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Chronic obstructive pulmonary disease (COPD) has become a global epidemic disease with an increased morbidity and mortality in the world. Inflammatory process progresses and contributes to irreversible airflow limitation. However, there is no available therapy to better control the inflammatory progression and therefore to reduce the exacerbations and mortality. Thus, the development of efficient anti-inflammatory therapies is a priority for patients with COPD. β(2) -Adrenoceptor agonists and anticholinergic agents are widely used as first line drugs in management of COPD because of their efficient bronchodilator properties. At present, many studies in vitro and some data obtained in laboratory animals reveal the potential anti-inflammatory effects of these bronchodilators but their protective role against chronic inflammation and the development of emphysema in patients with COPD remains to be investigated. The anti-inflammatory effects of theophylline at low doses have also been identified. Beneficial interactions between glucocorticoids and bronchodilators have been reported, and signaling pathways explaining these synergistic effects begin to be understood, especially for theophylline. Recent data demonstrating interactions between anticholinergics with β(2) -adrenoceptor agonists aiming to better control the pulmonary inflammation and the development of emphysema in animal models of COPD justify the priority to investigate the interactive effects of a tritherapy associating corticoids with the two main categories of bronchodilators.

International audience; The development of temperate deciduous and conifers forests stands usually results in accumulation of forest floor organic matter and a shift from mull to moder humus forms. It has been suggested that an increase in nutrient uptake by trees during their rapid growth phase leads to topsoil acidification, decrease in earthworm density and thereby a decrease in litter turnover. The focus of this paper was to examine if the mull-moder shift with forest ageing results from higher leaf litter production and/or lower litter decay rates. The objectives of this research were to determine (1) changes in macro-morphological properties of humus forms, leaf litter production, litter decay rates, soil nutrients content and pH along a 130-year pure beech (Fagus sylvatica L.) chronosequence in Normandy (Northwest France), (2) if humus form varied from mull to moder with increasing stand age, and (3) if a shift from mull to moder resulted from increased litter production, decreased litter decay rates, or both. Annual litter production did not change significantly along the chronosequence (mean 2.41 t ha−1). In contrast, litter decay rates decreased significantly during the rapid growth phase of trees. In consequence, the litter turnover time (1/k) was lower in the youngest stands (20 months) compared to the oldest ones (31 months). Even in the absence of a significant pattern of variation, litter production was positively correlated with the thickness of the OF (Oi) horizon. In contrast, litter decay was strongly negatively correlated with maximum thickness of the OH (Oa) horizon, suggesting that the appearance of the humification layer was mainly due to a decrease in litter decay rate. We did not find significant changes in the main properties of the organo-mineral horizon, suggesting that soil nutrient availability may not directly affect litter dynamics. We concluded that moder development along the chronosequence resulted in decreasing litter decay rates during the aggradation phase while litter production was stable. Further studies are required to identify the ecological factors responsible for moder development along forest ageing.

Staphylococcus (S.) aureus is a major pathogen involved in chronic bovine mastitis. Staphylococcal mastitis is difficult to control due to the ability of S. aureus to invade and survive within host cells. We therefore postulated that induction of CD8(+) cytotoxic T lymphocyte (CTL) responses leading to destruction of infected cells could help in the control of S. aureus mastitis. We demonstrate that immunization of mice with heat-killed S. aureus together with agonistic anti-CD40 monoclonal antibodies elicits strong CTL responses capable of reducing the severity of subsequent staphylococcal mastitis. Our study shows promise for CTL-dependent vaccination against S. aureus mastitis.

The objective of the current study was to investigate (i) the outcome of experimentally induced Escherichia coli mastitis in primiparous cows during early lactation in relation with production of eicosanoids and inflammatory indicators, and (ii) the validity of thermography to evaluate temperature changes on udder skin surface after experimentally induced E. coli mastitis. Nine primiparous Holstein Friesian cows were inoculated 24 ± 6 days (d) after parturition in both left quarters with E. coli P4 serotype O32:H37. Blood and milk samples were collected before and after challenge with E. coli. The infrared images were taken from the caudal view of the udder following challenge with E. coli. No relationship was detected between severity of mastitis and changes of thromboxane B2 (TXB2), leukotriene B4 (LTB4) and lipoxin A4 (LXA4). However, prostaglandin E2 (PGE2) was related to systemic disease severity during E. coli mastitis. Moreover, reduced somatic cell count (SCC), fewer circulating basophils, increased concentration of tumor necrosis factor-α (TNF-α) and higher milk sodium and lower milk potassium concentrations were related to systemic disease severity. The thermal camera was capable of detecting 2-3°C temperature changes on udder skin surface of cows inoculated with E. coli. Peak of udder skin temperature occurred after peak of rectal temperature and appearance of local signs of induced E. coli mastitis. Although infrared thermography was a successful method for detecting the changes in udder skin surface temperature following intramammary challenge with E. coli, it did not show to be a promising tool for early detection of mastitis.

La stabilité structurale est une propriété physique des sols importante, indicatrice de leur sensibilité à la battance et à l’érosion. Les sols limoneux, qui couvrent des surfaces importantes en France et dans le Nord de l’Europe ont, du fait de leur texture et de leur faible teneur en matières organiques, une stabilité structurale faible. Dans un contexte où l’on se préoccupe de la diminution de la teneur en matière organique des sols, où se développe l’apport de produits résiduaires organiques au sol et où se diversifient les pratiques culturales et les systèmes de culture, il apparaît nécessaire de développer des outils de prédiction de la stabilité structurale en fonction de la quantité et de la qualité des matières organiques. Nous avons développé une relation statistique qui relie la stabilité de la structure à la constitution des sols, en particulier à leur teneur en carbone organique (C), à partir d’une base de données AGRESTA de 480 sols assemblée pendant cette étude. Cependant sa faible capacité prédictive ne permet pas de l’utiliser comme fonction de pédotransfert. L’examen de 7 essais de longue durée ou réseaux de parcelles sur sols limoneux a montré des améliorations de la stabilité de la structure en quelques années avec des pratiques qui permettent une augmentation de la teneur en C dans l’horizon de surface (non labour, prairies temporaires, semis direct sous couvert végétal (SCV), apports répétés de composts). La teneur en C totale de l’horizon est souvent un meilleur prédicteur de la stabilité structurale que des fractions organiques (carbohydrates solubles à l’eau chaude, matières organiques particulaires) ou que la biomasse des microorganismes. Des expérimentations réalisées en laboratoire d’apport de matières organiques (résidus de culture, composts) à des sols limoneux, nous ont permis de confirmer que le rôle de ces apports était essentiellement indirect, par la stimulation des microorganismes du sol que leur décomposition entraîne. Les microorganismes agrégent les particules de sols par leur sécrétions ou mécaniquement. Nous avons proposé un modèle prédictif de la stabilité structurale suite à des apports organiques dont on connaît la qualité biochimique : CANTIS-STAB, qui couple un modèle de décomposition des matières organiques à une fonction statistique. Comme la gestion des matières organiques constitue un volant d’action majeur pour agir sur la sensibilité de sols limoneux à la battance et à l’érosion, les outils proposés doivent être développés, afin de servir de base à des préconisations agronomiques ou à l’apport raisonné de produits résiduaires organiques au sol., Aggregate stability is a major soil physical property, which is a good indicator of the sensitivity of soils to crusting and erosion. Silty soils, which cover large surface areas in France and Northern Europe, have a low aggregate stability, because of their texture and of their frequent low organic matter contents. In the present context of (i) soil organic matter content depletion, (ii) development of organic wastes application to soils and (iii) diversification of cropping systems, it is necessary to have tools to predict changes in soil aggregate stability with changes in soil organic matter content and quality. Using a data base of 480 soils, established during this study, we developed a statistical relationship which relates soil aggregate stability with soil characteristics, in particular with its organic carbon (C) content. However, its low predictive capacity does not allow to use it as a pedotransfer function. Based on 7 in situ long term experiments or cultivated plots series, we found rapid increased of soil aggregate stability when practices that increases the soil C content in the surface layer are implemented (no till, rotations with leys, permanent coverage of soil with plants, repeated compost additions). The total C content of soil was often better correlated to aggregate stability than other organic variables, such as hot water soluble polysaccharides, particulate organic matter contents, or microbial biomass C. Laboratory experiments, in which a wide range of organic materials (crop residues, composts) were added to soil showed that the effect of the organic materials was indirect, through the stimulation of the microbial decomposers, which aggregated the soil. We proposed a new model to simulate the temporal changes in aggregate stability after organic matter additions of given biochemical qualities and in controlled amounts. “CANTIS-STAB” couples a decomposition model with a statistical function. Managing organic matter in soils is a powerful option to decrease the sensitivity of silty cultivated soils to crusting and erosion. The different tools proposed in this program will be developed further, to serve as a basis to optimize agronomic practices and organic wastes addition in this perspective.

