Neutrophil degranulation differentially modulates phenotype and function of bovine monocyte subsets

Monocytes and neutrophils are important players in the innate immune response and cooperate during infection and inflammation. In our study we analyzed the effects of neutrophil degranulation products (polymorphonuclear granulocytes degranulation products, PMN-DGP) on the activation, the adhesion and the migration of three bovine monocyte subsets, as well as their effects on monocyte-macrophage differentiation. Cross-linking of surface CD18 molecules on bovine PMN resulted in the release of primary, secondary and tertiary granules as well as of secretory vesicles. PMN-DGP induced a significant Ca2+-influx in classical (classical monocytes, cM) and intermediate monocytes (intermediate monocytes, intM) but not in non-classical monocytes (non-classical monocytes, ncM). A selective and up-regulated expression induced by PMN-DGP was only seen for CD11a and CD31 on intM. PMN-DGP induced a selective migration of intM in vitro. The presence of PMN-DGP during the differentiation of cM or intM into macrophages resulted in increased expression of membrane CD163 and reduced expression of MHC-II molecules. PMN-DGP-derived macrophages produced more IL-12 and IL-10 and showed enhanced phagocytosis and ROS production capacities. In conclusion, PMN-DGP selectively attract bovine intM and skew the functional maturation of cM and intM

Mécanismes physiques et fondements théoriques de la récupération d'énergie micro-ondes ambiante pour les dispositifs sans fil à faible consommation

RÉSUMÉ La récupération d’énergie micro-ondes ambiantes (REMA) pour alimenter de circuits à faible consommation et faible rapport cyclique a été le sujet de plusieurs publications au cours des dernières années. L’intérêt par ce sujet a été poussé principalement par les diverses applications prévues par l’Internet des Objets, l’Immotique et les nouveaux développements des dispositifs pour les « Body Area Netwoks ». Un des besoins récurrents que l’on retrouve parmi plusieurs de ces applications est une source d’énergie intégrée, qui ne nécessiterait pas de maintenance régulière, qui serait de petite taille et d’un faible poids. Pour beaucoup d’applications à venir, les piles sont trop encombrantes et demandent un plan de maintenance pour les recharger ou les remplacer, ce qui n’est pas possible. Dans ce contexte, un autre type de source d’énergie est nécessaire. La récupération d’énergie ambiante est ici proposée comme une source alternative de puissance pour ces circuits à faible consommation. Ce travail explore plus spécifiquement la récupération d’énergie micro-ondes ambiantes par l’utilisation centrale de circuits redresseurs à diodes. Un modèle mathématique a tout d’abord été développé pour décrire les mécanismes qui contribuent au processus de redressement d’énergie micro-ondes aux niveaux de puissance que l’on retrouve dans l’ambiant. Ce modèle est évalué en utilisant des résultats de simulation et de mesures réalisées sur trois prototypes fabriqués dans le cadre de cette maîtrise. Le modèle développé inclut les pertes dans les composants parasitiques de l’élément non linéaire utilisé pour le redressement d’énergie ainsi que les pertes d’insertion du réseau d’adaptation d’impédance. Partant de ce modèle, deux possibilités sont explorées pour améliorer l’efficacité de conversion de puissance des redresseurs micro-ondes actuels, particulièrement pour ceux fonctionnant à des niveaux de puissance trouvés dans la REMA. On considèrera dans ce travail que la plage de puissance correspondant à la REMA se situe à une valeur de crête de -30 dBm, et à des niveaux de puissance moyenne bien en dessous de ce seuil. Un circuit hybride coopératif de récupération d’énergie ambiante va ensuite être proposé. Celui-ci présente la particularité de n’être basé que sur un seul composant non linéaire pour redresser l’énergie micro-ondes et l’énergie mécanique de façon coopérative.---------- ABSTRACT Powering low consumption and low duty cycle devices and circuits using Ambient Microwave Energy Harvesting (AMEH) has been the subject of several investigations in recent years. The interest for this research topic has been promoted mainly by various and new applications driven mainly by the Internet of things, Building Automation and new developments in devices for the Body Area Netwoks. A common characteristic among several of these applications is the need for a wireless source which does not require regular maintenance, and has a small size and low weight. Batteries are often too cumbersome and require a maintenance plan to recharge or replace them, which is not always possible. A new source of energy is thus necessary. Ambient energy harvesting is proposed as an alternative source of power to these low power consumption devices and circuits. This M.A.Sc. work is developed to explore the microwave ambient energy harvesting using diode rectifier circuits. A mathematical model is first developed to explain the mechanisms that contribute to the process of recovery of microwave energy in the range of power found in the ambient microwave energy harvesting applications. An evaluation of this model is made using simulation results and then measurements results from three prototypes developed under this M.A.Sc. program. The results show an excellent agreement between the three methods. The developed model includes losses in the parasitic components of the non-linear element used for the rectification of energy as well as the impedance matching network insertion losses. Based on this model, two possible ways of improving the efficiency of ambient microwave power rectifiers at the power levels found in the AMEH are explored. In this work, it is considered that the AMEH takes place within the range of powers with a peak value of -30 dBm, however at average power levels well below this threshold. First, a cooperative hybrid circuit of ambient energy harvesting is presented where collected microwave and mechanical energies are converted in a cooperative manner through a single non-linear component. Theory, simulations and measurements show that the total power recovered by the proposed scheme can provide up to twice the efficiency of a circuit combining the output of two independent harvesters

Estudo das frequencias de recombinaçao entre o loco da cadeia beta da hemoglobina (Hb beta)e os locos do sistema sangüíneo MNSs e da cadeia delta da hemoglobina (Hb delta)

