Rozprawy doktorskie na temat „Breath sounds”

Existing breath sound measurement systems and possible new methods have been critically investigated. The frequency response of each part of the measurement system has been studied. Emphasis has been placed on frequency response of acoustic sensors; especially, a method to study a diaphragm type air-coupler in contact use has been proposed. Two new methods of breath sounds measurement have been studied: laser Doppler vibrometer and mobile phones. It has been shown that these two methods can find applications in breath sounds measurement, however there are some restrictions. A reliable automatic wheeze detection algorithm based on auditory modelling has been developed. That is the human’s auditory system is modelled as a bank of band pass filters, in which the bandwidths are frequency dependent. Wheezes are treated as signals additive to normal breath sounds (masker). Thus wheeze is detectable when it is above the masking threshold. This new algorithm has been validated using simulated and real data. It is superior to previous algorithms, being more reliable to detect wheezes and less prone to mistakes. Simulation of cardiorespiratory sounds and wheeze audibility tests have been developed. Simulated breath sounds can be used as a training tool, as well as an evaluation method. These simulations have shown that, under certain circumstance, there are wheezes but they are inaudible. It is postulated that this could also happen in real measurements. It has been shown that simulated sounds with predefined characteristics can be used as an objective method to evaluate automatic algorithms. Finally, the efficiency and necessity of heart sounds reduction procedures has been investigated. Based on wavelet decomposition and selective synthesis, heart sounds can be reduced with a cost of unnatural breath sounds. Heart sound reduction is shown not to be necessary if a time-frequency representation is used, as heart sounds have a fixed pattern in the time-frequency plane.

Diversi studi hanno dimostrato i reciproci rapporti di influenza tra respiro e variabili psicologiche ma scarsa attenzione è stata rivolta all’indagine delle funzioni espressive del suono del respiro. Studio 1 è stato finalizzato alla costruzione di un modello di analisi multi-livello per la descrizione acustica del respiro. Sono state effettuate audio e video registrazioni del respiro di 2 coppie di soggetti in performance individuali e congiunte. Sui 1903 respiri raccolti sono state condotte analisi video e audio che hanno portato alla costruzione di tre categorie di indici: respiratori, acustici e interattivi (relativi alla relazione tra i comportamenti respiratori di due partners in un’azione congiunta). Studio 2 ha investigato cosa poteva essere inferito dall’ascolto del suono respiro relativamente all’identità, allo stato emotivo e all’attività svolta da una persona. Inoltre, ha verificato se l’imitazione facilitasse l’identificazione rispetto al solo ascolto. Sono state raccolte registrazioni ecologiche del respiro di persone coinvolte in 6 attività e 4 emozioni. Un campione di 90 persone ha ascoltato le tracce e risposto ad un questionario relativo alle suddette variabili. Altre 90 persone prima di rispondere imitavano il respiro. Il campione ha efficacemente inferito informazioni relative alle variabili indagate, con risultati migliori nel task di imitazione. Studio 3 mirava a descrivere acusticamente 6 condizioni di respiro emotivo (rabbia, paura, tristezza, disgusto, tenerezza, gioia) e a investigare se “respirare insieme” influenzasse il processo di sintonizzazione. 20 coppie di donne hanno partecipato. Per ogni emozione, entro ogni coppia una partecipante leggeva una storia emotivamente connotata e respirava come se si trovasse in quella situazione. L’altra esprimeva la propria vicinanza respirando come lei. Al termine rispondevano a un questionario. I respiri sono stati audio registrati e analizzati. E’ stato possibile ricondurre ciascuna delle emozioni a un differente pattern respiratorio. Inoltre il task ha influenzato diverse dimensioni della sintonizzazione: la sincronizzazione, il decoding emotivo, l’esperienza emotiva e la percezione di somiglianza interpersonale.
Several psycho-physiological studies have provided evidences about the reciprocal influence between respiration and psychological variables but few attentions have been brought to the investigation of the expressive function of breathing sounds. Study 1 aims to build a multilayer analysis model that enable an acoustic description of breathing sounds. Audio and video recordings of breathing of two pairs of participants engaged in individual and joint performance were conducted. 1903 were video and audio analyzed and 3 sets of indexes were derived: respiratory, acoustic and interactive (aimed to relate partner’s respiratory behaviour during joint performances). Study 2 investigates what could be inferred about a person’s identity, emotional state and activity from the sound of his/her breathing. Moreover, it aims to verify whether imitation of breathing patterns improve the identification of those features. Ecological recordings of breathing sounds of people engaged in 6 activities and 4 emotions. 90 participants listen to the tracks and answer a questionnaire about such variables. Other 90 mimicked the tracks before answering. Participants were able to infer valuable information about the aspects under investigation with better results in the imitation task. Study 3 aims to describe the acoustic features of 6 emotional breathing patterns (anger, fear, sadness, disgust, tenderness and joy) and to see whether breathing together could strengthen the attunement process. 20 pairs of women participated. For each emotion, within the pair one participant read a story emotionally connoted and breathes as if she actually was in that situation. Her partner had to convey her closeness breathing in the same way. Finally they filled in a questionnaire. Different respiratory patterns were related to each emotion. Moreover the task influenced several attunement dimensions: synchronization, emotional decoding, emotional experience and perception of interpersonal similarity.

Diversi studi hanno dimostrato i reciproci rapporti di influenza tra respiro e variabili psicologiche ma scarsa attenzione è stata rivolta all’indagine delle funzioni espressive del suono del respiro. Studio 1 è stato finalizzato alla costruzione di un modello di analisi multi-livello per la descrizione acustica del respiro. Sono state effettuate audio e video registrazioni del respiro di 2 coppie di soggetti in performance individuali e congiunte. Sui 1903 respiri raccolti sono state condotte analisi video e audio che hanno portato alla costruzione di tre categorie di indici: respiratori, acustici e interattivi (relativi alla relazione tra i comportamenti respiratori di due partners in un’azione congiunta). Studio 2 ha investigato cosa poteva essere inferito dall’ascolto del suono respiro relativamente all’identità, allo stato emotivo e all’attività svolta da una persona. Inoltre, ha verificato se l’imitazione facilitasse l’identificazione rispetto al solo ascolto. Sono state raccolte registrazioni ecologiche del respiro di persone coinvolte in 6 attività e 4 emozioni. Un campione di 90 persone ha ascoltato le tracce e risposto ad un questionario relativo alle suddette variabili. Altre 90 persone prima di rispondere imitavano il respiro. Il campione ha efficacemente inferito informazioni relative alle variabili indagate, con risultati migliori nel task di imitazione. Studio 3 mirava a descrivere acusticamente 6 condizioni di respiro emotivo (rabbia, paura, tristezza, disgusto, tenerezza, gioia) e a investigare se “respirare insieme” influenzasse il processo di sintonizzazione. 20 coppie di donne hanno partecipato. Per ogni emozione, entro ogni coppia una partecipante leggeva una storia emotivamente connotata e respirava come se si trovasse in quella situazione. L’altra esprimeva la propria vicinanza respirando come lei. Al termine rispondevano a un questionario. I respiri sono stati audio registrati e analizzati. E’ stato possibile ricondurre ciascuna delle emozioni a un differente pattern respiratorio. Inoltre il task ha influenzato diverse dimensioni della sintonizzazione: la sincronizzazione, il decoding emotivo, l’esperienza emotiva e la percezione di somiglianza interpersonale.
Several psycho-physiological studies have provided evidences about the reciprocal influence between respiration and psychological variables but few attentions have been brought to the investigation of the expressive function of breathing sounds. Study 1 aims to build a multilayer analysis model that enable an acoustic description of breathing sounds. Audio and video recordings of breathing of two pairs of participants engaged in individual and joint performance were conducted. 1903 were video and audio analyzed and 3 sets of indexes were derived: respiratory, acoustic and interactive (aimed to relate partner’s respiratory behaviour during joint performances). Study 2 investigates what could be inferred about a person’s identity, emotional state and activity from the sound of his/her breathing. Moreover, it aims to verify whether imitation of breathing patterns improve the identification of those features. Ecological recordings of breathing sounds of people engaged in 6 activities and 4 emotions. 90 participants listen to the tracks and answer a questionnaire about such variables. Other 90 mimicked the tracks before answering. Participants were able to infer valuable information about the aspects under investigation with better results in the imitation task. Study 3 aims to describe the acoustic features of 6 emotional breathing patterns (anger, fear, sadness, disgust, tenderness and joy) and to see whether breathing together could strengthen the attunement process. 20 pairs of women participated. For each emotion, within the pair one participant read a story emotionally connoted and breathes as if she actually was in that situation. Her partner had to convey her closeness breathing in the same way. Finally they filled in a questionnaire. Different respiratory patterns were related to each emotion. Moreover the task influenced several attunement dimensions: synchronization, emotional decoding, emotional experience and perception of interpersonal similarity.

The Break is a personal investigation into problems and possibilities of representing my specific transgender identity. Trans as a tactic to speak about a state of forever becoming, forever in between, outside of and in opposition to dominant social norms of being. Trans as a model for a different way of viewing and being in the world. Can we form a different kind of horizontal shared power though a collective refusal to play into existing structures from which we have been excluded? What are the potentials for modeling other ways of being, other ways of (dis)engaging, other ways to be in the world? What if we can disengage words from their established meanings? Can we re-see each other without the language that upholds the social conditions that maintain internalized categories? Can we collectively create the conditions to imagine the possibility of building other worlds in this world?