International audience; This study aims at describing the mechanisms of earthworm species assemblages in a temperate grassland ageing gradient. Earthworms were sampled by a combination of formaldehyde extraction and hand sorting. Density data were analysed by combining correspondence analysis (CA) and null model analyses of niche overlap patterns and morphological trait dispersion. The first axis of the CA arranged samples according to the pasture ageing gradient and separated “pioneer” (CA1−) from “old pasture” (CA1+) species assemblages. The second axis segregated two different assemblages (CA2− and CA2+) that were consistently represented along the ageing gradient and was assumed to represent intra-plot assemblage heterogeneity. Niche overlap according to soil organic C, C:N ratio and root biomass was higher than expected by chance (EBC) in most assemblages, and was higher when calculated for the whole regional species pool than for local assemblages. Morphological dispersion was random or lower than expected by chance for the regional species pool and both CA1− and CA1+, and higher than expected by chance for both CA2− and CA2+. These results indicate that: (1) habitat and dispersal constraints act as filters by allowing only those species with similar prerequisite traits into assemblages; (2) inter-specific competition limit composition in a further step by calling for a minimal level of overdispersion in morphological traits.

Zinc finger protein regulator of apoptosis and cell cycle arrest (Zac1) is a transcription factor able to induce apoptosis or cell cycle arrest through independent pathways. In spite of the important potential functions attributed to Zac1, little is known of its physiological regulation and biological function. We discovered that variant Zac1b was expressed in murine embryonic fibroblasts (MEFs) treated with polyriboinosinic polyribocytidylic acid [poly(I:C)], a synthetic double-stranded RNA. This regulation occurred mainly through Toll-Like Receptor 3 (TLR3)- and Interferon Regulatory Factor 3 (IRF3)-dependent pathways. As TLR3 and IRF3 are central activators of antiviral immunity, we hypothesized that Zac1 may be implicated in antiviral responses. In line with this notion, we observed that Zac1b was expressed in MEFs infected with Encephalomyocarditis virus (EMCV). We also observed that Zac1-deficient MEFs were less sensitive to EMCV-induced cell death than wild-type MEFs. However, Zac1 gene inactivation had no effect on the survival of mice infected with EMCV. In conclusion, this study describes for the first time a transcriptional regulation of Zac1b, induced by synthetic dsRNA and RNA viruses, the functional significance of which remains to be further investigated.

The anti-inflammatory properties of inhaled formoterol and ipratropium bromide, alone or in combination, were investigated in a rat model of chronic pulmonary inflammation with airspace enlargement induced by cadmium inhalation. At the end of the protocol, cadmium-induced increase of airway resistance was prevented by formoterol (4 mg/30 ml) or ipratropium (0.20 mg/20 ml). Formoterol elicited a significant decrease in total cell and neutrophil counts in bronchoalveolar lavage fluid as well as on the activity of gelatinase B (MMP-9), an enzyme strongly expressed in alveolar macrophages and epithelial cells. Additionally, a significant attenuation of the lung lesions characterized by inflammatory cell infiltration within the alveoli and the interstitium and a decrease in mean linear intercept were observed. Although ipratropium alone had no effects on the cadmium-induced pulmonary inflammation and emphysema, its combination with an inefficient concentration of formoterol (1 mg/30 ml) showed a synergistic inhibitory effect on neutrophil and total cell counts as well as on the mean linear intercept associated with a synergistic inhibition on the MMP-9 activity. Gelatinase A (MMP-2) activity was not influenced by drug pretreatments. Neither macrophage metalloelastase (MMP-12) activity nor levels of cytokines IL-1β, TNF-α and GM-CSF in bronchoalveolar lavage fluid were modified in rats chronically exposed to cadmium. No desensitization of β(2)-adrenoceptors or cholinergic receptors on airway smooth muscles and inflammatory cells during the protocol was observed. In conclusion, formoterol alone or combined with ipratropium bromide partially protects the lungs against the chronic inflammation and airspace enlargement by reducing neutrophilic infiltration possibly via the inhibition of MMP-9 activity.

The respiratory tract is continuously exposed to both innocuous airborne antigens and immunostimulatory molecules of microbial origin, such as LPS. At low concentrations, airborne LPS can induce a lung DC-driven Th2 cell response to harmless inhaled antigens, thereby promoting allergic asthma. However, only a small fraction of people exposed to environmental LIPS develop allergic asthma. What prevents most people from mounting a lung DC-driven Th2 response upon exposure to LPS is not understood. Here we have shown that lung interstitial macrophages (IMs), a cell population with no previously described in vivo function, prevent induction of a Th2 response in mice challenged with LPS and an experimental harmless airborne antigen. IMs, but not alveolar macrophages, were found to produce high levels of IL-10 and to inhibit LPS-induced maturation and migration of DCs loaded with the experimental harmless airborne antigen in an IL-10-dependent manner. We further demonstrated that specific in vivo elimination of IMs led to overt asthmatic reactions to innocuous airborne antigens inhaled with low doses of LPS. This study has revealed a crucial role for IMs in maintaining immune homeostasis in the respiratory tract and provides an explanation for the paradox that although airborne LPS has the ability to promote the induction of Th2 responses by lung DCs, it does not provoke airway allergy under normal conditions

International audience; The long pentraxin 3 (PTX3) plays an important role in host defence and its over-expression may contribute to airway injury. The aim of the present study was therefore to characterize in more detail PTX3 and its expression in the horses' airway. Six healthy horses and six horses affected by recurrent airway obstruction (R.A.O.) were submitted to a dusty environment challenge. PTX3 DNA and cDNA were cloned and sequenced. PTX3 expression was evaluated by RT-qPCR, Western blotting and immuno-histochemistry in bronchoalveolar lavage fluid (BALF) cells, BALF supernatant and bronchial epithelial cells. An alternative splicing of the second exon of PTX3 occurred, resulting in two forms of the protein: "spliced" (32 kDa) and "full length" (42 kDa). PTX3 was detected in BALF macrophages, neutrophils and bronchial epithelial cells. It was over-expressed in the BALF supernatant from R.A.O.-affected horses in crisis. However, dust was unable to induce PTX3 in BALF cells ex vivo, indicating that dust is an indirect inducer of PTX3. Dust exposure in-vivo induced PTX3 in BALF macrophages but there was no significant difference between healthy and R.A.O.-affected horses. Conversely, PTX3 was over-expressed in the bronchial epithelial cells from R.A.O.-affected horses in crisis. These data indicate a differential regulatory mechanism in inflammatory and bronchial epithelial cells and offer therapeutically interesting perspectives.

The conditions leading to the activation/differentiation of T-helper (Th) cells dedicated for B-cell antibody production are still poorly characterized. We now demonstrate that interleukin-6 (IL-6) promotes the differentiation of naive T lymphocytes into helper cells able to promote B-cell activation and antibody secretion. IL-6–driven acquisition of B-cell help capacity requires expression of the signal transducer and activator of transcription 3 (STAT3), but not STAT4 or STAT6 transcription factors, suggesting that the ability to provide help to B cells is not restricted to a well-defined Th1 or Th2 effector population. T cell–specific STAT3-deficient mice displayed reduced humoral responses in vivo that could not be related to an altered expansion of CXCR5-expressing helper T cells. IL-6 was shown to promote IL-21 secretion, a cytokine that was similarly found to promote the differentiation of naive T cells into potent B-cell helper cells. Collectively, these data indicate that the ability to provide B-cell help is regulated by IL-6/IL-21 through STAT3 activation, independently of Th1, Th2, Th17, or follicular helper T cell (TFH) differentiation.

