Tuning the immune response of dendritic cells to surface-assembled poly(I:C) on microspheres through synergistic interactions between phagocytic and TLR3 signaling.

The artificial dsRNA polyriboinosinic acid-polyribocytidylic acid, poly(I:C), is a potent adjuvant candidate for vaccination, as it strongly drives cell-mediated immunity. However, because of its effects on non-immune bystander cells, poly(I:C) administration may bear danger for the development of autoimmune diseases. Thus poly(I:C) should be applied in the lowest dose possible. We investigated microspheres carrying surface-assembled poly(I:C) as a two-in-one adjuvant formulation to stimulate maturation of monocyte-derived dendritic cells (MoDCs). Negatively charged polystyrene microspheres were equipped with a poly(ethylene glycol) corona through electrostatically driven surface assembly of a library of polycationic poly(l-lysine)-graft-poly(ethylene glycol) copolymers, PLL-g-PEG. Stable surface assembly of poly(I:C) was achieved by incubation of polymer-coated microspheres in an aqueous poly(I:C) solution. Surface-assembled poly(I:C) exhibited a strongly enhanced efficacy to stimulate maturation of MoDCs by up to two orders of magnitude, as compared to free poly(I:C). Multiple phagocytosis events were the key factor to enhance the efficacy. The cytokine secretion pattern of MoDCs after exposure to surface-assembled poly(I:C) differed from that of free poly(I:C), while their ability to stimulate T cell proliferation was similar. Overall, phagocytic signaling plays an important role in defining the resulting immune response to such two-in-one adjuvant formulations.

Role of β-catenin and γ-catenin in haematopoiesis : Generation of floxed Delta like 4 gene targeted mice

Résumé La voie de signalisation de Wnt est extrêmement conservée au cours de l'évolution. Les protéines Wnt sont des molécules sécrétées qui se lient à la famille de récepteurs Frizzled. Cette interaction mène à la stabilisation de la protéine β-caténine, qui va s'accumuler dans le cytoplasme puis migrer dans le noyau où elle peut s'hétérodimériser avec les facteurs de transcription de la famille TCF/LEF. Il a été démontré que cette voie de signalisation joue un rôle important durant la lymphopoïèse et de récents résultats suggèrent un rôle clé de cette voie dans le renouvellement des Cellules Souches Hématopoïétique (CSH). Des études se basant sur un système de surexpression de protéines montrent clairement que la voie Wnt peut influencer l'hématopoïèse. Cependant, le rôle de la protéine β-caténine dans le système hématopoïétique n'a jamais été testé directement. Ce projet de thèse se propose d'étudier la fonction de la protéine β-caténine par sa délétion inductible via le système Cre-loxP. De façon surprenante, nous avons pu démontrer que les progéniteurs de la moelle osseuse, déficients en β-caténine, ne montrent aucune altération dans leur capacité à s'auto-renouveler et/ou à reconstituer toutes les lignées hématopoïétiques (myéloïde, érythroïde et lymphoïde) dans les souris-chimères. De plus, le développement, la survie des thymocytes ainsi que la prolifération des cellules T périphériques induite par un antigène, sont indépendants de β-caténine. Ces résultats suggèrent soit que la protéine β-caténine ne joue pas un rôle primordial dans le système hématopoiétique, soit que son absence pourrait être compensée par une autre protéine. Un candidat privilégié susceptible de se substituer à β-caténine, serait plakoglobine, aussi connu sous le nom de γ-caténine. En effet, ces deux protéines partagent de multiples caractéristiques structurelles. Afin de démontrer que la protéine γ-caténine peut compenser l'absence de β-caténine, nous avons généré des souris dans lesquelles, le système hématopoïétique est déficient pour ces deux protéines. Cette déficience combinée de β- caténine et γ-caténine ne perturbe pas la capacité des Cellules Souche Hématopoïétique-Long Terme (CSH-LT) de se renouveler, par contre elle agit sur un progéniteur précoce déjà différencié de la moelle osseuse. Ces résultats mettent en évidence que la protéine γ-caténine est capable de compenser l'absence de protéine β-caténine dans le système hématopoïétique. Par conséquent, ce travail contribue à une meilleure connaissance de la cascade Wnt dans l'hématopoïèse. Summary The canonical Wnt signal transduction pathway is a developmentally highly conserved. Wnts are secreted molecules which bind to the family of Frizzled receptors in a complex with the low density lipoprotein receptor related protein (LRP-5/6). This initial activation step leads to the stabilization and accumulation of β-catenin, first in the cytoplasm and subsequently in the nucleus where it forms heterodimers with TCF/LEF transcription factor family members. Wnt signalling has been shown to be important during early lymphopoiesis and has more recently, been suggested to be a key player in self-renewal of haematopoietic stem cells (HSCs). Although mostly gain of function studies indicate that components of the Wnt signalling pathway can influence the haematopoietic system, the role of β-catenin has never been directly investigated. The aim of this thesis project is to investigate the putatively critical role of β-catenin in vivo using the Cre-loxP mediated conditional loss of function approach. Surprisingly, β-catenin deficient bone marrow (BM) progenitors arc not impaired in their ability to self-renew and/or to reconstitute all haematopoietic lineages (myeloid, erythroid and lymphoid) in both mixed and straight bone marrow chimeras. In addition, both thymocyte development and survival, and antigen-induced proliferation of peripheral T cells are β- catenin independent. Our results do not necessarily exclude the possibility of an important function for β-catenin mediated Wnt signalling in the haematopoietic system, it rather raises the question that β-catenin is compensated for by another protein. A prime candidate that may take over the function of β-catenin in its absence, is the close relative plakoglobin, also know as γ-catenin. This protein shares multiple structural features with β-catenin. In order to investigate whether γ-catenin can compensate for the loss of β-catenin we have generated mice in which the haematopoietic compartment is deficient for both proteins. Combined deficiency of β-catenin and γ-catenin does not perturb Long Term-Haematopoietic Stem Cells (LT-HSC) self renewal, but affects an already lineage committed progenitor population within the BM. Our results demonstrate that y-catenin can indeed compensate for the loss of β-catenin within the haematopoietie system.

