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The small GTPase RhoA is involved in a variety of fundamental processes in normal tissue. Spatiotemporal control of RhoA is thought to govern mechanosensing, growth, and motility of cells, while its deregulation is associated with disease development. Here, we describe the generation of a RhoA-fluorescence resonance energy transfer (FRET) biosensor mouse and its utility for monitoring real-time activity of RhoA in a variety of native tissues in vivo. We assess changes in RhoA activity during mechanosensing of osteocytes within the bone and during neutrophil migration. We also demonstrate spatiotemporal order of RhoA activity within crypt cells of the small intestine and during different stages of mammary gestation. Subsequently, we reveal co-option of RhoA activity in both invasive breast and pancreatic cancers, and we assess drug targeting in these disease settings, illustrating the potential for utilizing this mouse to study RhoA activity in vivo in real time.

Pancreatic cancer has a devastating prognosis, with an overall 5-year survival rate of ~8%, restricted treatment options and characteristic molecular heterogeneity. SerpinB2 expression, particularly in the stromal compartment, is associated with reduced metastasis and prolonged survival in pancreatic ductal adenocarcinoma (PDAC) and our genomic analysis revealed that SERPINB2 is frequently deleted in PDAC. We show that SerpinB2 is required by stromal cells for normal collagen remodelling in vitro, regulating fibroblast interaction and engagement with collagen in the contracting matrix. In a pancreatic cancer allograft model, co-injection of PDAC cancer cells and SerpinB2−/−mouse embryonic fibroblasts (MEFs) resulted in increased tumour growth, aberrant remodelling of the extracellular matrix (ECM) and increased local invasion from the primary tumour. These tumours also displayed elevated proteolytic activity of the primary biochemical target of SerpinB2—urokinase plasminogen activator (uPA). In a large cohort of patients with resected PDAC, we show that increasing uPA mRNA expression was significantly associated with poorer survival following pancreatectomy. This study establishes a novel role for SerpinB2 in the stromal compartment in PDAC invasion through regulation of stromal remodelling and highlights the SerpinB2/uPA axis for further investigation as a potential therapeutic target in pancreatic cancer.

A number of naturally occurring isoforms of the tumour suppressor protein p53 have been discovered, which appear to have differing roles in tumour prevention or promotion. We are investigating the tumour-promoting activities of the ?133p53 isoform using our mouse model of ?133p53 (?122p53). Here, we report that tumours from ?122p53 homozygous mice show evidence of invasion and metastasis and that ?122p53 promotes migration though a 3-dimensional collagen matrix. We also show that ?122p53 and ?133p53 promote cell migration in scratch wound and Transwell assays, similar to the ‘gain-of-function’ phenotypes seen with mutant p53. Using the well-defined B16 mouse melanoma metastatic model, we show that ?122p53 leads to faster generation of lung metastases. The increased migratory phenotypes are dependent on secreted factors, including the cytokine interleukin-6 and the chemokine CCL2. We propose that ?122p53 (and ?133p53) acts in a similar manner to ‘gain-of-function’ mutant p53 proteins to promote migration, invasion and metastasis, which may contribute to poor survival in patients with ?133p53-expressing tumours.

Although a wide number of breast cancer susceptibility alleles associated with various levels of risk have been identified to date, about 50% of the heritability is still missing. Although the major BRCA1 and BRCA2 genes are being extensively screened for truncating and missense variants in breast and/or ovarian cancer families, potential regulatory variants affecting their expression remain largely unexplored. In an attempt to identify such variants, we focused our attention on gene regulation mediated by microRNAs (miRs). We screened two genes, MIR146A and MIR146B, producing miR-146a and miR-146b-5p, respectively, that regulate BRCA1, and the 3′- untranslated regions (3′-UTRs) of BRCA1 and BRCA2 in the GENESIS French national case/control study (BRCA1- and BRCA2-negative breast cancer cases with at least one sister with breast cancer and matched controls). We identified one rare variant in MIR146A, four in MIR146B, five in BRCA1 3′-UTR and one in BRCA2 3′-UTR in 716 index cases and 619 controls. Among these 11 rare variants, 7 were identified each in 1 index case. None of the three relevant MIR146A/MIR146B variants affected the pre-miR sequences. The potential causality of the four relevant BRCA1/BRCA2 3′-UTRs variants was evaluated with luciferase reporter assays and co-segregation studies, as well as with bioinformatics analyses to predict miRs-binding sites, RNA secondary structures and RNA accessibility. This is the first study to report the screening of miR genes and of BRCA2 3′-UTR in a large series of familial breast cancer cases. None of the variant identified in this study gave convincing evidence of potential pathogenicity.

