摘要
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. .
Subject(s)
Humans , DNA-Directed RNA Polymerases/metabolism , Oncogene Proteins, Viral/genetics , RNA Polymerase III/metabolism , Sarcosine/analogs & derivatives , Transcription Factors, TFIII , Transcription, Genetic , Transcription Factors/metabolism , Adenovirus Early Proteins , Detergents , DNA-Binding Proteins/genetics , HeLa Cells , Kinetics , Sarcosine/pharmacology , Transcription Factor TFIIIB , Transcription Factors/genetics , Transcription, Genetic/drug effects摘要
RNA polymerase IV and V are principal players in the RdDM pathway, where their current study has shown interaction of several factors that control DNA silencing of intergenic regions and siRNA production. DNA silencing is an important process during cell differentiation, nuclear structure and viral control. However, RNA pol IV and V are yet to be study in model monocot systems like Oryza sativa that can provide further information on genetic silence mechanism in plats. We show the expression pattern of these polymerases in tissues extracts of Oryza sativa. Detectable amounts of these polymerases are found in specific adult plant tissues and particularly expressed during somatic embryogenesis but not during early stages of normal embryo development. The use of synthetic auxin leads to an induction of both RNA pol IV and V in scutellum tissue where nuclear localization may be required for genome reorganization and gene silencing.
Subject(s)
Infant , DNA-Directed RNA Polymerases/metabolism , Oryza/enzymology , Oryza/genetics , DNA-Directed RNA Polymerases/genetics , Blotting, Western , Embryonic Development , Fluorescent Antibody Technique , Gene Silencing , Real-Time Polymerase Chain Reaction , Seeds摘要
The type II restriction endonuclease, Bam HI, has been overexpressed in E. coli by cloning the Bam HI gene in frame with an E. coli Ribosome Binding Site (RBS) under the T7 promoter of an E. coli expression vector pRSET A. The expression level of Bam HI endonuclease using this construct was found to be higher than that reported of the overexpressing clone pAEK14. Our overexpressing clone, pAABRw in BL21 cells in presence of Bam HI methylase in pMAP6 following induction with IPTG yields about 9.2 x 10(6) units per gram wet cell paste. In vivo activity of the recombinant endonuclease could be confirmed by the SOS induction assay in JH139 cells even in the absence of T7 polymerase and cognate Bam HI methylase because of leaky expression in E. coli. This provides an alternate way to screen the active endonuclease and its variants.
Subject(s)
Binding Sites , Cell Division , Cells, Cultured , DNA-Directed RNA Polymerases/metabolism , Deoxyribonuclease BamHI/genetics , Mutation , Plasmids , Recombinant Proteins/isolation & purification , Viral Proteins摘要
Transcription is the foremost event in gene expression in which the enzyme RNA polymerase copies the genetic information from DNA to RNA. Much of our understanding of this process have come from studies carried out in Escherichia coli. A faithful and efficient transcription machinery of E. coli can be reconstituted in vitro with purified RNA polymerase and promoter-containing DNA. It is generally believed that in E. coli and most other organisms, the control of gene expression lies with the initiation of transcription. In this review, an attempt has been made to understand the mechanistic details of the initiation of transcription from the structural point of view of the promoter and the RNA polymerase. Allosteric nature of the enzyme has also been discussed at the end.
Subject(s)
Allosteric Regulation , DNA-Directed RNA Polymerases/metabolism , Escherichia coli/genetics , Models, Genetic , Promoter Regions, Genetic , Transcription, Genetic摘要
The expression of a native form of the foot-and-mouth disease virus RNA polymerase was obtained. Two oligonucleotides of 66 base pairs were used to renuild the 5' end of the gene and to introduce the first methionine codon. The expression of the active polymerase in E. coli was achieved by inserting the gene before the tac promoter of the pKK223-3 plasmid
Subject(s)
DNA-Directed RNA Polymerases/genetics , Hand, Foot and Mouth Disease/genetics , Escherichia coli/genetics , Gene Expression Regulation, Viral , Amino Acid Sequence , Aphthovirus/enzymology , Base Sequence , Cloning, Molecular , Codon/genetics , DNA-Directed RNA Polymerases/immunology , DNA-Directed RNA Polymerases/metabolism , Molecular Sequence Data , Oligonucleotides/chemistry , Plasmids , Recombinant Proteins/genetics , Recombinant Proteins/immunology , RNA, Viral/genetics摘要
A RNA-polimerase (RNA-pol) do vírus da Febre Aftosa foi expressa em Escherichia coli de uma maneira controlável. Para isso um vetor de expressäo foi construído, clonando-se um cDNA do gene contendo o primeiro codon (AGT) e as sequências de Shine & Delgarno foi adicionada no terminal 5' do gene. Após termoinduçäo da cepa recombinante, a RNA-pol do virus da Febre Aftosa foi expressa, chegando a atingir 10% da proteína total da célula. Esta RNA-pol recombinante apresentava um peso molecular esperado (50Kda) assim como reaçäo cruzada em Wstern blot com soros anti RNA-pol (natural) e de bovinos convalescentes de Febre Aftosa. Um soro policlonal contra a RNA-pol recombinante foi capaz de inibir a atividade enzimática da RNA-pol natural. A expressäo da RNA-pol do vírus da Febre Aftosa em E. coli facilitará a análise da atividade enzimática desta molécula assim como tornará possível o desenvolvimento de um método diagnóstico eficiente e seguro para a Febre Aftosa
Subject(s)
Cattle , Aphthovirus/genetics , DNA-Directed RNA Polymerases/metabolism , Escherichia coli/genetics , Aphthovirus/enzymology , Escherichia coli/enzymology摘要
La RNA polimerasa II es respondable de la síntesis de RNA mensajero en organismos eucariontes. Algunas preparaciones de esta enzima presentan 3 formas llamadas II0, IIa y IIB. Utilizamos la metodología de transferencia e inmunodetección de proteínas en filtro de nitrocelulosa, para conocer la proporción d elas distintas formas de esta enzima en células de riñon de mono en cultivo (CVI). Encontramos que la subespecie II0 (con una subúnidad mayor de 240.000 d) representa una fracción importante de la RNA polimerasa II de estas células, contrario a lo comúnmnete observado en preparaciones purificadas de la enzima, en donde las subespecies IIA y IIB son mayoritarias. Mediante el empleo de la mencionada metodológia detectamos subunidades específicas de la enzima, en preparaciones de células, núcleo y cromatina "parcialmente purificada". Esto nos permitió analizar directamente la estructura de la enzima, sin utilizar los procedimientos tradicionales de purificación, los cuales resultan en una pérdida selectiva de la suespecie IIo; la enzima II0 no solo parece representar una alta proporción de la RNA polimerasa II en estas células, sino que además la encontramos asociada principalmente con la cromatina celular