Differential effects of Ha-RasG12V, Ki-RasG12V and Ki-RasG13D on cell proliferation

Although differing only for the last 24 aminoacids, the three major isoforms of p21 Ras (Ha-, Ki- and N –Ras) can trigger alternative pathways of signal transduction, at least in part as a consequence of different post-translational modifications and subcellular localization. Ras mutations are a common event in tumorigenesis. In colorectal carcinomas (CRCs) the mutations affect almost exclusively Ki-Ras, while Ha-Ras mutations are mostly found in bladder carcinomas and N-Ras mutations in leukemia cells. In almost all cases, the genetic alteration is a point mutation in codons 12 or 13, and less frequently in codon 61. By affecting the GTPase activity of the protein, they always lead to a constitutively active protein. However, data obtained in different experimental systems or by analysis of primary and metastatic tumors show that mutations in different codons, different mutations in the same codon, and mutations of different isoforms of Ras may have diverse biological consequences. To shed more light on the molecular mechanisms responsible for the different effects of Ras mutations, we have obtained stable clones of HT-29 (a human colorectal adenocarcinoma cell line in which the endogenous Ras genes are wild type) transfected with cDNAs codifying Ha-RasG12V, Ki-RasG12V and Ki-RasG13D, under the control of an hormone-inducible promoter. The expression of each of these mutated Ras isoforms induces specific, different effects on cell morphology and growth rate. FACS analysis shows also a differential effect on the cell cycle. H-RasG12V expression, in addition, seems to determine apoptosis. Despite these differences, all three mutated isoforms of Ras increase the expression of the CDK inhibitor p21. Preliminary data reveal epigenetic changes in the p21 promoter region upon induction of H-RasG12V expression

Association between antibodies to carbamylated proteins and subclinical atherosclerosis in rheumatoid arthritis patients

BACKGROUND: Rheumatoid arthritis (RA) patients carry a high risk of cardiovascular morbidity and mortality. The excess of cardiovascular disease cannot be entirely explained by traditional risk factors and the immune system contributes to the development of atherosclerosis. Moreover, post-translational modifications such as citrullination and carbamylation have been linked to inflammation and atherosclerosis. Anti-carbamylated proteins antibodies (anti-CarP) are a new subset of autoantibodies identified in RA patients. This study aimed to investigate a possible association between anti-CarP and subclinical atherosclerosis in RA patients. METHODS: We enrolled RA patients and normal healthy controls (NHS) without known cardiovascular risk factors or heart disease. Cardiovascular risk was assessed using the Modified Systemic Coronary Risk Evaluation (mSCORE). Anti-CarP were investigated by a solid phase "home-made" ELISA. Anti-citrullinated protein antibodies (ACPA) and Rheumatoid Factor (RF) were investigated by ELISA assays. Subclinical atherosclerosis was evaluated by brachial artery Flow-Mediated Dilatation (FMD) and Carotid Intima-Media Thickness (c-IMT) while arterial stiffness by Ankle-Brachial Index (ABI) and Cardio-Ankle Vascular Index (CAVI). RESULTS: We enrolled 50 RA patients (34 F and 16 M, mean age 58.4 ± 13.1 years, mean disease duration 127 ± 96.7 months) and 30 age and sex matched NHS. According to the mSCORE, 58% of patients had a low risk, 32% a moderate and 8% a high risk for cardiovascular disease. FMD was significantly lower in RA patients than in NHS (5.6 ± 3.2 vs 10.7 ± 8.1%; p < 0.004) and CAVIs significantly higher in a RA patients compared to NHS (left CAVI 8.9 ± 1.7 vs 8.1 ± 1.5; p < 0.04 for and right CAVI 8.8 ± 1.6 vs 8.0 ± 1.4; p < 0.04 for the). ABI and c-IMT did not differ between the two populations. The multivariate regression analysis showed a significant association of anti-CarP antibodies with FMD, left and right CAVI and both c-IMT (r = 1.6 and p = 0.05; r = 1.7 and p = 0.04; r = 2.9 and p = 0.05; r = 1.5 and p = 0.03; r = 1.1 and p = 0.03 respectively). CONCLUSIONS: This study confirms that RA patients, without evidence of cardiovascular disease or traditional risk factors, have an impaired endothelial function. Moreover, we found an association with anti-CarP antibodies suggesting a possible contribution of these autoantibodies to endothelial dysfunction, the earliest stage of atherosclerosis. Besides ultrasound assessment, anti-CarP should be assessed in RA patients and considered an additional cardiovascular risk factor

