer of metals, and (ii) the establishment of high fluid pressure conditions, which coupled with the regionalcompressive crustal regime, triggered the opening of mineralized veins above the granite roof. Finally, this studyemphasizes that reactive hydrothermal fluids are able to generate their own pathways in initially impermeable rocks. Thisprocess represents an important mechanism to enhance fluid flow and promote the formation of large hydrothermaldeposits.

Einleitung: Die Vena jugularis interna (VJI) gilt als das wichtigste Gefäß der zerebralen Drainage. Es gibt jedoch Hinweise darauf, dass das vertebrale Venensystem in Abhängigkeit von der Körperposition, an der zerebralen venösen Drainage beteiligt ist. Im Rahmen dieser Arbeit soll die Bedeutung der VJI und des vertebralen Venensystems für die zerebralvenöse Drainage in unterschiedlichen Körperpositionen untersucht werden. Methode: Bei 23 gesunden Probanden wurde der Blutfluss in den VJI und den Vv. vertebrales (VV) duplexsonographisch bestimmt. Dazu wurde die Person auf einem Kipptisch gelagert. Die Messungen wurden in den Positionen -15° (Kopftieflage), 0° (horizontal), 15°, 30°, 45°, und 90° (Stehen) durchgeführt. Der arterielle zerebrale Blutfluss (CBFA) wurde in den Positionen 0° und 45° bestimmt. Ergebnisse: Der Blutfluss der VJI ging von 810 ? 360 ml/min in Kopftieflage (-15°) auf 70 ? 100 ml/min im Stehen zurück. Gleichzeitig stieg der Blutfluss VV von 20 ? 15 ml/min in Kopftieflage auf 210 ? 120 ml/min im Stehen an. Der CBFA betrug 800 ? 153 ml/min in der 0°-Position und 720 ? 105 ml/min in der 45°-Position. Diskussion: Es konnte eine deutliche Lageabhängigkeit der zerebralvenösen Drainage nachgewiesen werden. Es zeigte sich, dass die zentrale Bedeutung der VJI für die zerebrale venöse Drainage auf die liegende Position beschränkt ist. Im Stehen verläuft die zerebrale venöse Drainage weitgehend über das vertebrale Venensystem. / Background: The internal jugular veins (IJV) are considered to be the main outflow of cerebral venous blood. However, there is evidence that the vertebral venous system also forms part of the cerebral venous outflow, depending on the position of the body. This paper asseses the hemodynamic consequences of postural changes in cerebral venous drainage by color-coded duplex sonography. Methods: Volume-blood-flow-measurements were conducted in 23 healthy volunteers in supine position on a tilt table. Both IJV and VV were studied in -15° (head-down tilt), 0°, 15°, 30°, 45°, and 90° (upright position) tilt. Arterial cerebral blood flow (CBFA) was measured in 0° and 45°-position. Results: Bloodflow in the IJV dropped from 810 ? 360 ml/min in the head-down-position (-15°) to 70 ? 100 ml/min at 90°. Simultaneously blood flow in the VV increased from 20 ? 15 ml/min in -15°-position to 210 ? 120 ml/min in the 90°-position. Discussion: The results show, that the cerebral blood drainage pathways depend heavily on the inclination of the body. The role of the IJV as the main drainage pathway of the cerebral blood appears to be confined to the supine position. In the erect position, the vertebral venous system was found to be the major outflow pathway in humans.