This thesis analyses the current strategies to segment breast lesions in Ultra-Sound (US) data and proposes a fully automatic methodology for generating accurate segmentations of breast lesions in US data with low false positive rates. The proposed approach targets the segmentation as a minimization procedure for a multi-label probabilistic framework that takes advantage of min-cut/max- flow Graph-Cut (GC) minimization for inferring the appropriate label from a set of tissue labels for all the pixels within the target image. The image is divided into contiguous regions so that all the pixels belonging to a particular region would share the same label by the end of the process. From a training image dataset stochastic models are built in order to infer a label for each region of the image. The main advantage of the proposed framework is that it splits the problem of segmenting the tissues present in US the images into subtasks that can be taken care of individually
En aquest treball, es proposa un sistema automàtic per generar delineacions acurades de lesions de mama en imatges d’ultrasò. El sistema proposat planteja el problema de trobar la delineació corresponent a la minimització d’un sistema probabilístic multiclasse mitjançant el tall de mínim cost del graf que representa la imatge. El sistema representa la imatge com un conjunt de regions i infereix una classe per cada una d’aquestes regions a partir d’uns models estadístics obtinguts d’unes imatges d’entrenament. El principal avantatge del sistema és que divideix la tasca en subtasques més fàcils d’adreçar i després soluciona el problema de forma global

La formation de métastases osseuses implique de nombreuses interactions entre les cellules de cancer du sein et le microenvironnement osseux. Les gradients d’hypoxie et l’activation de HIF (hypoxia inducible factor) 1alpha sont essentiels au maintien de l’homéostasie osseuse. Le rôle de la signalisation HIF dans les ostéoblastes lors du processus métastatique n’a pourtant jamais été exploré. Dans cette étude, nous montrons que les cellules ostéoprogénitrices (OPC), se situent dans des niches hypoxique, et que l’activation de la signalisation HIF dans ces cellules augmente la masse osseuse et favorise les métastases osseuses du cancer sein. L’effet de la signalisation HIF dans les OPC n’est pas limité au squelette, en effet celle-ci stimule aussi la croissance des tumeurs mammaires et la dissémination tumorale dans les poumons et d’autres organes distants. Nous avons mis en évidence que la signalisation HIF dans les OPC induit l’augmentation de la concentration plasmatique de la chimiokine C-X-C motif ligand 12 (CXCL12), qui entraine une augmentation systémique de la prolifération et de la dissémination tumorale, via l’activation de son récepteur CXCR4 sur les cellules cancéreuses. Ainsi, nos résultats mettent en évidence le rôle protumorigénique de l’hypoxie dans le lignage ostéoblastique, lors de la formation de métastases osseuse, mais également par une action systémique sur les tumeurs mammaires et les métastases dans les tissus mous. Nous démontrons également que des altérations de l’anabolisme osseux peuvent affecter la progression du cancer du sein, révélant un nouveau rôle du squelette au sein du macroenvironnement tumoral
Bone metastasis involves dynamic interplay between tumor cells and thelocal stromal environment. In bones, local hypoxia and activation of the hypoxiainducible factor (HIF)-1alpha in osteoblasts are essential to maintain skeletalhomeostasis. However, the role of osteoblast-specific HIF signaling in cancermetastasis is unknown. Here we show that osteoprogenitor cells (OPC) are locatedin hypoxic niches in the bone marrow, and that activation of HIF signaling in thesecells increases bone mass and favors breast cancer metastasis to bone locally.Remarkably, HIF signaling in osteoblast lineage cells also promotes breast cancergrowth and dissemination remotely, in the lungs and in other tissues distant frombones. Mechanistically, we found that activation of HIF signaling in OPC increasesblood levels of the chemokine C-X-C motif ligand 12 (CXCL12), which leads to asystemic increase of breast cancer cell proliferation and dissemination, throughdirect activation of the CXCR4 receptor. Hence, our data reveal a previouslyunrecognized role of the hypoxic osteogenic niche in promoting tumorigenesisbeyond the local bone microenvironment. They also indicate that alterations inbone formation can affect breast cancer progression, and support the concept thatthe skeleton is an important regulator of the systemic tumor environment

This study explores how the design of women’s social space of action is presented in Hagar Olsson’s play S.O.S: Save our souls (1928). The focus of the study is on the female main character of the play, Maria. The analysis model consists of close reading with the play's melodramatic elements as a starting point. The theoretical framework for the study is the epic theater tradition where Bertolt Brecht has an important influence. The study shows that there is an existing deficiency condition regarding women´s social space of action. This design interprets as a space where women´s happiness and freedom experiences as difficult to access. There is also a critique towards this design through Maria and her actions. The critique is targeted towards long-standing social constructions. However, it should be pointed out that the play takes plays in a bourgeoise environment. Therefore, the design of the space as well as the critique targets towards this environment. The study also shows that Maria´s actions combined with the play opens up for a possible new space where contemporary bourgeoise women allows to act beyond their expectations. This design creates a possibility for women to follow their own way to happiness.

Le cancer du sein est le cancer est le plus fréquent chez la femme. Les patientes traitées par chirurgie, radiothérapie et/ou chimiothérapie peuvent souffrir de rechute et de métastases à plus ou moins long terme. Il est à présent admis qu’une sous-population de cellules particulières dénommées cellules souches cancéreuses (CSC) jouent un rôle important dans ces événements. Il est donc crucial d’isoler et de caractériser ces cellules pour comprendre leurs propriétés particulières d’autorenouvellement et de résistance aux traitements, ce qui permettra de les cibler pour obtenir des traitements plus efficaces. C’est dans ce contexte que s’inscrit ma thèse. Au laboratoire, j’ai mis en place un modèle de souris bi-transgéniques (bi-Tg) en croisant les souris C3(1)-Tag qui est un modèle de tumorigenèse mammaire (et prostatique) bien établi, avec les souris Tg s-SHIP-GFP qui ont déjà permis l’isolement de CS normales mammaires (et prostatiques). Dans ces souris, le promoteur de s-SHIP contrôle l’expression de la protéine fluorescente GFP ce qui permet de marquer et d’isoler les cellules. Dans les tumeurs mammaires développées par ces souris biTg, j’ai isolé une population rare de cellules s-SHIP/GFP+ possédant des propriétés de CSC, surtout une capacité à former des sphères en culture non adhérente et à générer des tumeurs par transplantation en série très supérieure à celle des autres cellules de la tumeur. Une analyse transcriptomique globale qui compare les gènes dérégulés dans les cellules GFP+ et GFP- a mise en évidence le rôle d’un composant de la voie Notch dans le maintien de la pluripotence.Nous avons également dérivé plusieurs lignées cellulaires dénommées MAM (pour mammary) à partir des tumeurs mammaires. L’une d’entre, MAM326 est une lignée de cellules épithéliales cancéreuses avec environ 10 % de cellules GFP+ et j’ai démontré que les cellules GFP+ sont plus résistantes à différentes drogues anti-cancéreuses ainsi qu’à l’irradiation. Une analyse transcriptomique a été réalisée pour déterminer la signature moléculaire de cette résistance. Cette analyse a mis en évidence une vingtaine de gènes significativement surexprimés dans les cellules GFP+, et dont la nature et/ou la fonction est pertinente dans le contexte d’une résistance aux traitements antitumoraux. L'un de ces gènes est la synucléine-gamma dont le rôle dans la radiorésistance du cancer du sein a été suggéré mais non démontré expérimentalement. Par la surexpression ectopique et l’inhibition par siRNA, nous avons démontré que la synucléine gamma peut induire la radiorésistance dans plusieurs lignées cellulaires de cancer du sein. En conclusion, ces résultats démontrent que l’expression de s-SHIP est un marqueur de CSC mammaires chez la souris et son intérêt dans l’étude du cancer du sein
Breast cancer is the most common cancer in women worldwide. The isolation and characterization of breast cancer stem cells (CSC) are crucial for understanding cancer biology and revealing potential therapeutic targets. One of the major issues in the study of CSC is the lack of reliable markers. A transgenic mouse model (Tg 11.5kb–GFP) was generated using the 11.5kb s-SHIP (stem-SH2-containing 5’-Inositol Phosphatase) promoter that specifically expressed enhanced green fluorescent protein (GFP) in embryonic and various tissue stem cells. In the mammary gland, previous experiments showed that GFP labels puberty cap cells and pregnancy basal alveolar bud cells, and it has been demonstrated that these mammary GFP+ cells are activated tissue stem cells. In order to determine if s-SHIP promoter expression could also mark mammary cancer stem cells, we generated a bi-transgenic mouse model by crossing Tg 11.5kb-GFP mice with Tg C3(1)/Tag mice. Tg C3(1)/Tag mice express SV40 T antigen under the regulatory control of the rat prostatic steroid binding protein C3(1) gene. In female mice, the transgene is expressed primarily in the mammary gland. Mice develop mammary hyperplasia by 3 months of age with subsequent development of mammary adenocarcinoma by 6 months of age.Here we show the presence of a rare population of GFP+ cells, which are also CD24+/CD49f+/CD29+ in mammary tumors of female bi-transgenic mice. As compared to GFP- cells, GFP+ cells exhibit both a higher tumor sphere-forming potential, and a higher tumorigenicity when transplanted into SCID and FVB recipient mice. Moreover, upon subsequent transplantation, the GFP+ cells generated heterogeneous tumors that displayed properties similar to the primary tumor. Transcriptomic analysis of these GFP+ vs GFP- cells revealed several differentially expressed genes including one protein implicated in the Notch pathway. In addition, from the murine mammary tumor, I have derived a cell line containing a s-SHIP/GFP+ subpopulation that shows resistance to chemotherapy and radiation. I have further studied this subpopulation and found that synuclein gamma could confer radiation resistance to breast cancer cells. Altogether, these results demonstrate that s-SHIP promoter expression is a marker of mammary CSC that enables their identification and isolation via a single consistent parameter

