Carboxylate isosteres for caspase inhibitors: the acylsulfonamide case revisited.

As part of an ongoing effort to discover inhibitors of caspase-1 with an optimized selectivity and biopharmaceutical profile, acylsulfonamides were explored as carboxylate isosteres for caspase inhibitors. Acylsulfonamide analogues of the clinically investigated caspase-1 inhibitor VRT-043198 and of the pan-caspase inhibitor Z-VAD-CHO were synthesized. The isostere-containing analogues with an aldehyde warhead had inhibitory potencies comparable to the carboxylate references. In addition, the conformational and tautomeric characteristics of these molecules were determined using 1H- and 13C-based NMR. The propensity of acylsulfonamides with an aldehyde warhead to occur in a ring-closed conformation at physiological pH significantly increases the sensitivity to hydrolysis of the acylsulfonamide moiety, yielding the parent carboxylate containing inhibitors. These results indicate that the acylsulfonamide analogues of the aldehyde-based inhibitor VRT-043198 might have potential as a novel type of prodrug for the latter. Finally, inhibition of caspase 1 and 11 mediated inflammation in mouse macrophages was found to correlate with the potencies of the compounds in enzymatic assays.

Análisis descriptivo de 10 años de broncoscopia terapéutica intervencionista / 10-year descriptive analysis of therapeutic interventional bronchoscopy

Objetivo: Describir nuestros casos en el campo de la broncoscopia terapéutica intervencionista después de 10 años de experiencia Materiales y métodos: Hemos recogido historia, tipo de patología traqueobronquial, técnicas, resultados y complicaciones Resultados: Fueron incluidos 186 pacientes (113 hombres y 73 mujeres), con edad media de 51,3 años (rango 8 - 86). Se han realizado 320 procedimientos o técnicas sobre ellos. Indicaciones del procedimiento: estenosis benigna de la vía aérea 45,69% (85/186), estenosis maligna 41,93 % (78/186), fistula traqueoesofágica y broncopleural 5,91% (11/186), extracción de cuerpo extraño con broncoscopio rígido 4,3% (8/186) y 3 casos de hemoptisis amenazante. La estenosis benigna secundaria a intubación seguida por la post traqueostomia fueron las más comunes, 35,29% (30) y 34,11% (29) respectivamente. Entre las lesiones malignas, 63 casos (80,76%) estaban localizados en el pulmón, de estos, el carcinoma broncogenico, 60 de 78 pacientes, era el más frecuente y 3 tumores carcinoides. El 19,23% restante (15/78) no eran tumores broncopulmonares. La técnica más usada ha sido el láser en 100 casos, dilatación neumática en 65, endoprótesis 53 (35 metálicas y 18 silicona), electrocirugía en 50, resección mecánica en 45 pacientes, 3 sellados de fístulas y 3 oclusiones con balón por hemoptisis. El procedimiento combinado más empleado fue la resección laser más dilatación neumática. Complicaciones intraoperatorias en 11 casos (4,8%) y tardías en 4 (2,15%). Todos los pacientes (100%) mejoraron de sus síntomas después del procedimiento. Conclusiones: La broncoscopia terapéutica intervencionista es un procedimiento seguro y efectivo en el manejo de las lesiones de la vía aérea

Objective: To describe our cases in the field of therapeutic Interventional Bronchoscopy after 10 years of experience. Materials and Methods: We collected history, type of Tracheobronchial Pathology, technique, results and complications. Results: Were they performed 186 (113 male and 73 female) patients with mean age 51.3 years (range: 8-86). Had 320 procedures or techniques performed on them. Indications for the procedure: benign airway stenosis 45,69% (85/186), malignant stenosis 41.93% (78/186), tracheoesophageal and bronchopleural fistulas 5.91% (11/186), foreign bodies extracted with rigid bronchoscope 4.3% (8/186), and 3 cases of threatening hemoptysis. Among the benign stenosis secondary to intubation followed by the post-tracheotomy were the most common, 35.29% (30) and 34.11% (29) respectively. Between malignant lesions, 63 cases (80,76%) had home lung, of these, carcinoma, bronchogenic, 60 of 78 patients, was the most frequent and 3 carcinoid tumors. The rest 19,23% (15 / 78) were non bronchopulmonary tumors. The most used techniques have been laser in 100 cases, pneumatic dilatation in 65, Endoprosthesis 53 (35 metal and silicone 18), electrosurgery in 50, mechanical resection in 45 patients, 3 fistulas sealments and 3 occlusion balloon by hemoptysis. The most employed combined procedure was resection laser and mechanics more pneumatic dilation. Intraoperative complications in 11 cases (4.8%) and late in 4 (2.15%). All patients (100%) improved their symptoms after the procedure. Conclusions: The therapeutic interventionist bronchoscopy is a safe and effective procedure in the management of injuries in the airways)

In-planta detection and monitorization of endophytic colonization by a Beauveria bassiana strain using a new-developed nested and quantitative PCR-based assay and confocal laser scanning microscopy.

Beauveria bassiana strain 04/01-Tip obtained from larvae of the opium poppy stem gall Iraella luteipes endophytically colonizes opium poppy plants and protect it against this pest. Development of a specific, rapid and sensitive technique that allows accurately determining the process and factors leading to the establishment of this strain in opium poppy plants would be essential to achieve its efficient control in a large field scale. For that purpose in the present study, species-specific primers that can be used in conventional or quantitative PCR protocols were developed for specifically identification and detection of B. bassiana in plant tissues. The combination of the designed BB.fw/BB.rv primer set with the universal ITS1-F/ITS4 primer set in a two-step nested-PCR approach, has allowed the amplification of up to 10fg of B. bassiana. This represented an increase in sensitivity of 10000- and 1000-fold of detection than when using the BB.fw/BB.rv primers in a single or single-tube semi-nested PCR approaches, respectively. The BB.fw and BB.rv primer set were subsequently optimized to be used in real time quantitative PCR assays and allowed to accurately quantify B. bassiana DNA in different plant DNA backgrounds (leaves and seeds) without losing accuracy and efficiency. The qPCR protocol was used to monitor the endophytic colonization of opium poppy leaves byB. bassiana after inoculation with the strain EABb 04/01-Tip, detecting as low as 26fg of target DNA in leaves and a decrease in fungal biomass over time. PCR quantification data were supported in parallel with CLMS by the monitoring of spatial and temporal patterns of leaf and stem colonization using a GFP-tagged transformant of the B. bassiana EABb 04/01-Tip strain, which enabled to demonstrate that B. bassiana effectively colonizes aerial tissues of opium poppy plants mainly through intercellular spaces and even leaf trichomes. A decline in endophytic colonization was also observed by the last sampling times, i.e. from 10 to 15days after inoculation, although fungal structures still remained present in the leaf tissues. These newly developed molecular protocols should facilitate the detection, quantification and monitoring of endophytic B. bassiana strains in different tissues and host plants and would help to unravel the factors and process governing the specific endophytic association between opium poppy and strain EABb 04/01-Tip providing key insights to formulate a sustainable strategy for I. luteipes management in the host.