Rápido método de selección de plantas resistentes a herbicidas

Rápido método de selección de plantas resistentes a herbicidasFil: Larran, A. S. Universidad Nacional de Rosario. Facultad de Ciencias Agrarias; Argentin

Stepwise Optimization of the Procedure for Assessment of Circulating Progenitor Cells in Patients with Myocardial Infarction

The number and functional activity of circulating progenitor cells (CPCs) is altered in diabetic patients. Furthermore, reduced CPC count has been shown to independently predict cardiovascular events. Validation of CPCs as a biomarker for cardiovascular risk stratification requires rigorous methodology. Before a standard operation protocol (SOP) can be designed for such a trial, a variety of technical issues have to be addressed fundamentally, which include the appropriate type of red blood cell lysis buffer, FMO or isotype controls to identify rare cell populations from background noise, optimal antibody dilutions and conditions of sample storage. We herein propose improvements in critical steps of CPC isolation, antigenic characterization and determination of functional competence for final application in a prospective investigation of CPCs as a biomarker of outcome following acute myocardial infarction.In this validation study, we refined the standard operating procedure (SOP) for flow cytometry characterisation and functional analysis of CPCs from the first 18 patients of the Progenitor Cell Response after Myocardial Infarction Study (ProMIS). ProMIS aims to verify the prognostic value of CPCs in patients with either ST elevation or non-ST elevation myocardial infarction with or without diabetes mellitus, using cardiac magnetic resonance imaging (MRI) for assessment of ventricular function as a primary endpoint. Results indicate crucial steps for SOP implementation, namely timely cell isolation after sampling, use of appropriate lysis buffer to separate blood cell types and minimize the acquisition events during flow cytometry, adoption of proper fluorophore combination and antibody titration for multiple antigenic detection and introduction of counting beads for precise quantification of functional CPC activity in migration assay.With systematic specification of factors influencing the enumeration of CPC by flow cytometry, the abundance and migration capacity of CPCs can be correctly assessed. Adoption of validated SOP is essential for refined comparison of patients with different comorbidities in the analysis of risk stratification

Safety perspectives on presently considered drugs for the treatment of COVID‐19

Intense efforts are underway to evaluate potential therapeutic agents for the treatment of COVID‐19. In order to respond quickly to the crisis, the repurposing of existing drugs is the primary pharmacological strategy. Despite the urgent clinical need for these therapies, it is imperative to consider potential safety issues. This is important due to the harm–benefit ratios that may be encountered when treating COVID‐19, which can depend on the stage of the disease, when therapy is administered and underlying clinical factors in individual patients. Treatments are currently being trialled for a range of scenarios from prophylaxis (where benefit must greatly exceed risk) to severe life‐threatening disease (where a degree of potential risk may be tolerated if it is exceeded by the potential benefit). In this perspective, we have reviewed some of the most widely researched repurposed agents in order to identify potential safety considerations using existing information in the context of COVID‐19

Suspensión del tratamiento con inhibidores de BCR-ABL en los pacientes con leucemia mieloide crónica en respuesta molecular profunda dentro de la práctica clínica asistencial: Experiencia española en un total de 236 casos

Oral Presentation [CO-092] Introducción: La mitad de los pacientes con leucemia mieloide crónica (LMC) en respuesta molecular profunda no pierde la respuesta molecular mayor (RMM) tras la suspensión del tratamiento con inhibidores de BCR-ABL (ITC). Esta estrategia ha demostrado ser segura en ensayos clínicos pero hay poca información acerca de su aplicabilidad en la práctica clínica asistencial. El objetivo del estudio fue analizar la experiencia con la suspensión del tratamiento fuera de ensayo clínico en España. Métodos: Se analizan los resultados de 236 pacientes con LMC en fase crónica que suspendieron el tratamiento fuera de ensayo clínico en 33 hospitales. Criterios de inclusión: a) tratamiento con ITC >3 años, b) respuesta molecular grado 4.5 durante >2 años (se permitió una única determinación de RM4 durante ese período). Se excluyeron los pacientes trasplantados. Resultados: Las características de la serie se muestran en la tabla 1. Los motivos principales para suspender el tratamiento fueron los efectos secundarios (n=66), lograr la remisión libre de tratamiento (n=166) y el embarazo (n=4). La mediana de seguimiento tras la suspensión fue de 21, 5 meses y 5 pacientes fallecieron por causas no relacionadas con la LMC. Durante este periodo, 67 pacientes reiniciaron el tratamiento por recaída molecular (pérdida de RMM: n=52, aumento de tránscritos >1 log en dos controles sucesivos sin pérdida de RMM: n=12), decisión del paciente (n=2) o síndrome de discontinuación (n=1). Un paciente perdió la RMM a los 20 meses y decidió no tratarse, recuperando la RMM espontáneamente. Cuarenta y nueve recaídas (75% del total) ocurrieron en los primeros 6 meses, 8 entre los meses 7-12, y 8 tras los 12 meses, produciéndose la pérdida de RMM más tardía a los 30 meses. La supervivencia libre de reinicio del tratamiento (figura 1) y de recaída molecular fue del 66, 8% y del 67, 5% a los 3 años, respectivamente. Los factores asociados a mayor supervivencia libre de recaída fueron la duración del tratamiento con ITC >5 años (p=0.01) y la RM4.5 >4 años antes de la suspensión (p=0.017). Un total de 51 pacientes (22%) desarrollaron dolor osteomuscular tras la suspensión. No se registró ningún caso de progresión a fases avanzadas. El valor mediano de la carga de BCR-ABL al reinicio del tratamiento fue del 0, 3% (>5% en 7 casos). La mediana de seguimiento tras reinicio del tratamiento fue de 20 meses; 46 de 52 casos (88%) recuperaron la RMM tras una mediana de tiempo de 3 meses; 50 de 64 recuperaron la RM4 (mediana 3, 5 meses) y 47 de 64 recuperaron la MR4.5 (mediana 5 meses). En el último control, el estado de la respuesta fue: RM4.5 (n=195), RM4 (n=15), RMM (n=14), respuesta citogenética completa (n=10), otros (n=2). Conclusiones: los resultados confirman que la suspensión del tratamiento es factible en la práctica clínica asistencial en España. La duración del tratamiento y de la respuesta molecular profunda se asociaron con la supervivencia libre de recaída. Esta información puede ser útil para establecer recomendaciones generales acerca de la discontinuación del tratamiento de la LMC en nuestro medio

