Characterization of BMP signaling in breast development and tumorigenesis

Résumé L'influence des hormones reproductives sur le développement du cancer du sein a été établie au travers de nombreuse études épidémiologiques. Nous avons précédemment démontré que le gène Wnt-4 est un médiateur essentiel de la progestérone dans le développement lobulo-alvéolaire de l'épithélium mammaire. De plus, le rôle de la voie de signalisation Wnt dans la tumorigénèse de la glande mammaire mutine est largement établi. Pour comprendre sa fonction dans le cancer du sein, nous avons activée cette voie en surexprimant le gène Wnt-1 dans des cellules épithéliales primaires de sein, au moyen d'un rétrovirus. Ceci a conduit à la transformation oncogénique de ces cellules et à l'obtention d'un modèle de carcinogénèse du sein dénommé Wnt-1 HMEC. L'analyse de l'expression des gènes induits par la surexpression de Wnt-1 dans ces cellules, a permis d'identifier les gènes BMP4 et 7. Alors que des analyses de RT-PCR ont montré leur forte expression dans les cellules Wnt-1-HMECs, la présence d'une grande quantité de la protéine BMP7 a été constatée dans les tumeurs dérivées de ces cellules. L'importante phosphorylation des Smad 1, 5, S dans les Wnt-1 HMECs indique l'activation de la voie BMP, possiblement due à la stimulation ce celle-ci par BMP7. L'activation de la voie Wnt par la ß-Caténine, conduit à la transcription de BMP7, identifiant ainsi ce gène comme un gène cible de la voie canonique. La pertinence de nos observations a par ailleurs été confirmée par le fait que BMP7 est surexprimé dans les tumeurs de seins humains. Afin d'élucider la fonction de la voie BMP dans le sein, nous avons utilisé le modèle mutin. L'expression du gène BMP7 dans les souris transgéniques MMTV Wnt-1 s'est avérée élevée, démontrant qu'il est aussi un gène cible de la voie Wnt in-vivo. L'expression de l'ARN messager .codant pour la protéine BMP7 est induite lors du développement lobulo-alvéolaire, qui se fait sous l'influence de la progestérone et de Wnt-4. Ensemble, ces observations corroborent le fait qu'une stimulation avec de la progestérone suffit à induire la transcription du gène dans les 24h. Nos résultats coïncident d'autre part avec le fait que BMP7 est exprimé dans la couche myoépithéliale de l'épithélium où la voie Wnt est activée. L'analyse de souris reportrices de l'activité de la voie BMP, suggère une activation dans la couche luminale de l'épithélium durant tout le développement de la glande mammaire. Curieusement, cette même voie est active dans le mésenchyme lors de la mammogénèse embryonnaire. Finalement, nos analyses d'immunofluorescence démontrent la capacité de prolifération des cellules ayant activé BMP, ainsi que leur nette ségrégation d'avec les cellules exprimant le récepteur à la progestérone. Nos résultats démontrent que le gène BMP7 est un gène cible de la voie Wnt canonique dans le sein. Son expression dans la couche myoépitheliale est induite par Wnt-4, lui-même sécrété par les cellules luminales sensibles à la progestérone. La sécrétion de la protéine BMP7 conduit finalement à l'activation de la voie BMP dans les cellules négatives pour le récepteur à la progestérone. Abstract Epidemiological studies highlight the repetitive exposure to circulating progesterone as a major risk in the development of breast cancer. Work in our laboratory showed that Wnt-4 is an essential mediator of progesterone-driven side-branch formation, while Wnt signaling has long been established as strongly oncogenic in the mouse mammary gland. To address the role of Wnt in breast tumorigenesis we activated the pathway in primary human breast epithelial cells by means of refroviral Wnt-1 expression. This resulted in a Wnt1-induced breast carcinogenesis model, being referred to as Wnt-1-HMECs. Gene expression profiling revealed the Bone Morphogenetic Protein 4 and 7 (BMP4 and 7) a mong the most upregulated gene by ectopic Wnt-1 expression in primary HMECs. RT-PCR analysis confirmed elevated BMP4 and 7 mRNA levels in Wnt-1-infected HMECs, as well as strong BMP7 expression in the tumors derived from these cells. Smad 1, 5, 8 phosphorylation was high in Wnt-1HMECs whereas below detection limit in primary HMECs suggesting that the increased expression of BMP-7 results in activation of downstream signaling. Ectopic expressíon of a stabilized form of ßcatenin in primary HMECs resulted in increased transcription of BMP-7 suggesting that it is a target of canonical Wnt signaling. The clinical relevance of our observations was confirmed by the finding of BMP7 being upregulated in human breast tumor samples. To elucidate the role of BMP ligands in the breast in-vivo, we made use of the mouse model. Expression of the BMP7 gene was found to be increased in MMTV-Wnt-1 transgenic animals, suggesting that BMP7 may also be a Wnt 1 target gene in vivo. Expression of BMP7 was upregulated in mid-pregnancy which coincides with progesterone/Wnt induced side branching. BMP7 was induced within 24 hours by progesterone. Consistent with it being a target of canonical Wnt signaling, we demonstrated preferential expression of this ligand in the myoepithelial cells, the target cells of Wnt signals. In-vivo analysis of BMP signaling using a reporter mouse revealed the activation of the pathway in the luminal layer of the epithelium throughout postnatal development. Interestingly, during embryonic mammogenesis the pathway was found to be active in the mesenchyme. Immunofluorescence studies demonstrated that cells with BMP activity can proliferate. They also revealed a clear segregation between progesterone receptor positive cells and cells with active BMP signaling. Together our observations suggest that BMP-7 is a canonical Wnt signaling target both in HMECs and in the mouse mammary gland in-vivo. It is expressed in the myoepithelium possibly in response to Wnt-4, which is secreted by steroid receptor positive cells in response to progesterone. BMP-7 in turn may impinge on lumina) epithelial cells and activate BMP signaling in PR negative cells.