En cas de pertes de substances osseuses de grande étendue, la capacité naturelle de réparation du tissu osseux n’est pas suffisante et nécessite d’être assistée. La greffe d’os autologue constitue actuellement la référence. Cependant, cette thérapeutique présente tout de même des inconvénients qui ont entrainé le développement de substituts osseux. Mais, aucun matériau à ce jour ne peut remplacer totalement l’os autologue, en raison notamment de la difficulté à recréer un système vasculaire fonctionnel au niveau du site lésé. Depuis quelques années, les espoirs se tournent vers la création d’un substitut osseux prévascularisé afin de pallier la principale limite des alternatives actuelles : l’établissement d’un réseau vasculaire au sein de ce biomatériau. Notre projet vise à évaluer l’effet stimulateur d’un composé naturel, les principes actifs de la nacre solubles dans l’éthanol (appelé Ethanol Soluble Matrix, ESM), à la fois sur les capacités angiogéniques de cellules de la lignée endothéliale et sur la différenciation ostéogénique de cellules souches mésenchymateuses (CSM) avec comme objectif le développement d’un substitut osseux prévascularisé. Dans un premier temps, nous avons montré que l’ESM stimulait les capacités angiogéniques des cellules de la lignée endothéliale : cellules endothéliales matures (HUVECs, cellules endothéliales issues de la veine ombilicale humaine) et cellules progénitrices endothéliales (CPEs) issues de sang de cordon. L'ESM, utilisé à la concentration de 200µg/mL, a permis de dépasser les résultats obtenus (expression génique et test fonctionnel) avec le milieu de culture de référence des CPEs : l’EGM-2 (Endothelial Growth Medium). Nous avons ensuite mis en évidence que l’ESM exerçait un effet stimulateur également sur les CSMs en augmentant l’expression de marqueurs spécifiques des chondrocytes et des chondrocytes hypertrophiques, suggérant une orientation de ces cellules vers une ossification endochondrale. En parallèle de ces travaux, nous avons étudié l’effet paracrine des CSMs sur les cellules de la lignée endothéliale, HUVECs et CPEs. Les vésicules extracellulaires de taille nanométrique (nEVs) ont montré leur capacité à induire une stimulation in vitro de la formation de réseaux vasculaires et de l’expression de gènes endothéliaux. Ces résultats encourageants soulignent la faisabilité de l’utilisation de l’ESM en tant que stimulus à la fois de l’angiogenèse des CPEs et de l’ostéogenèse des CSMs. Ce stimulus pourrait être associé aux nEVs issues de CSMs et aux CPEs au sein d’une matrice tridimensionnelle pour développer un substitut osseux prévascularisé / In case of critical-sized defects, the bone tissue ability of natural healing is not sufficient and needs to be assisted. The autologous bone graft is currently the gold standard. However, this solution has drawbacks that have led to the development of bone substitutes. Nowadays, no substitute is able to supply autogenous bone, due to the difficulties to mimic the vascular system. In recent years, the hopes are focusing on the creation of a prevascularized bone substitute to overcome the main limitation of current alternatives: the creation of a functional vascular network inside the substitute. Our project aims to evaluate the stimulating effect of a natural compound, the nacre extracts called Ethanol Soluble Matrix (ESM), both on the angiogenic abilities of endothelial cell lineage and on the osteogenic differentiation of mesenchymal stem cells (MSCs) to develop a pre-vascularized bone substitute. First, we showed that ESM stimulates the angiogenic potential of two types of endothelial cells: mature endothelial cells (HUVECs, human umbilical vein endothelial cells) and endothelial progenitor cells (EPCs) from cord blood. The ESM, used at the concentration of 200µg/mL, exceeded results obtained with the reference culture medium of EPCs: the EGM-2 (Endothelial Growth Medium). Then, we demonstrated that ESM also exerted a stimulating effect on MSC by increasing the expression of chondrocyte and hypertrophic chondrocyte specific markers, suggesting an orientation of these cells towards endochondral ossification. In line with this work, we studied the paracrine effect of MSCs on endothelial cell lineage, HUVECs and EPCs. Nanoscale extracellular vesicles (nEVs) have been shown to induce an in vitro stimulation of the vascular network formation and of the endothelial gene expression. These encouraging results highlight the feasibility of using ESM as a stimulus for both angiogenesis of EPCs and osteogenesis of MSCs. This stimulus could be associated with MSC-derived nEVs and EPCs within a three-dimensional matrix to develop a pre-vascularized bone substitute