Systemic Lupus Erythematosus with and without Anti-dsDNA Antibodies: Analysis from a Large Monocentric Cohort

Objectives. The anti-dsDNA antibodies are a marker for Systemic Lupus Erythematosus (SLE) and 70–98% of patients test positive. We evaluated the demographic, clinical, laboratory, and therapeutical features of a monocentric SLE cohort according to the antidsDNA status. Methods. We identified three groups: anti-dsDNA + (persistent positivity); anti-dsDNA ± (initial positivity and subsequent negativity during disease course); anti-dsDNA − (persistent negativity). Disease activity was assessed by the European Consensus Lupus Activity Measurement (ECLAM). Results. We evaluated 393 patients (anti-dsDNA +: 62.3%; anti-dsDNA ±: 13.3%; anti-dsDNA −: 24.4%). The renal involvement was signifiantly more frequent in anti-dsDNA + (30.2%), compared with antidsDNA ± and anti-dsDNA − (21.1% and 18.7%, resp.; = 0.001). Serositis resulted signifiantly more frequent in anti-dsDNA − (82.3%) compared to anti-dsDNA + and anti-dsDNA ± (20.8% and 13.4%, resp.; < 0.0001). Th reduction of C4 serum levels was identified significantly more frequently in anti-dsDNA + and anti-dsDNA ± (40.0% and 44.2%, resp.) compared with antidsDNA − (21.8%, = 0.005). We did not identify significant differences in the mean ECLAM values before and after modifiation of anti-dsDNA status ( = 0.7). Conclusion. Anti-dsDNA status influences the clinical and immunological features of SLE patients. Nonetheless, it does not appear to affect disease activity

Development of Drift-Reducing Spouts For Vineyard Pneumatic Sprayers: Measurement of Droplet Size Spectra Generated and Their Classification

Pneumatic spraying is especially sensitive to spray drift due to the production of small droplets that can be easily blown away from the treated field by the wind. Two prototypes of environmentally friendly pneumatic spouts were developed. The present work aims to check the effect of the spout modifications on the spray quality, to test the convenience of setting the liquid hose out of the spout in cannon-type and hand-type pneumatic nozzles and its effect on the droplet size, homogeneity and driftability in laboratory conditions. Laboratory trials simulating a real sprayer were conducted to test the influence of the hose insertion position (HP), including conventional (CP), alternative (AP), outer (OP) and extreme (XP), as well as the liquid flow rate (LFR) and the airflow speed (AS) on the droplet size (D50, D10 and D90), homogeneity and driftability (V100). Concurrently, the droplet size spectra obtained by the combination of aforementioned parameters (HP × LFR × AS) in both nozzles were also classified according to the ASABE S572.1. Results showed a marked reduction of AS outside the air spout, which led to droplet size increase. This hypothesis was confirmed by the droplet size spectra measured (D50, D10, D90 and V100). A clear influence of HP was found on every dependent variable, including those related with the droplet size. In both nozzles, the longer the distance to CP, the coarser the sprayed drops. Moreover, LFR and AS significantly increased and reduced droplet size, respectively. A higher heterogeneity in the generated drops was obtained in XP. This position yielded V100 values similar to those of the hydraulic low-drift nozzles, showing an effective drift reduction potential. The classification underlines that the variation of HP, alongside AS and LFR, allowed varying the spray quality from very fine to coarse/very coarse, providing farmers with a wide range of options to match the drift-reducing environmental requirements and the treatment specifications for every spray application