State of Iowa Juvenile Delinquency Annual Statistical Report, 2006

The Iowa Juvenile Court Services Offices are issuing their fourth annual statewide report. The the Iowa Division of Criminal and Juvenile Justice Planning (CJJP). This report would not be possible without the dedication of, and assistance from, all of the above-mentioned people. The eight Chief Juvenile Court Officers would like to take this opportunity to thank their staff for their dedication and their ability to enter accurate information on every youth referred to Juvenile Court Services; the staff at the Iowa Court Information System, without whom this report would not be possible; and CJJP for their maintenance of the Iowa Justice Data Warehouse and their support in preparing this document.

State of Iowa Juvenile Delinquency Annual Statistical Report, 2007

The Iowa Juvenile Court Services Offices are issuing their fourth annual statewide report. The the Iowa Division of Criminal and Juvenile Justice Planning (CJJP). This report would not be possible without the dedication of, and assistance from, all of the above-mentioned people. The eight Chief Juvenile Court Officers would like to take this opportunity to thank their staff for their dedication and their ability to enter accurate information on every youth referred to Juvenile Court Services; the staff at the Iowa Court Information System, without whom this report would not be possible; and CJJP for their maintenance of the Iowa Justice Data Warehouse and their support in preparing this document.

Darker eumelanic barn owls better withstand food depletion through resistance to food deprivation and lower appetite.

The intensity of selection exerted on ornaments typically varies between environments. Reaction norms may help to identify the conditions under which ornamented individuals have a selective advantage over drab conspecifics. It has been recently hypothesized that in vertebrates eumelanin-based coloration reflects the ability to regulate the balance between energy intake and expenditure. We tested two predictions of this hypothesis in barn owl nestlings, namely that darker eumelanic individuals have a lower appetite and lose less weight when food-deprived. We found that individuals fed ad libitum during 24 h consumed less food when their plumage was marked with larger black spots. When food-deprived for 24 h nestlings displaying larger black spots lost less weight. Thus, in the barn owl the degree of eumelanin-based coloration reflects the ability to withstand periods of food depletion through lower appetite and resistance to food restriction. Eumelanic coloration may therefore be associated with adaptations to environments where the risk of food depletion is high.

Ecological-economic optimization of biodiversity conservation under climate change

Substantial investment in climate change research has led to dire predictions of the impacts and risks to biodiversity. The Intergovernmental Panel on Climate Change fourth assessment report(1) cites 28,586 studies demonstrating significant biological changes in terrestrial systems(2). Already high extinction rates, driven primarily by habitat loss, are predicted to increase under climate change(3-6). Yet there is little specific advice or precedent in the literature to guide climate adaptation investment for conserving biodiversity within realistic economic constraints(7). Here we present a systematic ecological and economic analysis of a climate adaptation problem in one of the world's most species-rich and threatened ecosystems: the South African fynbos. We discover a counterintuitive optimal investment strategy that switches twice between options as the available adaptation budget increases. We demonstrate that optimal investment is nonlinearly dependent on available resources, making the choice of how much to invest as important as determining where to invest and what actions to take. Our study emphasizes the importance of a sound analytical framework for prioritizing adaptation investments(4). Integrating ecological predictions in an economic decision framework will help support complex choices between adaptation options under severe uncertainty. Our prioritization method can be applied at any scale to minimize species loss and to evaluate the robustness of decisions to uncertainty about key assumptions.