Le cancer du sein triple négatif (TNBC) est un sous-type de cancer du sein très agressif et hétérogénène qui ne bénéficie actuellement d'aucun traitement efficace. Le but de mon projet de Thèse était d'explorer les mécanismes qui conduisent au TNBC pour concevoir de nouvelles approches thérapeutiques.Dans notre équipe, nous avons généré un modèle de souris unique (souris MMTV-R26Met) dans lesquelles le récepteur tyrosine kinase MET est faiblement augmenté. Elles développent spontanément et exclusivement des TNBC. Ce modèle récapitule la formation de métastases pulmonaires, la résistance aux agents chimiothérapeutiques classiques et à un ensemble de thérapies moléculaires ciblées combinées.Une analyse protéomique des tumeurs MMTV-R26Met combinée à une approche de type "machine learning" ont montré que le modèle MMTV-R26Met récapitule largement l'hétérogénéité du TNBC observée chez les patients TNBC.Nous avons identifié deux nouvelles combinaisons d'agents très délétères pour les lignées cellulaires dérivées de tumeurs MMTV-R26Met et un panel de cellules TNBC humaines. La première cible le facteur anti-apoptotique BCL-XL et CDK1/2 (régulateurs du cycle cellulaire). La deuxième, validée in vivo, agit sur BCL-XL et WEE1 (régulateur du cycle cellulaire et épigénétique). Mécanistiquement, nous montrons que le ciblage combiné de WEE1 et de BCL-XL induit des dommages à l'ADN, une entrée prématurée en mitose, une catastrophe mitotique et l'apoptose.Nos travaux peuvent être très pertinents pour leur transposition potentielle à la clinique, compte tenu également des études prometteuses de monothérapie avec les inhibiteurs de BCL-XL et WEE1 dans les essais cliniques de phase II
Triple-negative breast cancer (TNBC) is a highly aggressive, heterogeneous breast cancer subtype, and has currently no effective treatment. The aim of my PhD project was to understand the mechanisms triggering TNBC and to develop new therapeutic approaches.In our lab, we have generated a unique mouse model (MMTV-R26Met mice) in which a subtle increase in the expression levels of the wild-type MET receptor tyrosine kinase leads to spontaneous, exclusive TNBC formation. This model recapitulates formation of lung metastasis, resistance to conventional chemotherapeutic agents and to a set of combined targeted molecular therapies.Proteomic profiling of MMTV-R26Met tumors and machine learning approach showed that the MMTV-R26Met model largely recapitulates TNBC heterogeneity observed in TNBC patients. We identified two new drug combinations highly deleterious for the MMTV-R26Met tumor-derived cell lines, and a panel of human TNBC cells. The first drug combination targets the anti-apoptotic factor BCL-XL and CDK1/2 (cell cycle regulators). The second drug combination, validated by in vivo studies, is based on a combinatorial targeting of BCL-XL and of WEE1 (cell cycle and epigenetic regulator). Mechanistically, we show that combined inhibition of WEE1 and BCL-XL leads to DNA damage, premature entry into mitosis, resulting in mitotic catastrophe and apoptosis.Our findings may be highly relevant for their potential translation to the clinic, also in view of promising studies of monotherapy with BCL-XL and WEE1 inhibitors in phase II clinical trials

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior
O cÃncer de mama (CM) à um problema de saÃde pÃblica mundial. Uma em cada oito mulheres terà cÃncer de mama, tornando-se assim a neoplasia mais freqÃente entre as mulheres. O cÃncer de mama està ligado à identidade feminina no tocante a sua potÃncia orgÃstica e a sua sensualidade. Elaborar estratÃgicas cirÃrgicas confiÃveis pouco agressivas e menos multilantes tem sido busca de respostas a muitas pesquisas. Objetivo: O presente estudo visou avaliar a concordÃncia do volume tumoral entre a avaliaÃÃo anatomopatolÃgica e os mÃtodos de diagnÃstico por imagem dedicados a mama: Mamografia (MX), Ultra-som (US) e RessonÃncia magnÃtica (RMM), em mulheres com cÃncer de mama localmente avanÃado, submetidas à quimioterapia neoadjuvante. Pacientes e MÃtodos: Foram analisadas 95 mulheres portadoras de neoplasia mamÃria oriundas dos ambulatÃrios da Maternidade Escola Assis Chateaubriant (MEAC), Centro Regional e Integrado de Oncologia (CRIO) e Instituto do CÃncer do Cearà (ICC), no perÃodo de marÃo de 2003 a marÃo de 2005. Estas pacientes foram divididas em dois grupos. Grupo Tratamento (GT) com n=46 e Grupo Controle (GC) com n=49. Realizou-se anÃlise nÃo paramÃtrica correlacionando as variÃveis estudadas com o padrÃo ouro, ou seja, a anatomia patolÃgica. Os resultados mostraram concordÃncia na anÃlise volumÃtrica dos mÃtodos imaginolÃgicos e o padrÃo-ouro no GT nos seguintes percentuais: RMM: 0,70; US: 0,63 e MX: 0,42. Quando comparado ao grupo controle os valores encontrados foram RMM: 0,72; US: 0,70 e MX: 0,41. ConclusÃo: A RMM deteve os maiores valores de concordÃncia volumÃtrica em percentuais semelhantes nos dois grupos (GT e GC) e a MX, mÃtodo de imagem mais difundido, ofereceu os menores Ãndices de concordÃncia imaginolÃgica quando comparado com a avaliaÃÃo anatomopatolÃgica. A US apresentou valores de concordÃncia volumÃtrica satisfatÃria quando, comparada a RMM, devendo ressaltar que se trata de mÃtodo operador dependente.
The cancer of breast (CB) it is a problem of world public health. One in each eight women will have cancer of breast, becoming like this the most frequent neoplasia among the women. CB it is tied up the feminine identity concerning its potency of orgasm and her sensuality. To elaborate surgical strategic reliable not very aggressive and less mutilate has been search of answers to many researches. Objective: The present study was to evaluate the agreement of the volume of tumor to the evaluation anatomopathological and the diagnosis methods for image dedicated to he/she suckles her: Mammography (MX), Ultra-sound (US) and magnetic Resonance (RMM), in women with cancer of breast locally advanced, submitted to the neoadjuvante chemotherapy. Patient and Methods: 95 women carriers of mammary neoplasia originating from of the national health clinics of the Maternity Escola Assis Chateaubriant (MEAC), Center Regional and Integrated of Oncology (CRIO) and Institute of Cancer of Cearà (ICC) were analyzed at the period of 2003 March to 2005 March. These patient ones were divided in two groups. Treatment Group (TG) with n=46 and Control Group (CG) with n=49. Non parametric analysis correlating the variables was compared with the standard gold, the anatomopathological analysis. Results: The data showed agreement in it analyzes at volumÃtrica of the imaginolÃgicos methods and the pattern-gold in TG in the following percentages: RMM: 0.70; US: 0.63 and MX: 0.42, when compared to the control group, where were found values of RMM: 0.72; US: 0.70 and MX: 0.41. Conclusion: RMM stopped the largest values of agreement volumÃtrica in percentile similar in the two groups (GT and GC) and MX, spread image method, offered the smallest indexes of agreement imaginolÃgica when compared with the evaluation anatomopathological. US presented values of agreement satisfactory volumÃtrica when, compared RMM, should stand out that is method dependent operator.

Les progrès de la recherche sur le cancer du sein dépendent de la disponibilité d’outils appropriés, comme les lignées cellulaires qui peuvent être implantées chez des souris immunocompétentes. La souche de souris C57Bl/6 est la plus étudiée et c’est la seule pour laquelle certaines variantes génétiques sont disponibles. Étant donné qu'aucune lignée cellulaire de carcinome mammaire à récepteurs hormonaux positifs de souche C57Bl/6 n'est disponible, nous avons décidé d'établir des lignées cellulaires de ce type. Nous avons induit des cancers du sein chez des souris C57BL/6 femelles en utilisant un analogue synthétique de la progestérone combiné à un agent endommageant l'ADN. Des lignées cellulaires ont été établies à partir de ces tumeurs et sélectionnées pour leur positivité au niveau du double récepteur (estrogène + progestérone), ainsi que pour leur transplantabilité chez les femelles C57BL/6. Parmi plusieurs lignées, une lignée cellulaire, que nous avons appelée MD5, remplissait ces critères et a permis l'établissement de tumeurs mal différenciées et très prolifératives. Ces tumeurs ont réduit leur croissance (sans toutefois régresser) lors du traitement par des antagonistes des récepteurs d’œstrogènes, ainsi que par une chimiothérapie à base d'anthracylines. Cependant, ce dernier effet n'a pas été influencé par la déplétion des lymphocytes T et, en outre, ces tumeurs n'ont pas répondu au blocage de PD-1, ce qui suggère que les tumeurs MD5 sont immunologiquement froides. En conclusion, les cellules MD5, dérivées des animaux C57BL/6, constituent un modèle de cancer du sein à récepteurs hormonaux positifs de mauvais pronostic
Progress in breast cancer research relies on the availability of suitable cell lines that can be implanted in immunocompetent laboratory mice. The best explored mouse strain, C57Bl/6, is also the only one for which multiple genetic variants are available. Driven by the fact that no hormone receptor-positive C57Bl/6-derived mammary carcinoma cell lines are available, we decided to establish such cell lines. Breast cancers were induced in female C57BL/6 mice using a synthetic progesterone analogue combined with a DNA damaging agent. Cell lines were established from these tumors and selected for dual (estrogen + progesterone) receptor positivity, as well as transplantability into C57BL/6 females. One cell line, which we called MD5,fulfilled these criteria and allowed for the establishment of poorly differentiated, highly proliferative, immune cold tumors. Such tumors reduced their growth (though did not regress) upon treatment with estrogen receptor antagonists, as well as with anthracyline-based chemotherapy. However, the latter effect was not influenced by T cell depletion and MD tumors failed to respond to PD-1 blockade, suggesting that they are immunologically cold. In conclusion, C57BL/6-derived MD5 cells constitute a model of poor prognosis hormone receptor-positive breast cancer

Cette étude porte sur les corrélats phonétiques des registres tonals yin vs. yang du shanghaïen parlé dans la région urbaine de Shanghai. Nos investigations acoustique, articulatoire et perceptive ont montré qu’en dehors du F0, des indices multi-dimensionnels comme le voisement (voisé pour yang et non-voisé pour yin), le pattern de durée (ratio C/V bas pour yang et élevé pour yin), et le type de phonation (soufflé pour yang et modal pour yin) participent tous à la définition du registre tonal. Parmi tous ces indices, nous tâchons de distinguer les traits redondants liés aux effets coarticulatoires de ceux qui sont des survivants de changements diachroniques. En particulier, la voix soufflée qui accompagne les tons yang est un trait anciennement distinctif et aujourd’hui redondant, issu d’une évolution tonale qui est la transphonologization de distinction de voisement vers la distinction de registre tonal, ou « bipartition tonale ». Nous proposons que la perte d’un trait redondant issu d’un changement diachronique peut être très lente si ce trait ne contrarie pas les effets coarticulatoires et/ou si le trait a une fonction perceptive.En nous basant sur les données synchroniques des locuteurs de deux générations (20-30 ans vs. 60-80 ans), nous constatons une tendance vers la disparition de cette phonation soufflée. Nous constatons également une évolution plus avancée chez les femmes que les hommes de leur âge. Dans notre étude, nous essayons d’expliquer ce changement tant par des causes internes que par des causes externes
This study bears on the phonetic correlates of the yin vs. yang tone registers of Shanghai Chinese as spoken in Shanghai urban area. Our acoustic, articulatory, and perceptual investigations showed that beside F0, multidimensional cues, such as voicing (voiced for yang vs. voiceless for yin), duration pattern (low C/V ratio for yang vs. high C/V ratio for yin), and phonation type (breathy for yang vs. modal for yin) enter in the specification of tone register. Among all these cues, we attempt to distinguish the redundant features related to coarticulatory effects from those that are remnants of diachronic changes. In particular, the breathy voice accompanying yang tones, which was a distinctive feature in a previous state of the language and has now become a redundant feature, arose from a tonal evolution, namely the transphonologization of a voicing contrast into a tone register contrast, that is, the “tone split.” We propose that the loss of a redundant arisen from a diachronic change may be very slow if that feature does not oppose to coarticulatory effects and/or if that feature has a perceptual function.Based on the synchronic data from the speakers of two generations (20-30 years vs. 60-80 years), we find a trend toward the loss of this breathy phonation. We also find that this evolution is more advanced in women than men of the same age. In our study, we try to explain this change by internal factors as well as by external factors