E-cadherin-mediated cell-cell junctions play a prominent role in maintaining the epithelial architecture. The disruption or deregulation of these adhesions in cancer can lead to the collapse of tumor epithelia that precedes invasion and subsequent metastasis. Here we generated an E-cadherin-GFP mouse that enables intravital photobleaching and quantification of E-cadherin mobility in live tissue without affecting normal biology. We demonstrate the broad applications of this mouse by examining E-cadherin regulation in multiple tissues, including mammary, brain, liver, and kidney tissue, while specifically monitoring E-cadherin mobility during disease progression in the pancreas. We assess E-cadherin stability in native pancreatic tissue upon genetic manipulation involving Kras and p53 or in response to anti-invasive drug treatment and gain insights into the dynamic remodeling of E-cadherin during in situ cancer progression. FRAP in the E-cadherin-GFP mouse, therefore, promises to be a valuable tool to fundamentally expand our understanding of E-cadherin-mediated events in native microenvironments.

An increasing number of breast cancer patients develop brain metastases (BM). Standard-of-care treatments are largely inefficient, and breast cancer brain metastasis (BCBM) patients are considered untreatable. Immunotherapies are not successfully employed in BCBM, in part because breast cancer is a “cold” tumor and also because the brain tissue has a unique immune landscape. Here, we generate and characterize immunocompetent models of BCBM derived from PyMT and Neu mammary tumors to test how harnessing the pro-senescence properties of doxorubicin can be used to prime the specific immune BCBM microenvironment. We reveal that BCBM senescent cells, induced by doxorubicin, trigger the recruitment of PD1-expressing T cells to the brain. Importantly, we demonstrate that induction of senescence with doxorubicin improves the efficacy of immunotherapy with anti-PD1 in BCBM in a CD8 T cell-dependent manner, thereby providing an optimized strategy to introduce immune-based treatments in this lethal disease. In addition, our BCBM models can be used for pre-clinical testing of other therapeutic strategies in the future.

Localizada sobre una zona plantar, la queratodermia dinámica difiere de los queratomas puntuales ligados al apoyo, al roce articular o a presiones indeterminadas. Durante la marcha, las células queratinocíticas se enmarañan, se deslizan las unas sobre las otras, forman columnas que se inclinan hacia uno u otro lado, pero permanecen en conjunto. La unión dermoepidérmica conecta la dermis con la epidermis y facilita su flexibilidad durante el paso. Las líneas de Langer, o dermatoglifos, marcan el anclaje epidérmico a lo largo de líneas paralelas, hundidas, que resisten las tracciones de la epidermis, lo que explica el pliegue de las capas superficiales entre las líneas de Langer, el revestimiento epidérmico. El mecanismo continúa en tres etapas: primero una cresta gruesa y seca, a continuación el espolón y, a nivel del corion, el banderín o la lengüeta. Se observa el banderín sobre todo en los dedos en supra o infraductus, la lengüeta bajo la arcada anterior de un pie cavo y el espolón a nivel del reborde posterior del talón. La piel plantar presenta los efectos del cambio del eje corporal en la estática del pie, produciendo una rotación dermoepidérmica alrededor del eje de una articulación. El callo dinámico, síntesis puntual del conjunto, presenta un eje oblicuo.