Symptoms of chronic stress at work in NGO workers

Working in a NGO often involves providing life saving resources (food, medicine, equipment, water, etc) to needy populations around the globe. Such duty requires highly dedicated employees and humanitarian workers are said to face a hign degree of pressure in their daily work. Despite the evidence of taxing work demands, and a high potential for stress related problems, very few studies on occupational chronic stress have specifically looked at NGO workers. Assuming that "field stress" can relay to workers at headquarters, we carried out an exploratory study about occupational health among employees of a NGO's headquarters. We sent a questionnaire to all employees (N=130) of a NGO headquarters located in Switzerland. We used the TST questionnaire (French version of the Langner's questionnaire on psychiatric symptoms) to identify cases with potential mental health problems. We also included in the questionnaire some items about motivation, acknowledgment, work-life balance, job demand, and autonomy. A total of 75 employees answered our questionnaire (57% response rate). 44% of our sample were men (n=33) and 56% were women (n=42). The mean age was of 40 years (SD=7.6). 56% were working at the headquarters of the NGO in questions as of 2 years or less. Not surprisingly, a majority of respondents reported to be highly motivated (74%) and the meaning of work was important for 80% of them. However, 35% indicated having problems in conciliating their private and professional life. Most frequent reported symptoms included feeling "weak all over" (81%), having "trouble getting asleep often" (35%), "clogging in nose" (35%), feeling "nervous often" (33%), and "memory not all right" (33%). The score for psychiatric symptoms was high in 8 (11%) employees whose health might therefore be at risk. In comparison, other sudies showed that this proportion was 9% for French teachers and 16% for sales personnel1. Results show that symptoms of mental health problems do occur among NGO workers. Some of these symptoms are known to be linked to occupational stress. Chronic stress manifests itself first in non-specific symptoms (e.g. fatigue) and later in specific pathologies. This could explain the relatively low proportion of cases with a high score in Langner's scale than was expected. Therefore, we hypothesize a healthy worker effect. The fact that our sample is 40 years old in average, and that the turnover is quite high can also support this hypothesis. Further research is needed in order to better understand occupational stress in this specific population. An upcoming study will investigate the role of organizational factors associated with health complaints. Therefore, a longitudinal survey including quantitative and qualitative methods is appropriate.