Changes in soil N mineralization pathways occurring along a full rotation cycle have received little attention to date, while tree uptake for N may change during forest ageing. The aims of this study were (i) to characterize changes in potential net N mineralization and potential net nitrification within organic layers and the topsoil (organo-mineral horizon) along a 100-year chronosequence for a temperate oak–hornbeam forest and (ii) to reveal covariances between potential net N mineralization pathways and the properties of the humic epipedon (defined as the sum of organic layers and topsoil). For that purpose, a space-for-time substitution procedure and aerobic laboratory incubation method for 28 days at 28 °C in the dark were used. In addition, acetylene and captan were used to discriminate between autotrophic and heterotrophic (bacterial and/or fungal) nitrification. Several humic epipedon properties were determined, e.g. pH, exchangeable cation concentrations, effective cation exchange capacity, total C and N, dissolved organic C and N, fungal and microbial biomass N. Potential net N mineralization and nitrification pathways changed greatly along the mixed forest chronosequence. Potential net N mineralization in the organic layers increased with stand maturation whereas potential net nitrification in the topsoil decreased significantly. Selective inhibitors revealed changes in nitrification pathways along the chronosequence, i.e. potential net nitrification was autotrophic in the topsoil while it was mainly heterotrophic within the organic layers. In the organic layer, potential net nitrification was autotrophic at the onset of the chronosequence while it appeared heterotrophic during the aggradation phase and finally fungal in mature stands. A Co-Inertia Analysis was used to reveal covariances between N mineralization pathways and humic epipedon properties. The analysis showed two functional temporal shifts within N cycling along the chronosequence, one probably controlled by organic matter quality and high competition for available N resulting in the autotrophic versus heterotrophic nitrification shift in the organic layers and one mainly controlled by (i) fine organic matter abundance, allowing high N mineralization in the organic layers and (ii) acidity inhibited autotrophic nitrification in the topsoil.

When a globiferous pedicellaria ofSphaerechinus granularis injects its venom, the head autotomizes whereas the stalk remains on the test and enters a regression process with concomitant resorption of its supporting ossicle (i.e. the rod). Scanning electron microscope investigations of the morphological changes undergone by the stereom of resorbing rods show that: (1) resorption proceeds both axially and laterally, and leads to a reduction of approximately 80% of the original length of the rod, (2) secondary growth of new stereom processes occurs concomitantly with resorption but never ensures even a partial regeneration of the rod, and (3) resorption and secondary growth stop before the rod is totally destroyed leaving a static stump that remains in place up to 190 days. Particular resorption figures result from either the axial or the lateral resorption of the rod shaft. These consist chiefly of terraced conical cupules, dense cylinders and concentric lamellae whose walls or edges are typically made of closely piled and/or aligned subprismatic crystallites. Whatever their location along the rod, these crystallites always organize strictly parallel to the rod axis. Whether the crystallites are mosaic blocks composing larger monocrystalline units or discrete monocrystals themselves is for the moment unclear. A growth model, which accounts for the observed resorption figures, is proposed for the shaft of pedicellarial rods. According to this model, the early growth of the shaft would produce elongated, interconnected trabeculae (initial trabeculae) made of densely piled and perfectly aligned crystallites. Thickening and coalescence of adjoining trabeculae would progressively occur by adjunction around the initial trabeculae of successive and concentric layers of similarly arranged crystallites. Coalescent trabeculae would then be cemented together in a perforate stereom layer by the final deposition of larger crystallite layers surrounding the whole shaft periphery. Growth of secondary stereom processes occurs both in the resorbing rod (here the newly formed processes are resorbed soon after they have been produced) and in rods where resorption has stopped. These are always irregular processes that localize near or on the actual sites of resorption. It is suggested these processes result from an uncontrolled activation of the skeleton-forming cells in areas where the concentration of calcium ions increases as a consequence of calcite resorption.

Cadmium (Cd) induces centrilobular emphysema and is suspected to contribute to tobacco related lung diseases as chronic obstructive pulmonary disease (COPD). This study aimed to check whether the inflammatory process observed in rats exposed to nebulised Cd is resistant to betamethasone as observed in COPD and to determine the influence of this drug on airspace enlargement together with the MMP-2-9/TIMP-1-2 imbalance. Our results showed that betamethasone induced emphysema by itself in healthy rats. Moreover, pre-treatment of rats with betamethasone could only partially modulate the increase in bronchoalveolar lavage fluid cell counts and the absence of preventive effect of this compound against emphysema development is associated with its inability to rebalance the MMP-2-9/TIMP-1-2 ratio.

Nicotinamide phosphoribosyl transferase (Nampt)/pre-B cell colony-enhancing factor (PBEF)/visfatin is a protein displaying multiple functional properties. Originally described as a cytokine-like protein able to regulate B cell development, apoptosis, and glucose metabolism, this protein also plays an important role in NAD biosynthesis. To gain insight into its physiological role, we have generated a mouse strain expressing a conditional Nampt allele. Lack of Nampt expression strongly affects development of both T and B lymphocytes. Analysis of hemizygous cells and in vitro cell lines expressing distinct levels of Nampt illustrates the critical role of this protein in regulating intracellular NAD levels. Consequently, a clear relationship was found between intracellular Nampt levels and cell death in response to the genotoxic agent MNNG (N-methyl-N′-nitro-N-nitrosoguanidine), confirming that this enzyme represents a key regulator of cell sensitivity to NAD-consuming stress secondary to poly(ADP-ribose) polymerases overactivation. By using mutant forms of this protein and a well-characterized pharmacological inhibitor (FK866), we unequivocally demonstrate that the ability of the Nampt to regulate cell viability during genotoxic stress requires its enzymatic activity. Collectively, these data demonstrate that Nampt participates in cellular resistance to genotoxic/oxidative stress, and it may confer to cells of the immune system the ability to survive during stressful situations such as inflammation.

International audience; Accurate estimates of forest soil organic matter (OM) are now crucial to predictions of global C cycling. This work addresses soil C stocks and dynamics throughout a managed beechwood chronosequence (28–197 years old, Normandy, France). Throughout this rotation, we investigated the variation patterns of (i) C stocks in soil and humic epipedon, (ii) macro-morphological characteristics of humic epipedon, and (iii) mass, C content and C-to-N ratio in physical fractions of humic epipedon. The fractions isolated were large debris (>2000 μm), coarse particular OM (cPOM, 200–2000 μm), fine particular OM (fPOM, 50–200 μm) and the mineral associated OM (MaOM

Neutrophils rapidly undergo spontaneous apoptosis following their release from the bone marrow. Although central to leukocyte homeostasis, the mechanisms that regulate neutrophil apoptosis remain poorly understood. We show here that apoptosis of cultured neutrophils is preceded by a substantial increase in the intracellular levels of 16 and 24 carbon atom (C16- and C24)-ceramides, which are lipid second messengers of apoptosis and stress signaling. Treatment of neutrophils with fumonisin B2, a selective inhibitor of the de novo pathway of ceramide synthesis, prevented accumulation of C16- and C24-ceramides. Moreover, fumonisin B2 significantly reduced caspase-3, -8, and -9 activation and apoptosis in these cells. Conversely, 3-O-methylsphingomyelin and fantofarone, which are specific inhibitors of neutral and acid sphingomyelinases, respectively, neither inhibited C16- and C24-ceramide production nor decreased the apoptosis rate in neutrophils, indicating that in these cells, ceramides are not generated from membrane sphingomyelin. Further experiments showed that increasing endogenous C16- and C24-ceramide levels by using DL-threo-1-phenyl-2-palmitoylamino-3-morpholino-1-propanol and (1S,2R)-D-erythro-2-(N-myristoylamino)-1-phenyl-1-propanol, two inhibitors of ceramide metabolism, enhances caspase-3, -8, and -9 activity and increases neutrophil apoptosis. Similarly, apoptosis was induced rapidly when synthetic C16- and/or C24-ceramides were added to neutrophil cultures. Finally, GM-CSF, a cytokine that delays neutrophil apoptosis, abrogated C16- and C24-ceramide accumulation totally in cultured neutrophils, whereas Fas ligation accelerated apoptosis in these cells without affecting de novo ceramide production. We conclude that de novo generation of C16- and C24-ceramides contributes to spontaneous neutrophil apoptosis via caspase activation and that GM-CSF exerts its antiapoptotic effects on neutrophils, at least partly through inhibition of ceramide accumulation.