La production sonore biologique, étudiée par acoustique passive, doit être évaluée comme méthode complémentaire pour l’étude des effets des perturbations anthropiques sur les milieux côtiers.Alors que les sons émis par les mammifères marins et les poissons sont largement documentés, ceux issus des invertébrés benthiques restent peu décrits, même si ces derniers peuvent assurer des fonctions-clés dans les systèmes marins. L’enjeu principal de cette thèse était d’évaluer la pertinence de l’utilisation de l’acoustique passive comme outil écologique en s’appuyant sur la biophonie des invertébrés benthiques d’habitats côtiers tempérés et plus particulièrement en prenant comme habitat-modèle les bancs de maërl de la rade de Brest. Nous avons mis en évidence l’existence d’espèces sonifères remarquables qui constituent de bons candidats pour leur suivi en milieu naturel. Ces travaux ont de plus montré l’importante contribution de certains invertébrés benthiques dans le paysage sonore sous-marin, encourageant à ne plus imputer systématiquement les sons benthiques uniquement aux « crevettes claqueuses ». Le deuxième volet de cette thèse a montré que les variations de production sonore des invertébrés benthiques, à l’échelle individuelle ou collective, offrent des informations précieuses sur la mise en évidence de stress tels qu’un bloom d’algues toxiques ou l’impact du dragage. Ainsi, le nombre de mouvements sonifères des coquilles Saint-Jacques est doublé en présence de fortes concentrations d’algues toxiques, et le paysage sonore des bancs de maërl fortement pêchés est trois fois plus silencieux et moins complexe que celui du maërl préservé. Les résultats de ce travail de thèse, discutés dans un contexte de développement d’outils pour l’évaluation de l’état de santé des écosystèmes marins, permettent l’émergence de nouvelles hypothèses de travail en écologie marine
Biological sound production, as studied by passive acoustics, should be considered as a complementary method to study the effect of anthropogenic disturbances on coastal systems. While sounds emitted by marine mammals and fish are well documented, those from benthic invertebrates are poorly described, although they can play key roles in marine ecosystems. The main goal of this PhD work was to evaluate the suitability of passive acoustics as an ecological tool based on sound production by benthic invertebrates living in temperate coastal habitats, and particularly in maerl beds of the Bay of Brest.We highlighted the existence of interesting soniferous species that appear to be good candidates to monitor in the field. This work also showed the important contribution of some benthic invertebrates to the underwater soundscape, suggesting that benthic sounds should not be attributed only to snapping shrimps. The second part of this PhD thesis emphasized that changes in benthic invertebrate sound production, at individual or collective scales, provides valuable information on the detection of stress such as a toxic algal blooms or the impact of dredging. Thus, the number of soniferous movements made by the great scallop doubles in the presence of high concentrations of toxic algae, and the soundscape of heavily fished maerl beds is three times quieter and less complex than preserved ones. The results of this PhD thesis, which are discussed in the context of tool development for the assessment of marine ecosystem health, raise new working hypotheses in marine ecology

.La consommation journalière de fructose est en augmentation croissante, jusqu'à plus de 50g par jour aux Etats-Unis et en Europe de l'Ouest. Le fructose est de plus en plus incorporé dans les boissons, les produits laitiers et les conserves, les produits cuisinés ou transformés. Le fructose peut déclencher ou aggraver les symptômes digestifs chez des volontaires sains, mais aussi dans le syndrome de l'intestin irritable. Le rôle de l'hypersensibilité viscérale dans le déclenchement des symptômes, lié à la prise de fructose a été suspecté mais n'a jamais été évalué. La prévalence de la malabsorption du fructose était mal documentée chez les patients souffrant d'un syndrome de l'intestin irritable, principalement en raison de l'hétérogénéité des méthodes diagnostiques. Le premier objectif de ce travail a été de définir, dans une population de patients souffrant d'un syndrome de l'intestin irritable, la prévalence de la malabsorption du fructose. Notre test de malabsorption du fructose a été défini par une dose de charge de 25g. Chez nos patients souffrant d'un syndrome de l'intestin irritable, 22% présentaient une malabsorption du fructose. Les patients jeunes et de sexe masculin étaient plus à risque de malabsorption du fructose. Nous avons également étudié l'association de la malabsorption du fructose avec d'autres anomalies physiopathologiques connues dans le syndrome de l'intestin irritable. Nous n'avons pas retrouvé d'association entre la présence d'une inflammation digestive et la présence ou non d'une malabsorption du fructose. En revanche, une association a été retrouvée entre malabsorption du fructose et hypersensibilité viscérale. L'efficacité du régime appauvri en fructose dans le syndrome de l'intestin irritable est connue. L'existence ou non d'une malabsorption du fructose associée pourrait être un facteur prédictif d'efficacité d'un tel régime. Le deuxième objectif de ce travail a été de déterminer si le test de dépistage de la malabsorption au fructose permettait de prédire l'efficacité du régime appauvri en fructose sur les symptômes digestifs des patients souffrant d'un syndrome de l'intestin irritable. Les résultats de notre étude ont confirmé l'efficacité du régime appauvri en fructose dans le syndrome de l'intestin irritable. En revanche, la présence ou non d'un test respiratoire au fructose positif n'impactait pas l'efficacité du régime. Le dernier objectif de ce travail était de modéliser la malabsorption du fructose sur des modèles murins, pour permettre d'identifier les mécanismes physiopathologiques sous-jacents. La modélisation sur 3 modèles murins de malabsorption du fructose (par régime riche en fructose, par délétion des gènes codant les transporteurs du fructose GLUT5 et GUT2) permettait d'induire une hypersensibilité viscérale associée à une augmentation de la perméabilité intestinale, deux anomalies déjà rapportées dans le syndrome de l'intestin irritable. L'étude des mécanismes physiopathologiques sous-jacents a permis d'écarter l'implication d'une inflammation de bas grade qui n'était pas retrouvée chez nos souris. L'augmentation d'activité élastase dans les selles de souris avec malabsorption du fructose était associée à l'hypersensibilité viscérale. Or il a déjà été démontré que l'activité protéasique pouvait être responsable d'une hypersensibilité viscérale et d'une augmentation de la perméabilité intestinale. Les récepteurs associés à la protéase-2 sont connus pour être associés à l'hypersensibilité viscérale et l'augmentation de la perméabilité intestinale. Les résultats obtenus dans le cadre de ce travail soulignent le rôle de la malabsorption du fructose, qui entraine la survenue d'une hypersensibilité viscérale et d'une augmentation de la perméabilité intestinale, dans le syndrome de l'intestin irritable. Un régime appauvri en fructose n'améliore pas de manière ciblée les symptômes des patients souffrant d'un syndrome de l'intestin irritable avec malabsorption du fructose
Fructose intake has increased by up to 50 g per day in the USA and Western Europe. Fructose is increasingly incorporated in beverages, dairy products and canned, baked or processed foods worldwide. Fructose has been shown to trigger or worsen digestive symptoms not only in healthy volunteers, but also in patients with irritable bowel syndrome. The involvement of visceral hypersensitivity has been suspected but has never been assessed. The prevalence of fructose malabsorption in patients with irritable bowel syndrome in Western Europe remains poorly documented, due to the heterogeneity of available tests. Therefore, the first objective of this present work was to assess the prevalence of fructose malabsorption in patients with irritable bowel syndrome. We assessed fructose malabsorption with a fructose breath test, after a 25 g load. We systematically ruled out small intestinal bacterial overgrowth which could promote false positive. In our irritable bowel syndrome patients, 22% had fructose malabsorption. Young, male patients were more likely to have fructose malabsorption. We also assessed the association between fructose malabsorption and other abnormalities. We did not observe any association between low-grade inflammation (with faecal calprotectin dosage) or fructose malabsorption. In contrast, an association between fructose malabsorption and visceral hypersensitivity was evidenced. Low fructose diet is known to improve symptoms in patients with irritable bowel syndrome. The presence of fructose malabsorption could be predictive of the efficacy of a low fructose diet. The second objective of this work was to determine if an abnormal fructose breath test was a predictor of symptomatic response to low fructose diet in irritable bowel syndrome. Our study has confirmed the efficacy of low fructose diet on irritable bowel syndrome. However, the results of the fructose breath test had no impact on its efficacy. One explanation for this result could be the presence of other abnormalities (including visceral hypersensitivity) that were not addressed only with a diet. The last objective of this work was to model fructose malabsorption in mice, in order to identify the underlying mechanisms. We used three models of fructose malabsorption (high fructose diet, invalidation of GLUT5 and GLUT2 coding gene). In these models, fructose malabsorption induced visceral hypersensitivity and increased intestinal permeability, the two abnormalities being reported in irritable bowel syndrome. In our models, there was no low-grade inflammation. Increased elastase activity in mice faeces was associated with visceral hypersensitivity. Protease-activated receptor-2 is known to be associated with visceral hypersensitivity and increases intestinal permeability. Further works are warranted to determine the involvement of protease-activated receptor-2 in fructose malabsorption-associated visceral hypersensitivity. The results of this work underlined the role of fructose malabsorption in irritable bowel syndrome, in the onset of visceral hypersensitivity and increased intestinal permeability. A low fructose diet is not helpful to improve symptoms of irritable bowel syndrome with fructose malabsorption