Dans le cadre d’un PHRC pour évaluer le dépistage par l’analyse protéomique plasmatique, nous avons inclus 82 femmes à risque héréditaire de cancer de l’ovaire. Nous rapportons ici les conséquences du dépistage avec les examens habituels. Un dosage du CA 125 et une échographie transvaginale étaient réalisés tous les six mois. Soixante-douze patientes sont évaluables. Deux cas incidents de carcinose péritonéale (tumeurs séreuses de haut grade de stade FIGO IIIB) ont été découverts lors des tests de dépistage chez deux femmes asymptomatiques mutées BRCA1 (2,7 %). Il n’y a pas d’autre cas de cancer primitif, mais une métastase ovarienne d’un cancer du sein. Quarante femmes sont sorties d’étude, 32 pour chirurgie annexielle préventive. En totale concordance avec la littérature, le dépistage par les examens actuels, même avec une fréquence rapprochée, est inefficace pour déceler précocement ces tumeurs pelviennes. Ce dépistage parait encore plus inopérant que dans les formes sporadiques. Aujourd’hui, il est important de rechercher les mutations délétères des deux gènes (BRCA1/2) associés au risque de cancer du sein et de l’ovaire pour expliquer aux femmes qui ont une mutation que la seule solution connue pour limiter les risques de décès par cancer est la chirurgie préventive dont il ne reste à discuter que le moment.

Face à un patient atteint d’une maladie psychiatrique telle qu’un trouble de la personnalité, le médecin oncologue ne peut l’occulter et doit adapter sa prise en charge. Certains traits de caractère pathologiques nécessitent des aménagements pour assurer la bonne compréhension par le patient du diagnostic et du traitement mais aussi pour obtenir la meilleure adhésion aux soins. Étant donné qu’il n’existe pas de traitement pharmacologique spécifique, la collaboration entre oncologues et équipe de psycho-oncologie s’avère indispensable. L’intrication de la maladie somatique et du trouble psychiatrique impose des temps d’échanges entre équipes soignantes, faits d’explications et de conseils. Des exemples cliniques viendront illustrer chaque trouble de la personnalité et mettront en perspective les problématiques soulevées par le trouble psychiatrique. Les principes de traitement seront évoqués dans un deuxième temps.

Purpose: To investigate the medical and psychosocial factors determining the time to prophylactic surgery of unaffected women carriers of a deleterious BRCA1/2 mutation.Methods: Prospective study on a French national cohort of unaffected BRCA1/2 carriers (N = 244); multivariate Cox proportional hazard modeling.Results: Median follow-up time was 2.33 years (range, 0.04–6.84 years). Time to surgery was shorter when the psychological impact of BRCA1/2 result disclosure was stated to be higher (P = 0.01). Those who intended to opt for prophylactic surgery before being tested did so faster and more frequently after test disclosure than those who were undecided/opposed. The older the women were, the faster their uptake of risk-reducing salpingo-oophorectomy (adjusted hazard ratio >2.95; P < 0.001) was; the uptake of those with at least two children was also faster (adjusted hazard ratio = 2.51; [1.38–4.55]). Those who opted most quickly for risk-reducing mastectomy more frequently had a younger child at the time of testing (adjusted hazard ratio = 4.63 [1.56–13.74]). Time to surgery was shorter when there was a first-degree relative with ovarian/breast cancer (P = 0.01).Conclusion: Time to prophylactic surgery depends on the stated psychological impact of disclosure and on women's cognitive anticipation of surgery after adjusting on sociodemographic characteristics.

To investigate the medical and psychosocial factors determining the time to prophylactic surgery of unaffected women carriers of a deleterious BRCA1/2mutation.

A 380 m thick Aptian platform to basin transition has been studied along a 16 km long transect of excellent and continuous outcrops in NE Spain. The series has been dated using biostratigraphy (foraminifera and ammonites) and carbon-isotope stratigraphy, and has been subdivided at four scales of depositional sequences. The Aptian marine succession is subdivided into two-large scale sequences separated by a middle Aptian sub-aerial exposure surface. A characteristic trend of the floral-faunal fossil assemblages is present, which evolves from orbitolinid-ooid dominated ramps in Sequence I-1, to a coral-stromatoporoid-microbialite dominated platform in Sequence I-2, to a rudist-dominated platform top in Sequence II-1, and finally to a second episode of orbitolinid-ooid dominated ramp system in Sequence II-2. There was an influx of siliciclastic sediments at the base and at the top of this succession. The detailed carbon-isotope curve measured along the Miravete section and covering almost the complete Aptian succession, is compared with published Aptian curves recorded in both basinal and carbonate platform settings along the northern and southern NeoTethys margins. It shows that the Galve sub-basin curve represents all the major isotope excursions of the lower and upper Aptian, in a dominantly shallow-water succession.