BACKGROUND: Although STAT5 promotes survival of hematopoietic progenitors, STAT5-/- mice develop mild neutrophilia. METHODOLOGY/PRINCIPAL FINDINGS: Here, we show that in STAT5-/- mice, liver endothelial cells (LECs) autonomously secrete high amounts of G-CSF, allowing myeloid progenitors to overcompensate for their intrinsic survival defect. However, when injected with pro-inflammatory cytokines, mutant mice cannot further increase neutrophil production, display a severe deficiency in peripheral neutrophil survival, and are therefore unable to maintain neutrophil homeostasis. In wild-type mice, inflammatory stimulation induces rapid STAT5 degradation in LECs, G-CSF production by LECs and other cell types, and then sustained mobilization and expansion of long-lived neutrophils. CONCLUSION: We conclude that STAT5 is an ambivalent factor. In cells of the granulocytic lineage, it exerts an antiapoptotic function that is required for maintenance of neutrophil homeostasis, especially during the inflammatory response. In LECs, STAT5 negatively regulates granulopoiesis by directly or indirectly repressing G-CSF expression. Removal of this STAT5-imposed brake contributes to induction of emergency granulopoiesis., Journal Article, Research Support, Non-U.S. Gov't, info:eu-repo/semantics/published

1. Denitrification in floodplain soils is one of the main biological processes emitting and reducing nitrous oxide, a greenhouse gas, and the main process responsible for the buffering capacity of riparian zones against diffuse nitrate pollution. 2. The aim of this study was to measure denitrification rates under a wide range of current climatic conditions and hydrological regimes in Europe (from latitude 64°N to latitude 42°N and from longitude 2°W to longitude 25°E), in order to determine the response patterns of this microbial process under different climatic and hydrological conditions, and to identify denitrification proxies robust enough to be used at the European scale. 3. Denitrification activity was significant in all the floodplain soils studied whatever the latitude. However, we found an increase in rates of an order of magnitude from high to mid latitudes. Maximum rates (above 30 g N m?2 month?1) were measured in the maritime conditions of the Trent floodplain. These rates are similar to mineralisation rates measured in alluvial soils and of the same order of magnitude as the amount of N stored in herbaceous plants in alluvial soils. 4. We used Multivariate Adaptative Regression Splines to relate the response variable denitrification with five relevant predictors, namely soil moisture, temperature, silt plus clay, nitrate content and herbaceous plant biomass. 5. Soil moisture, temperature, and nitrate were the three main control variables of microbial denitrification in alluvial soils in decreasing order of importance. 6. The model developed for denitrification with interaction effects outperformed a pure additive model. Soil moisture was involved in all interactions, emphasising its importance in predicting denitrification. 7. These results are discussed in the context of scenarios for future change in European hydrological regimes.

This work addresses the impact of eight detritivorous species of soil macro-invertebrates (three millipedes, two woodlice and three earthworms) on short-term carbon mineralization and mechanical breakdown of beech leaves. The production rate, size class distribution and OM content of invertebrate faeces were also measured. Hierarchical clustering (HC) and multivariate analysis were performed to find relevant functional groups among the species studied. Our results identified three groups of macro-invertebrates on the basis of their impacts on beach leaf degradation (hierarchical clustering): (1) invertebrates that produce fresh faeces with high N contents compared with other species (i.e. polydesmidae and a single species of lumbricidae); (2) other lumbricidae that fragment litter into fine particles in their faeces and actively stimulate CO2 release; (3) other arthropods that fragment litter into coarse particles and have weak impacts on OM mineralization. These groups over-ride taxonomy, and are proposed as a tentative functional classification of litter dwelling invertebrates. On the other hand, an idiosyncratic impact of species was observed in each group, highlighting how much empirical data are still needed to propose a robust functional classification of litter invertebrates. # 2006 Elsevier B.V. All rights reserved.

We sought to determine whether prolactin (PRL) could influence the neutrophilic inflammation that characterizes chronic mastitis. Most of the genes encoding inflammatory proteins depend on the nuclear factor kappaB (NF-kappaB) for their expression. We addressed the hypothesis that immunomodulatory activities of PRL might arise from an increase in NF-kappaB activity. MAC-T cells, a bovine mammary epithelial cell line, were stimulated with increasing concentrations of bovine PRL (1, 5, 25, 125, and 1,000 ng/mL). Level of NF-kappaB binding activity was measured and mRNA was evaluated for IL-1beta, IL-6, IL-8, granulocyte-macrophage colony-stimulating factor (GMCSF), IFN-gamma, and tumor necrosis factor (TNF)-alpha, cytokines known to require NF-kappaB for their maximal transcription. Prolactin activated NF-kappaB; maximal NF-kappaB activation was weaker with PRL than with TNF-alpha at 30 or 180 min poststimulation. In addition, PRL significantly amplified, in a dose-dependent manner, mRNA expression of IL-1beta, IL-6, IL-8, GMCSF, and TNF-alpha. We measured PRL concentrations in blood and milk from healthy and chronic mastitis-infected cows, and studied the relationship between the PRL concentration and the degree of inflammation in the mammary gland as indirectly assessed by somatic cell counts (SCC). Plasma PRL did not differ significantly between healthy and chronic mastitis-affected cows (63.7 and 67.5 ng/mL, respectively). Milk PRL concentration was significantly increased in chronic mastitis-affected quarters with the highest SCC, and had a positive significant correlation between SCC, as well as between the number of neutrophils present in milk samples. The present findings show that PRL promotes an inflammatory response in bovine mammary epithelial cells via NF-kappaB activation, and suggest a role for PRL in the pathogenesis of chronic mastitis.

Glucocorticoid (GC) therapy is recognized to be effective for the treatment of recurrent airway obstruction (RAO) in horses. Anti-inflammatory properties of GC are thought to be mediated by suppression of inflammatory gene expression via inhibition of transcription factors such as nuclear factor-kappaB (NF-kappaB) and activator protein-1 (AP-1). The purpose of this study was to evaluate the effect of low-dose inhaled beclomethasone dipropionate and injectable dexamethasone 21-isonicotinate on clinical signs, pulmonary function, airway cytology, and activity of NF-kappaB and AP-1 in bronchial cells of RAO-affected horses. Seven horses with RAO were exposed to moldy hay until they developed airway obstruction on 3 separate occasions. In a crossover design, they were then treated with a placebo (injection on day 1), inhaled beclomethasone (500 microg q12h for 10 days), or dexamethasone (0.06 mg/kg, IM on day 1) and monitored for 10 days. Pulmonary function, bronchoalveolar lavage fluid cytology, and NF-kappaB and AP-1 activity in bronchial brushing cells were measured before (day 1) and after treatment (day 10). Treatment with beclomethasone resulted in significantly improved pulmonary function of RAO-affected horses compared with placebo and dexamethasone treatments. However, none of the treatments had an effect on bronchoalveolar lavage fluid cytology or NF-kappaB and AP-1 activity. These findings reveal that, in a model of severe RAO, the benefits of low-dose inhaled beclomethasone on pulmonary function are not accompanied by a decrease in airway inflammatory cells or a suppression of transcription factors NF-kappaB and AP-1 DNA-binding activity.

Staphylococcus ( S. ) aureus is a major udder pathogen causing bovine mastitis. Some pro-inflammatory cytokines, including tumor necrosis factor-α (TNF-α), enhance extracellular and intracellular growth of S. aureus , indicating that the inflammatory process favors S. aureus infection. Helenalin is a sesquiterpene lactone with potent anti-inflammatory properties. This study was designed to evaluate the effects of helenalin on S. aureus infection. First, in vitro experiments were conducted. These studies revealed that proliferation of S. aureus in bovine mammary epithelial MAC-T cells treated in the presence or absence of TNF-α was markedly reduced in the presence of helenalin. Secondly, in vivo effects of helenalin were investigated. Lactating mice treated in the presence or absence of helenalin were challenged by the intramammary route with S. aureus and the bacteria in the mammary glands were counted 12 h after infection. Significantly less numbers of bacteria were recovered from the infected glands of helenalin-treated mice compared with untreated mice. Moreover, histological examination of mammary tissue from helenalin-treated mice that were challenged with S. aureus indicated that helenalin is able to significantly reduce leukocyte infiltration in the mammary gland following S. aureus inoculation. Our results show that helenalin reduces S. aureus intracellular growth and experimental S. aureus infection. We conclude that helenalin may be of potential interest in the treatment of S. aureus -induced mastitis in the bovine species.