Les lymphocytes T Vγ9Vδ2 sont impliqués dans la réponse immunitaire contre de nombreux pathogènes et contre les cellules tumorales. Ils reconnaissent des antigènes solubles, non peptidiques, ayant une faible masse moléculaire, qui sont appelés phosphoantigènes. Dans ce contexte, l’isopentenyl pyrophosphate (IPP) et un métabolite de l’ATP (ApppI) sont des phosphoantigènes qui s’accumulent dans les cellules suite à l’inhibition par les amino-bisphosphonates de l’activité d’une enzyme clé de la voie du mévalonate, la farnesyl pyrophosphate synthase (FPPS). Les bisphosphonates sont utilisés en clinique dans le traitement et la prévention des complications liées aux ostéolyses malignes. Les études pré-cliniques montrent que les bisphosphonates (et en particulier les amino-bisphosphonates) ont également une activité anti-tumorale qui peut être directe et/ou indirecte. Ils peuvent bloquer différentes fonctions des cellules tumorales (adhésion, invasion, prolifération) en inhibant l’activité de la FPPS. Ils agissent aussi sur les cellules endothéliales et inhibent l’angiogenèse tumorale. Les amino-bisphosphonates activent aussi l’activité cytotoxique des lymphocytes T Vγ9Vδ2 vis-à-vis des cellules tumorales. Les mécanismes moléculaires sous-jacents responsables de cette activation sont par contre méconnus. Les travaux réalisés dans cette thèse montrent tout d’abord que les aminobisphosphonates (zolédronate et risédronate) induisent l’accumulation d’IPP et d’ApppI dans différentes lignées humaines de cancer du sein et qu’il existe une corrélation entre le niveau de production d’IPP/ApppI et la capacité des lymphocytes T Vγ9Vδ2 à détruire ces cellules tumorales in vitro. Nous avons ensuite montré que le traitement de souris immunodéficientes NOD-SCID avec des aminobisphosphonates stimule la différenciation des lymphocytes T Vγ9Vδ2 humains à partir des cellules du sang périphérique lorsque celles-ci sont injectées par voie intrapéritonéale aux animaux. Par ailleurs, ces lymphocytes T vont ensuite venir infiltrer des tumeurs mammaires sous-cutanées qui ont été préalablement greffées chez les animaux traités. Cette infiltration lymphocytaire survient uniquement lorsque les tumeurs produisent de l’IPP/ApppI. Il en résulte alors une régression des tumeurs chez les souris NOD-SCID. L’infiltration des tumeurs par les lymphocytes T Vγ9Vδ2 humains s’explique par le fait que l’IPP et l’ApppI stimulent la migration des lymphocytes T. Les lymphocytes T Vγ9Vδ2 interagissent alors avec les cellules tumorales par le biais de mécanismes faisant intervenir ICAM-1, puis les lymphocytes sécrètent des facteurs (interféron γ, perforine) qui sont cytotoxiques pour les cellules tumorales. L’ensemble de nos travaux montre donc que les amino-bisphosphonates pourraient être utilisés en immunothérapie dans le traitement des cancers du sein
Vγδ9Vδ2 T lymphocytes are involved in the immune response against several pathogens and tumoral cells. They recognize non-peptidic, low molecular weight soluble antigens, so called phosphoantigens. In this context, isopentenyl pyrophosphate (IPP) and a metabolite of ATP (ApppI) are phosphoantigens which accumulate in many cells following the inhibition by amino-bisphosphonates of farnesyl pyrophosphate synthase (FPPS), a key enzyme in the mevalonate pathway. Bisphosphonates are commonly used in the clinic for the treatment and prevention of complications associated with malignant osteolysis. Preclinical studies demonstrate that bisphosphonates (in particular amino-bisphosphonates) also have direct and indirect anti-tumor activity. They are able to block different functions in tumor cells, such as adhesion, invasion and proliferation, by inhibiting FPPS activity. Bisphosphonates also act on endothelial cells and inhibit tumor angiogenesis. Amino-bisphosphonates also activate the cytotoxic activity of Vγδ9Vδ2 T lymphocytes against tumor cells. However, the underlying molecular mechanisms responsible for this activation are still unknown. In the present thesis, we first demonstrate that amino-bisphosphonates (e.g. zoledronate and risedronate) induce the accumulation of IPP and ApppI in different human breast cancer cell lines and that a correlation exists between the levels of IPP/ApppI production and the capacity of Vγδ9Vδ2 T lymphocytes to destroy tumor cells in vitro. We then demonstrate that the treatment of immunodeficient NOD-SCID mice with amino-bisphosphonates stimulates the differentiation of Vγδ9Vδ2 T lymphocytes from human peripheral blood mononuclear cells injected intraperitoneally in animals. Following their expansion, the T lymphocytes are then able to infiltrate subcutaneous mammary tumors, which were grafted in treated animals. This infiltration of γδ T lymphocytes arises only when the tumors produce IPP/ApppI, resulting in the regression of tumor growth in NOD-SCID mice. The infiltration of tumors by human Vγδ9Vδ2 T lymphocytes is explained by the capacity of IPP and ApppI to stimulate the migration of γδ T lymphocytes. Vγδ9Vδ2 T lymphocytes then interact with tumor cells through a mechanism involving ICAM-I and several secretion factors (i.e. interferon γ, perforine) which are cytotoxic for tumor cells. Thus, our work demonstrates that amino-bisphosphonates could be useful for breast cancer immunotherapy

La progression tumorale est un processus multi-étapes dépendant notamment des interactions entre les cellules cancéreuses et le stroma environnant. Le développement du cancer du sein implique une communication étroite entre les cellules épithéliales mammaires, les cellules inflammatoires, les myofibroblastes et les cellules endothéliales. Ainsi, le microenvironnement tumoral apparaît comme une cible de choix dans le traitement anti-tumoral. L’utilisation de modèles précliniques est une étape clé dans le développement et la validation de nouvelles thérapies. Néanmoins, peu d’études sont disponibles sur le rôle du stroma péri-tumoral dans ces modèles.Dans le but d’étudier le stroma péri-tumoral des modèles précliniques de cancers du sein, nous avons combiné une analyse par cytométrie en flux à une analyse par immunohistochimie afin d’identifier, puis de quantifier, les différentes populations stromales hématopoïétiques (lymphocytes, monocytes/macrophages, polynucléaires) et non hématopoïétiques (myofibroblastes, cellules endothéliales). Vingt et un modèles de xénogreffe de tumeurs humaines de cancers du sein ainsi que 2 modèles transgéniques (MMTV-PyMT et MMTV-ErbB2), ainsi que leurs allogreffes respectives, furent utilisés lors de ce travail.Les analyses des tumeurs humaines et murines ont montré un infiltrat stromal très hétérogène d’une tumeur à l’autre, avec pour composante majoritaire les macrophages. Un infiltrat important en polynucléaires a également été détecté dans les modèles de PDX, caractéristique d’une inflammation locale importante dans ces modèles. L’analyse phénotypique de macrophages a montré une expression variable de marqueurs M1 et M2 dans les modèles de PDX. Les macrophages issus de tumeurs murines transgéniques, spontanées ou allogreffées, présentaient quant à eux un profil majoritairement M1. L’étude transcriptomique de macrophages triés, a permis à la fois de valider les résultats obtenus au niveau protéique mais a également mis en évidence des différences majeures dans l’expression de nombreux gènes, impliqués dans des voies de signalisation variées telles que la croissance tumorale, l’invasion et la métastase.Cette étude nous a permis de mettre en évidence le rôle de la tumeur sur son microenvironnement. En effet, celle-ci est à la fois capable d’attirer un panel de cellules stromales qui lui et propre et ensuite de l’activer de façon spécifique
Tumor development is a multi-step process influencing by interactions between tumor cells and surrounding stroma. Breast cancer development involves a high level of communication between mammary epithelial cells, inflammatory cells, myofibroblasts and endothelial cells. So, the tumoral microenvironment appears as a prime target for anti-tumoral treatment. The use of preclinical models is a critical step in development and validation processes of new therapies. Nevertheless, the role of stroma in these models is poorly understood.In order to evaluate stromal cell populations in breast cancer preclinical models, we combined flow cytometry analysis and immunohistochemistry to identify, and then quantify, various stromal populations as hematopoietic cells (lymphocytes, monocytes/macrophages, polymorphonuclear leukocytes) and non-hematopoietic cells (myofibroblasts, endothelial cells). Twenty-one breast cancer patient-derived xenografts as well as 2 transgenic mouse models (MMTV-PyMT and MMTV-ErbB2), and their respective allografts, were studied.Analysis of human and murine tumors showed a strong heterogeneity between tumors regarding infiltrating stroma-cells, with a high proportion of macrophages. A significant amount of polymorphonuclear leukocytes was also detected in PDXs, indicating a local inflammation in these models. The phenotypic analysis of macrophages showed a variable expression of M1 and M2 markers in PDXs. Macrophages infiltrating transgenic mouse tumors, spontaneous or allografted, were mainly M1. Transcriptomic analyses of sorted macrophages, allowed us to validate previous results but also highlighted major differences in the expression of numerous genes implicated in various pathways as tumor growth, invasion and metastasis.Finally, this study highlighted the impact of tumor cells on their surrounding stroma. Indeed, we demonstrate that cancer cells are able to attract a specific panel of stromal cells and activate them in a specific way

Current flow estimation methods use tracheal sounds in all except one step of the process: ‘breath phase detection’, is done by assuming alternating breath phases or using a second acoustic channel. The alternating assumption is unreliable in long recordings; non-breathing events (apnea, swallow or cough) change the alternating pattern. Although phases can be detected using lung sounds intensity, the additional channel and associated labor is clinically impractical. We present a method using breath sound parameters to differentiate between the two respiratory phases. The novel method is independent of flow level, requiring only one prior- and one post- breath segment to identify the phase. This was tested on data from 93 healthy individuals, without any history of pulmonary diseases, at 4 different flow levels. The most prominent features were duration, volume and shape of the sound envelope. This method showed accuracy of 95.6%, 95.5% sensitivity and 95.6% specificity.

