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Figure S1. Levels of pro-inflammatory mediators in cell supernatants in vitro (adapted from [13]). Figure S2. Levels of proteins, LDH (cytotoxicity) and intracellular ROS (oxidative stress) in BALF. Figure S3. Examples of critical effect doses (CED) and dose intervals (CEDL/BMDL and CEDU/BMDU) determined using benchmark dose response modeling. Figure S4. Size distribution of the NMs in the suspensions used to expose rats. Figure S5. Initial lung burden in vivo assessed by ICP-MS 3 h after instillation (n = 2). Table S1. Doses deposited in vitro in submerged conditions in function of nominal concentrations in suspensions (First published in [13]). Table S2. LOAELs (in μg/cm2) determined in vitro with the pro-inflammatory effects for each exposure method used (First published in [13]). Table S3. Dose intervals (in μg/106 macrophages) determined for each NM and each methodology. Table S4. Dose intervals normalized by primary surface areas (in cm2/106macrophages) for each NM and methodology. Table S5. Dose intervals normalized by agglomerate surface areas (in cm2/106 macrophages) for each NM and methodology. Table S6. Characterization of mass deposited in vitro on cells after 3 h exposure at the ALI to aerosols of NMs (Adapted from [13]). (DOCX 831 kb)

Our understanding of how manufactured nanoparticles affect pulmonary immunological response is poor. The aim of the studywas to evaluate, in healthy rats, the acute lung toxicity of intratracheally instilled nanoscale TiO2 (15 nm) and carbon black nanoparticles (Printex 60 and FW2, Degussa). Groups of six rats were instilled once with 100 (micro)g of particles suspended in rat concentrated bronchoalveolar lavage fluid (BALF) as a vehicle. Rats were sacrificed 3, 7 and 21 days post-exposure. ffect assessment included lung histology and markers of lung inflammatory and immunological response. Exposure to FW2 particles induced a significant increase in total alveolar cells after 3 days (4.2×10(6) cells/rat versus 1.8×10(6) cells in control rats). This increase was no more visible at 7 and 21 days. For all groups, alveolar cells consisted mainly in macrophages, with only a slight influx of neutrophils after 7 days exposure to TiO2 and 7 and 21 days to FW2. One day after exposure, the percentage of phagocytosing macrophages was about 14% for TiO2 and P60 and 8% for FW2. The number of macrophages with particles inside remained constant (up to 21 days) for the carbon-based nanoparticles but decreased with time for TiO2 (4.8% at day 21). No TNF-(alpha) was detected either in serum or in BALF of control and exposed-rats. No visible alteration of the lung tissue was noted. As a conclusion, all these particles showed little toxicity in healthy rats in our experimental conditions. FW2 was the most potent to induce a slight but significant inflammatory response.

L’allergie est un problème de santé majeur dans la plupart de nos sociétés développées. Il est unanimement reconnu que la prévalence de certaines allergies augmente depuis plusieurs dizaines d’années dans de nombreux pays industrialisés. Les causes de cette augmentation sont encore inconnues, mais, en raison de la rapidité du phénomène, il est admis que des facteurs environnementaux plutôt que génétiques soient impliqués. Parmi ceuxci, la pollution de l’air semble avoir un rôle non négligeable. Les polluants atmosphériques particulaires (en particulier les particules diesel) et gazeux (NO2, O3 et SO2 notamment) sont des facteurs adjuvants connus de la réaction allergique respiratoire. Ils exacerbent la nocivité des allergènes, dont celle des aéroallergènes extérieurs, représentés en majorité par les pollens. En plus de cet effet adjuvant, divers travaux ont montré que l’interaction pollens-polluants aboutissait également à une modification qualitative et quantitative des allergènes de pollens et à une agrégation des pollens avec les polluants particulaires. Durant trois ans, une étude a été menée par l’INERIS, en collaboration avec le centre de recherche RIVM aux Pays-Bas et le laboratoire Environnement et Chimie Analytique de l’École Supérieure de Physique Chimie Industrielle (ESPCI) à Paris, pour déterminer l’impact de la pollution gazeuse sur le potentiel allergisant des pollens de graminées