Cinema and the Work of Doré

According to Ray Harryhausen, a special effects expert in the film industry, "Gustave Doré would have made a great director of photography . . . He saw things from the point of view of the camera." Doré's work has had a permanent impact on the imaginative realm of film since its very early days. In return, the silver screen has etched Doré into the 20th century imagination. Almost every film about the Bible since The Life and Passion of Jesus Christ produced by Pathé in 1902 refers to his illustrations, and every film adaptation of Dante or Don Quixote has used him as a model, from Georg Wilhelm Pabst and Orson Welles to Terry Gilliam. All films dealing with life in London in the Victorian era by directors ranging from David Lean, to Roman Polanski and Tim Burton draw on the visions in London: a pilgrimage for their sets. A large number of dream fantastical or phantasmagorical scenes take their inspiration from Doré's graphic world, beginning with Georges Méliès' A Trip to the Moon in 1902. In the realm of cartoons and animation, Walt Disney owes a huge debt to Doré. Doré primal forests, from Atala in particular, were also used in the various versions of King Kong from 1933 to the 2005 film by Peter Jackson, who had already drawn on Doré for The Lord of the Rings. Jean Cocteau was also indebted to the illustrations for Perrault's Fairy Tales for his Beauty and the Beast (1945), as was George Lucas for the character Chewbacca in Star Wars (1977) and even the Harry Potter film series. Through his influence on film history, Doré shaped the mass culture imagination.

The INTERMED questionnaire for predicting return to work after a multidisciplinary rehabilitation program for chronic low back pain.

OBJECTIVES: To evaluate the performance of the INTERMED questionnaire score, alone or combined with other criteria, in predicting return to work after a multidisciplinary rehabilitation program in patients with non-specific chronic low back pain. METHODS: The INTERMED questionnaire is a biopsychosocial assessment and clinical classification tool that separates heterogeneous populations into subgroups according to case complexity. We studied 88 patients with chronic low back pain who followed an intensive multidisciplinary rehabilitation program on an outpatient basis. Before the program, we recorded the INTERMED score, radiological abnormalities, subjective pain severity, and sick leave duration. Associations between these variables and return to full-time work within 3 months after the end of the program were evaluated using one-sided Fisher tests and univariate logistic regression followed by multivariate logistic regression. RESULTS: The univariate analysis showed a significant association between the INTERMED score and return to work (P<0.001; odds ratio, 0.90; 95% confidence interval, 0.86-0.96). In the multivariate analysis, prediction was best when the INTERMED score and sick leave duration were used in combination (P=0.03; odds ratio, 0.48; 95% confidence interval, 0.25-0.93). CONCLUSION: The INTERMED questionnaire is useful for evaluating patients with chronic low back pain. It could be used to improve the selection of patients for intensive multidisciplinary programs, thereby improving the quality of care, while reducing healthcare costs.

Meanings and perceptions of patient-centeredness in social work, nursing and medicine: A comparative study.

Answering patients' evolving, more complex needs has been recognized as a main incentive for the development of interprofessional care. Thus, it is not surprising that patient-centered practice (PCP) has been adopted as a major outcome for interprofessional education. Nevertheless, little research has focused on how PCP is perceived across the professions. This study aimed to address this issue by adopting a phenomenological approach and interviewing three groups of professionals: social workers (n = 10), nurses (n = 10) and physicians (n = 8). All the participants worked in the same department (the General Internal Medicine department of a university affiliated hospital). Although the participants agreed on a core meaning of PCP as identifying, understanding and answering patients' needs, they used many dimensions to define PCP. Overall, the participants expressed value for PCP as a philosophy of care, but there was the sense of a hierarchy of patient-centeredness across the professions, in which both social work and nursing regarded themselves as more patient-centered than others. On their side, physicians seemed inclined to accept their lower position in this hierarchy. Gieryn's concept of boundary work is employed to help illuminate the nature of PCP within an interprofessional context.

Estimating the contribution of occupational solar ultraviolet exposure to skin cancer

BACKGROUND: Exposure to solar ultraviolet (UV) light is the main causative factor for skin cancer. Outdoor workers are at particular risk because they spend long working hours outside, may have little shade available and be bound to take their lunch at their workplace. Despite epidemiological evidence of a doubling in risk of squamous cell carcinoma in outdoor workers, the recognition of skin cancer as an occupational disease remains scarce. OBJECTIVE: To assess occupational solar UV doses and its contribution to skin cancer risk. METHODS: A numerical model (SimUVEx) was used to assess occupational and lunch break exposures, characterize exposure patterns and anatomical distribution. Risk of squamous cell carcinoma (SCC) was estimated from an existing epidemiological model. RESULTS: Horizontal body locations received 2.0-2.5 times more UV than vertical locations. Dose associated to lunch outdoor every day was similar to outdoor work one day per week but only half of a seasonal worker. Outdoor workers are associated with an increased risk of SCC but also of frequent acute episodes. CONCLUSION: Occupational solar exposure contributes largely to the overall lifetime UV dose, resulting in an excess risk of SCC. The magnitude of the estimated excess in risk supports the recognition of SCC as an occupational disease.

