RESUMO
In nature, dynamic interactions between enzymes play a crucial role in defining cellular metabolism. By controlling the spatial and temporal organization of these supramolecular complexes called metabolons, natural metabolism can be tuned in a highly dynamic manner. Here, we repurpose the CRISPR-Cas6 family proteins as a synthetic strategy to create dynamic metabolons by combining the ease of RNA processing and the predictability of RNA hybridization for protein assembly. By disturbing RNA-RNA networks using toehold-mediated strand displacement reactions, on-demand assembly and disassembly are achieved using both synthetic RNA triggers and mCherry messenger RNA. Both direct and 'Turn-On' assembly of the pathway enzymes tryptophan-2-monooxygenase and indoleacetamide hydrolase can enhance indole-3-acetic acid production by up to ninefold. Even multimeric enzymes can be assembled to improve malate production by threefold. By interfacing with endogenous mRNAs, more complex metabolons may be constructed, resulting in a self-responsive metabolic machinery capable of adapting to changing cellular demand.
Assuntos
Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas , Endonucleases , Sistemas CRISPR-Cas/genética , Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas/genética , Endonucleases/metabolismo , RNA/metabolismo , Processamento Pós-Transcricional do RNARESUMO
La transfusión masiva es la transfusión de más del 50% del volumen sanguíneo total en 3 horas; 100% en 24 horas o reemplazar la pérdida de sangre en curso de más de 10% del volumen sanguíneo total por minuto. Los pacientes pediátricos requieren transfusión masiva en una variedad de entornos. Existen datos limitados para apoyar estrategias óptimas de transfusión masiva en niños, los datos son extrapolados de adultos y no es lo más conveniente, sin embargo alrededor de 15% de los niños la requieren. La evidencia actual sugiere que en centros donde se realizan transfusiones masivas basadas en protocolos, la morbimortalidad es mucho menor.
Massive transfusion is the transfusion of more than 50 % of the total blood volume in 3 h; 100 % in 24 h or to replace the ongoing blood loss of more than 10% total blood volume per minute. Pediatric patients require massive transfusion in a variety of settings. There are limited data to support optimal mass transfusion strategies in children, the data is extrapolated from adults and is not the most convenient, however, about 15% of children require it. Current evidence suggests that in centers where massive protocol-based transfusions are performed, morbidity and mortality is much lower.