Around 334 million people have asthma worldwide. Asthma is an inflammatory disease of the airways which causes cough, breathlessness, chest tightness, and other peculiar sounds during breathing. The golden standard test spirometry is used to diagnose and monitor asthma. Spirometry is a lung function test that measures the time and volume of air a person can exhale after a deep inhalation. In general, patients repeat the test multiple times to get accurate readings, making it very time-consuming, and strenuous, especially for children and older people. A spirometer is also an expensive and bulky device that is not suitable for a home monitoring setup. Hence, a need for an easy and fast method exists. Sound-based analysis can be one such method. The motivation behind using sounds for monitoring and diagnosis of asthma originates from the sound production mechanism. In literature, non-speech sounds, namely, cough and breath recorded at the chest, have been explored. However, the analysis of speech and non-speech sounds recorded at the mouth is least investigated. Therefore, the work done in this thesis addresses this problem. For the analysis, we started with two tasks, namely classification and spirometry prediction for each sound category. Therefore, the thesis is divided into two parts.\\ Analysis of speech sounds For speech sound analysis, classification between the asthmatic and healthy subjects and spirometry prediction tasks have been performed. Results of the classification and spirometry prediction task show that \textipa{/oU/} (as in 'Home') is the best for the classification task, whereas \textipa{/i:/} (as in 'Meet') is best for the spirometry prediction. Results of the classification task suggest that Mel-frequency cepstral coefficients (MFCC) statistics carry maximum information for the discrimination in the case of all speech sounds considered in this work. Spirometry variables prediction task results show that the low frequency carries more information in best-performing sound \textipa{/i:/}.\\ Analysis of non-speech sounds Classification performed with non-speech sounds, cough, and breath, indicates that MFCC's are best, but interestingly, static MFCC coefficients are more informative than the velocity and acceleration coefficients. Breath performs the best for the classification task in the non-speech sound group. Further analysis of breath signal shows that discriminative information for classification is not uniform in the entire breath signal. Interestingly, the middle 50\% of the breath signal carries maximum information for the classification. To extract the middle 50\% of a breath, prior knowledge of breath boundaries is required. Therefore, we developed an unsupervised breath segmentation algorithm using dynamic programming. Classification results using predicted boundaries are found to be on par with the ground truth boundaries. Comparison of speech sounds and breath for the classification tasks shows that breath sound outperformed speech sounds. Experiments conducting for spirometry prediction tasks using non-speech sound indicate that breath performance is better than cough. In the spirometry prediction task, speech sounds outperformed the non-speech sounds. As asthma is not curable, patients needs to be on continuous medication known as bronchodilaotrs to reduce inflammation in the airways. But what kind of changes introduced due to airway change is not known. A linear discriminant-based analysis has been carried out to determine what kind of spectral changes occur in an asthmatic patient's sound before and after taking a bronchodilator. For this task, breath sounds have been chosen. It has been observed that frequency bands 400Hz-500Hz and 1480Hz -1900Hz are more sensitive to obstruction change in breath sound.

碩士
國立臺北科技大學
電子工程系
106
This work aims to identify patients by their body sounds acquired with stethoscopes. Bronchial breath sounds are one type of body sounds, which can be picked up by a stethoscope. They are formed by glottic or tracheal or main bronchi and are inhaled or exhaled through the mouth and nose. Individuals produce distinctive bronchial breath sounds, which therefore can be used for identifying and distinguishing patients. On the other hand, considering that stethoscopes can also capture the patient’s voice, and thus both bronchial sounds and human voices can be used together to perform patient identification, when the equipment is placed in the bronchial position. In this study, respiratory and articulation sounds of the neck bronchi were collected. By calculating their Mel-frequency cepstral coefficients and using the Gaussian mixture model to determine the identity, we further used “I-Vector & LDA” method to improve the accuracy of identity recognition. This method can achieve a recognition accuracy of 96.87% under the database test of 16 subjects.

碩士
國立臺北科技大學
電資國際專班
107
This study aims to develop a person identification (PID) system based on combined use of bronchial breath sounds and speech signals acquired by stethoscope. Two major methods, including support vector machines, and artificial neural networks are evaluated in the task of breath-sound-based PID. With the consideration of convenience, the amount of sound data collected from each person should be as small as possible, and hence the performance of PID may be limited, when the sound data for training the system is insufficient. To boost the performance of PID with sparse training data, this work studies data augmentation (DA) techniques that avoid the system training process from the overfitting problem. In addition, Feature engineering techniques are utilized to find the informative subset of breath sound features which is beneficial for PID. Our experimental data is provided by 16 volunteers, including equal number of male and female participants. In test phase, Both Support vector machine in combined with feature selection and Artificial Neural Networks approaches yielded the comparable accuracies of 98%.

"Black Sacred Breath: Historicity, Performance and the Aesthetics of BlackPentecostalism" considers are the aesthetic practices found in BlackPentecostalism, a multiracial, multi-class, multi-national Christian sect that began in Los Angeles, California in 1906 to argue that the aesthetic practices are the condition of possibility for a performative assessment and antiphonal criticism of normative theology and philosophy. Indeed, the history of these performances is an atheological-aphilosophical project, produced against the grain of liberal logics of subjectivity. By showing that theology and philosophy were abstractions of thought that produced the conceptual body as the target of racialization, the atheological-aphilosophical couplet indexes modes of intellectual practice that engulf and exceed such reductivism. BlackPentecostalism is a social, musical, intellectual form of new life, predicated upon the necessity of ongoing new beginnings. The religious practices I analyze produce a range of common sensual experiences: of "shouting" as dance; "testimony" and "tarry service" as song and praise noise; "whooping" (ecstatic, eclipsed breath) that occurs in praying and preaching; as well as, finally, "speaking in tongues." I ultimately argue that these aesthetic practices and sensual experiences are not only important objects of study for those interested in alternative modes of social organization, but they also yield a general hermeneutics, a methodology for reading culture.

During the antebellum era, both clergy and scholars alike levied incessant injunctions against loud singing and frenzied dancing in religion and popular culture. Calling for the relinquishment of these sensual spiritual experiences, I argue that these framing injunctions led to a condition where BlackPentecostal aesthetics, even in the much later institutional Black Studies, were and are thought as excessive performances. "Black Sacred Breath" investigates how discourses that emerged within the cauldron of spatiotemporal triangular trades in coffee, tea, sugar and human flesh of Transatlantic slavery necessitated a theology and philosophy of race, and consequently, the racializing of aesthetic practices. Over and against this discursive theology-philosophy were the performance practices of BlackPentecostalism, an atheology-aphilosophy. These sensual experiences were not merely performed through duress but were the instantiation and sign of love, of life. As love and life, these performative dances, songs, noises and tongues illustrate how enjoyment, desire and joy are important for the historicity - the theory of history found in these practices - that antiphonally speaks back against aversion, embarrassment and abandonment, against the debasement and denigration of blackness. Fundamentally, "Black Sacred Breath" is about the possibility for Black Study (as opposed to and differentiated from university institutional Black Studies), about the capacity for aesthetic practices typically deemed excessive can be constitutive, can provide new models for collective intellectual practice.

Dissertation

碩士
臺灣大學
電機工程學研究所
98
Recently, breast cancer has been one of the leading causes of death from cancer in females. Therefore, the study of its early detection has become an important issue now. Mammography has always been considered as the most common and the most effective screening method for detecting breast cancer because it can detect non-palpable and small tumors. However, the issue about ionization radiation is disputable. Besides, the images of dense breast region cover the images of small tumor region so that misdiagnosis happens. Toward this drawback, ultrasonic breast imaging can provide some aid. In short, ultrasonic breast imaging has many advantages, including noninvasive method, without ionizing radiation, portable, not limited to dense breast, and real-time. However, the use of conventional ultrasonic pulse-echo B-mode imaging to find breast tumors is also often limited by the image distortion caused by sound-velocity inhomogeneities in the breast tissue. Using other characteristics of tissue, such as sound velocity and attenuation coefficient, would provide diagnosticians additional information to increase the accuracy of diagnosis. The aim of the first part of this study was to determine the efficacy of using sound velocity and tissue attenuation to clinically discriminate breast cancer from healthy tissues. The method requires only raw channel data acquired by a linear transducer array and can therefore be implemented on existing clinical systems. In this study, these methods were tested on clinical data. A total of 19 biopsy-proven cases were evaluated. A imaging setup consisting of a 5-MHz, 128-channel linear array was used to simultaneously obtain B-mode image data, time-of-flight data and attenuation data. The sound velocity and attenuation coefficient can be reconstructed inside and outside a region of interest manually selected in the B-mode image. To reduce distortion caused by tissue inhomogeneities, an optimal filter derived from pulse-echo data—with water replacing the breast tissue—is applied. These results indicate that carcinoma (CA) can be discriminated from fibroadenoma (FA) and fat by choosing an appropriate threshold for the relative sound velocity (i.e., 18.5 m/s). However, the large variations in the attenuation within the same type of tissue make simple thresholding ineffective. Nevertheless, the method described in this study has the potential to reduce negative biopsies and to improve the accuracy of breast cancer detection in clinics. In the second part of this study, we implemented three different approaches to evaluate attenuation and one combined method in order to help the differentiation of breast cancer. Performance of these approaches is investigated based on simulation data. The three approaches are: video signal analysis (VSA), spectral estimation using periodogram (PER), and minimum side difference (MSD). Note that all approaches can readily be implemented using current B-mode imaging setup. First, VSA is to observe the gray-level gradient on a B-mode image. Second, PER is implemented by estimating the center frequency from the periodogram of the beamform data. Third, MSD is calculated using gray-level values of areas posterior to the region of interest, and left and right posterior to the ROI. In VSA, an effective frequency is required and typically the nominal center frequency is used. However, with a broadband pulse an accurate estimate of the effective frequency is needed to avoid large errors in VSA. In this study, we propose a modified VSA method in which PER is used to estimate the effective frequency. It is shown that accuracy of the VSA method can be improved by the proposed method particularly when the transmit bandwidth is large.

