The value of selected in vitro and in silico methods to predict acute oral toxicity in a regulatory context: results from the European Project ACuteTox.

ACuteTox is a project within the 6th European Framework Programme which had as one of its goals to develop, optimise and prevalidate a non-animal testing strategy for predicting human acute oral toxicity. In its last 6 months, a challenging exercise was conducted to assess the predictive capacity of the developed testing strategies and final identification of the most promising ones. Thirty-two chemicals were tested blind in the battery of in vitro and in silico methods selected during the first phase of the project. This paper describes the classification approaches studied: single step procedures and two step tiered testing strategies. In summary, four in vitro testing strategies were proposed as best performing in terms of predictive capacity with respect to the European acute oral toxicity classification. In addition, a heuristic testing strategy is suggested that combines the prediction results gained from the neutral red uptake assay performed in 3T3 cells, with information on neurotoxicity alerts identified by the primary rat brain aggregates test method. Octanol-water partition coefficients and in silico prediction of intestinal absorption and blood-brain barrier passage are also considered. This approach allows to reduce the number of chemicals wrongly predicted as not classified (LD50>2000 mg/kg b.w.).

Fractures non traumatiques liées à la grossesse et à l'allaitement : s'agit-il toujours d'une ostéoporose? A propos de 2 cas

Introduction : L'ostéoporose et/ou les fractures liées à la grossesse sont souvent sous-diagnostiquées. Nous rapportons 2 cas de fractures diagnostiquées peu après l'accouchement de 2èmes grossesses. Patientes. Cas 1. Patiente de 30 ans présentant des dorso-lombalgies à la fin de sa 2ème grossesse. Une IRM après l'accouchement montre 2 fractures vertébrales (L1 et L2). Densité minérale osseuse (DMO): T-score colonne: -3.9 DS, col fémoral -1.7 DS et hanche totale -0.6 DS. Cas 2. Patiente de 32 ans, présentant des douleurs fessières à la fin de sa 2ème grossesse. L'IRM pelvienne après l'accouchement montre une fracture de l'aile sacrée droite S1-S3 et de l'aile sacrée gauche S1.DMO: T-score colonne -1.4 DS, col fémoral 0.2 DS et hanche totale 0.0 DS. La microarchitecture est normale (TBS 1.429). Nous retenons dans le 1er cas le diagnostic d'une ostéoporose fracturaire liée à la grossesse. De l'ibandronate trimestriel iv est prescrit. Dans le 2ème cas, au vu de la DMO quasi normale, de la trabéculométrie normale et du site atypique de la fracture, nous concluons à une fracture non ostéoporotique sur augmentation du stress mécanique lié à la grossesse. Aucun traitement à visée osseuse n'est prescrit. Discussion : " L'ostéoporose " liée à la grossesse et à l'allaitement se manifeste le plus souvent par des fractures vertébrales non traumatiques pendant le 3ème trimestre de la 1ère grossesse ou durant le post-partum. Une DMO et un bilan à la recherche d'une cause secondaire sont indispensables. Cette pathologie est sous-diagnostiquée, car les douleurs dorsolombaires sont souvent mises sur le compte d'une hyperlaxité ligamentaire physiologique liée aux hormones. Les facteurs de risque sont les mêmes que pour une ostéoporose post-ménopausique. Les apports bas en calcium et en vitamine D3 ainsi qu'un capital osseux moindre à la fin de l'adolescence seraient des facteurs prédisposants. La DMO lombaire diminue de 7.6 +/-0.1%, celle du corps entier de 3.9 +/-0.1% pendant la grossesse et l'allaitement. Habituellement on assiste à une récupération de la DMO dans les mois qui suivent la fin de l'allaitement. Conclusion : Devant des douleurs rachidiennes en fin de grossesse il faut évoquer une fracture ostéoporotique liée à la grossesse. La densitométrie osseuse peut aider au diagnostic même s'il faut l'interpréter prudemment dans les mois qui suivent l'accouchement. Il n'y a pas de consensus concernant le traitement spécifique.

