Purification of native HBHA from Mycobacterium avium subsp. paratuberculosis.

International audienceBACKGROUND: Paratuberculosis remains today a major global problem in animal health, especially for dairy cattle. However, the diagnosis of its etiologic agent, Mycobacterium avium subsp. paratuberculosis (Map), still lacks sensitivity because of the lack of available antigens. Little is known about the virulence factors for this pathogen. In this study we have developed a method to produce and purify the heparin-binding hemagglutinin (HBHA), a major adhesin of Mycobacteria, from a culture of Map. FINDINGS: For this extremely slow-growing Mycobacterium, a culture was established in a 3-liter bioreactor. Using the bioreactor the amount of the Map biomass was increased 5-fold compared to a classical culture in flasks. The map-HBHA was purified from a Map lysate by heparin-Sepharose chromatography on HiTrap columns. Binding of map-HBHA onto heparin-Sepharose can be reduced in the presence of salt. Consequently, all steps of sample preparation and column equilibration were carried out in 20 mM Tris-HCl (pH 7.2). The map-HBHA was eluted by a linear NaCl gradient. High resolution mass spectrometry analyses revealed that the native form of map-HBHA has posttranslational modifications, including the removal of the initiation methionine, acetylation of the alanine residue at the N-terminal extremity and the presence of methylated lysines in the C-terminal domain of the protein. CONCLUSIONS: An optimized culture of Map in a bioreactor was established to purify the native map-HBHA from a Map lysate by heparin-Sepharose chromatography. The availability of this antigen offers the possibility to study the structure of the protein and to examine its role in pathogenicity, in particular to better understand the specific interactions of Map with the intestinal tissue. The map-HBHA obtained in its native immunogenic form may also be useful to improve the diagnostic test, especially for the development of a new T-cell-based interferon gamma release assays

A mineralogical study in contrasts: highly mineralized whale rostrum and human enamel

The outermost enamel of the human tooth and the rostrum of the whale Mesoplodon densirostris are two highly mineralized tissues that contain over 95wt.% mineral, i.e., bioapatite. However, the same mineral type (carbonated hydroxylapatite) does not yield the same material properties, as revealed by Raman spectroscopy, scanning electron microscopy, electron microprobe analysis, and synchrotron X-ray diffraction analysis. Overall, the outermost enamel of a tooth has more homogeneous physical and chemical features than the rostrum. Chemical comparison of rostrum and enamel shows bioapatite in the rostrum to be enriched in Na, Mg, CO3, and S, whereas the outermost enamel shows only a slightly enriched Cl concentration. Morphologically, mineral rods (at tens of μm scale), crystallites and prisms (at μm and sub-μm scale), and platelets (at tens of nm scale) all demonstrate less organized texture in the rostrum than in enamel. Such contrasts between two mineralized tissues suggest distinct pathways of biomineralization, e.g., the nature of the equilibrium between mineral and body fluid. This study illustrates the remarkable flexibility of the apatite mineral structure to match its chemical and physical properties to specific biological needs within the same animal or between species.The work was partially funded by NIH grant 1R21AR055184-01A2 and SRF for ROCS, SEM

Molecular characterization of Coxiella burnetii isolates by infrequent restriction site-PCR and MLVA typing

BACKGROUND: Coxiella burnetii, the causative agent of Q fever, has a wide host range. Few epidemiological tools are available, and they are often expensive or not easily standardized across laboratories. In this work, C. burnetii isolates from livestock and ticks were typed using infrequent restriction site-PCR (IRS-PCR) and multiple loci variable number of tandem repeats (VNTR) analysis (MLVA). RESULTS: By applying IRS-PCR, 14 C. burnetii isolates could be divided into six groups containing up to five different isolates. Clustering as deduced from MLVA typing with 17 markers provided an increased resolution with an excellent agreement to IRS-PCR, and with the plasmid type of each strain. MLVA was then applied to 28 additional C. burnetii isolates of different origin and 36 different genotypes were identified among the 42 isolates investigated. The clustering obtained is in agreement with published Multiple Locus Sequence Typing (MLST) data. Two panels of markers are proposed, panel 1 which can be confidently typed on agarose gel at a lower cost and in any laboratory setting (10 minisatellite markers with a repeat unit larger than 9 bp), and panel 2 which comprises 7 microsatellites and provides a higher discriminatory power. CONCLUSION: Our analyses demonstrate that MLVA is a powerful and promising molecular typing tool with a high resolution and of low costs. The consistency of the results with independent methods suggests that MLVA can be applied for epidemiological studies. The resulting data can be queried on a dedicated MLVA genotyping Web service