The kinetics of pulmonary inflammatory response were investigated in male Balb/c mice (25-27 g) exposed to 1 ppm O3 or 50 ppm NO2 for 3h. At various times after beginning of exposure (3 /24 /48 / 72/ 96 /120 h), airway responsiveness to metacholine was assessed using whole-body plethysmography. Animals were then anesthetized and several inflammatory mediators (proteins, LDH, cytokines, eicosanoids) were measured in the bronchoalveolar lavage (BAL). Alveolar and peritoneal macrophages were tested for phagocytosis. Proliferative response of splenocytes was also evaluated. Exposed animals had mainly effects at the pulmonary level, except for a decrease in thymus weight at 24 to 72h after exposure to NO2 and at 48h after exposure to O3. Mice exposed to NO2 showed airway hyperresponsiveness till 72h post exposure. Response to ozone was mild and transient with bronchial hyperreactivity only just after exposure. NO2 exposure induced an important pulmonary inflammatory response characterized by a neutrophil influx at 48h, a persistent alteration of epithelial permeability and a transient increase in LDH levels in BAL. TNF-, PGF2 and IL-13 were unchanged in BAL. IL-6 and LTB4 levels temporarily increased just after exposure, IL-1 level peaked at 48h and persistent increased levels of PGE2 and LTC4/D4/E4 were noted. AM phagocytosis was also decreased between 3 and 48h after NO2 exposure. Ozone-induced inflammatory response was less intense, with only a neutrophil influx after 48h and a transient increase in IL-6 level in BAL just after the end of exposure. This study is the first step in studying the effects of exposure to a combination of these two gases.

The pulmonary effects of two environmentally relevant aldehydes were investigated in ovalbumin (OA)-sensitized guinea-pigs (GP). Four week-old male Hartley GP, weighing about 300 g, were intraperitoneally injected with 1 ml of a NaCl solution containing 100 µg OA and 100 mg Al(OH)3. They were then exposed either to acetaldehyde or benzaldehyde (500 ppb each) during 4 weeks (6 hours/day, 5 days/week). At the end of exposure, GP were challenged with an OA aerosol (0.1% in NaCl) and pulmonary functions were measured. The day after, guinea-pigs were anesthetized and several endpoints related to inflammatory and allergic responses were assessed in : blood (numeration, formula, biochemistry), whole lung (histology), and bronchoalveolar lavage (BAL) (differential cell count, protein and eicosanoid concentrations). Sensitized non-exposed GP (n = 8) had an increased number of eosinophils in blood and BAL, together with an increase in total protein and leukotrien (LTB4 and LTC4/D4/E4) contents in BAL. In non-sensitized GP (n = 8), exposure to acetaldehyde or benzaldehyde did not induce any change in the tested parameters, with the exception of a slight irritation of the respiratory tract as detected by histology and an increased number of alveolar macrophages in animals exposed to acetaldehyde (21.9 +- 1.6 and 42.2 +- 5.4 x 106 cells in control and exposed GP respectively). In sensitized GP, exposure to acetaldehyde induced a moderate irritation of the respiratory tract but no change in biological parameters linked to the inflammatory and allergic responses. In contrast, exposure to benzaldehyde induced : a decrease in both OA-induced bronchoconstriction and neutrophil number in BAL, an increase in the bronchodilatator mediator PGE2 and a decrease in the bronchoconstrictor mediators LTC4/D/E4. Further investigations are needed to determine if the attenuated response observed in sensitized GP exposed to benzaldehyde is due to an alteration of the mechanism of sensitization or to a more direct effect on various mechanisms of the allergic response.

International audience; A two step purification protocol was developed to purify rainbow trout (Oncorhynchus mykiss) Vitellogenin (Vtg) and was successfully applied to Vtg of chub (Leuciscus cephalus) and gudgeon (Gobio gobio). Capture and intermediate purification were performed by anion exchange chromatography on a Resource Q column and a polishing step was performed by gel permeation chromatography on Superdex 200 column. This method is a rapid two step purification procedure that gave a pure solution of Vtg as assessed by silver staining electrophoresis and immunochemical characterisation.; L'induction et la purification de la vitellogénine chez la truite, le chevaine et le goujon a été réalisée. Le but de ces expériences est de disposer de la protéine purifiée utilisable dans la méthode ELISA pour quantifier la vitellogénine présente dans les échantillons à partir d'une calibration spécifique pour chaque espèce. Le travail réalisé a permis la mise au point d'une méthode de purification à deux étape chromatographiques

The toxicity of pentachlorophenol (PCP), a polluting substance believed to exert a narcotic effect, was assayed using the Caco-2 cell line as a model. In order to assess this toxicity as fully as possible, several viability tests, each examining different endpoints, have been used. Neutral red uptake was found to be more sensitive to PCP than MTT and Alamar Blue tests. Transepithelial electrical resistance (TEER) was shown to be the most sensitive to PCP at concentrations and exposure times where the Alamar Blue, LDH leakage and Blue Dextran passage did not evidence any effect. Blue Dextran passage and optical microscopy revealed cellular detachment at concentrations where LDH and Alamar Blue showed little or no cytotoxicity. Thus, PCP seems to affect the integrity of the intestinal barrier at levels where no cytotoxicity is seen. Our results support the notion that TEER can be used as a very sensitive method for evaluating membrane-perturbing toxicants