Orientadora: Profa. Dra. Eleidi A. Chautard Freire MaiaDissertação (mestrado) -Universidade Federal do Paraná, Setor de Ciências Biológicas, Curso de Pós-Graduação em Genética HumanaInclui referências: p. 63-7

TLR ligands, but not modulators of histone modifiers, can induce the complex immune response pattern of endotoxin tolerance in mammary epithelial cells

Excessive stimulation of the TLR4 axis through LPS reduces the expression of some cytokine genes in immune cells, while stimulating the expression of immune defense genes during a subsequent bacterial infection. This endotoxin tolerance (ET) is mediated via epigenetic mechanisms. Priming the udder of cows with LPS was shown to induce ET in mammary epithelial cells (MEC), thereby protecting the udder against reinfection for some time. Seeking alternatives to LPS priming we tried to elicit ET by priming MEC with either lipopeptide (Pam2CSK4) via the TLR2/6 axis or inhibitors of histone-modifying enzymes. Pre-incubation of MEC with Pam2CSK4 enhanced baseline and induced expression of bactericidal (beta-defensin;SLPI) and membrane protecting factors (SAA3, TGM3), while reducing the expression of cytokine-and chemokine-encoding genes (TNF, IL1 beta) after a subsequent pathogen challenge, the latter, however, not as efficiently as after LPS priming. Pre-treating MEC with various inhibitors of histone H3 modifiers (for demethylation, acetylation or deacetylation) all failed to induce any of the protective factors and only resulted in some dampening of cytokine gene expression after the re-challenge. Hence, triggering immune functions via the TLR axis, but not through those histone modifiers, induced the beneficial phenomenon of ET in MEC

Lipopolysaccharide priming enhances expression of effectors of immune defence while decreasing expression of pro-inflammatory cytokines in mammary epithelia cells from cows

Background: Udder infections with environmental pathogens like Escherichia coli are a serious problem for the dairy industry. Reduction of incidence and severity of mastitis is desirable and mild priming of the immune system either through vaccination or with low doses of immune stimulants such as lipopolysaccharide LPS was previously found to dampen detrimental effects of a subsequent infection. Monocytes/macrophages are known to develop tolerance towards the endotoxin LPS (endotoxin tolerance, ET) as adaptation strategy to prevent exuberant inflammation. We have recently observed that infusion of 1 mu g of LPS into the quarter of an udder effectively protected for several days against an experimentally elicited mastitis. We have modelled this process in primary cultures of mammary epithelial cells (MEC) from the cow. MEC are by far the most abundant cells in the healthy udder coming into contact with invading pathogens and little is known about their role in establishing ET. Results: We primed primary MEC cultures for 12 h with LPS (100 ng/ml) and stimulated three cultures either 12 h or 42 h later with 10(7)/ml particles of heat inactivated E. coli bacteria for six hours. Priming-related alterations in the global transcriptome of those cells were quantified with Affymetrix microarrays. LPS priming alone caused differential expression of 40 genes and mediated significantly different response to a subsequent E. coli challenge of 226 genes. Expression of 38 genes was enhanced while that of 188 was decreased. Higher expressed were antimicrobial factors (beta-defensin LAP, SLPI), cell and tissue protecting factors (DAF, MUC1, TGM1, TGM3) as well as mediators of the sentinel function of MEC (CCL5, CXCL8). Dampened was the expression of potentially harmful pro-inflammatory master cytokines (IL1B, IL6, TNF-alpha) and immune effectors (NOS2, matrix metalloproteases). Functional network analysis highlighted the reduced expression of IL1B and of IRF7 as key to this modulation. Conclusion: LPS-primed MEC are fitter to repel pathogens and better protected against misguided attacks of the immune response. Attenuated is the exuberant expression of factors potentially promoting immunopathological processes. MEC therefore recapitulate many aspects of ET known so far from professional immune cells

Escherichia coli infection induces distinct local and systemic transcriptome responses in the mammary gland

Background: Coliform bacteria are the most common etiologic agents in severe mastitis of cows. Escherichia coli infections are mostly restricted to a single udder quarter whereas neighboring quarters stay clinically inapparent, implicating the presence of a systemic defense reaction. To address its underlying mechanism, we performed a transcriptome study of mammary tissue from udder quarters inoculated with E. coli (6 h and 24 h post infection), from neighboring quarters of the same animals, and from untreated control animals. Results: After 6 h 13 probe sets of differentially expressed genes (DEG) were detected in infected quarters versus control animals. Eighteen hours later 2154 and 476 DEG were found in infected and in neighboring quarters vs. control animals. Cluster analysis revealed DEG found only in infected quarters (local response) and DEG detected in both infected and neighboring quarters (systemic response). The first group includes genes mainly involved in immune response and inflammation, while the systemic reaction comprises antigen processing and presentation, cytokines, protein degradation and apoptosis. Enhanced expression of antimicrobial genes (S100A8, S100A9, S100A12, CXCL2, GNLY), acute phase genes (LBP, SAA3, CP, BF, C6, C4BPA, IF), and indicators of oxidative stress (GPX3, MT1A, MT2A, SOD2) point to an active defense reaction in infected and neighboring healthy quarters. Its early onset is indicated by increased transcription of NFIL3 at 6 h. NFIL3 is a predicted regulator of many genes of the systemic response at 24 h. The significance of our transcriptome study was evidenced by some recent findings with candidate gene based approaches. Conclusions: The discovery and holistic analysis of an extensive systemic reaction in the mammary gland significantly expands the knowledge of host-pathogen interactions in mastitis which may be relevant for the development of novel therapies and for genetic selection towards mastitis resistance