Diversité virale à travers des gradients de salinité nordiques en évolution

Les communautés microbiennes dominent la plupart des écosystèmes aquatiques en termes de biomasse et de productivité, particulièrement dans les environnements nordiques hostiles à la faune et la flore macroscopiques. Les virus y contrôlent l'abondance des populations microbiennes et peuvent modifier le métabolisme des cellules infectées par l'utilisation de gènes métaboliques auxiliaires. Le court-circuit viral affecte aussi le transfert des nutriments et de l'énergie dans les réseaux trophiques en emprisonnant l'énergie dans la boucle microbienne, ne la rendant accessible qu'aux micro-organismes. Ces manipulations de l'activité de la prépondérante biomasse microbienne peuvent se répercuter jusque dans les cycles biogéochimiques des environnements aquatiques nordiques. La salinité est une caractéristique importante de ces écosystèmes puisqu'elle influence directement le métabolisme des micro-organismes. Chez les virus, ces effets se traduisent dans leur distribution, leur infectivité et leurs interactions avec leurs hôtes. L'objectif de cette thèse est d'offrir une connaissance approfondie de l'écologie des virus aquatiques en examinant plus spécifiquement les changements de diversité selon les gradients de salinité dans les milieux nordiques. La composition des communautés virales a donc été étudiée dans trois sites nordiques sélectionnés pour leur importance écologique et la spécificité de leur zone de transition saline. D'abord, une approche par amplification PCR de marqueurs conservés pour les Phycodnaviridae et les Picornavirales a permis de dresser un portrait des virus infectant le phytoplancton dans la zone de turbidité maximale du fleuve Saint-Laurent, important cours d'eau d'Amérique du Nord. La détection de ces virus dans six sites allant de l'habitat dulcicole à l'habitat marin a révélé peu de phylotypes viraux communs entre les sites d'eau douce et d'eau salée. La salinité semble ainsi y être le premier déterminant de la composition de la communauté virale. De plus, nos analyses indiquent que les virus découverts dans l'estuaire du Saint-Laurent divergent de ceux qui ont déjà été classifiés et des virotypes d'environnements comparables publiés. Ces résultats suggèrent que des virus à ADN et à ARN infectant le phytoplancton seraient actifs dans la zone de turbidité maximale et que cette région possède des assemblages viraux uniques. Ensuite, une approche viromique a permis de caractériser les communautés virales d'un lac arctique hautement stratifié, le lac A, dont les eaux profondes issues de l'océan Arctique ancien se sont révélées riches en virus nouveaux. Ce type de lacs à stratification pérenne est dépendant de la couverture de glace qui maintient la structure chimique de sa colonne d'eau et la distribution des communautés microbiennes qui y sont associées. Les communautés virales des trois strates du lac étaient remarquablement distinctes, ce qui suggère qu'il y a peu d'échange de virotypes entre elles. L'abondance virale et les ratios virus/procaryotes étaient plus élevés en profondeur et la division nette des communautés virales correspondait avec celle d'hôtes potentiels. Les communautés virales du lac A se distinguaient aussi de données de l'océan Arctique et du lac méromictique Ace Lake en Antarctique. Enfin, la même approche viromique peu invasive développée pour les travaux de la présente thèse a permis de documenter la diversité virale du dernier lac épiplateforme de l'Arctique, le lac épiplateforme du fjord Milne. Contrairement aux résultats obtenus au lac A, les assemblages viraux de surface étaient les plus diversifiés et une communauté virale propre à la chémocline du lac n'a pas pu être détectée. La division nette entre les assemblages dulcicoles et marins s'expliquait principalement par la conductivité et la concentration en oxygène dissout. La caractérisation de certains groupes de virotypes cooccurents a aussi permis de prévoir leurs hôtes potentiels. Les zones de transition étudiées dans cette thèse sont maintenues par des équilibres hydriques et glaciaires fragiles, déjà perturbés par les changements climatiques et l'activité humaine. Avec le réchauffement accéléré des régions nordiques, la fonte est accrue dans la plateforme glacière du Groenland et les glaciers continentaux. Ce fort débit d'eau douce qui se jette dans les océans Arctique et Atlantique Nord risque de déséquilibrer la circulation thermohaline et de bouleverser la surface des écosystèmes marins, réservoirs de productivité primaire microbienne irremplaçables. Connaissant les effets de l'activité virale dans les écosystèmes aquatiques, il est urgent de décrire les communautés virales d'environnements changeants, comme les zones de transition saline, afin de décrire les fonctions uniques qu'elles pourraient détenir et mieux comprendre leurs effets à long terme sur ces écosystèmes en évolution.Microbial communities dominate most aquatic ecosystems in terms of biomass and productivity, particularly in high latitude areas where conditions are hostile to macro fauna and flora. There, viruses have many roles in population control, for example, through their auxiliary metabolic genes that modify the metabolism of infected host cells. They also alter the transfer of nutrients and energy through trophic webs. The viral shunt, for example, traps energy inside the microbial loop, making it more accessible to microorganisms. These effects on aquatic trophic webs can have repercussions that affect the biogeochemical cycles of northern and arctic aquatic environments where metabolic activity from the abundant microbial biomass contributes to most processes. Just as salinity restrains the distribution of aquatic pluricellular organisms, it also directly affects the metabolism of microbes and their impact on ecosystem functioning. In viruses, these effects can translate into changes in infectivity and distribution but can also modify their interactions with their hosts. The main objective of this thesis was to characterize the ecology of aquatic viruses by examining the changes in their diversity through salinity gradients in northern aquatic habitats. We studied the composition of viral communities in three northern sites selected for their ecological relevance and the uniqueness of their saline transition zones. First, PCR amplification of conserved genetic markers of Phycodnaviridae and Picornavirales allowed us to describe the viruses that infect phytoplankton in the estuarine transition zone of the Saint Lawrence River, an important North American waterway. Detection of these viruses at six sites ranging from freshwater to marine habitats revealed few shared viral phylotypes between fresh and saline waters. This suggests that salinity might be the main factor influencing viral community composition. In addition, our analyses indicate that the virotypes found in the St. Lawrence estuary differ from published viruses and those found in comparable environments. These results suggest that DNA and RNA viruses infecting phytoplankton are active in the estuarine transition zone and that this zone harbours its own unique viral assemblages. Second, a viromic approach (that was developed during this research) allowed for the characterization of the viral communities in a highly stratified arctic lake. Lake A, with deeper waters that originate from the ancient Arctic Ocean revealed an abundance of novel viruses. This type of perennially stratified lake is dependant on ice cover which maintains the chemical structure of the water column and the distribution of the associated microbial communities. The viral communities in the three strata of the lake were remarkably distinct, suggesting that there is little exchange of virotypes. Viral abundance and virus/prokaryote ratios were highest in the deeper water and the clean separation between the viral communities matched that of potential hosts. The viral communities of Lake A also differed from those of the Arctic Ocean and the meromictic Ace Lake in Antarctica. Lastly, the same minimally invasive viromic approach allowed us to document the viral diversity in the last Arctic epishelf lake, the Milne Fiord epishelf lake. Contrary to what was observed in Lake A, the surface viral assemblages were more diverse, and no specific viral community was detected based on the chemocline. The clear division between the fresh and marine assemblages was mostly correlated with conductivity and dissolved oxygen concentration. Finally, the characterization of co-occurring virotypes allowed us to predict potential hosts. The transition zones examined in this research are maintained by a fragile balance of hydrological and glacial conditions which are already affected by climate change, but also by other human activities. Knowing how viruses affect aquatic ecosystems, a thorough description of the viral communities in changing environments is urgent and essential to gain a better understanding of their potential long-term effects within aquatic ecosystems. As the northern regions continue to warm at an accelerated rate, the melting increases in the Greenland platform and continental glaciers. The large amount of freshwater discharge into the Arctic Ocean and the North Atlantic may disturb thermohaline circulation and drastically change the surface of marine ecosystems, a crucial site for microbial primary production. Understanding how viruses and microbes thrive in the saline transition and their dependence on potentially unique functions and interactions may be key to anticipating the long-term effects they could have in these irreversibly and continuously changing ecosystems