Ce document est un guide pour les entreprises utilisant ou réalisant pour des clients des Contrôles ou Examens Non Destructifs. Il reprend l'ensemble des étapes de ce type de prestations (sur la base d'un travail déjà réalisé au niveau de la région PACA) et identifie la répartition des responsabilités entre : entreprise utilisatrice (EU)/ donneur d'ordre, ensemblier ou maître d'oeuvre (MOE) et entreprise intervenante (EI) en radiographie. Il rappelle certaines obligations et propose de " bonnes pratiques " recueillies auprès d'industriels du domaine. Il convient d'abord de préciser les points forts qui définissent cette répartition de responsabilités. Celle-ci s'appuie en premier lieu sur une relation contractuelle entre les 2 ou 3 parties prenantes. L'entreprise utilisatrice confie ainsi à l'ensemblier/maître d'oeuvre et luimème à l'entreprise intervenante les responsabilités de faire réaliser et/ou de réaliser les prestations dans le respect des exigences réglementaires et dans les conditions de sécurité optimum. Chaque entité assure donc ses responsabilités, ceci étant la base fondamentale et incontournable de leur implication pertinente dans la tenue des objectifs opérationnels et de sécurité. Cette relation contractuelle se construit en particulier à certaines étapes clés : l'identification précise des objectifs et conditions de réalisation (déclinaison des exigences) lors de l'élaboration du cahier des charges ; identification des éléments devant être précisés pour maîtriser l'intervention lors de la revue d'offre ; l'acceptation ou l'identification des réserves à lever lors de la revue de commande ; l'identification nominative des personnes responsables du contrat et des travaux dans chaque entité ; et enfin l'élaboration du plan de prévention. Dans certains cas, pour la recherche des informations nécessaires au cahier des charges, l'élaboration du plan de prévention, la détermination des conditions radiologiques, la définition de la zone d'opération, etc., l'entité concernée peut, faute de compétences existantes ou disponibles, faire appel au conseil et support de sociétés compétentes. Cela ne la dégage pas de ses responsabilités mais peut lui permettre de répondre efficacement aux exigences à satisfaire. NOTA : en fonction des conseils proposés, si l'entité ne souhaite pas suivre les recommandations, elle doit le signifier clairement et la société d'assistance lui rappeler ses obligations. De plus, l'entité qui sous-traite une partie de ses activités s'assure de la bonne réalisation par la surveillance qu'elle juge utile. Enfin, le retour d'expérience est une démarche qui, faite par l'entreprise intervenante ou capitalisée par l'entreprise utilisatrice, est de nature à améliorer les conditions de réalisation dans le temps dès qu'il est réutilisé comme données d'entrée pour de nouvelles interventions.