Epicoccum nigrum P16, a Sugarcane Endophyte, Produces Antifungal Compounds and Induces Root Growth

Background: Sugarcane is one of the most important crops in Brazil, mainly because of its use in biofuel production. Recent studies have sought to determine the role of sugarcane endophytic microbial diversity in microorganism-plant interactions, and their biotechnological potential. Epicoccum nigrum is an important sugarcane endophytic fungus that has been associated with the biological control of phytopathogens, and the production of secondary metabolites. In spite of several studies carried out to define the better conditions to use E. nigrum in different crops, little is known about the establishment of an endophytic interaction, and its potential effects on plant physiology. Methodology/Principal Findings: We report an approach based on inoculation followed by re-isolation, molecular monitoring, microscopic analysis, plant growth responses to fungal colonization, and antimicrobial activity tests to study the basic aspects of the E. nigrum endophytic interaction with sugarcane, and the effects of colonization on plant physiology. The results indicate that E. nigrum was capable of increasing the root system biomass and producing compounds that inhibit the in vitro growth of sugarcane pathogens Fusarium verticillioides, Colletotrichum falcatum, Ceratocystis paradoxa, and Xanthomomas albilineans. In addition, E. nigrum preferentially colonizes the sugarcane surface and, occasionally, the endophytic environment. Conclusions/Significance: Our work demonstrates that E. nigrum has great potential for sugarcane crop application because it is capable of increasing the root system biomass and controlling pathogens. The study of the basic aspects of the interaction of E. nigrum with sugarcane demonstrated the facultative endophytism of E. nigrum and its preference for the phylloplane environment, which should be considered in future studies of biocontrol using this species. In addition, this work contributes to the knowledge of the interaction of this ubiquitous endophyte with the host plant, and also to a better use of microbial endophytes in agriculture.State of Sao Paulo Research Foundation (FAPESP)FAPESP (State of Sao Paulo Research Foundation) [02/14143-3, 10/08286-2]Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq)Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq

Perspectives for biocatalytic lignin utilization: cleaving 4-O-5 and C??-C?? bonds in dimeric lignin model compounds catalyzed by a promiscuous activity of tyrosinase

Background: In the biorefinery utilizing lignocellulosic biomasses, lignin decomposition to value-added phenolic derivatives is a key issue, and recently biocatalytic delignification is emerging owing to its superior selectivity, low energy consumption, and unparalleled sustainability. However, besides heme-containing peroxidases and laccases, information about lignolytic biocatalysts is still limited till date. Results: Herein, we report a promiscuous activity of tyrosinase which is closely associated with delignification requiring high redox potentials (>1.4 V vs. normal hydrogen electrode [NHE]). The promiscuous activity of tyrosinase not only oxidizes veratryl alcohol, a commonly used nonphenolic substrate for assaying ligninolytic activity, to veratraldehyde but also cleaves the 4-O-5 and C??-C?? bonds in 4-phenoxyphenol and guaiacyl glycerol-??-guaiacyl ether (GGE) that are dimeric lignin model compounds. Cyclic voltammograms additionally verified that the promiscuous activity oxidizes lignin-related high redox potential substrates. Conclusion These results might be applicable for extending the versatility of tyrosinase toward biocatalytic delignification as well as suggesting a new perspective for sustainable lignin utilization. Furthermore, the results provide insight for exploring the previously unknown promiscuous activities of biocatalysts much more diverse than ever thought before, thereby innovatively expanding the applicable area of biocatalysis