abstract: Until the second half of the 20th century, publications on breathing techniques for woodwinds have been scarce and often failed to adequately address this aspect of performance and pedagogy. It is through various sensory experiences and because of recent technological advances that academics recognize a gap in the existing literature and have since included studies using various methods, as well as modern technical devices and experiments into the woodwind literature and teaching. These studies have proven to be of great importance to confirm ideas and hypotheses on the matter. The aim of this project is to collect woodwind journal publications into a meta-analysis, focusing specifically on the breathing techniques for woodwind instruments and provide a comprehensive annotated bibliography on the topic and its application. The project is limited to journal articles on breathing techniques applied for woodwinds only, and will not review literature discussing breathing from other perspectives or in a broader sense. Major findings show that misconceptions and contradictions on the subject still exist. At the same time, they also highlight unique approaches used to help the learner overcome general and specific challenges while mastering the art of breathing. The project highlights areas where future research on breathing would be encouraged and should be complemented by measured data. Such studies might include a woodwind specific examination of the relationship between the tongue and the soft palate, or analysis of how tension in the torso muscles influences the movement of the diaphragm, or how rhythmical breathing affects breath control and capacity, and finally a discussion on how larynx influences the air stream.
Dissertation/Thesis
Doctoral Dissertation Music 2019

碩士
國立清華大學
電機工程學系所
106
The mortality rate of breast cancer among women in Taiwan ranks the top five in 2017 according to the statistics of the cause of death of the Ministry of Health and Welfare, Taiwan. Hence early screening/diagnosis via mammogram or ultrasound of breast cancer is vital. Due to the painful process of taking mammogram and fear of excess X-ray radiation exposure, ultrasound breast image examination become cheaper and well-accepted in breast diagnosis. Tumor segmentation of ultrasound breast (USB) images becomes important. Usually, traditional image segmentation methods are applied to obtain tumor segmentation of USB images. In this thesis, an AI-deep learning technique, namely the Mask-RCNN is adopted to solve the USB tumor segmentation problem. In addition to the speed advantage of the Faster-RCNN, the Mask-RCNN can also provide instance segmentation, which is suitable for tumor segmentation of USB images. Several error measures (ME, RFAE, MHD, TPR, TNR, ACC) are used to evaluate the segmentation results of this thesis. Comparisons with the other four methods (retracting DRLSE, expansion DRLSE, the algorithm proposed by Tsai-Wen Niu, and the algorithm proposed by Hsun Hsieh) are made. It is found that the segmentation result of this thesis is superior to the other four methods in terms of ME, RFAE, MHD.

碩士
國立清華大學
電機工程學系
104
Lesion segmentation for breast ultrasound images has been studied by many people, but it is hard to achieve using traditional edge detection because ultrasound images don’t have sharp contours and exist a lot of noise. In this thesis, we propose an automatic segmentation method for breast ultrasound images which combines morphological image processing and distance-regularized level-set evolution method (DRLSE), and improve combined DRSLE (cDRLSE) proposed by Yung-Hsuan Hsu. The most serious problem of cDRLSE is that it spend a lot of time on obtaining initial contour through the operation which applied the texture features (e.g., gray level co-occurrence matrix (GLCM)) for support vector machine (SVM), therefore we propose a new method based on morphological operations to search for initial contour which is effective and efficient. Using the new initial contours, we can capture the tumor area more precisely after applying DRLSE. To evaluate the result of segmentation, we compare it with expansion DRLSE method, contraction DRLSE method and cDRLSE method using three evaluation metrics, including misclassification error (ME), relative foreground area error (RFAE) and modified Hausdorff distance (MHD). We find that the proposed method is basically better than expansion DRLSE method and contraction DRLSE method which confirms the importance of initial contour to DRLSE. However, it is better than cDRLSE method in RFAE but worse than cDRLSE in ME and MHD, probably due to ranking failure in the proposed method or improved segmentation accuracy of post processing in cDRLSE method; even so, the proposed method not only spends less time obviously but also has no need to apply post processing. The proposed method has the following properties: 1.A fully automatic segmentation method for breast ultrasound images which has no need to set initial contour manually. 2.The way to obtain initial contour is efficient, and moreover if the tumor is smooth, the initial contour will be close to the tumor’s real boundaries. 3.Compared with other methods, the segmentation result of the proposed method is truly closer to ground truth image if the proposed initial contour mostly lies inside the ground truth.

碩士
國立臺灣海洋大學
環境生物與漁業科學學系
105
Climate-type disasters led by over emission of greenhouse gas are becoming increasingly serious and many countries have been actively developing renewable energy. As Taiwan Strait is good for wind power development, the Government actively promotes the construction of offshore wind farm. Among many negative and positive effects that may caused by the project, the fish-harvesting effect of wind turbine should be the most positive effects that we expected, but this effect must be evaluated objectively. Scientific echo sounder can provide fishery-independent evaluation for such effects in the wind farm areas. However, due to the spatial confinement of the wind field, the change of the scientific echo sounder detection in the horizontal or tilt detection can keep both detection effect and navigation safety. However, it may also cause changes in the target strength, volume backscattering strength and abundance evaluation. So, in this study three tank experiments were designed to explore these changes. Suspended single fish, free swimming single fish and fish shoal were used to measure TS in different attacking beam angle in order to analyze the problems that may be encountered in the future field survey in wind farm areas. Major results are summarized below: (1)The TS value of the change is less obvious in the pitching plane, the higher the TS is in the back; in the rolling plane and yawing plane will have a change of nearly 10 dB, and the two rotating plane are in the body side has a high TS. (2)In the single fish free swim, the linear relationship between average TS and average fish lengths were no more existed while the beam angle tilted to near horizontal(< 30 degree). The loss of the linear relationship was mainly due to the large TS variation in the yawing plane and also due to more frequent changes of TS while detecting near-horizontally toward a fish with changing swimming direction. (3)In the experiment free swimming fish shoals, we found that 0 degree and 90 degree although estimated body length varied greatly, the density estimated were very close. Therefore, tilt detection beam angle may cause variation of TS and result in bias of estimated body length; however the density estimation were not influenced at all.

The topic of this thesis is nomination of justices of the Supreme Court, the highest federal court in the United States. The Constitution gives the power to appoint new federal justices to the president, with "advice and consent" of the Senate. This clause is based on one of the primary constitutional principles used in the United States - the system of checks and balances. The first part describes the general aspects of all nominations, from court vacancies to the final vote on Senate floor. The primary roles of the president and the Senate are defined. Focus is also set on the evolution of the nomination process throughout the history, as well as the criteria for selecting new justices, who shape the constitutional development of one of the oldest functioning democracies. The structure of the first part is then applied for a description of the nomination of Brett M. Kavanaugh, who was appointed to the Supreme Court by President Donald Trump in 2018. The course of this nomination was strongly affected by sexual assault accusations against the nominee and fierce partisan battles that resulted in the closest confirmation vote in history. The final part applies quantitative research methods to discover the influence of partisan difference between the president and the Senate majority on the...

L’obligation de disponibilité du travailleur met en lumière un temps durant lequel le travailleur demeure à la disposition de son employeur malgré le fait qu’il n’accomplit pas concrètement sa prestation de travail, le temps de disponibilité. Conséquence de plusieurs phénomènes, notamment la mondialisation des marchés, la financiarisation de l’économie et l’essor des nouvelles technologies de l’information et de la communication, l’obligation de disponibilité est mal définie par les règles juridiques actuelles au Québec. Considérant cet état du droit, notre mémoire cherche à évaluer dans quelle mesure la Loi sur les accidents du travail et les maladies professionnelles capte l’obligation de disponibilité du travailleur. Pour ce faire, nous avons effectué une analyse quantitative et qualitative de la jurisprudence rendue par la Commission d’appel en matière de lésions professionnelles et la Commission des lésions professionnelles entre 1995 et 2014. Notre mémoire comporte trois volets. Sous un angle plus sociologique, nous voulions examiner si notre population de décisions nous permettait, d’une part, de constituer différentes manifestations-types de l’obligation de disponibilité et d’autre part, de discerner une progression dans le temps du nombre de décisions portant sur une obligation de disponibilité. Enfin, nous désirions examiner dans quelle mesure un travailleur victime d’une lésion durant une période de disponibilité peut se prévaloir des droits accordés par la L.a.t.m.p.. À l’aune de notre mémoire, plusieurs manifestations-types de l’obligation de disponibilité peuvent être en constituées en fonction du moment durant lequel elle se manifeste. L’obligation de disponibilité peut également être rattachée au statut de travailleur autonome. En outre, il semble que la reconnaissance de la lésion professionnelle du travailleur varie en fonction de ces manifestations-types. Cependant, nos résultats ne révèlent pas une progression dans le temps du nombre de décisions portant sur une obligation de disponibilité.
The concept of worker availability requirements refers to instances in which workers are required to be available to their employers outside of standard work hours (off-duty). Due to several factors, especially the internationalization of markets, financialisation of the economy, and the development of information and communication technology, this normative requirement is inadequately regulated by current legislation in Quebec. Therefore, our research evaluates the extent that the Loi sur les accidents du travail et les maladies professionnelles addresses availability requirements imposed on workers in the province. To this end, we carried out qualitative and quantitative analyses of decisions made by the Commission d’appel en matière de lésions professionnelles and Commission des lésions professionnelles between 1995 and 2014. From a sociological perspective, we attempted to produce different categories of off-duty availability requirements, and to discern whether an augmentation in the number of decisions concerning availability requirements has taken place over time. Finally, we examined the extent that workers benefit from the rights provided by the L.a.t.m.p. when injuries occur during off-duty hours in which workers are required to be available. Our results demonstrate that many categories of availability requirements can be developed related to the period of time during when such obligation occurs. Furthermore, these requirements are also linked to individuals’ status as self-employed workers. Finally, it seems that the recognition of workers’ employment injuries vary along these categories. However, our results also suggest that there exists no positive trend in the number of decisions concerning worker availability requirements over time.