Für das Kambrium der Iberischen Ketten wird eine neu überarbeitete Synthese der Ökosysteme und der Veränderungen der benthischen Lebensgemeinschaften vorgestellt. Sie basiert auf der Anwendung von Konzepten zur Biodiversität, Sequenz- und Eventstratigraphie. Die Entwicklung der Iberischen Plattform im Kambrium wurde im Wesentlichen von fünf Faktoren beeinflusst: regionaler Tektonik, lokalem Klima, Meeresspiegelschwankungen, terrigener Schüttung und Karbonatbildung. Die Süddrift des Westrandes von Gondwana und die daraus folgende Tendenz zur Abkühlung bewirkten einen allmählichen Wechsel der Zusammensetzung der Plattformsedimente und der benthischen Lebensgemeinschaften. Dieser Trend von arid subtropischem zu temperiertem Klima dokumentiert sich in der Abnahme und dem Verschwinden von (i) Ooiden, Evaporitrelikten und mikrobiellen Karbonaten, (ii) Eocrinoiden-Schwamm Rasen, (iii) hochenergetisch bioklastischen Karbonaten in schmalen Zonen auf tektonischen Hochgebieten, (iv) Trilobiten-dominierten Lebensgemeinschaften auf tonig-siltigen Böden und (v) einer Tendenz zu weniger fossilreichen Lebensgemeinschaften und dem Vorherrschen linguliformer Brachiopodengemeinschaften in sandigen Flachwassergebieten. Die kambrischen Gesteine der Iberischen Ketten werden in vier sedimentäre Megasequenzen eingeteilt (zusammengesetzt aus hochfrequenten Zyklen und niedrigfrequenten Stapelungsmustern). Die Änderungen der Biodiversität, insbesondere die der Trilobiten, entsprechen ungefähr den Großschwankungen der Sedimentzufuhr. Jede Schwankung der Diversität (Expansion/Kontraktion) stimmt im Großen und Ganzen überein mit dem Intervall, das einer transgressiv-regressiven Abfolge einer Megasequenz entspricht. Lediglich das Valdemiedes Extinction Event (an der Grenze Unter-Mittelkambrium) ist kein rein ökologisches Phänomen, sondern spiegelt die Zuwanderung und Entstehung völlig neuer Gattungen und Familien wider.

In the carbonate platforms of the western Gondwana margin, the extinction recorded at the Lower–Middle Cambrian boundary is accompanied by a profound change in the style of carbonate deposition. The Láncara Formation of the Esla nappe (Cantabrian Mountains, northern Spain) contains a distinct sedimentary turnover due to a combination of tectonism, eustatic fluctuations, and immigration and colonization of new benthic communities, such as the youngest archaeocyathan assemblage of the entire Iberian Peninsula. During latest Early Cambrian times, a regressive trend is recorded in the Láncara Formation. This regression was recorded on a peritidal-dominant, homoclinal ramp that is topped by a tectonically induced discontinuity (D1). The latter surface marks the beginning of a last prograding, regressive tendency recorded on an intra-shelf ramp with ooidal/bioclastic shoals protecting archaeocyathan-microbial patch reefs. The overlying discontinuity (D2) corresponds to a major erosive unconformity, which coincides with the Lower–Middle Cambrian boundary in the Cantabrian Mountains. The subsequent, long-term, earliest Middle Cambrian rise in relative sea-level allowed deposition of low-relief, bioclastic shoals bearing a diverse and cosmopolitan assemblage of benthic fauna. Finally, the previous evolution is bounded by a third discontinuity (D3), which marks the beginning of a rhythmic sedimentation indicative of a major phase of tectonic breakdown and drowning of platforms recognised throughout southwestern Europe. Two associations of calcimicrobes occur in the latest Early Cambrian regressive trend of the Láncara Formation: (i) Proauloporaand Subtifloraare identified in peritidal, high-energy settings, lacking self-supported structures, whereas (ii) intergrowths of Epiphyton, Renalcisand Girvanellaencrusted branching colonies and solitary archaeocyaths in protected (back-shoal) patch reefs. The latest Early Cambrian regression is correlated in southwestern Europe in both siliciclastic (Iberian Chains and Ossa–Morena) and carbonate-dominant platforms (Cantabrian Mountains, Montagne Noire and Sardinia). Its tops are recognised as diachronous unconformities ranging in age from early Bilbilian to the Bilbilian–Leonian or Lower–Middle Cambrian boundary.