Demographic, clinical, and service-use characteristics related to the clinician’s recommendation to transition from child to adult mental health services

Purpose: The service configuration with distinct child and adolescent mental health services (CAMHS) and adult mental health services (AMHS) may be a barrier to continuity of care. Because of a lack of transition policy, CAMHS clinicians have to decide whether and when a young person should transition to AMHS. This study describes which characteristics are associated with the clinicians’ advice to continue treatment at AMHS. Methods: Demographic, family, clinical, treatment, and service-use characteristics of the MILESTONE cohort of 763 young people from 39 CAMHS in Europe were assessed using multi-informant and standardized assessment tools. Logistic mixed models were fitted to assess the relationship between these characteristics and clinicians’ transition recommendations. Results: Young people with higher clinician-rated severity of psychopathology scores, with self- and parent-reported need for ongoing treatment, with lower everyday functional skills and without self-reported psychotic experiences were more likely to be recommended to continue treatment. Among those who had been recommended to continue treatment, young people who used psychotropic medication, who had been in CAMHS for more than a year, and for whom appropriate AMHS were available were more likely to be recommended to continue treatment at AMHS. Young people whose parents indicated a need for ongoing treatment were more likely to be recommended to stay in CAMHS. Conclusion: Although the decision regarding continuity of treatment was mostly determined by a small set of clinical characteristics, the recommendation to continue treatment at AMHS was mostly affected by service-use related characteristics, such as the availability of appropriate services

Cohort profile : demographic and clinical characteristics of the MILESTONE longitudinal cohort of young people approaching the upper age limit of their child mental health care service in Europe

Purpose: The presence of distinct child and adolescent mental health services (CAMHS) and adult mental health services (AMHS) impacts continuity of mental health treatment for young people. However, we do not know the extent of discontinuity of care in Europe nor the effects of discontinuity on the mental health of young people. Current research is limited, as the majority of existing studies are retrospective, based on small samples or used non-standardised information from medical records. The MILESTONE prospective cohort study aims to examine associations between service use, mental health and other outcomes over 24 months, using information from self, parent and clinician reports. Participants: Seven hundred sixty-three young people from 39 CAMHS in 8 European countries, their parents and CAMHS clinicians who completed interviews and online questionnaires and were followed up for 2 years after reaching the upper age limit of the CAMHS they receive treatment at. Findings to date: This cohort profile describes the baseline characteristics of the MILESTONE cohort. The mental health of young people reaching the upper age limit of their CAMHS varied greatly in type and severity: 32.8% of young people reported clinical levels of self-reported problems and 18.6% were rated to be ‘markedly ill’, ‘severely ill’ or ‘among the most extremely ill’ by their clinician. Fifty-seven per cent of young people reported psychotropic medication use in the previous half year. Future plans: Analysis of longitudinal data from the MILESTONE cohort will be used to assess relationships between the demographic and clinical characteristics of young people reaching the upper age limit of their CAMHS and the type of care the young person uses over the next 2 years, such as whether the young person transitions to AMHS. At 2 years follow-up, the mental health outcomes of young people following different care pathways will be compared. Trial registration number: NCT03013595

Les effets d'âge et de génération sur le niveau et la structure de la consommation