L'ús dels clítics pronominals del català i la seva adquisició per parlants de romanès i de tagal

El nostre estudi ha analitzat els mecanismes de transferència de la L1 en dos grups de parlants, un que té el romanès i l’altre el tagal com a llengües mare, en el procés d’aprenentatge del català com a L2. En concret ens hem centrat en l’estudi de l’ús dels clítics pronominals, una de les parts de la gramàtica catalana més apassionant per les seves idiosincràsies i la seva diversitat formal. L’ús d’aquest tipus de pronoms és un aspecte que resulta complicat d’adquirir per parlants de llengües en què no existeixen o on l’ús que en fan és diferent. En el nostre estudi, ens hem basat inicialment en la descripció de les característiques fonamentals dels clítics pronominals del català, del romanès i del tagal, a partir de bibliografia especialitzada i més general, i, en el cas del català, fent servir també la pròpia competència. Després d’aquest apartat més descriptiu, hem utilitzat un corpus d’entrevistes fetes en català a dos grups de parlants, el primer amb aprenents de català que tenen el romanès com a L1 i el segon amb aprenents de català que tenen el tagal com a L1, per analitzar quantitativament i qualitativament l’ús dels clítics pronominals. Un altre grup d’entrevistes, aquesta vegada a persones autòctones (català L1), ens ha servit com a grup de control. El nostre estudi dóna suport a la hipòtesi que atorga una importància especial a la transferència de la L1 en l’adquisició d'una segona llengua en general, i en la dels clítics pronominals d’una llengua romànica, en particular. Els resultats mostren que hi ha diferències entre els dos grups d'aprenents que són estadísticament significatives (ús del programa SPSS) i poden ser atribuïdes a les característiques de la L1.

Peroxisome proliferator-activated receptors mediate host cell proinflammatory responses to Pseudomonas aeruginosa autoinducer.

The pathogenic bacterium Pseudomonas aeruginosa utilizes the 3-oxododecanoyl homoserine lactone (3OC(12)-HSL) autoinducer as a signaling molecule to coordinate the expression of virulence genes through quorum sensing. 3OC(12)-HSL also affects responses in host cells, including the upregulation of genes encoding inflammatory cytokines. This proinflammatory response may exacerbate underlying disease during P. aeruginosa infections. The specific mechanism(s) through which 3OC(12)-HSL influences host responses is unclear, and no mammalian receptors for 3OC(12)-HSL have been identified to date. Here, we report that 3OC(12)-HSL increases mRNA levels for a common panel of proinflammatory genes in murine fibroblasts and human lung epithelial cells. To identify putative 3OC(12)-HSL receptors, we examined the expression patterns of a panel of nuclear hormone receptors in these two cell lines and determined that both peroxisome proliferator-activated receptor beta/delta (PPARbeta/delta) and PPARgamma were expressed. 3OC(12)-HSL functioned as an agonist of PPARbeta/delta transcriptional activity and an antagonist of PPARgamma transcriptional activity and inhibited the DNA binding ability of PPARgamma. The proinflammatory effect of 3OC(12)-HSL in lung epithelial cells was blocked by the PPARgamma agonist rosiglitazone, suggesting that 3OC(12)-HSL and rosiglitazone are mutually antagonistic negative and positive regulators of PPARgamma activity, respectively. These data identify PPARbeta/delta and PPARgamma as putative mammalian 3OC(12)-HSL receptors and suggest that PPARgamma agonists may be employed as anti-inflammatory therapeutics for P. aeruginosa infections.

Product differentiation, competitive pressure and the effects of innovation on the success and survivability of firms in oligopoly markets

In a recent paper Tishler and Milstein (2009) fi…nd that increased competition may increase aggregate R&D spending while market output decreases. Therefore, they obtain the surprising result that R&D spending is excessive when competition becomes intense. Their result is based on the standard linear demand function for differentiated products introduced by Bowley (1924) where decreased product differentiation is interpreted as more competitive pressure. In this paper I show that at an aggregate level this interpretation is problematic because equilibrium effects are dominated by a demand reduction effect. A slight modifi…cation of the standard demand function eliminates this effect. For the Tishler and Milstein (2009) setting it is shown that then increased competition increases both R&D spending and aggregate market output. Therefore, at least for consumers, more intense competition increases welfare. Journal of Economic Literature Classi…fication Numbers: D43, L1, O3. Keywords: Oligopoly markets, Product differentiation, Competitive pressure.