Predicting Non Return to Work after Orthopaedic Trauma: The Wallis Occupational Rehabilitation RisK (WORRK) Model.

BACKGROUND: Workers with persistent disabilities after orthopaedic trauma may need occupational rehabilitation. Despite various risk profiles for non-return-to-work (non-RTW), there is no available predictive model. Moreover, injured workers may have various origins (immigrant workers), which may either affect their return to work or their eligibility for research purposes. The aim of this study was to develop and validate a predictive model that estimates the likelihood of non-RTW after occupational rehabilitation using predictors which do not rely on the worker's background. METHODS: Prospective cohort study (3177 participants, native (51%) and immigrant workers (49%)) with two samples: a) Development sample with patients from 2004 to 2007 with Full and Reduced Models, b) External validation of the Reduced Model with patients from 2008 to March 2010. We collected patients' data and biopsychosocial complexity with an observer rated interview (INTERMED). Non-RTW was assessed two years after discharge from the rehabilitation. Discrimination was assessed by the area under the receiver operating curve (AUC) and calibration was evaluated with a calibration plot. The model was reduced with random forests. RESULTS: At 2 years, the non-RTW status was known for 2462 patients (77.5% of the total sample). The prevalence of non-RTW was 50%. The full model (36 items) and the reduced model (19 items) had acceptable discrimination performance (AUC 0.75, 95% CI 0.72 to 0.78 and 0.74, 95% CI 0.71 to 0.76, respectively) and good calibration. For the validation model, the discrimination performance was acceptable (AUC 0.73; 95% CI 0.70 to 0.77) and calibration was also adequate. CONCLUSIONS: Non-RTW may be predicted with a simple model constructed with variables independent of the patient's education and language fluency. This model is useful for all kinds of trauma in order to adjust for case mix and it is applicable to vulnerable populations like immigrant workers.

Etude systématique, cladistique et biogéographique des Baetidae (Ephemeroptera) de Madagascar