Extreme viral partitioning in a marine-derived High Arctic lake

ABSTRACT High-latitude, perennially stratified (meromictic) lakes are likely to be especially vulnerable to climate warming because of the importance of ice in maintaining their water column structure and associated distribution of microbial communities. This study aimed to characterize viral abundance, diversity, and distribution in a meromictic lake of marine origin on the far northern coast of Ellesmere Island, in the Canadian High Arctic. We collected triplicate samples for double-stranded DNA (dsDNA) viromics from five depths that encompassed the major features of the lake, as determined by limnological profiling of the water column. Viral abundance and virus-to-prokaryote ratios were highest at greater depths, while bacterial and cyanobacterial counts were greatest in the surface waters. The viral communities from each zone of the lake defined by salinity, temperature, and dissolved oxygen concentrations were markedly distinct, suggesting that there was little exchange of viral types among lake strata. Ten viral assembled genomes were obtained from our libraries, and these also segregated with depth. This well-defined structure of viral communities was consistent with that of potential hosts. Viruses from the monimolimnion, a deep layer of ancient Arctic Ocean seawater, were more diverse and relatively abundant, with few similarities to available viral sequences. The Lake A viral communities also differed from published records from the Arctic Ocean and meromictic Ace Lake in Antarctica. This first characterization of viral diversity from this sentinel environment underscores the microbial richness and complexity of an ecosystem type that is increasingly exposed to major perturbations in the fast-changing Arctic. IMPORTANCE The Arctic is warming at an accelerating pace, and the rise in temperature has increasing impacts on the Arctic biome. Lakes are integrators of their surroundings and thus excellent sentinels of environmental change. Despite their importance in the regulation of key microbial processes, viruses remain largely uncharacterized in Arctic lacustrine environments. We sampled a highly stratified meromictic lake near the northern limit of the Canadian High Arctic, a region in rapid transition due to climate change. We found that the different layers of the lake harbored viral communities that were strikingly dissimilar and highly divergent from known viruses. Viruses were more abundant in the deepest part of the lake containing ancient Arctic Ocean seawater that was trapped during glacial retreat and were genomically unlike any viruses previously described. This research demonstrates the complexity and novelty of viral communities in an environment that is vulnerable to ongoing perturbation