Tradiciones, monstruos y misterio en <i>Maruja</i> de Ema Wolf

Traemos a estas jornadas un análisis de la nouvelleMaruja (1989) de Ema Wolf a partir de la poética de su autora, quien construye sus personajes y su voz narrativa, moldeando diversos subgéneros narrativos sin perder de vista su preocupación por la realidad argentina, en especial por los lectores infantiles, inquietudes que resuelve magistralmente en sus escritos. Dice la autora: “busco información para el libro, de manera de poder ver mejor lo que cuento, aunque esa información no concurra directamente al texto. La información corre como una hebra entre dos ejes, y allí se sostiene. Va y viene. Y la información es altamente estimulante para resolver ciertas situaciones”. Para realizar este análisis centraremos nuestra mirada en especial en los personajes de Maruja y Veremundo, verdaderos monstruos hacedores y víctimas al mismo tiempo de una serie de eventos que entrelazan elementos propios de lo fantástico, lo policial, lo gótico, el terror y el humor negro.Facultad de Humanidades y Ciencias de la Educació

Tradiciones, monstruos y misterio en <i>Maruja</i> de Ema Wolf

Traemos a estas jornadas un análisis de la nouvelleMaruja (1989) de Ema Wolf a partir de la poética de su autora, quien construye sus personajes y su voz narrativa, moldeando diversos subgéneros narrativos sin perder de vista su preocupación por la realidad argentina, en especial por los lectores infantiles, inquietudes que resuelve magistralmente en sus escritos. Dice la autora: “busco información para el libro, de manera de poder ver mejor lo que cuento, aunque esa información no concurra directamente al texto. La información corre como una hebra entre dos ejes, y allí se sostiene. Va y viene. Y la información es altamente estimulante para resolver ciertas situaciones”. Para realizar este análisis centraremos nuestra mirada en especial en los personajes de Maruja y Veremundo, verdaderos monstruos hacedores y víctimas al mismo tiempo de una serie de eventos que entrelazan elementos propios de lo fantástico, lo policial, lo gótico, el terror y el humor negro.Facultad de Humanidades y Ciencias de la Educació

TNFα expressed on the surface of microparticles modulates endothelial cell fate in rheumatoid arthritis

Background: Rheumatoid arthritis (RA) is associated with a high prevalence of atherosclerosis. Recently increased levels of microparticles (MPs) have been reported in patients with RA. MPs could represent a link between autoimmunity and endothelial dysfunction by expressing tumor necrosis factor alpha (TNFα), a key cytokine involved in the pathogenesis of RA, altering endothelial apoptosis and autophagy. The aim of this study was to investigate TNFα expression on MPs and its relationship with endothelial cell fate. Methods: MPs were purified from peripheral blood from 20 healthy controls (HC) and from 20 patients with RA, before (time (T)0) and after (T4) 4-month treatment with etanercept (ETA). Surface expression of TNFα was performed by flow cytometry analysis. EA.hy926 cells, an immortalized endothelial cell line, were treated with RA-MPs purified at T0 and at T4 and also, with RA-MPs in vitro treated with ETA. Apoptosis and autophagy were then evaluated. Results: RA-MPs purified at T0 expressed TNFα on their surface and this expression significantly decreased at T4. Moreover, at T0 RA-MPs, significantly increased both apoptosis and autophagy levels on endothelial cells, in a dose-dependent manner. RA-MPs did not significantly change these parameters after 4 months of in vivo treatment with ETA. Conclusions: Our data demonstrate that MPs isolated from patients with RA exert a pathological effect on endothelial cells by TNFα expressed on their surface. In vivo and in vitro treatment with ETA modulates this effect, suggesting anti-TNF therapy protects against endothelial damage in patients with RA