Tn5-directed cloning of pqq genes from Pseudomonas fluorescens CHA0: mutational inactivation of the genes results in overproduction of the antibiotic pyoluteorin.

Pseudomonas fluorescens CHA0 produces several secondary metabolites, e.g., the antibiotics pyoluteorin (Plt) and 2,4-diacetylphloroglucinol (Phl), which are important for the suppression of root diseases caused by soil-borne fungal pathogens. A Tn5 insertion mutant of strain CHA0, CHA625, does not produce Phl, shows enhanced Plt production on malt agar, and has lost part of the ability to suppress black root rot in tobacco plants and take-all in wheat. We used a rapid, two-step cloning-out procedure for isolating the wild-type genes corresponding to those inactivated by the Tn5 insertion in strain CHA625. This cloning method should be widely applicable to bacterial genes tagged with Tn5. The region cloned from P. fluorescens contained three complete open reading frames. The deduced gene products, designated PqqFAB, showed extensive similarities to proteins involved in the biosynthesis of pyrroloquinoline quinone (PQQ) in Klebsiella pneumoniae, Acinetobacter calcoaceticus, and Methylobacterium extorquens. PQQ-negative mutants of strain CHA0 were constructed by gene replacement. They lacked glucose dehydrogenase activity, could not utilize ethanol as a carbon source, and showed a strongly enhanced production of Plt on malt agar. These effects were all reversed by complementation with pqq+ recombinant plasmids. The growth of a pqqF mutant on ethanol and normal Plt production were restored by the addition of 16 nM PQQ. However, the Phl- phenotype of strain CHA625 was due not to the pqq defect but presumably to a secondary mutation. In conclusion, a lack of PQQ markedly stimulates the production of Plt in P. fluorescens.

Using spatially distributed parameters and multi-response objective functions to solve parameterization of complex applications of semi-distributed hydrological models

Application of semi-distributed hydrological models to large, heterogeneous watersheds deals with several problems. On one hand, the spatial and temporal variability in catchment features should be adequately represented in the model parameterization, while maintaining the model complexity in an acceptable level to take advantage of state-of-the-art calibration techniques. On the other hand, model complexity enhances uncertainty in adjusted model parameter values, therefore increasing uncertainty in the water routing across the watershed. This is critical for water quality applications, where not only streamflow, but also a reliable estimation of the surface versus subsurface contributions to the runoff is needed. In this study, we show how a regularized inversion procedure combined with a multiobjective function calibration strategy successfully solves the parameterization of a complex application of a water quality-oriented hydrological model. The final value of several optimized parameters showed significant and consistentdifferences across geological and landscape features. Although the number of optimized parameters was significantly increased by the spatial and temporal discretization of adjustable parameters, the uncertainty in water routing results remained at reasonable values. In addition, a stepwise numerical analysis showed that the effects on calibration performance due to inclusion of different data types in the objective function could be inextricably linked. Thus caution should be taken when adding or removing data from an aggregated objective function.

Adolescent medicine : from clinical practice to public health

In most countries, adolescent health problems have shifted from pure medical conditions to more psychosocial burdens such as injuries and violence, substance use, unsafe sex and chronic conditions including under nutrition or obesity. This new situation requires specific actions which have to take into account the specificities of the bio psychosocial development of the adolescent. Youth friendly services offering adequate environment and policies as well as carefully trained physicians represent one answer to the health needs of adolescents. Another lies in the development of school prevention and health promotion. Finally, policies aiming at securing a safe environment represent an effective mean to improve the health of adolescents.

Organismos filantrópicos y no gubernamentales en el desarrollo de nuevas ciencias y tecnologías: el caso de las micro y nanotecnologías en México