A value-based comparison of the management of ambulatory respiratory diseases in walk-in clinics, primary care practices, and emergency departments : protocol for a multicenter prospective cohort study

Background: In Canada, 30%-60% of patients presenting to emergency departments are ambulatory. This category has been labeled as a source of emergency department overuse. Acting on the presumption that primary care practices and walk-in clinics offer equivalent care at a lower cost, governments have invested massively in improving access to these alternative settings in the hope that patients would present there instead when possible, thereby reducing the load on emergency departments. Data in support of this approach remain scarce and equivocal. Objective: The aim of this study is to compare the value of care received in emergency departments, walk-in clinics, and primary care practices by ambulatory patients with upper respiratory tract infection, sinusitis, otitis media, tonsillitis, pharyngitis, bronchitis, influenza-like illness, pneumonia, acute asthma, or acute exacerbation of chronic obstructive pulmonary disease. Methods: A multicenter prospective cohort study will be performed in Ontario and Québec. In phase 1, a time-driven activity-based costing method will be applied at each of the 15 study sites. This method uses time as a cost driver to allocate direct costs (eg, medication), consumable expenditures (eg, needles), overhead costs (eg, building maintenance), and physician charges to patient care. Thus, the cost of a care episode will be proportional to the time spent receiving the care. At the end of this phase, a list of care process costs will be generated and used to calculate the cost of each consultation during phase 2, in which a prospective cohort of patients will be monitored to compare the care received in each setting. Patients aged 18 years and older, ambulatory throughout the care episode, and discharged to home with one of the aforementioned targeted diagnoses will be considered. The estimated sample size is 1485 patients. The 3 types of care settings will be compared on the basis of primary outcomes in terms of the proportion of return visits to any site 3 and 7 days after the initial visit and the mean cost of care. The secondary outcomes measured will include scores on patient-reported outcome and experience measures and mean costs borne wholly by patients. We will use multilevel generalized linear models to compare the care settings and an overlap weights approach to adjust for confounding factors related to age, sex, gender, ethnicity, comorbidities, registration with a family physician, socioeconomic status, and severity of illness. Results: Phase 1 will begin in 2021 and phase 2, in 2023. The results will be available in 2025. Conclusions: The end point of our program will be for deciders, patients, and care providers to be able to determine the most appropriate care setting for the management of ambulatory emergency respiratory conditions, based on the quality and cost of care associated with each alternative

Polyphenols modulate calcium-independent mechanisms in human arterial tissue-engineered vascular media

Background: In the present study, an arterial tissue-engineered vascular media (TEVM) was produced from cultured human smooth muscle cells of the umbilical artery and we took advantage of this model to evaluate the regulation of contraction and the signalling pathways of polyphenols in arteries. Methods: Cultured human smooth muscle cells of the umbilical artery were used to produce arterial TEVMs. Contraction experiments were performed to determine intracellular targets involved in the modulation of contraction by polyphenols extract from red wine, Provinols (SEPPIC Groupe Air Liquide, Paris, France). Results: Smooth muscle cells in arterial TEVM displayed a differentiated phenotype as demonstrated by the expression of -smooth muscle actin, a vascular smooth muscle-specific marker, and tissue contraction in response to vasoconstrictor and vasodilator agents. Contractions caused by histamine were associated with an increase in [Ca2]i and a Ca2- independent signalling pathway. The latter pathway involved mechanisms sensitive to protein kinase C, myosin light chain kinase, and Rho-associated protein kinase inhibitors. The regulation of contraction induced by Provinols shows that treatment of arterial TEVM with this compound significantly decreased histamine-induced contraction. This effect was associated with the inhibition of the Rho-associated protein kinase pathway and the decrease in -smooth muscle actin expression. Conclusion: The use of arterial TEVM, brings new insights into the mechanisms by which polyphenols regulate vascular contraction in the human artery

