Diagnosis of bronchial hyperresponsiveness in sport by PC20 value

En este trabajo se exponen los criterios de positividad del Comité Olímpico Internacional versus a la Sociedad Española de Neumología y Cirugía Torácica. Participaron en este estudio ochenta deportistas de alto rendimiento, realizando una historia clínica, una espirometría de reposo y un test de metacolina. Se analiza la sensibilidad y especificidad del test de metacolina mediante curvas ROC. El comité Internacional antidopaje (WADA) requiere que la disminución sea con un PC20 < a 4mg/ml, mientras que para la práctica clínica este descenso debe presentar un PC20 < 8mg/ml. Los resultados fueron: 25% tuvieron un PC20 > de 8mg/ml; el 61% obtuvieron un PC20 < 4mg/ml y un 14% presentaron un PC20 entre 4 y 8mg/ml, correspondiendo el mejor punto de corte a PC20 de 7,6mg/ml con especificidad de 98,3 y sensibilidad de 100%. Se tendría que determinar los mismos criterios para el diagnóstico de los deportistas y los que no lo son.In this work criteria of the International Olympic Committee versus the Spanish Society of Pneumology and Thoracic Surgery are exposed. A study was conducted in eighty high performance athletes of several sports . Who underwent a medical history, resting spirometry and a methacholine challenge test. You get the sensitivity and specificity of methacholine challenge test using Receiver Operating Curves (ROC curves). International anti-doping Committee requires that the decline is a PC20 < 4mg/ml, while clinical practice this fall must submit a PC20 < 8mg/ml. Twenty five percent of those studied had a PC20 > of 8mg/ml, 61% had a PC20 < 4mg/ml and 14% had a 4 to 8mg/ml PC20. The best cutoff point was found for a PC20 of 7.6 mg/ml with a specificity of 98.3 and a sensitivity of 100%. It would have to determine the same criteria for positive diagnosis of athletes and those who are not

Multi-scenario modelling of learning

International audienceDesigning an educational scenario is a sensitive and challenging activity because it is the vector of learning. However, the designed scenario may not correspond to some learners’ characteristics (pace of work, cognitive styles, emotional factors, prerequisite knowledge, …). To personalize the learning task and adapt it gradually to each learner, several scenarios are needed. Adaptation and personalization are difficult because it is necessary on the one hand to know in advance the profiles and on the other hand to produce the multiple scenarios corresponding to these profiles. Our model allows to design many scenarios without knowing the learner profiles beforehand. Furthermore, it offers each learner opportunities to choose a scenario and to change it during their learning process. The model ensures that all announced objectives have enough resources for acquiring knowledge and activities for evaluation

Depolarization-repolarization synchrony after right ventricular and left bundle branch area pacing

Introduction: Left bundle branch area pacing (LBBAP) has been recently proposed to overcome the limitations associated with right ventricular pacing (RVP) and has been suggested as a new physiological pacing form with high feasibility and safety. A greater difference between QRS complex and T-wave angle directions has been proposed as a marker of abnormal electrical activity in several patient populations, but a comparison between these two pacing modalities has never been performed. The total cosine R to T (TCRT) is an ECG descriptor that accounts for depolarization-repolarization synchrony by measuring the difference between their directions. The purpose of this study was to compare TCRT in patients referred for RVP and LBBAP pacing as anti-bradycardia therapy. Methods: ECG recordings from 134 patients (82 LBBAP, 52 RVP) were classified into two groups, narrow QRS and wide QRS, depending on the patient’s QRS duration prior to implantation. In the post-implantation state, the TCRT index was calculated from a median beat calculated for each patient. Singular value decomposition was applied to the median beat in the eight independent ECG leads (I, II, V1, V2, V3, V4, V5, V6). The QRS complex and T wave loops in a three-dimensional space were determined from the first three components of the decomposition. TCRT was computed as the average of the cosines of the angles between the QRS complex directions and the maximum T wave direction. More positive values corresponded to more synchronized depolarization and repolarization processes while more negative values indicated larger differences in the orientation of the QRS and T wave loops and, therefore, greater dyssynchronization. Results: showed that TCRT took negative values for both techniques, RVP and LBBAP, and both groups, narrow and wide QRS, indicating that pacing generated dyssynchronization between ventricular depolarization and repolarization. Nevertheless, TCRT values for both groups were significantly more negative (p<0.01) for RVP than for LBBAP. We hypothesize that cardiac memory induced by pacing could account for these negative TCRT values. In any case, LBBAP did not increase the difference in the QRS complex and T wave loop orientations as much as RVP. Conclusion: LBBAP induces less dyssynchrony than RVP in the depolarization-repolarization process

Gene therapy for liver diseases: recent strategies for treatment of viral hepatitis and liver malignancies

Gene therapy has emerged as a powerful and very plastic tool to regulate biological functions in diseased tissues with application in virtually all medical fields. An increasing number of experimental and clinical studies underline the importance of genes as curative agents in the future. However, intense research is needed to evaluate the potential of gene therapy to improve efficacy and minimise the toxicity of the procedure