Invertebrates play significant, but largely ignored, roles in the delivery of ecosystem services by soils at plot and landscape scales. They participate actively in the interactions that develop in soil among physical, chemical and biological processes. We show that soils have all the attributes of self-organized systems as proposed by Perry (Trends Ecol. Evol. 10 (1995) 241) and detail the scales at which invertebrates operate and the different kinds of ecosystem engineering that they develop. This comprehensive analysis of invertebrate activities shows that they may be the best possible indicators of soil quality. They should also be considered as a resource that needs to be properly managed to enhance ecosystem services provided by agro-ecosystems. © 2006 Published by Elsevier Masson SAS.

International audience; We investigate the influence of tree canopy composition on humus form variability and heterogeneity by comparing a pure beech stand and a mixed beech-hornbeam one (70% beech and 30% hornbeam). Macro-morphological humus form descriptors were recorded using a spatially explicit sample design at stand level. Leaf litter composition and light intensity accounting for stand management as well as bulk density for harvesting practices (soil compaction) were also recorded. A multiple correspondence analysis (MCA) and geostatistics were used to assess humus form variability and heterogeneity, as well as the spatial correlation between stand characteristics and humus forms. Humus form variability and activity were greater under the mixed stand than under the pure one. Geostatistics revealed that humus form patchiness was greater under the mixed stand than under the pure one and the improvement in decomposition processes seemed to be confined to spatial distribution of hornbeam litterfall. From a practical viewpoint, these results could provide ideas on the way of mixing tree species at stand level. We assume that with a given percentage of mull-forming tree species, a dispersed tree mixture provides a more extensive improvement than a clumped mixture.; Variabilité et hétérogénéité des formes d'humus à l'échelle du peuplement : comparaison entre peuplement pur de hêtre et forêt mélangée hêtre-charme. Nous avons étudié l'influence de la composition d'un peuplement sur la variabilité qualitative et l'hétérogénéité spatiale des formes d'humus en comparant un peuplement pur de hêtre et un peuplement mélangé (70 % hêtre - 30 % charme). Les caractéristiques macro-morphologiques des formes d'humus ainsi que la composition spécifique de la litière, l'éclairement relatif sous le peuplement et la densité apparente du sol ont été mesurées à l'aide d'un échantillonnage spatialement explicite. Une analyse des correspondances multiples (ACM) couplée à des géostatistiques a été utilisée pour caractériser la variabilité et l'hétérogénéité des formes d'humus ainsi que leur corrélation spatiale avec les caractéristiques de peuplement. La variabilité, l'hétérogénéité et l'activité des formes d'humus sont plus fortes sous peuplement mixte. Sous ce peuplement, l'amélioration de l'activité des formes d'humus est liée à la distribution spatiale des retombées de feuilles de charme. Ces résultats semblent indiquer que, pour un pourcentage donné d'une espèce ligneuse à litière améliorante, un mélange d'arbre par pied a un meilleur impact sur les formes d'humus qu'un mélange par bouquet.

Horses with recurrent airway obstruction (RAO) present many similarities with human asthmatics including airway inflammation, hyperresponsiveness, reversible obstruction, and increased NF-kappaB expression. Studies in experimental asthma models have shown that transcriptions factors such as activator protein-1 (AP-1), GATA-3, cyclic AMP response element binding protein (CREB) and CAAT/enhancer binding protein (C/EBP) may also play an important role in airway inflammation. The purpose of this study was to measure DNA binding activity of these transcription factors in the airways of horses with RAO and to compare it to pulmonary function and bronchoalveolar lavage fluid (BALF) cytology. Seven horses with RAO and six control animals were studied during a moldy hay challenge and after 2 months at pasture. Pulmonary function, BALF cytology and transcription factors' activities in bronchial brushings were measured during hay and pasture exposures. During moldy hay challenge, RAO-affected horses developed severe airway obstruction and inflammation and a significantly higher airway AP-1 binding activity than in controls. After 2 months on pasture, pulmonary function and airway AP-1 binding activity were not different between RAO and control horses. The DNA binding activity of CREB in airways of RAO-affected horses increased significantly after 2 months at pasture and became higher than in controls. A significant positive correlation was detected between AP-1 binding activity and indicators of airway obstruction and inflammation. Airway GATA-3, CEBP and CREB binding activities were negatively correlated with indices of airway obstruction. However, contrarily to CREB binding activity, GATA-3 and CEBP binding activities were not different between RAO and control horses and were unaffected by changes in environment. These data support the view that AP-1 and CREB play a role in modulating airway inflammation in horses with RAO.

In response to DNA damage, p53 activates a G1 cell cycle checkpoint, in part through induction of the cyclin-dependent kinase inhibitor p21(Waf1/Cip1). Here we report the identification of a new direct p53 target, Ptprv (or ESP), encoding a transmembrane tyrosine phosphatase. Ptprv transcription is dramatically and preferentially increased in cultured cells undergoing p53-dependent cell cycle arrest, but not in cells undergoing p53-mediated apoptosis. This observation was further confirmed in vivo using a Ptprv null-reporter mouse line. A p53-responsive element is present in the Ptprv promoter and p53 is recruited to this site in vivo. Importantly, while p53-dependent apoptosis is intact in mice lacking Ptprv, Ptprv-null fibroblasts and epithelial cells of the small intestine are defective in G1 checkpoint control. Thus, Ptprv is a new direct p53 target and a key mediator of p53-induced cell cycle arrest. Finally, Ptprv loss enhances the formation of epidermal papillomas after exposure to chemical carcinogens, suggesting that Ptprv acts to suppress tumor formation in vivo.

Rationale: Asthma is associated with increased expression of a typical array of genes involved in immune and inflammatory responses, including those encoding the prototypic Th2 cytokines interleukin (IL) 4, IL-5, and IL-13. Most of these genes contain binding sites for activator protein-1 (AP-1) within their promoter and are therefore believed to depend on AP-1 for their expression, suggesting that this transcription factor could be of particular importance in asthma pathophysiology. Objective: To clarify the role of AP-1 in the effector phase of pulmonary allergy. Methods: Ovalbumin (OVA)-sensitized mice were intratracheally given decoy oligodeoxyribonucleotides (ODNs) specifically directed to AP-1 or scrambled control ODNs before challenge with aerosolized OVA. Twenty-four hours after the last OVA challenge, airway hyperresponsiveness was measured and allergic airway inflammation was evaluated quantitatively. AP-1 decoys were localized using flow cytometry and immunohistochemistry. AP-1 activity in the lung was assessed using electrophoretic mobility shift assay. Measurements and Main Results: Intratracheally delivered AP-1 decoys efficiently targeted airway immune cells, thus precluding AP-1 activation on OVA challenge. Decoy-mediated local inhibition of AP-1 resulted in significant attenuation of all the pathophysiologic features of experimental asthma-namely, eosinophilic airway inflammation, airway hyperresponsiveness, mucous cell hyperplasia, production of allergen-specific immunoglobulins, and synthesis of IL-4, IL-5, and IL-13. Scrambled control ODNs had no detectable effects. Conclusions: Our results reveal a key role for AP-1 in the effector phase of pulmonary allergy and indicate that specific AP-1 inhibition in the airways may have therapeutic value in the control of established asthma.

The soil macrofauna of a pure beech (PS) and a mixed beech-hornbeam (MS) stand was recorded using a sample design spatially explicit at stand level. Humic epipedon morphological and chemical properties, relative irradiance, soil bulk density, and the specific composition of the litter in MS were also investigated. The taxonomic diversity is nearly similar on both sites, but the average by sample is greater under PS. The main factors controlling soil macrofauna spatial variability were: litter quality under MS and humus form activity under PS. These results suggest a distal control (i.e. external to humic epipedon) of macrofauna spatial variability when the litter quality is variable and a proximal control (i.e. internal to humic epipedon) when it is uniform at the spatial scale of the study.