碩士
國立清華大學
電機工程學系所
105
Due to the speckle noise and low contrast in breast ultrasound images, it is hard to locate the contour of the tumor by using a single method. In this thesis, a new method for finding an initial contour is proposed, which can improve the result of DRLSE on the segmentation of BUS images. The new method focuses on improving the algorithm proposed by Tsai-Wen Niu, which is a way to search an initial contour based on the local minimum in the images. When the BUS images contain calcification, it is possible to fail in searching of initial contour through such algorithm, hence leading to a poor segmentation result when the initial contour is on the wrong place. Therefore, we acquire a bigger initial contour by using a series of image smoothing methods and binarization, which can eliminate the weak edges and adjust the contrast in BUS images. In addition, some images without local minimum can be successfully detected by using the proposed method. However, the pixel value in these images are similar. It might be hard to accurately separate the tumor region from non-tumor region by the difference of pixel values. These obstacles are conquered by calculating the difference of length and pixel value in the suspect region. The ranking outcome is improved by using the morphological features. After applying DRLSE, our initial contour can reach the tumor region more accurately. To evaluate the result of segmentation, it is compared with the outcome of DRLSE obtained from different initial contours proposed by Tsai-Wen Niu, expansion DRLSE method, and contraction DRLSE method using three evaluation metrics, including ME, RFAE and MHD. The experimental results indicate that the proposed method is basically better than the other methods. However, the initial contour might contain non-tumor region when the edge of the tumor’s boundary is too ambiguous; even so, the proposed method drastically reduce the number of DRLSE iteration and computation time. According to the experimental results, the proposed method has three advantages over the other methods. First, it sets the initial contour automatically which is more efficient than setting the initial contour manually. Second, the region of the initial contour is much bigger than those obtained by the other methods, which can reduce the computation time and the number of DRLSE iteration. Third, if the tumor boundary is distinct, the new initial contour can improve the segmentation result of DRLSE.

Les récepteurs couplés aux protéines G (RCPG) constituent la plus grande famille de protéines membranaires du génome humain. Ils transmettent les signaux extracellulaires provenant de plusieurs stimuli comme les odeurs, les ions, les hormones et les neurotransmetteurs, à l'intérieur des cellules. En se liant aux RCPGs, ces molécules contribuent à la stabilisation des changements conformationnels activateurs qui se propagent jusqu'au domaine intracellulaire des récepteurs. Ces derniers engagent ensuite un ou plusieurs effecteurs, comme les protéines G hétérotrimériques et les β-arrestines (βarrs), qui activent une cascade d'événements moléculaires menant à la réponse cellulaire.Récemment, la publication de structures cristallines de RCPGs liant des ligands diffusibles a offert une opportunité de raffiner à une résolution atomique les modèles des mécanismes de transduction des signaux. Dans la première partie de cette thèse, nous avons donc exploré les déterminants de la signalisation du récepteur prototypique β2-adrénergique (β2AR), induite par les β-bloqueurs. En ne tenant compte que de leur efficacités sur le β2AR dans les voies de l'adénylate cyclase (AC) et des protéines kinases activées par les facteurs mitogéniques (MAPK), les β-bloqueurs peuvent être classés en 3 groupes distincts (agoniste inverse AC / agoniste MAPK, antagoniste neutre AC / agoniste MAPK et agoniste inverse AC / agoniste inverse MAPK). Afin de déterminer le lien entre leur efficacité et leur mode de liaison, nous avons réalisé des expériences d'arrimages moléculaires in silico entre des β-bloqueurs de chacun des groupes et la structure cristalline du β2AR liée au carazolol. De manière intéressante, les ligands à l'intérieur d'un groupe partagent un mode de liaison, alors que ceux des ligands entre les groupes divergent, suggérant que le mode de liaison des β-bloqueurs pourrait être utilisé pour prédire leur l'efficacité. En accord avec cette hypothèse, nous avons prédit et confirmé l'efficacité agoniste MAPK du carazolol, un inverse agoniste AC du β2AR se liant au récepteur de manière similaire au groupe inverse agoniste AC / agoniste MAPK. De manière intéressante, le groupement aryl des ligands agonistes inverses agonistes AC / agoniste MAPK, le seul groupement chimique variable de ce groupe, est prédite pour lier la région des 3e et 5e hélices transmembranaires (TM3 et TM5). Nous avons donc émis l'hypothèse que cette région pourrait être un déterminant de l'efficacité de ces ligands. En accord avec cette dernière, la mutation de 2 résidus (T118I, S203A) localisés proches du site de liaison des groupements aryls des β-bloqueurs, prévient l'efficacité agoniste inverse de l'ICI-118551 sur la voie de l'AC sans affecter l'efficacité d'un agoniste, indiquant que cette région est importante pour la transmission de l'effet agoniste inverse, du moins sur la voie de l'AC. Les βarrs sont des protéines d'échafaudage qui coordonnent la formation de complexes avec plusieurs dizaines d'effecteurs de signalisation. Originalement identifiées pour leur rôle dans la désensibilisation et l'internalisation des RCPGs, elles sont aussi d'importants effecteurs de la signalisation des RCPGs indépendante des protéines G hétérotrimériques. Cependant, contrairement aux protéines G hétérotrimériques, il n'existe que peu d'outils pour les étudier. Ainsi, la deuxième partie de la thèse est dédiée au développement d'outils pour l'étude des βarrs. À cette fin, nous avons d'abord tenté de transposer une méthode de mesure de l'interaction entre 2 protéines par la technologie de transfert d'énergie de bioluminescence par résonance (BRET) en microscopie et chez des souris transgéniques afin de mesurer de manière subcellulaire et dans un contexte natif l'engagement de la βarr à des RCPGs. Ainsi, nous avons établi les preuves de principe que le BRET peut être utilisé pour localiser l'interaction entre la βarr et le récepteur de la vasopressine de type 2 (V2R) sur une cellule au microscope et pour détecter l'interaction entre la βarr et le β2AR sur des tissus de souris transgéniques exprimant ces protéines fusionnées avec des partenaires BRET. Finalement, il n'existe aucun inhibiteur pharmacologique ciblant les βarrs. Ainsi, grâce à la combinaison d'approches de criblage virtuel sur un modèle de la structure des βarrs et d'essais de validation cellulaire, nous avons développé un inhibiteur pharmacologique des βarrs. À l'aide de cet outil, nous avons confirmé l'implication des βarrs dans l'activation des MAPK par le V2R, mais aussi montré un nouveau rôle des βarrs dans le recyclage du β2AR. Les connaissances et outils développés dans cette thèse permettront de mieux comprendre les déterminants moléculaires de la signalisation des RCPGs et entre autres, grâce à des nouvelles approches pour étudier le rôle cellulaire et physiologique des βarrs.
G Protein-Coupled Receptors (GPCR) are members of the largest family of membrane protein in the human genome. They transduce the signal from a variety of stimuli like odors, ions, hormones and neurotransmitters, inside the cells. By binding directly to the receptors, these molecules stabilize activating conformational changes that are allosterically propagated through transmembrane to intracellular domains. Effectors like heterotrimeric G protein and β-arrestins (βarrs) are then engaged by activated receptors and trigger a cascade of signalling events leading to a cellular response. Recently, the resolution of the crystal structure of GPCR that bind to freely diffusible ligands provided the opportunity to refine at an atomic level the models describing the mecanisms of receptor signal transduction. In the first section of this thesis, we have explored the determinants of the prototypical β2-adrenergic receptor (β2AR) signalling induced by β-blockers. Given their efficacy on Adenylate Cyclase (AC) and Mitogen-Activated Protein Kinase (MAPK) pathways, β-blockers can be classified within 3 signalling groups (AC inverse agonist / MAPK agonist, AC neutral antagonist / MAPK agonist and inverse agonist for AC and MAPK). In order to gain insight on the relation between their efficacy and binding mode, we performed in silico binding experiments between β-blockers from each group and the β2AR crystal structure bound to carazolol. Interestingly, ligands within a group share similar binding mode in contrast to those of different groups, suggesting that β-blockers binding mode could be used to predict their efficacy. In accordance to this hypothesis, we have predicted and confirmed that carazolol, an AC inverse agonist that bind to β2AR in a similar way than the AC inverse agonist / MAPK agonist group, is indeed an agonist for MAPK pathway. Moreover, aryl chemical function from AC inverse agonist / MAPK agonist ligands, barely the only variable structure feature of this group, was predicted to bind β2AR nearby the transmembrane helices 3 and 5 (TM3 and TM5). We thus have predicted that this region would be a determinant of the AC inverse agonist / MAPK agonist ligand efficacy. Accordingly, we found that mutation of 2 residues (T118I, S203A) close to the aryl moiety binding site prevents inverse agonist efficacy of ICI-118551 on AC pathway, without affecting agonist efficacy, indicating that this receptor region is important for the efficacy of these group of β-blockers, at least on AC inverse agonism.βarrs are scaffolding proteins that coordinate protein complex formation with dozen of signalling effectors. First identified for their role on GPCR desensitization and internalization, βarrs are also an important heterotrimeric G protein independent GPCR signalling effectors. However, in contrast to heterotrimeric G protein, only a few tools are available for their study. Thus, the second section of this thesis aim at developing tools for the study of βarrs. For this purpose, we had attempted to transpose a method to measure protein-protein interaction that use Bioluminescence Resonance Energy Transfer (BRET) technology, in microscopy and in transgenic mice, in order to detect subcellular localization and in a native context the engagement of βarr to RCPGs. Thus, we have established a proof of principle that BRET can be combined with microscopy to locate an interaction between βarr and the type 2 vasopressin receptor (V2R) within a cell. Moreover, we have established a second proof of principle that we can detect βarrs recruitment to β2AR on cells extracted from tissues of transgenic mice expressing these proteins fused to BRET partner. Finally, there is no pharmacological inhibitor of βarrs. Thus, using a combination of virtual screening and cellular validation approches, we have developed the first pharmacological βarrs inhibitor. With this novel tool, we have confirmed the implication of βarrs in V2R-mediated MAPK activation, but also showed a new role of βarrs in β2AR recycling.The finding and the tools presented in this thesis should allow to better understand the molecular determinants of GPCR signalling, and among other things, by proposing new tools to study βarrs cellular and physiological roles.

v anglickém jazyce - English abstract Protection of Right to Fair Trial in Relation to Preliminary Ruling Proceedings Right to fair trial is one of fundamental human rights, which enables individuals to effectively invoke their rights and freedoms before a lawful, impartial and independent court. In Europe, the protection of this right is guaranteed at a multi-layer level, whose layers interact: first layer is formed by constitutional orders of individual States, second layer by institutions of European Union, in particular the Court of Justice of the European Union, and a third one, that of the European Court for Human Rights. In the European Union, the Treaties have introduced the mechanism of preliminary ruling with the aim of preserving unity within the Union and of ensuring coherent interpretation and application of European law by the courts of the Member States. Preliminary ruling proceedings ensure effective cooperation between national courts and the Court of Justice of the European Union. Art. 267 TFEU provides for an obligation to request a preliminary ruling for national courts of last instance (if none of the CILFIT case law conditions is met). Breach of this obligation may entail violation of right to fair trial at all three layers of human rights protection in Europe. Each European...