Comparative genomics suggests that the human pathogenic fungus Pneumocystis jirovecii acquired obligate biotrophy through gene loss.

Pneumocystis jirovecii is a fungal parasite that colonizes specifically humans and turns into an opportunistic pathogen in immunodeficient individuals. The fungus is able to reproduce extracellularly in host lungs without eliciting massive cellular death. The molecular mechanisms that govern this process are poorly understood, in part because of the lack of an in vitro culture system for Pneumocystis spp. In this study, we explored the origin and evolution of the putative biotrophy of P. jirovecii through comparative genomics and reconstruction of ancestral gene repertoires. We used the maximum parsimony method and genomes of related fungi of the Taphrinomycotina subphylum. Our results suggest that the last common ancestor of Pneumocystis spp. lost 2,324 genes in relation to the acquisition of obligate biotrophy. These losses may result from neutral drift and affect the biosyntheses of amino acids and thiamine, the assimilation of inorganic nitrogen and sulfur, and the catabolism of purines. In addition, P. jirovecii shows a reduced panel of lytic proteases and has lost the RNA interference machinery, which might contribute to its genome plasticity. Together with other characteristics, that is, a sex life cycle within the host, the absence of massive destruction of host cells, difficult culturing, and the lack of virulence factors, these gene losses constitute a unique combination of characteristics which are hallmarks of both obligate biotrophs and animal parasites. These findings suggest that Pneumocystis spp. should be considered as the first described obligate biotrophs of animals, whose evolution has been marked by gene losses.

La recherche et la gestion des liens dans l'investigation criminelle : le cas de la criminalité organisée.

Cette recherche se concentre sur l'enquête judiciaire qui porte sur la criminalité organisée : Cette notion doit être ici entendue comme un phénomène criminel formé par l'ensemble des organisations criminelles au sens large, soit comme tout groupement criminel, sans distinction a priori de son degré d'organisation, dont le crime organisé ou la mafia représente la forme extrême. Ce projet propose une méthode de travail basée sur un modèle d'organisation du flux de données dans l'enquête judiciaire. L'architecture de ce modèle est fondée sur deux composants interconnectés et informatisés constituant un système d'aide aux investigations criminelles. Le premier composant (banque de données nationale JANUS) est alimenté et essentiellement partagé par l'ensemble des polices Suisses. Il répertorie des informations sur les personnes et des événements qui relèvent de la criminalité organisée. Le second composant est temporaire et est sollicité lorsque des affaires particulières nécessitent l'intégration d'un extrait d'informations provenant de JANUS, ainsi que l'absorption rapide d'informations spécifiques éventuellement pas encore confirmées (par exemple provenant d'informateurs) et qui ne présentent pas un intérêt général immédiat.