Bovine subclinical mastitis can be defined as a moderated inflammatory disease characterized by a persistent accumulation of neutrophils in milk. As GMCSF-mediated delay of neutrophil apoptosis contributes to the accumulation of inflammatory cells at the site of inflammation in many human diseases, we sought to determine whether subclinical mastitis in cows is also associated with a GMCSF-dependent increase in milk-neutrophil survival. We first addressed the hypothesis that GMCSF delays bovine neutrophil apoptosis by activation of the signal transducer and activator of transcription (STAT) family members STAT3 and STAT5, which are critical regulators of the expression of various Bcl-2 family proteins. Granulocyte-macrophage colony-stimulating factor significantly delayed apoptosis of blood neutrophils obtained from healthy cows. In these cells, GMCSF activated STAT5, but not STAT3, and induced an increase in the mRNA of the antiapoptotic Bcl-2 member, Bcl-xL. Granulocyte-macrophage colony-stimulating factor-dependent STAT5 activation and up-regulation of Bcl-xL mRNA were blocked by the Jak inhibitor, AG-490. This inhibition was associated with abrogation of the prosurvival effect of GMCSF, demonstrating a key role for STAT5 in delayed neutrophil apoptosis. We further found that GMCSF expression was increased in milk cells from cows affected with subclinical mastitis. Neutrophils from these cows demonstrated a significant delay of apoptosis as compared with neutrophils obtained from healthy cows and were unresponsive to GMCSF. Active STAT5 complexes were detected in these neutrophils. Finally, in the presence of AG-490, apoptosis was induced and a time-dependent down-regulation of Bcl-xL mRNA was observed in milk neutrophils from mastitis-affected cows. These results indicate that neutrophil survival is enhanced in milk of subclinical mastitis-affected cows and suggest a role for a GMCSF-activated STAT5 signaling pathway in this phenomenon. This pathway could thus represent a target for the control of persistent accumulation of neutrophils in the bovine mammary gland.

Heat shock transcription factor (HSF)-1 is recognized as a central component of the heat shock response, which protects against various harmful conditions. However, the mechanisms underlying the protection and the role of HSF-1 in these mechanisms have not yet been clearly elucidated. Using HSF-1 knockout mice ( Hsf1−/−), we examined whether heat shock response-mediated lung protection involved an inhibition of the proinflammatory pathway via an interaction between HSF-1 and NF-κB, in response to cadmium insult. The HSF-1-dependent protective effect against intranasal instillation of cadmium (10 and 100 μg/mouse) was demonstrated by the higher protein content (1.2- and 1.4-fold), macrophage (1.6- and 1.9-fold), and neutrophil (2.6- and 1.8-fold) number in bronchoalveolar fluids, higher lung wet-to-dry weight ratio, and more severe lung damage evaluated by histopathology in Hsf1−/−compared with wild-type animals. These responses were associated with higher granulocyte/macrophage colony-stimulating factor (GM-CSF; 1.7-fold) but not TNF-α concentrations in bronchoalveolar fluids of Hsf1−/−mice compared with those of wild-type animals, indicating that HSF-1 behaved as a repressor of specific cytokine production in our model. To further investigate the mechanism of GM-CSF repression, we analyzed the NF-κB activity and IκB stability. The DNA binding NF-κB activity, in particular p50 homodimer activity, was higher in Hsf1−/−mice than in wild-type mice after cadmium exposure. These results provide a first line of evidence that mechanisms of lung protection depending on HSF-1 involve specific cytokine repression via inhibition of NF-κB activation in vivo.

Thiazolidinediones (TZDs) are pharmacological ligands of the peroxisome proliferator-activated receptor (PPAR)-gamma that are extensively used in the treatment of type II diabetes. Recently, an anti-inflammatory potential for TZDs has been suggested, based on observations that these compounds may inhibit pro-inflammatory cytokine expression in vitro and may attenuate the inflammatory response in vivo. Here, we show that the TZDs rosiglitazone (RSG) and troglitazone (TRO) do not inhibit the inflammatory response to tumor necrosis factor (TNF)-alpha in various epithelial cell types. On the contrary, both RSG and TRO significantly potentiated TNF-alpha-induced production of granulocyte/macrophage-colony-stimulating factor, interleukin (IL)-6 and/or IL-8 in these cells. This increase in pro-inflammatory cytokine expression was functionally significant as supernatants from cells co-treated with TNF-alpha and TZDs displayed increased neutrophil pro-survival activity when compared with supernatants from cells treated with TNF-alpha alone. Additionally, it was shown that TZDs enhance cytokine expression at the transcriptional level, but that the pro-inflammatory effects of TZDs are independent on PPARgamma, nuclear factor kappaB or mitogen-activated protein kinase activation. Our study shows that TZDs may potentiate the inflammatory response in epithelial cells, a previously unappreciated effect of these compounds.

International audience; Using a space-for-time substitution procedure, we assessed the patterns of variation during stand development of the humic epipedon and vegetation diversity in a pure European beech (Fagus sylvatica L.) forest and in a mixed beech–hornbeam (Carpinus betulus L.) forest. The humic epipedon was investigated through macro morpho logical characteristics (thickness of the OL, OF, OH, and A horizons) and pH measurements (pHKCl, pHH20, and ΔpH). Vegetation diversity was assessed using correspondence analysis and hierarchical cluster analysis to identify the main plant community succession groups. The groups were compared using three diversity measures (species richness, Shannon index, and evenness index) and three indices describing the community composition (similarity index between stands and index of heterogeneity at the stand and herbaceous layer levels). The pH measurements revealed an acidification of the A horizon during pure beech stand maturation. Macromorphological characteristics showed an alternation of an organic matter accumulation phase and organic matter disappearance along the two silvicultural cycles. Vegetation diversity measures showed small differences between the two forests. Indices accounting for community composition revealed a homogenizing effect of the pure beech silvicultural type on species composition of plant communities occurring during stand development. They also revealed a strong heterogeneity at the herbaceous level. Comparison of species composition indicated an increasing occurrence of acidophilous species in the pure beech forest. We discuss the interest of these results for future sustainable management decisions; Au moyen d'une approche synchronique, nous avons observé la variabilité de l'épisolum humifère et de la diversité végétale dans une hêtraie (Fagus sylvatica L.) pure et une hêtraie–charmaie (Carpinus betulus L.). L'épisolum humifère a été décrit à l'aide de variables morphologiques (épaisseurs des horizons OL, OF, OH et A) et chimiques (pHKCl, pHH20, et ΔpH). La diversité végétale fut appréhendée au moyen d'une analyse factorielle des correspondances associée à une classification ascendante hiérarchique afin de caractériser les principales communautés de plantes se succédant au cours des cycles sylvicoles. Les groupes furent comparés à l'aide de trois mesures de diversité (richesse spécifique, indices de Shannon et indice d'équitabilité) et trois indices rendant compte de la composition des communautés (indice de similarité et indices d'hétérogénéité à l'échelle du peuplement et à l'échelle de la strate herbacée). L'étude de l'épisolum humifère montre l'alternance de phases d'accumulation et d'incorporation de la matière organique dans les deux forêts ainsi qu'une acidification de l'horizon A au cours de la maturation des peuplements purs. Les patrons de variation de la diversité floristique sont sensiblement équivalents pour les deux forêts. La composition spécifique des communautés végétales se succédant au cours du cycle sylvicole de la hêtraie pure varie peu. Les indices rendant compte de la composition des communautés indiquent également une très forte hétérogénéité à l'échelle de la strate herbacée pour les deux forêts. La comparaison de la composition floristique révèle une plus forte occurrence des espèces acidiphiles dans la hêtraie pure. L'intérêt de ces résultats pour l'élaboration de règles de gestion sylvicole durable est discuté