Analysis of genetic variability and mapping of point mutations in influenza virus by the RNase A mismatch cleavage method

We have applied the RNase A mismatch cleavage method to analyze genetic variability in RNA viruses by using influenza virus as a model system. Uniformly labeled RNA probes synthesized from a cloned hemagglutinin gene of a given viral strain were hybridized to RNA isolated from other strains of characterized or uncharacterized genetic composition. The RNA.RNA heteroduplexes containing a variable number of base mismatches were digested with RNase A, and the resistant products were analyzed by denaturing polyacrylamide gel electrophoresis. We show that many of these single base mismatches are cleaved by RNase A, generating unique and characteristic patterns of resistant RNA fragments specific for each of the different viral strains. Comparative analysis of the cleavage patterns allows a qualitative estimation of the genetic relatedness and evolution of field strains. We also show that cleavage by RNase A at single base mismatches can readily detect and localize point mutations present in monoclonal antibody-resistant variants. This method should have wide applications in the study of RNA viruses, not only for epidemiological analysis but also in some diagnostic problems, such as characterization of phenotypic mutants.This work was supported by National Institutes of Health Grant CA33021 awarded by the Nationa l Cancer Institute to M.P. and by grants from the Comision Asesora de Investigacion Cientificay Tecnica (Grant 608/438) and Fondo de Investigaciones Sanitarias to J.O. and J.A.M.C.L.-G. was a recipient of a NATO short-term post doctoral fellow-ship while on leave from the Centro Nacional de Microbiologia, Majadahonda, Madri

Immunotherapeutic synergy between anti-CD137 mAb and intratumoral administration of a cytopathic Semliki Forest virus encoding IL-12

Intratumoral injection of Semliki Forest virus encoding interleukin-12 (SFV-IL-12) combines acute expression of IL-12 and stressful apoptosis of infected malignant cells. Agonist antibodies directed to costimulatory receptor CD137 (4-1BB) strongly amplify pre-existing cellular immune responses toward weak tumor antigens. In this study, we provide evidence for powerful synergistic effects of a combined strategy consisting of intratumoral injection of SFV-IL-12 and systemic delivery of agonist anti-CD137 monoclonal antibodies (mAbs), which was substantiated against poorly immunogenic B16 melanomas (B16-OVA and B16.F10) and TC-1 lung carcinomas. Effector CD8(β)(+) T cells were sufficient to mediate complete tumor eradications. Accordingly, there was an intensely synergistic in vivo enhancement of cytotoxic T lymphocytes (CTL)-mediated immunity against the tumor antigens OVA and tyrosine-related protein-2 (TRP-2). This train of phenomena led to long-lasting tumor-specific immunity against rechallenge, attained transient control of the progression of concomitant tumor lesions that were not directly treated with SFV-IL-12 and caused autoimmune vitiligo. Importantly, we found that SFV-IL-12 intratumoral injection induces bright expression of CD137 on most tumor-infiltrating CD8(+) T lymphocytes, thereby providing more abundant targets for the action of the agonist antibody. This efficacious combinatorial immunotherapy strategy offers feasibility for clinical translation since anti-CD137 mAbs are already undergoing clinical trials and development of clinical-grade SFV-IL-12 vectors is in progress

Biliary Bicarbonate Secretion Constitutes a Protective Mechanism against Bile Acid-Induced Injury in Man

Background: Cholangiocytes expose a striking resistance against bile acids: while other cell types, such as hepatocytes, are susceptible to bile acid-induced toxicity and apoptosis already at micromolar concentrations, cholangiocytes are continuously exposed to millimolar concentrations as present in bile. We present a hypothesis suggesting that biliary secretion of HCO(3)(-) in man serves to protect cholangiocytes against bile acid-induced damage by fostering the deprotonation of apolar bile acids to more polar bile salts. Here, we tested if bile acid-induced toxicity is pH-dependent and if anion exchanger 2 (AE2) protects against bile acid-induced damage. Methods: A human cholangiocyte cell line was exposed to chenodeoxycholate (CDC), or its glycine conjugate, from 0.5 mM to 2.0 mM at pH 7.4, 7.1, 6.7 or 6.4, or after knockdown of AE2. Cell viability and apoptosis were determined by WST and caspase-3/-7 assays, respectively. Results: Glycochenodeoxycholate (GCDC) uptake in cholangiocytes is pH-dependent. Furthermore, CDC and GCDC (pK(a) 4-5) induce cholangiocyte toxicity in a pH-dependent manner: 0.5 mM CDC and 1 mM GCDC at pH 7.4 had no effect on cell viability, but at pH 6.4 decreased viability by >80% and increased caspase activity almost 10- and 30-fold, respectively. Acidification alone had no effect. AE2 knockdown led to 3- and 2-fold enhanced apoptosis induced by 0.75 mM CDC or 2 mM GCDC at pH 7.4. Discussion: These data support our hypothesis of a biliary HCO(3)(-) umbrella serving to protect human cholangiocytes against bile acid-induced injury. AE2 is a key contributor to this protective mechanism. The development and progression of cholangiopathies, such as primary biliary cirrhosis, may be a consequence of genetic and acquired functional defects of genes involved in maintaining the biliary HCO(3)(-) umbrella. Copyright (C) 2011 S. Karger AG, Base