Les récents développements en RMN et IRM ont eu un impact spectaculaire sur l’imagerie médicale et aident à appréhender de manière fonctionnelle et non invasive de nombreux mécanismes cérébraux. L’imagerie par RMN de perfusion a notamment une importance primordiale en neuroimagerie clinique dans la caractérisation de nombreux désordres cérébrovasculaires (tumeurs cérébrales, AVC). Cependant il n’existe pour le moment aucune technique qui quantifie de manière non-invasive le bon fonctionnement du réseau microvasculaire cérébral. Cette thèse se concentre sur l’utilisation de la séquence FENSI en imagerie préclinique à haut et ultra haut champ magnétique pour caractériser et quantifier la dynamique des flux microvasculaires dans le cerveau du rat. Nous présentons les enjeux de l’IRM de perfusion en neuroimagerie, ainsi que les contraintes associées aux méthodes conventionnelles – méthode de marquage de spin artériel (ASL) et IRM dynamique de contraste de susceptibilité magnétique (DSC-MRI) – dont la technique FENSI peut d’affranchir. Cependant d’autres problèmes sont adressés. Les images acquises avec FENSI et pondérées en flux peuvent être contaminées par des effets de transferts d’aimantation (MT) qui empêchent de quantifier le débit sanguin cérébral. Une première technique de correction de ces effets par post-traitement est proposée. Nous dérivons les premières cartes de flux sanguin cérébral chez le rat à 7 tesla. Une seconde approche est considérée, où les effets MT dans les images acquises avec et sans saturation des spins circulant dans le réseau capillaire (tag/control) se compensent. Cette seconde approche permet une vraie quantification non-invasive du flux sanguin cérébral (CBFlux) in-vivo. Le réseau microvasculaire est caractérisé via FENSI à différents stages du développement tumoral dans un modèle de gliosarcome cérébral (9L) chez le rat. Les mesures de flux mettent en évidence une forte hétérogénéité de développement vasculaire des gliosarcomes peu avancés (diamètre inférieur à 3 mm). A un stade avancé, le flux sanguin est significativement plus faible dans la tumeur (-40 %) que dans le sous cortex, en bon accord avec la littérature sur ce type de gliosarcome. De plus, les cartes paramétriques de flux permettent de distinguer différents compartiments dans la tumeur. Une comparaison avec une méthode de marquage endothélial sur coupes histologiques suggère que le flux sanguin calculé avec FENSI est corrélé avec la concentration locale en micro-vaisseaux. Nous avons également tentés d’évaluer la dynamique temporelle de la réponse vasculaire à un stimulus et d’appliquer FENSI à l’IRM fonctionnelle (IRMf). Nous avons mis en place un protocole d’IRMf chez le rat à 7 tesla et caractérisé la réponse hémodynamique obtenue par contraste BOLD. A 7 tesla la technique FENSI souffre d’un faible SNR temporel et semble plus adaptée pour quantifier des changements métaboliques associées à de longues plages temporelles. L’implémentation de la séquence à ultra-haut champ (17.2 tesla) donne lieu à de sérieux espoirs en IRMf. De plus, nous mettons en évidence à 17.2 tesla des contrastes spécifiques à l’utilisation de différents anesthésiques utilisés en routine à l’hôpital. La méthode que nous avons mise en place peut être sensibilisée à de nombreuses vitesses, augmentant le nombre de ses applications potentielles. Ainsi, un choix judicieux de paramètres permet d’explorer le volume sanguin ou l’orientation du débit ciblé. Les forces et faiblesses de la méthode sont détaillées. L’utilisation de FENSI n’est en général pas justifiée par un gain de signal par rapport à des séquences ASL optimisées, mais bénéficie de l’actuelle montée en champ des imageurs cliniques pour être étudiée. Les applications potentielles varient de l’IRMf et l’imagerie de diffusion au suivi pharmacologique et diagnostic de désordres cérébrovasculaires, dont l’étude via ASL est limitée dû à l’allongement des temps de transit sanguins. / The discoveries, implementations and developments of NMR and MRI have had a major impact in medical imaging. Compared to other imaging modalities (PET, SPECT, CT), current MRI research helps to further and better understand the inner mechanisms of the human body in a less invasive manner. In clinical neuroimaging, perfusion MRI is of spectacular importance to study cerebrovascular diseases and cancer. However, at the moment, there is no perfusion MRI sequence that allows for a complete, non-invasive and precise quantification of microvascular flow dynamics. This work focuses on the use of the recently introduced Flow Enhanced Signal Intensity method (FENSI) to characterize and quantify vasculature at capillary level, at high and ultra high magnetic field (7 and 17.2 tesla). For that purpose, the possible quantification of blood flux with FENSI is explored in vivo. The combination of flux quantification and flow-enhanced signal (compared to Arterial Spin Labeling) can make of FENSI an ideal method to characterize in a complete non-invasive way the brain microvasculature. After removal of magnetization transfer (MT) effects, the blood flow dynamics are studied with FENSI in a very aggressive and propagative rat brain tumor model: the 9L gliosarcoma. The objective is to assess whether FENSI is suitable for a longitudinal non-invasive characterization of microvascular changes associated with tumor growth. The results obtained with FENSI are compared with literature on 9L perfusion and immuno-histochemistry. In the first paper published on FENSI, a first glance was also casted on the potential of the flow enhanced technique when applied to fMRI. The results obtained at the time were contaminated by MT effects. With the implementation of a new MT-free FENSI technique, the possibility to map the brain cerebral functioning based on a quantitative physiological parameter (CBFlux) more directly related to neuronal activity than the usual BOLD signal is within reach. At ultra high field, the influence of different anesthetics on the rat brain microvascular network and BOLD contrast is also considered. After many developments around the FENSI technique, the method is compared to classical ASL and DSC perfusion MRI sequences. The strengths and weaknesses of the FENSI method, its characteristics, ‘precautions for use’, and potential main applications are detailed and discussed.