Among the factors produced at inflammatory sites and those capable of modulating dendritic cell (DC) functions, PGD2 may be important in the outcome of immune responses. The biological roles for PGD2 are in part effected through two plasma membrane G protein-coupled receptors: the D prostanoid (DP) receptor and the chemoattractant receptor-homologous molecule expressed on Th2 lymphocytes (CRTH2). In this report, we studied the effects of PGD2 and of its major physiological metabolite, 15-deoxy-Δ12,14-PGJ2 (15d-PGJ2), on the functions of human monocyte-derived DC. First, we show that PGD2 exerts in vitro chemotactic effects on monocytes via CRTH2 activation while it inhibits the chemokine-driven migration of monocyte-derived DC through DP. We also report that PGD2 and 15d-PGJ2 alter the LPS- and allergen-induced DC maturation and enhance the CD80/CD86 ratio on mature DC in a DP- and CRTH2-independent manner. Moreover, PGD2 and 15d-PGJ2 strongly reduce the secretion of the Th1 promoting cytokine IL-12 and affect the synthesis of chemokines involved in Th1 cell chemotaxis, particularly CXCL10. Inhibition of cytokine/chemokine secretion implicates at least in part DP, but not CRTH2. The effects exerted by PGD2 are associated with the phosphorylation of CREB, but do not parallel with the deactivation of the NF-κB and mitogen-activated protein kinase pathways. In contrast, 15d-PGJ2 seems to target other cellular proteins. Finally, in a model of Th CD45RA+ differentiation induced by allergen- and superantigen-pulsed DC, PGD2 impacts on the orientation of the immune response by favoring a Th2 response.

Prostaglandin (PG) D(2), and its metabolites, are known to be important mediators during acute and chronic inflammation. However, their functions during the early phases of the immune response are poorly documented. In the present study, we show that PGD(2 )inhibits, in a dose-dependent manner, the CD40- and LPS-induced secretion of the Th1-driving factor IL-12 by murine splenic dendritic cells (DC), the most potent antigen-presenting cells. The inhibition of IL-12 production is mediated only in part by the cell surface G alpha s protein-coupled D prostanoid receptor (termed DP1) but not by the G alpha i protein-coupled DP receptor, DP2. We show that recruitment of DP1 in DC results in the activation of a cyclic AMP/protein kinase A pathway that is partially responsible for the inhibition of IL-12 production. We also suggest that the DP1-independent effects exerted by PGD(2) on IL-12 production may be due to the action of ist PGJ(2), but not PGF(2)alpha, metabolites. Electrophoretic mobility shift assays demonstrated that PGD(2) affects NF-kappa B activation through (the) DP1-independent pathway(s). Together these data suggest that PGD(2), by interacting with DP1 and by binding to other target cellular proteins, may regulate immune responses by affecting IL-12 production in DC.

Background : CD40 engagement enhances eosinophil survival, suggesting a role for this receptor in the development of eosinophilia. Objective : We examined whether CD40 enhances eosinophil survival by inducing the expression of antiapoptotic proteins. Three members of the inhibitor of apoptosis protein (IAP) family, namely cellular (c)-IAP1, c-IAP2, and XIAP, and 2 antiapoptotic proteins of the Bcl-2 family, namely Bcl-x L and Bfl-1/A1, were investigated. Methods : Blood and sputum were obtained from healthy subjects and atopic asthmatic patients. Blood eosinophils were isolated by means of magnetic selection. Expression of CD40, IAPs, and Bcl-2 proteins was investigated by using flow cytometry, immunoblotting, or both. CD40 stimulation was achieved with agonistic antibodies or soluble ligands. Apoptosis was assessed by staining with propidium iodide and FITC-conjugated annexin-V. c-IAP2 expression was inhibited with antisense oligonucleotides. Results : Freshly isolated eosinophils from healthy and asthmatic patients did not express CD40. Conversely, eosinophils expressed CD40 spontaneously when cultured for 48 hours. At this time point, CD40 stimulation significantly delayed eosinophil apoptosis. Inhibition of eosinophil apoptosis was accompanied by induction of c-IAP2 but not c-IAP1, XIAP, Bcl-x L , or Bfl-1/A1 expression. Antisense knockdown of c-iap2 abolished CD40-induced enhancement of eosinophil survival. Sputum cells from asthmatic patients, unlike those from healthy subjects, substantially expressed CD40 and c-IAP2. Moreover, a strong correlation was found between the percentage of eosinophils in the sputum from asthmatic patients and the sputum level of CD40 and c-IAP2 expression. Conclusion : The results demonstrate that CD40 engagement enhances eosinophil survival through induction of c-IAP2 expression and suggest a role for this mechanism in allergic inflammation. (J Allergy Clin Immunol 2002;110:443-9.)

The aim of this study was to determine whether high spirometric performances in calves are associated with low economic losses due to the bovine respiratory disease complex (BRDC). Five spirometric variables (SV) were measured in 909 double-muscled Belgian Blue calves from 15 to 60 d of age. Afterward, calves were monitored for 6 mo to determine whether they developed BRDC and to determine the costs due to BRDC (i.e., medicine costs and veterinarians' fees, plus estimated financial losses due to mortality in case of death). To analyze the effects of spirometric performances on BRDC cost, a fixed linear model was used for each SV. In addition to SV, each model included the effects of sex, dam's parity, vaccination status, muscular development score, herd-period, and BW. Only herd-period and the maximal ventilation and the vital capacity had significant effects on costs due to BRDC, indicating that these two SV are major physiological determinants of economic losses associated with BRDC. Accordingly, it is assumed that an amelioration of maximal ventilation and vital capacity could result in increased resistance to BRDC in calves.

Genetic parameters and environmental effects for spirometric variables (SV) in calves were estimated using 734 Belgian Blue calves (15 to 297 d of age), sired by 20 AI bulls. For each calf, the following SV were measured: 1) the average ventilation (l/min) recorded during the 15 s of maximal ventilatory changes induced by lobeline administration (0.25 mg/kg, i.v.) (15-s MV(L)); 2) the vital capacity, and the maximal peak expiratory and inspiratory flows recorded after lobeline administration; and 3) the ventilatory reserve (15-s MV(L) - ventilation at rest). Analysis of environmental factors showed age of calf, herd, sex, and vaccination status had significant effects on SV. A sire model and a multiple-trait derivative-free REML procedure were used to estimate genetic parameters for SV, body weight, and muscling score. Heritabilities for SV ranged from 0.28 +/- 0.11 to 0.44 +/- 0.16. Genetic correlations among SV varied from 0.76 to 0.98 and environmental correlations from 0.69 to 0.80. Genetic correlations of SV with body weight (0.25 to 0.56) and with muscling score (0.21 to 0.76) were positive, as were environmental correlations of SV with body weight (0.44 to 0.70) and muscling score (0.09 to 0.25). These results suggest that selection may improve SV without impairing other traits of economic importance.

Solute analysis in bronchoalveolar lavage fluid involves the use of dilutional markers to correct for variable recovery of pulmonary epithelial lining fluid (PELF). Urea is the best characterised endogenous marker, whereas inulin appears to meet the requirements of an exogenous marker. In horses, the use of inulin has never been investigated and the impact of lower airway diseases such as heaves, on PELF recovery is unknown. In this study, five healthy and five heaves-affected horses underwent airway endoscopy and bronchoalveolar lavage. PELF recovery from bronchoalveolar lavage was calculated by the inulin and the urea method. The inulin method was compared to the urea method and differences between healthy and heaves-affected horses were analysed. From a technical and analytical point of view, inulin fulfilled the requirements of a marker of dilution as well as urea. When both healthy and heaves-affected horses groups were pooled together, PELF recovery calculated by the inulin method was significantly higher than by the urea method (6.43+/-4.08% versus 0.789+/-0.299%, P < 0.005). No significant differences were observed between healthy and heaves-affected horses, neither by the inulin nor by the urea method. Inulin did not present major advantages over urea, but the combined use of both markers can improve the standardisation of studies comparing PELF compounds, by providing upper limits (inulin dilution) and lower limits (urea dilution) of PELF recovery.