Pea PSII-LHCII supercomplexes form pairs by making connections across the stromal gap

In higher plant thylakoids, the heterogeneous distribution of photosynthetic protein complexes is a determinant for the formation of grana, stacks of membrane discs that are densely populated with Photosystem II (PSII) and its light harvesting complex (LHCII). PSII associates with LHCII to form the PSII-LHCII supercomplex, a crucial component for solar energy conversion. Here, we report a biochemical, structural and functional characterization of pairs of PSII-LHCII supercomplexes, which were isolated under physiologically-relevant cation concentrations. Using single-particle cryo-electron microscopy, we determined the three-dimensional structure of paired C2S2M PSII-LHCII supercomplexes at 14 angstrom resolution. The two supercomplexes interact on their stromal sides through a specific overlap between apposing LHCII trimers and via physical connections that span the stromal gap, one of which is likely formed by interactions between the N-terminal loops of two Lhcb4 monomeric LHCII subunits. Fast chlorophyll fluorescence induction analysis showed that paired PSII-LHCII supercomplexes are energetically coupled. Molecular dynamics simulations revealed that additional flexible physical connections may form between the apposing LHCII trimers of paired PSII-LHCII supercomplexes in appressed thylakoid membranes. Our findings provide new insights into how interactions between pairs of PSII-LHCII supercomplexes can link adjacent thylakoids to mediate the stacking of grana membranes

Intratumoral injection of bone-marrow derived dendritic cells engineered to produce interleukin-12 induces complete regression of established murine transplantable colon adenocarcinomas

Stimulation of the antitumor immune response by dendritic cells (DC) is critically dependent on their tightly regulated ability to produce interleukin-12 (IL-12). To enhance this effect artificially, bone marrow (BM)-derived DC were genetically engineered to produce high levels of functional IL-12 by ex vivo infection with a recombinant defective adenovirus (AdCMVIL-12). DC-expressing IL-12 injected into the malignant tissue eradicated 50-100% well established malignant nodules derived from the injection of two murine colon adenocarcinoma cell lines. Successful therapy was dependent on IL-12 transfection and was mediated only by syngeneic, but not allogeneic BM-derived DC, indicating that compatible antigen-presenting molecules were required. The antitumor effect was inhibited by in vivo depletion of CD8+ T cells and completely abrogated by simultaneous depletion with anti-CD4 and anti-CD8 mAbs. Mice which had undergone tumor regression remained immune to a rechallenge with tumor cells, showing the achievement of long-lasting systemic immunity that also was able to reject simultaneously induced concomitant untreated tumors. Tumor regression was associated with a detectable CTL response directed against tumor-specific antigens probably captured by DC artificially released inside tumor nodules. Our results open the possibility of similarly treating the corresponding human malignancies

Upregulation of natural killer cells functions underlies the efficacy of intratumorally injected dendritic cells engineered to produce interleukin-12

OBJECTIVE: Injection of dendritic cells (DC) engineered with recombinant adenoviral vectors to produce interleukin-12 (IL-12) inside experimental murine tumors frequently achieves complete regressions. In such a system the function of CD8(+) T cells has been shown to be an absolute requirement, in contrast to observations made upon depletion of CD4(+) T cells, which minimally affected the outcome. The aim of this work was to study the possible involvement of natural killer (NK) cells in this setting. MATERIALS, METHODS, AND RESULTS: Depletions with anti-AsialoGM1 antiserum showed only a small decrease in the proportion of complete regressions obtained that correlated with induction of NK activities in lymphatic tissues into which DC migrate, whereas combined depletions of CD4(+) and NK cells completely eliminated the antitumor effects. Likewise in vivo neutralization of interferon-gamma (IFN-gamma) also eliminated those therapeutic effects. Trying to define the cellular role played by NK cells in vivo, it was observed that injection of cultured DC inside the spleen of T- and B-cell-deficient (Rag1(-/-)) mice induced upregulation of NK activity only if DC had been adenovirally engineered to produce IL-12. In addition, identically transfected fibroblasts also activated NK cells, indicating that IL-12 transfection was the unique requirement. Equivalent human DC only activated in vitro the cytolytic and cytokine-secreting functions of autologous NK cells if transfected to express human IL-12. CONCLUSIONS: Overall, these results point out an important role played by NK cell activation in the potent immunotherapeutic effects elicited by intratumoral injection of IL-12--secreting DC and that NK activation under these conditions is mainly, if not only, dependent on IL-12