ABSTRACT
Leishmania major, a protozoan parasite that diverged early from the main eukaryotic lineage, exhibits unusual mechanisms of gene expression. Little is known in this organism about the transcription factors involved in the synthesis of tRNA, 5S rRNA, and snRNAs, transcribed by RNA Polymerase III (Pol III). Here we identify and characterize the TFIIIB subunit Bdp1 in L. major (LmBdp1). Bdp1 plays key roles in Pol III transcription initiation in other organisms, as it participates in Pol III recruitment and promoter opening. In silico analysis showed that LmBdp1 contains the typical extended SANT domain as well as other Bdp1 conserved regions. Nevertheless, LmBdp1 also displays distinctive features, including the presence of only one aromatic residue in the N-linker region. We were not able to produce null mutants of LmBdp1 by homologous recombination, as the obtained double replacement cell line contained an extra copy of LmBdp1, indicating that LmBdp1 is essential for the viability of L. major promastigotes. Notably, the mutant cell line showed reduced levels of the LmBdp1 protein, and its growth was significantly decreased in relation to wild-type cells. Nuclear run-on assays demonstrated that Pol III transcription was affected in the mutant cell line, and ChIP experiments showed that LmBdp1 binds to 5S rRNA, tRNA, and snRNA genes. Thus, our results indicate that LmBdp1 is an essential protein required for Pol III transcription in L. major.
Subject(s)
Leishmania major/genetics , RNA Polymerase III/genetics , Transcription Factor TFIIIB/genetics , Transcription, Genetic , Computer Simulation , Conserved Sequence/genetics , Gene Expression Regulation/genetics , Homologous Recombination/genetics , Mutant Proteins/genetics , Promoter Regions, Genetic , Protein Domains/genetics , Protein Subunits/genetics , RNA, Ribosomal, 5S/biosynthesis , RNA, Small Nuclear/biosynthesis , RNA, Transfer/biosynthesisABSTRACT
OBJECTIVES: to identify the number of electro-medical pieces of equipment in a coronary care unit, characterize their types, and analyze implications for the safety of patients from the perspective of alarm fatigue. METHOD: this quantitative, observational, descriptive, non-participatory study was conducted in a coronary care unit of a cardiology hospital with 170 beds. RESULTS: a total of 426 alarms were recorded in 40 hours of observation: 227 were triggered by multi-parametric monitors and 199 were triggered by other equipment (infusion pumps, dialysis pumps, mechanical ventilators, and intra-aortic balloons); that is an average of 10.6 alarms per hour. CONCLUSION: the results reinforce the importance of properly configuring physiological variables, the volume and parameters of alarms of multi-parametric monitors within the routine of intensive care units. The alarms of equipment intended to protect patients have increased noise within the unit, the level of distraction and interruptions in the workflow, leading to a false sense of security. .
OBJETIVOS: identificar o número de alarmes dos equipamentos eletromédicos numa unidade coronariana, caracterizar o tipo e analisar as implicações para a segurança do paciente na perspectiva da fadiga de alarmes. MÉTODO: trata-se de estudo quantitativo observacional descritivo, não participante, desenvolvido numa unidade coronariana de um hospital de cardiologia, com capacidade para 170 leitos. RESULTADOS: registrou-se o total de 426 sinais de alarmes, sendo 227 disparados por monitores multiparamétricos e 199 alarmes disparados por outros equipamentos (bombas infusoras, hemodiálise, ventiladores mecânicos e balão intra-aórtico), nas 40h, numa média total de 10,6 alarmes/hora. CONCLUSÃO: os resultados encontrados reforçam a importância da configuração de variáveis fisiológicas, do volume e dos parâmetros de alarmes dos monitores multiparamétricos à rotina das unidades de terapia intensiva. Os alarmes dos equipamentos destinados a proteger os pacientes têm conduzido ao aumento do ruído na unidade, à fadiga de alarmes, a distrações e interrupções no fluxo de trabalho e à falsa sensação de segurança. .
OBJETIVOS: identificar el número de alarmas de los equipamientos electromédicos en una unidad coronariana, caracterizar el tipo y analizar las implicaciones para la seguridad del paciente en la perspectiva de fatiga de alarmas. MÉTODO: se trata de un estudio cuantitativo, observacional, descriptivo, no participante, desarrollado en una unidad coronariana de un hospital de cardiología, con capacidad de 170 camas. RESULTADOS: se registró un total de 426 señales de alarmas, siendo 227 disparadas por monitores multiparamétricos y 199 disparadas por otros equipamientos (bombas de infusión, hemodiálisis, ventiladores mecánicos y balón intraaórtico), durante 40h, con un promedio total de 10,6 alarmas/hora. CONCLUSIÓN: los resultados encontrados refuerzan la importancia de la configuración de las variables fisiológicas, del volumen y de los parámetros de alarma de los monitores multiparamétricos, a la rutina de las unidades de terapia intensiva. Las alarmas de los equipamientos destinados a proteger a los pacientes, han llevado al aumento del ruido en la unidad, a la fatiga de alarmas, a las distracciones e interrupciones en el flujo de trabajo y a una falsa sensación de seguridad. .