Communities of Phytoplankton Viruses across the Transition Zone of the St. Lawrence Estuary

The St. Lawrence hydrographic system includes freshwater, brackish, and marine habitats, and is the largest waterway in North America by volume. The food-webs in these habitats are ultimately dependent on phytoplankton. Viral lysis is believed to be responsible for a major part of phytoplankton mortality. To better understand their role, we characterized the diversity and distribution of two viral taxa infecting phytoplankton: the picornaviruses and phycodnaviruses. Our study focused on the estuary transition zone, which is an important nursery for invertebrates and fishes. Both viral taxa were investigated by PCR amplification of conserved molecular markers and next-generation sequencing at six sites, ranging from freshwater to marine. Our results revealed few shared viral phylotypes between saltwater and freshwater sites. Salinity appeared to be the primary determinant of viral community composition. Moreover, our analysis indicated that the viruses identified in this region of the St. Lawrence diverge from classified viruses and homologous published environmental virotypes. These results suggest that DNA and RNA viruses infecting phytoplankton are likely active in the estuary transition zone, and that this region harbors its own unique viral assemblages

Liderazgo que ejercen los directores de dos establecimientos Municipales con distintos resultados SIMCE

Tesis (Magister en Dirección y Liderazgo para la Gestión Educacional)Esta investigación sobre el liderazgo que ejercen los directores de dos establecimientos municipales con distintos resultados SIMCE, es un estudio descriptivo de las prácticas de los líderes educativos, quiénes influyen en la vida de muchos niños y niñas de establecimientos con las mismas características y niveles de vulnerabilidad. El informe comienza con el problema generador del presente estudio, sintetizado en la pregunta ¿Cuáles son las características o tipo de liderazgo que ejercen los directores de ambos establecimientos educacionales y cómo influyen en los resultados de aprendizajes de todos sus estudiantes?. Además, abarca los antecedentes del problema, como el impacto del liderazgo en los aprendizajes de todos los estudiantes; definición y justificación del problema en sí. Nos hemos permitido revisar la bibliografía y contrastar las opiniones de diferentes autores relacionadas con el problema planteado en el presente estudio (sustento teórico) y definir brevemente cada uno de los estilos de liderazgo más recurrentes en los diferentes textos seleccionados para nuestra investigación. Además, se considera importante rescatar los hallazgos presentes en otras investigaciones relacionadas con el tema de estudio

TLR4/IFNγ pathways induce tumor regression via NOS II-dependent NO and ROS production in murine breast cancer models

International audienceToll-like receptor (TLR) 4 agonists have emerged as a new group of molecules used for cancer therapy. They have been exploited to enhance the immunogenicity of current chemotherapeutic regimens. However, their effects on cancer cells remain elusive. Here, we showed that a TLR4 agonist, namely a synthetic lipid A analog (ALA), OM-174, exhibits antitumor effects in several mammary tumor mouse models. We also showed that immune components are involved in such effects, as attested to by the failure of ALA to induce tumor regression or an increase of animal survival in mice knocked-out for interferon g (IFNg) or TLR4. TLR4 and IFNg receptor (INFR2) expressed by cancer cells are involved in the antitumor efficacy of ALA since this last did not inhibit tumor growth in mice bearing a tumor but lacking TLR4 or IFNg receptor 2 (IFNR2). Mechanistic investigations revealed that nitric oxide (NO), superoxide and peroxynitrite produced by uncoupling of inducible NO synthase (NOS II) in cancer cells are key mediators of ALA and IFNg-mediated tumor growth inhibition. We present here a comprehensive picture of tumor cell death induction, in vivo and in vitro, by immunotherapy and for the first time the involvement of the TLR4/IFNg/NOS II pathway in immunotherapy was investigated

Development of a Patient-Reported Experience Measure Tool for Ambulatory Patients With Acute Unexpected Needs: The APEX Questionnaire

Background: The aim of this study was to develop a patient-reported experience measure (PREM) for comparing the experience of care received by ambulatory patients with acute unexpected needs presenting in emergency departments (EDs), walk-in clinics, and primary care practices. Methods: The Ambulatory Patient EXperience (APEX) questionnaire was developed using a 5-phase mixed-methods approach. The questionnaire was pretested by asking potential users to rate its clarity, usefulness, redundancy, content and face validities, and discrimination on a 9-point scale (1 = strongly disagree to 9 = strongly agree). The pre-final version was then tested in a pilot study. Results: The final questionnaire is composed of 61 questions divided into 7 sections. In the pretest (n = 25), median responses were 8 and above for all dimensions assessed. In the pilot study, 63 participants were enrolled. Adjusted results show that access, cleanliness, and feeling treated with respect and dignity by nurses and physicians were significantly better in the clinics than in the ED. Conclusion: We developed a questionnaire to assess and compare experience of ambulatory care in different clinical settings