Microtubule-depolymerizing agents used in antibody-drug conjugates induce antitumor immunity by stimulation of dendritic cells.

Antibody-drug conjugates (ADC) are emerging as powerful treatment strategies with outstanding target-specificity and high therapeutic activity in patients with cancer. Brentuximab vedotin represents a first-in-class ADC directed against CD30(+) malignancies. We hypothesized that its sustained clinical responses could be related to the stimulation of an anticancer immune response. In this study, we demonstrate that the dolastatin family of microtubule inhibitors, from which the cytotoxic component of brentuximab vedotin is derived, comprises potent inducers of phenotypic and functional dendritic cell (DC) maturation. In addition to the direct cytotoxic effect on tumor cells, dolastatins efficiently promoted antigen uptake and migration of tumor-resident DCs to the tumor-draining lymph nodes. Exposure of murine and human DCs to dolastatins significantly increased their capacity to prime T cells. Underlining the requirement of an intact host immune system for the full therapeutic benefit of dolastatins, the antitumor effect was far less pronounced in immunocompromised mice. We observed substantial therapeutic synergies when combining dolastatins with tumor antigen-specific vaccination or blockade of the PD-1-PD-L1 and CTLA-4 coinhibitory pathways. Ultimately, treatment with ADCs using dolastatins induces DC homing and activates cellular antitumor immune responses in patients. Our data reveal a novel mechanism of action for dolastatins and provide a strong rationale for clinical treatment regimens combining dolastatin-based therapies, such as brentuximab vedotin, with immune-based therapies. Cancer Immunol Res; 2(8); 741-55. ©2014 AACR.

Study on the Elimination of Angiostrongylus costaricensis First Stage Larvae in the Experimental Infection of Swiss Mice

Abdominal angiostrongylosis is a nematode infection of wild rodents. Human infection may result in severe abdominal disease and has been reported from several countries in the Americas. The domestic mouse, Mus musculus, has not been found with natural infection and, like other urban rodents, should not be considered a natural host for Angiostrongylus costaricensis. Quantification of parasitic forms released for transmission may better express the coevolutionary status in parasite-host relationship. With this objective, five groups of experimentally infected Swiss mice were followed for up to 155 days post-infection (PI) days and the quantification of first stage larvae (L1) output revealed: an irregular elimination of L1 and a huge variation in the patency period (1 to 114 days) and in the number of L1 eliminated daily by individual animals (1 to 6340 L1/g). Overall mortality was 72% (range: 28% to 100%) at seven weeks PI. In conclusion, abdominal angiostrongylosis in M. musculus presents high mortality and a very variable and irregular elimination of L1 in feces.

SHP-1 phosphatase activity counteracts increased T cell receptor affinity.

Anti-self/tumor T cell function can be improved by increasing TCR-peptide MHC (pMHC) affinity within physiological limits, but paradoxically further increases (K(d) < 1 μM) lead to drastic functional declines. Using human CD8(+) T cells engineered with TCRs of incremental affinity for the tumor antigen HLA-A2/NY-ESO-1, we investigated the molecular mechanisms underlying this high-affinity-associated loss of function. As compared with cells expressing TCR affinities generating optimal function (K(d) = 5 to 1 μM), those with supraphysiological affinity (K(d) = 1 μM to 15 nM) showed impaired gene expression, signaling, and surface expression of activatory/costimulatory receptors. Preferential expression of the inhibitory receptor programmed cell death-1 (PD-1) was limited to T cells with the highest TCR affinity, correlating with full functional recovery upon PD-1 ligand 1 (PD-L1) blockade. In contrast, upregulation of the Src homology 2 domain-containing phosphatase 1 (SHP-1/PTPN6) was broad, with gradually enhanced expression in CD8(+) T cells with increasing TCR affinities. Consequently, pharmacological inhibition of SHP-1 with sodium stibogluconate augmented the function of all engineered T cells, and this correlated with the TCR affinity-dependent levels of SHP-1. These data highlight an unexpected and global role of SHP-1 in regulating CD8(+) T cell activation and responsiveness and support the development of therapies inhibiting protein tyrosine phosphatases to enhance T cell-mediated immunity.

Com conduïm després d’un programa formatiu de seguretat viària?