[fre] Les effets d'âge et de génération sur le niveau et la structure de la consommation La population française ne cesse de vieillir et l'accent est fortes incertitudes sur le niveau de la consommation des souvent mis sur le rôle que pourrait tenir, à l'avenir, la ménages au cours des décennies avenir., consommation de ceux que l'on nomme désormais les « nouveaux seniors ». En effet, le passage à la retraite . n'est plus, en général, synonyme de perte de niveau de : vie : déjà en 1990, le niveau de vie des ménages de plus de 60 ans était, en moyenne, de 1 0 % plus élevé que celui des ménages de moins de 60 ans. Or jusqu'à présent, les études sur la consommation suggèrent que celle-ci , diminue avec l'âge. La déformation de la structure de la population au profit des catégories de niveau de vie élevé, mais a priori sous-consommatrices, fait donc peser de Toutefois, ces études comparent des générations différentes à une même date. Suivre des échantillons des mêmes cohortes de ménages au fur et à mesure de leur - vieillissement permet, en revanche, de bien différencier, l'effet propre de l'âge de celui de l'appartenance à une génération spécifique. L'effet du vieillissement de la population sur la consommation apparaît alors plus faible. Une déformation notable de la structure de la consommation avec l'âge est la croissance de la part de consommation de loisirs après la retraite. [eng] The Effects of Age and Generation on the The French population is steadily ageing and the focus is often on the future potential role of the consumption of what are now termed the "new seniors". Retirement is by and large no longer synonymous with a decrease in the standard of living: In 1990, the standard of living of households aged over 60 was already 10% higher on average than that of households under 60 years old. Yet" consumption studies have hitherto suggested that consumption decreases with age. The change in the population structure towards categories with high- standards of living," but normally low consumption- Level and Structure of Consumption' therefore brings considerable uncertainties to bear on the ; level of household consumption over the coming decades. These studies compare different generations on the same ; date. However, tracking samples of given household cohorts as they get older is a good way of differentiating the specific age effect from the effect of belonging to a - given generation. Population ageing hence appears to have less of an effect on consumption. One striking change in the consumption structure with age is the growth in the share of leisure consumption after retirement. [ger] Alters- und generationsbedingte Auswirkungen auf das Konsumniveau und die Konsumstruktur Die franzôsische Bevôlkerung wird immer alter. Betont . wird in diesem Zusammenhang oftmals die Rolle, die der Konsum der sogenannten «neuen Senioren» kùnftig spielen kônnte. Denn der Eintritt in den Ruhestand ist im - allgemeinen nicht mehr mit einem Rùckgang des Lebensstandards gleichzusetzen: bereits 1990 lag der Lebensstandard der Haushalte von uber 60 Jahren im Durchschnitt urn 1 0% uber demjenigen der Haushalte von unter 60 Jahren.' Bislang gelangte man jedoch bei den Studien uber den Konsum zu dem SchluB, daB dieser mit dem Alter abnimmt. Aufgrund der Verformung der Bevôlkerungsstruktur zugunsten der Kategorien mit hôherem Lebensstandard, die aber a priori weniger ' konsumieren, herrscht hinsichtlich des Konsumniveaus der Haushalte in den kommenden Jahrzehnten Unsicherheit. Bei diesen Studien werden jedoch unterschiedliche Generationen zu ein und demselben Zeitpunkt miteinander verglichen. Dagegen ermôglicht die Verfolgung.von Stichproben ein und derselben' Haushaltskohorten entsprechend ihrer Alterung eine gute Differenzierung zwischen dem Effekt des Alters selbst und dem Effekt der Zugehôrigkeit zu einer bestimmten- Generation. Die Auswirkung der Bevôlkerungsalterung auf den Konsum scheint dann geringer zu sein. Eine nennenswerte Verformung der Konsumstruktur aufgrund des Alters ist darauf zurùckzufûhren, daB nach dem Eintritt in den Ruhestand der Anteil des Freizeitkonsums zunimmt. [spa] Los efectos de edad y de generacion sobre La poblaciôn francesa no cesa de envejecer y se hace . énfasis en el papel que podrîa desempenar en el futuro el ; consumo de los que se Maman ahora los « nuevos seniors ». La jubilaciôn ya no es sinônimo de pérdida de nivel de vida : en 1990, el nivel de vida de los hogares de ' mâs de 60 anos era en un promedio un 10 % superior al de los hogares de menos de 60 anos. Pero, hasta hoy dfa *. los estudios sobre consumo sugieren que este disminuye - con la edad. La deformaciôn de la estructura de la , poblaciôn en favor de las categorfas de alto nivel de vida pero a priori infraconsumidoras es una amenaza al nivel de consumo de los hogares para los decenios venideros. el nivel y la estructura del consumo Ahora bien, estos estudios comparan unas generaciones , diferentes en una misma fecha. Seguir unas muestras de ; las mismas cohortes de hogares a medida que van envejeciendo, permite en cambio separar el efecto propio de la edad del de la pertenencia a una generacion especifica. El efecto del envejecimiento de la poblaciôn ; sobre el consumo résulta en este caso mâs tenue. Una deformaciôn notable de la estrutura del consumo con la : edad es el crecimiento de la cuota de consumo de ocios después de la jubilaciôn.