Pregnancy-associated changes in host permissiveness to metastasis

THESIS SUMMARY : Metastasis is a multistep process involving tumour cell-autonomous features, the host tissue stroma of the primary tumour, the blood or lymphatic system as well as a receptive target organ. Most studies on factors influencing metastasis have concentrated on the characteristics of the disseminating tumour cell and on early steps of metastasis including invasion and angiogenesis. Although these steps are necessary for tumour cells to disseminate, it is the challenges encountered in the later steps of metastasis -survival while in the circulation and engraftment and outgrowth in the target organ -that account for the inefficiency of circulating tumour cells in establishing secondary lesions. Full understanding of the metastatic process therefore requires elucidation of the mechanisms that regulate these late steps, and in particular that determine what makes any given tissue permissive for metastatic tumour growth. To address this issue, we assessed the mechanisms whereby a physiological situation -pregnancy -can alter host permissiveness toward metastasis. We show that pregnant NOD/SCID mice -injected intravenously with tumour cells -develop more metastases than their non-pregnant counterparts irrespective of the tumour cell type. There was no direct effect of pregnancy-related circulating factors on tumour cell proliferation, and subcutaneous tumour growth does not vary between pregnant and nonpregnant animals. However, decreased elimination of tumour cells from the lung microvasculature was observed in pregnant mice, prompting us to assess whether pregnancy-related adaptations in innate immunity could account for this differential clearing. We found that natural killer (NK) cell fractions are decreased in blood and spleen of pregnant mice and that NK cell cytotoxicity is impaired, as reported previously. The use of NK-deficient mice or tumour cell lines resistant to NK killing abrogates the difference in metastasis load between pregnant and virgin mice. CD11 b+ Gr-1+ myeloid-derived suppressor cells (MDSC) have previously been shown to accumulate in tumour-bearing mice and to down-modulate NK activity. Accordingly, we show an increase in MDSC in pregnant mouse blood, spleen, lungs and liver. Depletion of MDSC prior to tumour cell injection decreased metastasis load in pregnant NOD/SCID mice but had no effect on virgin mice. Similarly, adoptive transfer of MDSC extracted from pregnant mice into virgin mice lead to increased metastasis take. In parallel, we investigated whether the lung and liver microenvironments are modified during pregnancy thereby providing a more "permissive soil" for the establishment of metastases. A comparative analysis of microarray data of pregnant mouse lungs and liver with "premetastatic niche" gene expression profiles of these organs shows that similar mechanisms could mediate an increase in lung and liver metastasis in pregnant mice and in mice harbouring an aggressive primary tumour. Several commonly up-regulated genes point towards the recruitment of myeloid cells, consistent with the accumulation of MDSC observed in pregnant mice. MDSC have never been evoked in the context of pregnancy before. Although the role of MDSC in pregnancy requires further investigation we suggest that MDSC accumulation constitutes an important and hitherto unrecognised common denominator of maternal immune tolerance and cancer immune escape. RESUME DE THESE : La métastatisation est un processus en plusieurs étapes qui implique des compétences particulières chez les cellules tumorales, le stroma de la tumeur primaire, les vaisseaux sanguins ou lymphatiques ainsi qu'un organe cible' réceptif. Jusqu'alors, la recherche s'est principalement intéressée aux facteurs qui influencent les étapes précoces de la métastatisation donc aux caractéristiques de la cellule métastatique, et aux processus tels que l'invasion et l'angiogenèse, tandis que peu d'études traitent des étapes tardives tel que la survie dans la circulation sanguine et l'établissement d'une lésion dans l'organe cible. En particulier, l'élucidation des facteurs qui déterminent la permissivité d'un tissu à la greffe de cellules disséminantes est indispensable à la compréhension de ce processus complexe qu'est la métastatisation. Nous proposons ici un modèle de souris récapitulant les étapes tardives de la métastatisation dans un contexte d'une permissivité accrue aux métastases chez la souris gravide, et nous évaluons les mécanismes impliqués. Les souris gestantes développent plus de métastases après l'injection intraveineuse de cellules tumorales, indépendamment du type de tumeur d'origine. Les taux élevés d'hormones et de facteurs de croissance chez la souris gravide n'inflúencent pas la prolifération des cellules tumorales et fa croissance de tumeurs sous-cutanées n'est pas non plus accélérée par la gestation. En revanche, une fois injectées, les cellules tumorales sont éliminées ` moins rapidement des vaisseaux pulmonaires chez la souris gravide que chez les contrôles. Cette observation est compatible avec un effet de la gestation sur l'immunité innée et nous avons mis en évidence une diminution des proportions de cellules NK (natural killer) dans le sang et la rate en particulier, ainsi qu'une cytotoxicité moindre envers des cellules tumorales. En utilisant des souris déficientes en cellules NK ou en injectant des cellules résistantes à l'attaqué par des cellules NK, la différence entre souris gestantes et non-gestantes disparaît. Il a été démontré chez des souris porteuses de tumeurs, que l'accumulation de cellules immunosuppressives de la lignée myélo-monocytaire (ou MDSC pour myeloid-derived suppressor tells) pouvait être responsable d'une inhibition de l'activité de cellules NK. Des nombres augmentés de ces cellules, caractérisées par les marqueurs de surface CD11b et Gr-1, ont été trouvés dans le sang, la rate, les poumons et le foie de souris gravides. Leur rôle dans la métastatisation est démontré par le fait que leur dépletion diminue le nombre de lésions secondaires chez la souris gestante, tandis que leur transfert dans des souris non-gestantes augmente le taux de métastases. L'utilisation de puces à ADN sur les foies et poumons de souris gravides a permis de mettre en évidence des différences d'expression génique proches de celles observées dans l'établissement de niches pré-métastatiques. Ceci suggère que des mécanismes similaires pourraient être responsables d'une permissivité accrue aux métastases chez la souris gravide et chez la souris porteuse d'une tumeur primaire agressive, telle que, en particulier, l'accumulation de cellules immunosuppressives dans les organes cibles. C'est la première fois que l'accumulation de MDSC est évoquée chez la souris gravide et nous proposons ici que celles-ci jouent un rôle dans la tolérance immunitaire envers le foetus et sont responsables de l'échappement de cellules tumorales injectées à la surveillance immunitaire par des cellules NK.