Els programes formatius de seguretat viària són una mesura que s’aplica des de l’execució penal a la comunitat en delictes de trànsit i tenen com a finalitat reduir la reincidència dels conductors de major risc. L’objectiu principal del treball és identificar les característiques comunes dels infractors de trànsit que fan aquest tipus d’intervenció, conèixer els factors de risc associats a aquests infractors i en quina mesura l’estat psicològic és un factor de risc en l’estil de conducció. També es volia determinar si hi havia diferències entre les entitats que impartien la formació i avaluar l’efectivitat d’aquests programes en l’estil de conducció dels participants en finalitzar el curs. En l’estudi, hi van participar 278 voluntaris del total de 354 infractors de trànsit que van fer el programa formatiu entre l’1 d’abril de 2009 i el 13 de febrer de 2010. How do we drive after a driving educational program? The study analyzes the results of educational programs that are applied as a community sanction for those convicted for driving offenses, in majority drunk driving, in Catalonia. Between April 1, 2009 and February 13, 2010 a total of 278 offenders participated in these mandatory educational programs. ¿Cómo conducimos después de un programa formativo de educación vial? El estudio analiza los resultados de los programas formativos que se aplican como medida penal alternativa a la prisión a los condenados por delitos de tráfico en Cataluña. En total han participado 278 infractores que realizaron el programa formativo entre el 1 de abril de 2009 y el 13 de febrero de 2010.

La inserció laboral dels ex-interns dels centres penitenciaris de Catalunya

L’estudi parteix d’un repàs de la literatura, 11 entrevistes a experts, 25 a ex-interns o interns i l’anàlisi de dades sòcio-demogràfiques, penitenciàries i d’activitats d’inserció i trajectòria laboral de 3.225 ex-interns espanyols que han sortit en llibertat definitiva a Catalunya entre l’1 de gener 2004 i el 31 de desembre 2007 Es constata que el 43,6% d’ex-interns obté ocupació amb alta a la Seguretat Social un cop en llibertat definitiva. La inserció laboral sol ser fràgil: contractes temporals, feines manuals, poc qualificades i de breu duració, la qual cosa els fa constituir-se com a un col·lectiu vulnerable. En general participen d’una cultura del treball no arrelada i de trajectòries laborals prèvies a la reclusió breus o inexistents. D’aquí, que existeixi una línia de continuïtat entre els vincles amb el mercat de treball abans i després de la reclusió. El treball productiu a presons repercuteix moderadament en més inserció laboral. Però treball productiu i formació professional són poc professionalitzadors, més útils per als que no han treballat, sense estudis ni habilitats laborals. Els estudis són un factor clau en la inserció laboral; també els vincles socials, sobretot els més propers, parella per als homes, fills per a les dones. La major predisposició de les dones per a la inserció no reverteix en altes a la S.S., explicable per factors culturals, dificultats d’obtenció de feina regularitzada o de conciliació d’horaris laborals i familiars. Finalment, l’estudi constata que els joves accedeixen més a una ocupació, no tant per motivació personal sinó per tenir més fàcil accés que els adults.

La inserción laboral de los ex internos de los centros penitenciarios de Catalunya

El estudio analiza –a partir de datos de la Seguridad Social- el impacto del trabajo productivo y la formación profesional en la inserción laboral de los exinternos de nacionalidad española que, en Catalunya, han obtenido la libertad definitiva entre el 2004 y el 2007.

Penetration Sites and Migratory Routes of Angiostrongylus costaricensis in the Experimental Intermediate Host (Sarasinula marginata)

The intermediate hosts of Angiostrongylus costaricensis are terrestrian molluscs, mostly of the family Veronicellidae. The present work aimed at clarifying more accurately the sites of penetration and the migratory routes of A. costaricensis in the tissue slugs and at verifying the pattern of the perilarval reaction at different times of infection. Slugs were individually infected with 5,000 L1, and killed from 30 min to 30 days after infection. From 30 min up to 2 hr after infection, L1 were found within the lumen of different segments of the digestive tube having their number diminished in more advanced times after exposition until complete disappearance. After 30 min of exposition, percutaneous infection occurred, simultaneously to oral infection. Perilarval reaction was observed from 2 hr of infection around larvae in fibromuscular layer, appearing later (after 6 hr) around larvae located in the viscera. A pre-granulomatous reaction was characterized by gradative concentration of amebocytes around larvae, evolving two well-organized granulomas. In this work we confirmed the simultaneous occurrence of oral and percutaneous infections. Perilarval reaction, when very well developed, defined typical granulomatous structure, including epithelioid cell transformation. The infection also caused a systemic mobilization of amebocytes and provoked amebocyte-endothelium interactions.