Submitted by Ana Lúcia Torres (bfmhuap@gmail.com) on 2017-09-28T13:11:50Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) DISSERTAÇÃO KLAUS STEINBRUCK.pdf: 1704112 bytes, checksum: 061dda187540e95c23369cc5a57f8c98 (MD5) / Approved for entry into archive by Ana Lúcia Torres (bfmhuap@gmail.com) on 2017-09-28T13:12:05Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) DISSERTAÇÃO KLAUS STEINBRUCK.pdf: 1704112 bytes, checksum: 061dda187540e95c23369cc5a57f8c98 (MD5) / Made available in DSpace on 2017-09-28T13:12:05Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) DISSERTAÇÃO KLAUS STEINBRUCK.pdf: 1704112 bytes, checksum: 061dda187540e95c23369cc5a57f8c98 (MD5) Previous issue date: 2012 / Hospital Federal de Bonsucesso, Serviço de Cirurgia Hepato-Biliar / O objetivo deste estudo foi avaliar as complicações associadas às reconstruções vasculares nos receptores de Transplante Hepático Intervivos operados no Hospital Federal de Bonsucesso, no período de dezembro de 2001 e fevereiro de 2011. Foram levantados dados referentes aos receptores, seus respectivos doadores e relacionados ao procedimento cirúrgico, visando identificar possíveis fatores de risco ao desenvolvimento das complicações vasculares. Cento e quarenta e quatro transplantes foram realizados em 141 receptores, 76 adultos e 65 crianças. Foram identificadas sete complicações (4,9% do total) da artéria hepática, cinco complicações (3,5%) da veia porta e uma complicação (0,7%) da veia hepática. Devido ao pequeno número de complicações, não foi possível realizar análise estatística de fatores de risco. A sobrevida em um ano dos pacientes com e sem complicação vascular foi de 26% e 82%, respectivamente. A sobrevida em um ano dos enxertos utilizados em pacientes com e sem complicação vascular foi de 15% e 82%, respectivamente. Houve diferença estatística (p < 0,001) na sobrevida de pacientes e enxertos, que foi menor no grupo que apresentou complicações vasculares. Concluiu-se que as técnicas de reconstrução vascular utilizadas no Hospital Federal de Bonsucesso apresentam resultados comparáveis aos grandes centros internacionais. A presença de complicação na reconstrução vascular diminui a sobrevida do receptor e do enxerto / The objective of this study was to evaluate the complications associated with vascular reconstruction in recipients of living donor liver transplantation at Bonsucesso Federal Hospital, between December 2001 and February 2011. Data associated to recipients, their donors and surgical procedure were collected to identify possible risk factors for vascular complications development. One hundred and forty-four transplants were performed in 141 recipients, 76 adults and 65 children. We identified seven hepatic artery complications (4.9% of total), five complications (3.5%) of portal vein and one complication (0.7%) of hepatic vein. Due to the small number of complications, statistical analysis of risk factors could not be performed. The 1-year survival of patients with and without vascular complications was 26% and 82%, respectively. The 1-year survival of grafts in patients with and without vascular complications was 15% and 82%, respectively. There was statistical difference (p <0.001) on survival of patients and grafts, which was lower in the group with vascular complications. It was concluded that techniques used in vascular reconstruction at Bonsucesso Federal Hospital showed results comparable to major international centers. The presence of vascular complications in the reconstruction decreases survival of recipients and grafts

During embryogenesis and post-embryonic patterning, active transport of the phytohormone auxin, reflected in the expression of the Arabidopsis PIN family of auxin efflux mediators, generates local auxin distributions that are crucial for correct organ and tissue specification. Polar auxin transport routes have also long been postulated to regulate vein formation in the leaf. The molecular identification of PIN proteins has made it possible to investigate this hypothesis further by visualizing auxin transport routes in developing leaves. In Arabidopsis leaf primordia, PIN1 is expressed before the earliest known markers of vascular identity, in domains that are gradually restricted