Informed probands are key actors for disclosing genetic information to their relatives when a mutation has been identified in the family. The objectives were to study women's attitudes towards the family disclosure of positive breast cancer genetic testing results and to determine the predictive factors of the diffusion patterns observed. A national multi-center cross-sectional survey was carried out at five French cancer genetic clinics during a 1-year period. Self-administered questionnaires were completed after the consultation by 84.5% (398/471) of women attending breast cancer genetic clinics for the first time. Among the 383 respondents who had at least one living first-degree relative to inform, 8.6% would inform none, 33.2% would inform at least one of them, and 58.2% would inform all of them. The sibship would be the most frequently informed blood relatives, sisters in 86.9% and brothers in 79% compared with mother in 71.4%, children in 70.4%, and father in 64.9%. Women of the family would be more frequently informed than men (P &lt; 0.05). After multivariate adjustment, age, the fact to be affected by cancer, the number of daughters, and the emotional disturbance due to cancer in a close relationship were the main determinants (P &lt; 0.05) of the diffusion patterns observed. The first step of the relatives' attendance to genetic counseling and the proband's willingness to disclose breast cancer genetic tests results was high in this study and was clearly dependent on the women's personal and emotional characteristics. Am. J. Med. Genet. 94:13–18, 2000. © 2000 Wiley-Liss, Inc.

Objective.The aim of this study was to evaluate the impact of platinum dose intensity on pathological response rate and overall survival in patients with advanced ovarian adenocarcinoma.

Early stage epithelial ovarian carcinoma is defined pathologically as a tumor strictly limited to one or both ovaries without any extra-ovarian disease (i.e., Stage IA or B of the International Federation of Gynecology and Obstetrics (FIGO) classification). This diagnosis can be obtained only after an exhaustive surgical staging procedure, performed as soon as the diagnosis of epithelial invasive ovarian carcinoma is established. This staging surgery currently encompasses a peritoneal cytology, the thorough inspection of all the visceral and parietal peritoneal surfaces with biopsy of any abnormality, total abdominal hysterectomy and bilateral salpingo-oophorectomy (TAH + BSO), random peritoneal biopsies, omentectomy, appendectomy and bilateral pelvic and para-aortic lymphadenectomies, up to the left renal vein. The results of this staging procedure and its indications are discussed. In all of the cases, the radical removal of the pathologic adnexa is indicated, along with the complete peritoneal and retroperitoneal staging. While fertility-sparing surgery (avoiding hysterectomy and contralateral adnexectomy, if possible) seems to be safe for young women, a TAH + BSO is the rule for the others. Adjuvant chemotherapy can be omitted in well-differentiated tumors with a negative staging operation, but currently it remains indicated in all other cases. Indeed, the ultimate goal in early stage ovarian carcinoma is to not impair by inadequate management the high chance of a cure. Semin. Surg. Oncol. 19:36–41, 2000. © 2000 Wiley-Liss, Inc.

Since there is a lack of common family profile among BRCA1-gene carriers, and since the risk of being a mutation carrier is not limited to women with a family history of breast or ovarian cancer, multivariate statistical analysis using the logistic-regression model was carried out, to discriminate between sporadic cases and BRCA1-breast cancers (BRCA1-BCs), especially when information about the family history of breast/ovarian cancer and ethnicity are irrelevant or unavailable, in order to offer specific medical treatment to this population. We examined 32 BRCA1-BCs selected at cancer genetic clinics and 200 consecutive controls without family history of breast cancer for age at onset and current morphological parameters. Following the multivariate analysis, 3 parameters only, namely, early age at cancer onset [odds ratio (OR) for each year = 1.16; p &lt; 0.0001], estrogen-receptor negativity (OR = 5.7; p = 0.01) and poor differentiation (OR = 5; p = 0.03) were found significant factors for predicting BRCA1-carrier status. The expected impact in BRCA1 screening of our model was estimated using data on 5 700 breast-cancer cases from a hospital-based registry. Only 50 and 15% of tumours with early age at onset below 35 years present one or the other 2 discriminant parameters respectively. Consequently, whereas the probability of finding a BRCA1 mutation is rated low (6.2%) when the sole criterion of early onset up to the age of 35 years is used, based on our model, in the sub-group of women with a tumor that is both estrogen-receptor-negative and poorly differentiated the mutation-detection rate is predicted to be above the 10% chance level recommended by the ASCO guidelines. This sub-group of women, representing about 1% of all breast-cancer cases in Western countries, consequently deserves to be tested. Int. J. Cancer (Pred. Oncol.) 84:263–267, 1999. © 1999 Wiley-Liss, Inc.