RESUMEN: La emergencia de nuevas ciencias y tecnologías vienen acompañadas de nuevas dinámicas en la producción, el uso y la diseminación de nuevos conocimientos científicos y tecnológicos. Estas nuevas dinámicas se reflejan en la reorganización de las actividades científicas, en la creación o la reorientación de nuevas temáticas en la investigación, en la emergencia de nuevas preocupaciones y debates sobre los riesgos e implicaciones sociales, y en la participación de nuevos actores en el desarrollo de la ciencia y la tecnología, entre otros factores. En este artículo se presentan los resultados del estudio sobre las funciones de un organismo con orígenes filantrópicos y no gubernamental que ha sido central en el desarrollo de los sistemas microelectromecánicos (MEMS) en México. Las funciones que se analizan han sido y son desarrolladas en las diferentes etapas del desarrollo de esta tecnología emergente en este país. Además de mostrar las funciones desempeñadas por este tipo de organismo, este texto se cuestiona sobre la modelización de las relaciones entre las diferentes entidades presentes en el desarrollo e instalación de nuevas tecnologías. En las conclusiones intentamos avanzar algunos elementos para tomar en cuenta estos organismos y enriquecer los estudios sociales sobre las nuevas ciencias y tecnologías. ABSTRACT: The emergence of new sciences and technologies come with new dynamics in the production, use and dissemination of new scientific and technological knowledge. These new dynamics are reflected in the reorganization of scientific activities, in creating or redirecting new topics in research, in the emergence of new concerns and debates about the risks and social implications, and the participation of new actors in the development of science and technology, among other factors. This article presents the results of the study on the functions of an organism with philanthropic and non-governmental sources that has been central to the development of microelectromechanical systems (MEMS) in Mexico. The functions have been discussed and are developed at different stages of development of this emerging technology in this country. In addition to showing the functions of such a body, this text is questioned on modeling the relationships between the various entities in the development and deployment of new technologies. In the conclusions we try moving some elements to take into account these organisms and enrich social studies on the new sciences and technologies.

Time for change: a roadmap to guide the implementation of the World Anti-Doping Code 2015

A medical and scientific multidisciplinary consensus meeting was held from 29 to 30 November 2013 on Anti-Doping in Sport at the Home of FIFA in Zurich, Switzerland, to create a roadmap for the implementation of the 2015 World Anti-Doping Code. The consensus statement and accompanying papers set out the priorities for the antidoping community in research, science and medicine. The participants achieved consensus on a strategy for the implementation of the 2015 World Anti-Doping Code. Key components of this strategy include: (1) sport-specific risk assessment, (2) prevalence measurement, (3) sport-specific test distribution plans, (4) storage and reanalysis, (5) analytical challenges, (6) forensic intelligence, (7) psychological approach to optimise the most deterrent effect, (8) the Athlete Biological Passport (ABP) and confounding factors, (9) data management system (Anti-Doping Administration & Management System (ADAMS), (10) education, (11) research needs and necessary advances, (12) inadvertent doping and (13) management and ethics: biological data. True implementation of the 2015 World Anti-Doping Code will depend largely on the ability to align thinking around these core concepts and strategies. FIFA, jointly with all other engaged International Federations of sports (Ifs), the International Olympic Committee (IOC) and World Anti-Doping Agency (WADA), are ideally placed to lead transformational change with the unwavering support of the wider antidoping community. The outcome of the consensus meeting was the creation of the ad hoc Working Group charged with the responsibility of moving this agenda forward.

CHO cell engineering to prevent polypeptide aggregation and improve therapeutic protein secretion.

The ability to efficiently produce recombinant proteins in a secreted form is highly desirable and cultured mammalian cells such as CHO cells have become the preferred host as they secrete proteins with human-like post-translational modifications. However, attempts to express high levels of particular proteins in CHO cells may consistently result in low yields, even for non-engineered proteins such as immunoglobulins. In this study, we identified the responsible faulty step at the stage of translational arrest, translocation and early processing for such a "difficult-to-express" immunoglobulin, resulting in improper cleavage of the light chain and its precipitation in an insoluble cellular fraction unable to contribute to immunoglobulin assembly. We further show that proper processing and secretion were restored by over-expressing human signal receptor protein SRP14 and other components of the secretion pathway. This allowed the expression of the difficult-to-express protein to high yields, and it also increased the production of an easy-to-express protein. Our results demonstrate that components of the secretory and processing pathways can be limiting, and that engineering of the secretory pathway may be used to improve the secretion efficiency of therapeutic proteins from CHO cells.

Which change talk dimensions are associated with subsequent drinking among young men receiving a brief motivational intervention in a non-clinical setting?