Wireworm (Coleoptera, Elateridae) risk factors within potato cultivation

Invisible damage caused by the wireworm up to the time of harvesting make it one of the most feared pests, especially in potato cultivation. As there is no curative management possible, the only way to fight against it is preventively. To achieve this goal, it is necessary to examine which factors make the risk of wireworm attacks more prevalent. This study aims to get a better understanding of risk of wireworm attacks within potato cultivation and a better understanding of potato producer views about wireworm management. Two types of enquiries were combined within the same survey: one about technical aspects and the other one about social aspects of the problem. They were completed by going in situ meeting farmers. Information collected was processed using statistical tools: MCA, PCA, and HCPC analyses. The technical part was analyzed within seven themes: previous crops, interculture, wireworm damage, supply of organic matter, plot environment, chemical use, and mechanical passes. The social part was analyzed using the three first steps of the ARDI method: Actor, Resource, and Dynamic

Les divergences d'évolution des marchés du travail allemand et européens

[fre] Depuis 1993, dernier creux conjoncturel, le taux de chômage de l’Allemagne a sensiblement augmenté et la situation du marché du travail s’est dégradée, alors qu’elle s’améliorait chez ses partenaires européens. Les problèmes du marché du travail se sont concentrés sur les actifs les moins qualifiés, et le chômage de longue durée a progressé, au contraire de presque toute l’Europe des 15. Ces divergences tiennent pour une part à la faiblesse de la croissance liée à la resynchronisation de la conjoncture allemande après le «boom» de l’unification. Pour une autre part, le fonctionnement du marché du travail s’est détérioré. En effet, l’économie allemande a dû faire face à plusieurs chocs de coûts au cours de la première moitié des années 90: fortes hausses salariales jusqu’en 1993, hausses successives de cotisations sociales, passage de 36 à 35 heures dans la métallurgie en 1995 avec pleine compensation salariale. L’effort d’ajustement qu’a mené l’industrie après 1995 pour faire face à ces chocs a appauvri la croissance en emplois: sur le dernier cycle économique, le seuil au-delà duquel l’économie allemande crée des emplois a été supérieur à 2 %, contre environ 1,25% en France. Cet effort de productivité s’est accompagné de plusieurs années de modération salariale, si bien que fin 1999, l’Allemagne avait regagné douze points sur les vingt de compétitivité-coût perdus entre 1987 et 1995. Ainsi, la résorption des chocs de coût a principalement émané des acteurs privés, des entreprises et des syndicats. Jusqu’en 1999, l’État n’a pas enrayé la montée des taux de cotisations sociales, n’a traité que partiellement le problème de l’emploi faiblement qualifié, et a introduit des mesures de flexibilité de façon limitée. En revanche, le gouvernement actuel souhaite dans le cadre de l’UEM renforcer les conditions internes nécessaires à une stabilité durable des prix. Il a aussi revitalisé la tradition de «pactes pour l’emploi», mais en y introduisant des discussions sur les salaires. Ces évolutions ont mené à la signature de textes non contraignants qui, au vu des accords salariaux signés depuis le début 2000, semblent ouvrir la voie à une modération salariale plus durable.

Facteurs de risque de taupin (Coleoptera, Elateridae) en culture de pomme de terre

Invisible damage caused by the wireworm up to the time of harvesting make it one of the most feared pests, especially in potato cultivation. As there is no curative management possible, the only way to fight against it is preventively. To achieve this goal, it is necessary to examine which factors make the risk of wireworm attacks more prevalent. This study aims to get a better understanding of risk of wireworm attacks within potato cultivation and a better understanding of potato producer views about wireworm management. Two types of enquiries were combined within the same survey: one about technical aspects and the other one about social aspects of the problem. They were completed by going in situ meeting farmers. Information collected was processed using statistical tools: MCA, PCA, and HCPC analyses. The technical part was analyzed within seven themes: previous crops, interculture, wireworm damage, supply of organic matter, plot environment, chemical use, and mechanical passes. The social part was analyzed using the three first steps of the ARDI method: Actor, Resource, and Dynamic.ARVALIS-Institut du végéta