to sites of vein formation. PIN1 polarity indicates that auxin is directed towards distinct “convergence points” (CPs) in the marginal epidermis, from which it defines the sites of major vein formation. Within incipient veins, PIN1 polarity indicates drainage of auxin into preexisting veins, such that veins connected at both ends display two divergent polarities. Local auxin application triggers the formation of ectopic CPs and new veins, demonstrating the sufficiency of auxin as a vein-specifying signal. However, not all PIN1-labeled auxin transport routes differentiate as veins: Minor veins are initially unstable, suggesting local competition for auxin transport. Expression of ATHB8, a marker of vascular cell selection, correlates with enhanced PIN1 expression domain (PED) stability and vascular differentiation. Auxin application and auxin transport inhibition reveal that both CP formation in the epidermis and subepidermal PED dynamics are auxin-dependent and self-organizing. Furthermore, normal auxin perception through the ARF-Aux/IAA signaling pathway is required for the restriction of PIN1-mediated auxin transport to narrow subepidermal domains. ARF-Aux/IAA signaling is known to control auxin transport through the regulation of PIN1 dynamics, but the mechanism of this regulation is unclear. It is here shown that two redundantly acting AUXIN RESPONSE FACTOR (ARF) transcription factors, ARF5/MONOPTEROS (MP) and ARF7/NPH4, jointly regulate both PIN1 expression and localization during lateral root patterning in Arabidopsis, in part through the direct transcriptional activation of PIN1 by MP. Taken together, these results indicate that feedback between PIN-mediated auxin transport and ARF-Aux/IAA signaling regulates the patterning of root and shoot organs.

During embryogenesis and post-embryonic patterning, active transport of the phytohormone auxin, reflected in the expression of the Arabidopsis PIN family of auxin efflux mediators, generates local auxin distributions that are crucial for correct organ and tissue specification. Polar auxin transport routes have also long been postulated to regulate vein formation in the leaf. The molecular identification of PIN proteins has made it possible to investigate this hypothesis further by visualizing auxin transport routes in developing leaves. In Arabidopsis leaf primordia, PIN1 is expressed before the earliest known markers of vascular identity, in domains that are gradually restricted to sites of vein formation. PIN1 polarity indicates that auxin is directed towards distinct “convergence points” (CPs) in the marginal epidermis, from which it defines the sites of major vein formation. Within incipient veins, PIN1 polarity indicates drainage of auxin into preexisting veins, such that veins connected at both ends display two divergent polarities. Local auxin application triggers the formation of ectopic CPs and new veins, demonstrating the sufficiency of auxin as a vein-specifying signal. However, not all PIN1-labeled auxin transport routes differentiate as veins: Minor veins are initially unstable, suggesting local competition for auxin transport. Expression of ATHB8, a marker of vascular cell selection, correlates with enhanced PIN1 expression domain (PED) stability and vascular differentiation. Auxin application and auxin transport inhibition reveal that both CP formation in the epidermis and subepidermal PED dynamics are auxin-dependent and self-organizing. Furthermore, normal auxin perception through the ARF-Aux/IAA signaling pathway is required for the restriction of PIN1-mediated auxin transport to narrow subepidermal domains. ARF-Aux/IAA signaling is known to control auxin transport through the regulation of PIN1 dynamics, but the mechanism of this regulation is unclear. It is here shown that two redundantly acting AUXIN RESPONSE FACTOR (ARF) transcription factors, ARF5/MONOPTEROS (MP) and ARF7/NPH4, jointly regulate both PIN1 expression and localization during lateral root patterning in Arabidopsis, in part through the direct transcriptional activation of PIN1 by MP. Taken together, these results indicate that feedback between PIN-mediated auxin transport and ARF-Aux/IAA signaling regulates the patterning of root and shoot organs.