Transobturator surgery for female stress incontinence: a comparative anatomical study of outside-in vs inside-out techniques.

OBJECTIVE To assess the specific risks of injury to neural and vascular structures inherent in two approaches to transobturator surgery for inserting a suburethral sling, i.e. the outside-in (standard technique) and inside-out approaches. MATERIALS AND METHODS The study comprised seven cadavers, providing 14 obturator regions. Five specimens had a tape inserted outside-in on one side, and inside-out on the other; of the remaining two cadavers, one had an inside-out tape and one an outside-in tape, bilaterally. After tape insertion, the cadavers were dissected. Particular attention was paid to the distances between the tape and the deep external pudendal vessels, and between the tape and the posterior branch of the obturator nerve. RESULTS With the inside-out technique, the safety margins were reduced, and the external pudendal vessels and the posterior branch of the obturator nerve were at greater risk of injury. CONCLUSION The two techniques are not equivalent, with a lower risk of injury to vascular and nerve structures with the outside-in technique.

A comparative study between crosslinked sodium hyaluronate (healaflow glaucoma implant) and high molecular weight sodium hyaluronate (healon 5) as subconjunctival spacers injected into failed filtration blebs during needling with MMC

Purpose: To compare the efficacy and safety of a slowly resorbable glaucoma implant, Healaflow (HF) and Healon 5 (H5) as spacers injected into failing filtration blebs at the time of bleb needlings, augmented with MMC Methods: Eighty-four glaucoma patients with filtration bleb failure following glaucoma surgery (deep sclerectomy/ trabeculectomy) underwent bleb needling using either HF (n=44) or H5 (n=40) as an intrableb viscoelastic spacer. All needlings were augmented by MMC and performed in the operating room, by a single surgeon. The choice between HF and H5 was randomised. Postoperative data for IOP, number of glaucoma medications (GMs), antimetabolite injections (AMIs), bleb morphology and all complications were recorded at day 1 (D1), weeks 1 (W1), 4 (W4), month 6 (M6) and last visit. Success was defined as IOP≤21mmHg and 20% reduction in IOP from baseline Results: Age was comparable between groups (HF vs H5; 66 vs 72 years; p=0.13*). There were no differences in mean postoperative IOP or GMs at any time point between groups (mean IOP = HF vs H5; prior to needling=19.8mmHg vs 18.7mmHg, p=0.48*; D1=10.4mmHg vs 10.2mmHg, p=0.85*; W1=12.3mmHg vs 13.8mmHg, p=0.33*; W4=15.7mmHg vs 15.1mmHg, p=0.69*; M6=14.3mmHg vs 13.8mmHg, p=0.69*. Mean GMs= HF vs H5; prior to needling=1.7 vs 1.7, p=0.96*; M6=0.9 vs 1.1, p=0.66*). Success rates were comparable between HF and H5 groups (74% vs 71%). Requirements for AMIs were similar between groups; post filtration surgery and prior to needling (mean AMIs 1.54 vs 1.65 p=0.82*) and within the first six months following needling ( 0.77 vs 0.65 p=0.70*). Complication rates were infrequent in both groups (9%, HF; 3%, H5) * two-sample t-test Conclusions: The early to mid-term success rates of needlings using intrableb spacers were high. However there were no differences observed between the use of crosslinked and high molecular weight sodium hyaluronate

Self reported physical activity, public health, and perceived environment: results from a comparative European study

The focus of physical activity promotion is moving from methods for increasing health enhancing physical activity on the individual level to higher level strategies including environmental and policy approaches. Scientific inquiry, traditionally related to individual-based strategies, requires adaptation and refinement when environmental and policy changes become more relevant. The objective of this study is to investigate the significance for behaviour and health of community-based environments that encourage physical activity. DESIGN AND SETTING The article presents data and results from a cross sectional comparative survey of the general population in six European countries (Belgium, Finland, Germany (East and West), Netherlands, Spain, Switzerland). Specifically, the relation between perceived community-based opportunities for physical activity, self reported physical activity, and self rated health status is investigated. PARTICIPANTS Representative samples of general populations (adults 18 years or older). Overall response rate: 53.5%. Sample sizes realised: Belgium: n=389; Finland: n=400; Germany (East): n = 913; Germany (West): n=489; Netherlands: n=366; Spain: n=380; Switzerland: n=406. MAIN RESULTS Analyses show that best opportunities are reported by people who are lightly to moderately physically active. People's self rated health is moderately, but significantly associated with both perceived opportunities, and physical activity itself. These predictors interact in that especially for women, the health impact of physical activity is more pronounced in case of good opportunities. CONCLUSIONS The paper shows the potential of opportunities within residential and community environments with regard to physical activity, both for behaviour and health. Opportunities may enable the population, especially women, to develop an active lifestyle, and thus improve their health. Future studies with objective indicators for physical activity related environments should test the findings that are based on perceptions.