Les Ephéméroptères constituent un ordre très archaïque d?insectes ailés, comprenant un nombre réduit d?espèces (actuellement environ 2500 espèces). Les larves sont aquatiques; la durée de ce stade est en général d?une année. Le stade adulte est par contre extrêmement bref: de quelques heures à quelques jours. La fonction quasi unique de ce stade est la reproduction. Par sa superficie, Madagascar est la quatrième île du monde. Elle est située dans la partie occidentale de l?Océan Indien à plus de 300 km de la côte africaine. Madagascar faisait partie du super-continent Gondwana. Elle s?est séparée de l?Afrique (-165 M.a.), puis a migré vers le Sud (-125 M.a.) avant de se détacher du sous-continent indien (-65 M.a.). La connaissance des Ephéméroptères malgaches était, jusqu?à très récemment, extrêmement limitée. Grâce au programme Biodiversité et biotypologie des eaux continentales malgaches, lancé conjointement par l?ORSTOM (actuel IRD, France) et le CNRE (Madagascar), un inventaire à large échelle de la macrofaune benthique malgache a été entrepris. La systématique de plusieurs familles d?Ephéméroptères (Tricorythidae, Polymitarcyidae, Palingeniidae,?), ainsi que d?autres groupes d?invertébrés (Trichoptères, Simuliidae, macrocrustacés) a fait l?objet d?études approfondies. La présente étude consistue un des volets de ce programme. Jusqu?au milieu des années 1990, seules quatre espèces valides appartenant à trois genres différents étaient décrites de Madagascar. En 6 ans, ce ne sont pas moins de 25 articles qui sont consacrés à la systématique des Baetidae, permettant de décrire 50 espèces et 8 genres nouveaux. La faune malgache des Baetidae compte actuellement 22 genres et 54 espèces. Malgré sa taille, Madagascar possède une richesse, tant générique que spécifique équivalente à celle d?un continent. Notre connaissance des Baetidae est suffisamment avancée pour mener une étude cladistique et biogéographique. La reconstruction phylogénétique a permis de mettre en évidence cinq lignées principales à Madagascar et de préciser, pour chacune d?elles, les genres inclus et les caractères propres. La faune des Baetidae malgaches présente un taux d?endémicité très élevé: 53 des 54 espèces et un tiers des genres sont endémiques. Elle montre des affinités extrêmement fortes avec la faune africaine, puisque 90% des genres présents à Madagascar ou en Afrique ont une répartition strictement restreinte à cette région. Les autres composantes, notamment orientales et océaniennes, sont négligeables; ces régions n?ont en commun avec Madagascar qu?un nombre restreint de genres cosmopolites. Ces affinités sont en contradiction avec les données géologiques de la dislocation du Gondwana. Plusieurs explications peuvent être données pour résoudre cette contradiction. La plus vraisemblable est que le pouvoir de dispersion des Ephéméroptères, et des Baetidae en particulier, est nettement sous-estimé. L?étude des faunes des îles volcaniques récentes, telles que les Comores, démontre clairement que les Baetidae sont capables de dispersion sur une distance de plus de 300 km. Il est donc possible d?envisager une colonisation de Madagascar à partir de l?Afrique continentale postérieure à la séparation des deux plaques. Nous avons établi des scénarios retraçant l?histoire biogéographique de chacune des cinq lignées. Pour quatre d?entre elles, l?Afrique continentale est le centre d?origine. La cinquième lignée aurait une origine paléarctique; l?Afrique représenterait un centre secondaire de spéciation. Ces lignées auraient secondairement colonisé Madagascar à partir de l?Afrique continentale. Ce travail ouvre donc d?importantes perspectives. Il rend possible l?utilisation à un niveau générique, voire spécifique, des Baetidae pour des travaux de faunistique ou d?écologie, en particulier pour des études liées à la dégradation de la qualité de l?eau. Il devrait également pouvoir servir de base pour l?étude et la compréhension des phénomènes de dispersion et colonisation dans les îles et archipels de l?Ouest de l?Océan Indien.<br/><br/>Mayflies (Ephemeroptera) are among the oldest known flying insects and encompass a very small number of species (ca 2500 species). Larvae are strictly freshwater inhabitants; this stage lasts generally one year. The imaginal stage is extremely short, from few hours to few days, and is devoted almost entirely to reproduction. Madagascar is the fourth largest island in the world by area. It is situated in the western part of the Indian Ocean, at a distance of more than 300 km from the African coast. Madagascar belonged to Gondwana. It was first separated from the African plate (-165 M.y.), then moved to the South (-65 M.y.), before the break-off with the Indian plate (-65 M.y.). Knowledge of the Malagasy mayflies was until recently extremely poor. The program Biodiversity and Biotypology of Malagasy Freshwaters, jointly run by the French ORSTOM and the Malagasy CNRE, began a global survey of the freshwater macroinvertebrates. The systematics of several mayfly families (Tricorythidae, Polymitarcyidae, Palingeniidae,?), and other invertebrate groups (Caddisflies, Blackflies,?) was the subject of ground studies. Our present study is one part of this global program. Until the middle of the nineties, only four baetid species belonging to three different genera had been described from Madagascar. During the last six years, 25 papers were dedicated to the systematics of the Baetidae, allowing the description of 50 new species and 8 new genera. The Malagasy fauna encompasses now 22 genera and 54 species. Despite its size, Madagascar has the same diversity, at specific and generic level, as a continent. Our knowledge of the Baetidae is sufficient to perform a cladistic and biogeographical study. Our phylogenetic reconstruction allows us to propose five main lineages and to indicate, for each of them, the genera included and their features. The Malagasy fauna of Baetidae possesses a high level of endemicity: 53 of the 54 species and one third of the genera are endemic. It shows extremely strong affinities with the African fauna, as more than 90% of the genera present in Madagascar or in Africa have a distribution restricted to this area. Other components, especially Oriental and Oceanian, are negligible. These areas share with Madagascar only a few widespread genera. These African affinities are in contradiction with the geological events, especially the break-off history of Gondwana. Some explanations can be given to solve this contradiction. The most likely is that the dispersal power of the mayflies, especially of the Baetidae, is greatly underestimated. The study of recent volcanic islands, particularly of the Comoros, clearly demonstrates that the Baetidae are able to disperse over more than 300 km. Consequently, a colonisation by the Baetidae, of Madagascar from the continental Africa, after the break-off must be considered as possible. We have established scenarios explaining the biogeographical history of each of the five lineages. For four of them, Africa has to be regarded as the centre of origin. The fifth lineage probably has a Palearctic origin; Africa should be considered as a secondary centre of speciation. These lineages should have secondarily colonised Madagascar from continental Africa. This work opens up new perspectives. It allows the use of the Baetidae for faunistic and ecological studies, especially for problems related to water quality. It must be also considered as a first step for understanding the dispersion and colonisation of the islands of the western part of the Indian Ocean.