Background: Patient change talk (CT) during brief motivational interventions (BMI) has been linked with subsequent changes in drinking in clinical settings but this link has not been clearly established among young people in non-clinical populations. Objective: To determine which of several CT dimensions assessed during an effective BMI delivered in a non-clinical setting to 20-year old men are associated with drinking 6 months later. Methods: Of 125 individuals receiving a face-to-face BMI session (15.8 ± 5.4 minutes), we recorded and coded a subsample of 42 sessions using the Motivational Interviewing Skill Code 2.1. Each patient change talk utterance was categorized as `Reason´, `Ability´, `Desire´, `Need´, `Commitment´, `Taking steps´, or `Other´. Each utterance was graded according to its strength (absolute value from 1 to 3) and direction (i.e. towards (positive sign) or away (negative sign) from change/in favor of status quo). `Ability´, `Desire´, and `Need´ to change (`ADN´) were grouped together since these codes were too scarce to conduct analyses. Mean strength scores over the entire session were computed for each dimension and later dichotomized in towards change (i.e. mean core > 0) and away from change/in favor of status quo. Negative binomial regression models were used to assess the relationship between CT dimensions and drinking 6 months later, adjusting for drinking at baseline. Results: Compared to subjects with a `Taking steps´ score away from change/in favor of status quo, subjects with a positive `Taking steps´ score reported significantly less drinking 6 months later (Incidence Rate Ration [IRR] for drinks per week: 0.56, 95% Confidence Interval [CI] 0.31, 1.00). IRR (95%CI) for subjects with a positive `ADN´ score was 0.58, (0.32, 1.03). For subjects with a positive `Reason´, `Commitment´, and `Other´ scores, IRR (95%CI) were 1.28 (0.77; 2.12) 1.63 (0.85; 3.14) and 1.03 (0.61; 1.72), respectively. Conclusion: A change talk dimension reflecting steps taken towards change (`Taking steps´) is associated with less drinking 6 months later among young men receiving a BMI in a non-clinical setting. Encouraging patients to take steps towa change may be a worthy objective for clinicians and may explain BMI efficacy.

Differentiated chemical reactivity of nanoparticles toward DTT

RATIONALE: Induction of oxidative stress and impairment of the antioxidant defense are considered important biological responses following nanoparticle (NP) exposure. The acellular in vitro dithiothreitol (DTT) assay is proposed to measure the oxidative potential of NP. In addition, DTT can be considered as a model compound of sulfur containing antioxidants. The objective of this work is to evaluate the surface reactivity in solution of a NP panel toward DTT. METHOD: The NP panel was composed of four carbonaceous particles, six types of metal oxides and silver with primary size ranged from 7 to 300 nm. Suspensions were prepared in surfactant solution with 30 min sonication. DTT was used as reductant to evaluate the oxidative properties of the different NP. The determination of the NP ability to catalyze electron transfer from DTT to oxygen was carried out as described in Sauvain et al., Nanotoxicology, 2008, 2:3, 121−129. RESULTS: All the carbonaceous NP catalyzed the oxidation of DTT by oxygen following the mass based order: carbon black > diesel exhaust particle > nanotubes > fullerene. A contrasting reactivity was observed for the metallic NP. Except for nickel oxide and metallic silver, which reacted similarly to the carbonaceous NP, all other metal oxides hindered the oxidation of DTT by oxygen, with ZnO being the most effective one. CONCLUSIONS : DTT was stabilized against oxidation in the presence of metal oxide NP in the solution. This suggests that different chemical interactions take place compared with carbonaceous NP. To explain these differences, we hypothesize that DTT could form complexes with the metal oxide surface (or dissolved metal ions), rendering it less susceptible to oxidation. By analogy, such a process could be thought to apply in biological systems with sulfur−containing antioxidants, reducing their buffer capacity. Such NP could thus contribute to oxidative stress by an alternative mechanism.

Construction of a scale to evaluate the diurnal impact of insomnia

The objective of this study was to develop and validate a scale to assess the diurnal impact of insomnia. The Insomnia Diurnal Impact Scale (IDIS) comprises six items designed to evaluate the daytime effects of insomnia. The sychometric properties of the original scale were analysed in a sample of 172 students, while its ability to differentiate insomniacs and non-insomniacs (according to the International Classification of Diseases (ICD)-10 criteria) was examined in a sample of 79 psychiatric patients and 82 individuals from the community. The psychometric properties of the English version were then analysed in a sample of 44 Englishspeaking participants. The results showed the internal consistency coefficient to be very good (0.86), with testretest reliability at 1 month being 0.79. A single factor explained almost 60% of the variance. Correlation of the IDIS with other scales varied between moderate and high values. Sensitivity was 78% and specificity 57% in the community sample, while the corresponding figures for the psychiatric population were 83% and 63%. Cronbach's ¿ coefficient for the English version reached a value of 0.93. These results indicate that the IDIS shows adequate reliability and validity with both general and psychiatric populations, and also that it can discriminate between the presence and absence of insomnia. The English version presents good preliminary results regarding item-corrected total correlation and internal consistency. In conclusion, the IDIS appears to be a useful tool in the primary care and mental health contexts for assessing insomnia-related diurnal dysfunction.