Oncogenic Ras inhibits Fas ligand-mediated apoptosis by downregulating the expression of Fas.

Tumor growth is the result of deregulated tissue homeostasis which is maintained through the delicate balance of cell growth and apoptosis. One of the most efficient inducers of apoptosis is the death receptor Fas. We report here that oncogenic Ras (H-Ras) downregulates Fas expression and renders cells of fibroblastic and epitheloid origin resistant to Fas ligand-induced apoptosis. In Ras-transformed cells, Fas mRNA is absent. Inhibition of DNA methylation restores Fas expression. H-Ras signals via the PI 3-kinase pathway to downregulate Fas, suggesting that the known anti-apoptotic effect of the downstream PKB/Akt kinase may be mediated, at least in part, by the repression of Fas expression. Thus, the oncogenic potential of H-ras may reside on its capacity not only to promote cellular proliferation, but also to simultaneously inhibit Fas-triggered apoptosis.

Monitoring of vaccine-specific gamma interferon induction in genital mucosa of mice by real-time reverse transcription-PCR.

Monitoring of T-cell responses in genital mucosa has remained a major challenge because of the absence of lymphoid aggregates and the low abundance of T cells. Here we have adapted to genital tissue a sensitive real-time reverse transcription-PCR (TaqMan) method to measure induction of gamma interferon (IFN-gamma) mRNA transcription after 3 h of antigen-specific activation of CD8 T cells. For this purpose, we vaccinated C57BL/6 mice subcutaneously with human papillomavirus type 16 L1 virus-like particles and monitored the induction of CD8 T cells specific to the L1(165-173) H-2D(b)-restricted epitope. Comparison of the responses induced in peripheral blood mononuclear cells and lymph nodes (LN) by L1-specific IFN-gamma enzyme-linked immunospot assay and TaqMan determination of the relative increase in L1-specific IFN-gamma mRNA induction normalized to the content of CD8b mRNA showed a significant correlation, despite the difference in the readouts. Most of the cervicovaginal tissues could be analyzed by the TaqMan method if normalization to glyceraldehyde-3-phosphate dehydrogenase mRNA was used and a significant L1-specific IFN-gamma induction was found in one-third of the immunized mice. This local response did not correlate with the immune responses measured in the periphery, with the exception of the sacral LN, an LN draining the genital mucosa, where a significant correlation was found. Our data show that the TaqMan method is sensitive enough to detect antigen-specific CD8 T-cell responses in the genital mucosa of individual mice, and this may contribute to elaborate effective vaccines against genital pathogens.

Language and syntactic impairment following stroke in late bilingual aphasics.

PURPOSE: Bilingual aphasia generally affects both languages. However, the age of acquisition of the second language (L2) seems to play a role in the anatomo-functional correlation of the syntactical/grammatical processes, thus potentially influencing the L2 syntactic impairment following a stroke. The present study aims to analyze the influence of late age of acquisition of the L2 on syntactic impairment in bilingual aphasic patients. METHODS: Twelve late bilingual participants (speaking French as L2 and either English, German, Italian or Spanish as L1) with stroke-induced aphasia participated in the study. The MAST or BAT aphasia batteries were used to evaluate overall aphasia score. An auditory syntactic judgement task was developed and used to test participants syntactic performance. RESULTS: The overall aphasia scores did not differ between L1 and L2. In a multiple case analysis, only one patient had lower scores in L2. However, four patients presented significantly lower performances in syntactic processing in the late L2 than in their native language (L1). In these four patients the infarct was localized, either exclusively or at least partially, in the pre-rolandic region. CONCLUSION: This pilot study suggests that, in late bilingual aphasics, syntactic judgment abilities may be more severely impaired in L2, and that this syntactic deficit is most likely to occur following anterior lesions.