The NF-kappaB transcription factor is ubiquitously expressed and controls the expression of a large number of genes. Experimental data clearly indicate that NF-kappaB is a major regulator of the inflammatory reaction by controlling the expression of pro-inflammatory molecules in response to cytokines, oxidative stress and infectious agents. We demonstrated that NF-kappaB activation by IL-1beta follows three distinct cell-specific pathways. Moreover, our studies indicated that in one model of inflammatory diseases, horse recurrent airway obstruction (RAO), the extent of NF-kappaB basal activity correlates with pulmonary dysfunction. Another role of NF-kappaB activity protects cancer cells against apoptosis and could participate in the resistance to cancer treatment. However, we did not observe any increased cytotoxicity after treatment with anticancer drugs or TNF-alpha of cells expressing a NF-kappaB inhibitor. Therefore, we can conclude that the inhibition of apoptosis by NF-kappaB is likely to be cell type and stimulus-dependent. Further studies are required to determine whether NF-kappaB could be a target for anticancer treatments.

Arterial blood gases, acid-base balance and respiratory function tests using impulse oscillometry (IOS) were performed on 40 clinically healthy newborn calves during the first 24 hours of life to evaluate their respiratory adaptation to extrauterine life. Gas exchange efficiency of the lung was significantly improved with time and was accompanied by the correction of the mixed acidosis observed at birth and by significant changes in respiratory mechanics. Major changes were detected within the first 6 hours. The significant decrease in resistance (R) and the increase in reactance (X) with time, demonstrate the improvement in respiratory mechanics of both upper and lower airways, and reflect the increase in lung volume, the improved lung tissue elasticity and/or distribution of the ventilation. Respiratory mechanical, arterial blood gases and acid-base balance data provided in this study describe a successful respiratory adaptation to extrauterine life in healthy newborn calves.

In human medicine, pulse oximetry is widely used to measure non-invasively and accurately the percentage of oxygen saturation of arterial haemoglobin (SpO(2)). Recently, pulse oximetry has been used in calves, but its accuracy has not been evaluated in newborn calves. The purpose of this study was to evaluate the accuracy of a pulse oximeter in newborn calves by comparing SpO(2) with arterial oxyhaemoglobin saturation (SaO(2)) obtained by use of a blood gas analyser. Fifty-five newborn calves were investigated from birth to 20 days old. Pulse oximetry readings and arterial blood samples were performed 5, 15, 30, 45, 60 min, 2, 3, 6, 12, 24 h and 1 and 3 weeks after birth. The transmission-type sensors of the pulse oximeter were fixed at the recommended site in the bovine species (at the base of the calf tail, where the skin had been shaved and was not pigmented) and arterial blood samples were withdrawn from the subclavian artery and analysed for SaO(2). Five-hundred paired data of SaO(2) and mean SpO(2)(mSpO(2)) were collected. Linear regression of the pooled data indicated a highly significant correlation of mSpO(2) with SaO(2) (r = 0.87;P0.001; mSpO(2) = 15.8 + 0.84 SaO(2)). The overall data bias value was positive (+2.1%), which indicated that the pulse oximeter tended to overestimate the SaO(2). The bias value for each SaO(2) category tended to become higher for lower ranges of SaO(2). Precision was also lower when SaO(2) values were low. The lower the SaO(2) value, the higher the positive bias (overestimation) and the lower the precision. These results suggest that pulse oximetry provides a relatively accurate non-invasive, immediate and portable method to monitor SaO(2) and to evaluate objectively the pulmonary function effectiveness in newborn calves during their adaptation to extra-uterine life.

The purposes of the present study were: (1) to develop original equations to predict spirometric variables (SV) in healthy Friesian and Belgian White and Blue (BWB) calves1 year of age; and (2) to determine the effects of somatic growth on SV. Sixty-seven Friesian and 500 BWB calves were investigated. For each calf, the following SV were calculated: (1) the average minute volume derived using all the ventilatory cycles recorded during the 15 s of maximal ventilatory changes induced by lobeline administration (0.25 mg/kg, i.v.) (15-s LMV); (2) the maximal tidal volume, the maximal peak expiratory flow and the maximal peak inspiratory flow recorded from single breaths after lobeline administration (MVTL, MPEFLand MPIFL, respectively); (3) the ventilatory reserve (VRL= 15-s LMV - VEr; VEr= minute volume at rest). Mass specific (s) values were also calculated. All SV changed linearly with somatic growth in both Friesian and BWB calves. Since the rise in SV was more related to body weight than the age of calves, equations for reference values of SV always had body weight as the only independent variable. In the youngest calves, s SV were lower in the BWB breed. In the oldest calves (i.e. 1 year of age), s MPEFLand s MPIFLremained small in BWB calves whereas s MVTL, s 15-s LMV and s VRLwere almost equal in both breeds. These results could be related to the lower resistance to respiratory disorders in BWB calves1 year of age than in Friesian calves1 year of age.

In order to define a reliable method for estimating maximal ventilation in cattle, 12 healthy calves underwent a rebreathing trial and injections of increasing doses of lobeline, a respiratory analeptic. The effects of these tests on the main ventilatory parameters (tidal volume, VT; respiratory frequency, fRand minute volume, ·VE) recorded during the 15 s of maximal response were studied and compared. The sharp rise in ·VE(4.8 times higher than the resting value) observed during the rebreathing trial was mainly due to an increase in VT. This rise in ventilation was the highest ever reported in calves. Lobeline dose-dependently enhanced ventilation up to a threshold dose of 0.25 mg/kg, which always produced a maximal response. This maximal response (3.7 times higher than the resting value), reflecting both an increase in fRand VT, was reproducible at an interval of 12 h and was highly correlated with that observed during the rebreathing trial (R= 0.98,P< 0.001). These results suggest that: (1) rebreathing trial is a reliable method to induce and measure maximal ventilation in calves; and (2) lobeline administration (0.25 mg/kg) is a reliable means of accurately estimating this variable. Lobeline administration, unlike the rebreathing trial, is safe and easy to standardize, and the test therefore seems to be the preferred way of studying maximal ventilation in calves.

various combinations of recombinant fusion proteins as vaccines against strangles. Ponies were vaccinated by a combination of intranasal and subcutaneous route with different sets of recombinant fusion proteins. Ponies were then subjected to experimental infection with S. equi to induce early stages of strangles. The ponies were followed clinically and were pathologically analysed at post mortem. ELISA was used to follow antibody response. The best combination of fusion proteins, Strangvac was used on 16 ponies. Of these, 15 did not become pyrexic or had any clinical signs of infection. Nine had no pathological signs on post mortem analysis and 5 had reduced signs. All nonvaccinated control ponies (n1⁄416) became infected and with clear pathological scoring on post mortem. Antibodies were obtained against all subcomponents in the fusions. Large scale and cost effective production of Strangvac could be obtained by optimizing various conditions during fermentation and downstream processing. The stability of Strangvac appears, so far as the study has gone on, to be good. One day after immunization, pyrexia was observed as the only adverse reaction. We therefore conclude that Strangvac is a safe and efficient vaccine against strangles and will now be taken through additional clinical trials for a registration.

Background Pattern-recognition receptors (PRRs) are critically involved in the pathophysiology of airway allergy, yet most of the signaling pathways downstream of PRRs implicated in allergic airway sensitization remain unknown. Objective We sought to study the effects of genetic depletion of interferon response factor (IRF) 3 and IRF7, important transcription factors downstream of various PRRs, in a murine model of house dust mite (HDM)–induced allergic asthma. Methods We compared HDM-induced allergic immune responses in IRF3-deficient (IRF3 −/− ), IRF7 −/− , and wild-type mice. Results Parameters of airway allergy caused by HDM exposure were strongly attenuated in IRF3 −/− , but not IRF7 −/− , mice compared with those in wild-type mice. Indeed, in HDM-exposed IRF3 −/− mice HDM-specific T H 2 cell responses did not develop. This correlated with impaired maturation and migration of IRF3 −/− lung dendritic cells (DCs) on HDM treatment. Furthermore, adoptive transfer of HDM-loaded DCs indicated that IRF3 −/− DCs had an intrinsic defect rendering them unable to migrate and to prime HDM-specific T H 2 responses. Intriguingly, we also show that DC function and allergic airway sensitization in response to HDM were independent of signaling by type I interferons, the main target genes of IRF3. Conclusion Through its role in DC function, IRF3, mainly known as a central activator of antiviral immunity, is essential for the development of T H 2-type responses to airway allergens.