Comparative functional analysis of two fibroblast growth factor receptor 1 (FGFR1) mutations affecting the same residue (R254W and R254Q) in isolated hypogonadotropic hypogonadism (IHH).

FGFR1 mutations have been identified in both Kallmann syndrome and normosmic HH (nIHH). To date, few mutations in the FGFR1 gene have been structurally or functionally characterized in vitro to identify molecular mechanisms that contribute to the disease pathogenesis. We attempted to define the in vitro functionality of two FGFR1 mutants (R254W and R254Q), resulting from two different amino acid substitutions of the same residue, and to correlate the in vitro findings to the patient phenotypes. Two unrelated GnRH deficient probands were found to harbor mutations in FGFR1 (R254W and R254Q). Mutant signaling activity and expression levels were evaluated in vitro and compared to a wild type (WT) receptor. Signaling activity was determined by a FGF2/FGFR1 dependent transcription reporter assay. Receptor total expression levels were assessed by Western blot and cell surface expression was measured by a radiolabeled antibody binding assay. The R254W maximal receptor signaling capacity was reduced by 45% (p<0.01) while R254Q activity was not different from WT. However, both mutants displayed diminished total protein expression levels (40 and 30% reduction relative to WT, respectively), while protein maturation was unaffected. Accordingly, cell surface expression levels of the mutant receptors were also significantly reduced (35% p<0.01 and 15% p<0.05, respectively). The p.R254W and p.R254Q are both loss-of-function mutations as demonstrated by their reduced overall and cell surface expression levels suggesting a deleterious effect on receptor folding and stability. It appears that a tryptophan substitution at R254 is more disruptive to receptor structure than the more conserved glutamine substitution. No clear correlation between the severity of in vitro loss-of-function and phenotypic presentation could be assigned.

Comparative phylogeography and postglacial colonization routes in Europe.

The Quaternary cold periods in Europe are thought to have heavily influenced the amount and distribution of intraspecific genetic variation in both animals and plants. The phylogeographies of 10 taxa, including mammals (Ursus arctos, Sorex spp., Crocidura suaveolens, Arvicola spp.), amphibians (Triturus spp.), arthropods (Chorthippus parallelus), and plants (Abies alba, Picea abies, Fagus sylvatica, Quercus spp.), were analysed to elucidate general trends across Europe. Only a small degree of congruence was found amongst the phylogeographies of the 10 taxa, but the likely postglacial colonization routes exhibit some similarities. A Brooks parsimony analysis produced an unrooted area phylogram, showing that: (i) the northern regions were colonized generally from the Iberic and Balkanic refugia; and (ii) the Italian lineages were often isolated due to the presence of the Alpine barrier. The comparison of colonization routes highlighted four main suture-zones where lineages from the different refugia meet. Some of the intraspecific genetic distances among lineages indicated a prequaternary divergence that cannot be connected to any particular cold period, but are probably related mainly to the date of arrival of each taxon in the European continent. As a consequence, molecular genetics so far appears to be of limited use in dating Quaternary events.

The influence of work context on demand for master's studies: a comparative analysisi between Brasil and Spain

The communication presents the results of an investigation of exploratory and comparative character which objective is to analyze the influence of the actual labour situation into the demand of official master studies in the field of education. The study has been developed in two countries with a very different labour situation: Brasil, country of economic expansion and Spain, in recession due to the actual economic crisis. In that sense, the study provides data for deep thinking about the influence of the constriction or expansion of employment on the behaviour and demand of the students who access master studies and on how the previous formative and labour trajectory affects their expectations, demands and future projects. The working methodology is qualitative and the strategy for data collection the “focus group”. As a first approach, two groups of discussion have been formed with master students. A first one with students from Universidad de Barcelona- España and another one with members of Universidade do Vale do Itajaí- Brasil. Then, we constituted a mixed group of discussion in order to analyze differences and similarities.