RESUMO
Fundamento: el Acinetobacter baumannii se ha convertido en uno de los patógenos nosocomiales más relevantes.Objetivo: caracterizar pacientes con diagnóstico de neumonía por Acinetobacter baumannii asociada a la ventilación mecánica en las Unidades de Cuidados Progresivos. Métodos: estudio de serie de casos realizado desde diciembre del 2013 a diciembre del 2014 en Unidades de Cuidados Progresivos del Hospital General Dr. Gustavo Aldereguía Lima, de Cienfuegos, que incluyó pacientes con diagnóstico de neumonía asociada a la ventilación mecánica con aislamiento de Acinetobacter baumannii. Variables analizadas: edad, sexo, comorbilidad, enfermedad que motivó el ingreso, duración de la ventilación, estadía, tipo de antibiótico usado y estado al egreso. Resultados: se estudiaron 39 pacientes, el 69, 2 % de las Unidades de Cuidados Progresivos. La edad media fue de 55,7 años. Predominó el sexo masculino. El 64, 2 % de los pacientes refirió uno o varios antecedentes patológicos personales. Predominó el ingreso en los servicios de urgencia por situaciones clínicas (51, 3 %). El 69,2 % de pacientes fue ventilado entre 3 y 21 días. La estadía media fue de 14,7 días. El 71, 8 % tuvo tratamiento combinado y la mayor parte de ellos egresaron vivos (64,1 %). La mortalidad general fue de 35,9 %.Conclusiones: en la serie analizada predominó el sexo masculino, edad superior a los 60 años y los casos clínicos; necesitaron ventilación mecánica de mediana duración y tratamiento antimicrobiano combinado. La mayor parte de los pacientes egresaron vivos y la mortalidad observada está en el rango de lo reportado en la literatura científica.
Background: Acinetobacter baumannii has become one of the most important nosocomial pathogens. Objective: to characterize the patients diagnosed with ventilator-associated pneumonia due to Acinetobacter baumannii in the Progressive Care Units. Methods: a case series study of patients diagnosed with ventilator-associated pneumonia due to Acinetobacter baumannii was conducted in the Progressive Care Units of the Dr. Gustavo Aldereguía Lima Hospital of Cienfuegos from December 2013 through December 2014. The study variables included: age, sex, comorbid conditions, cause of admission, duration of ventilation, length of stay, antibiotic used, and status at discharge. Results: a total of 39 patients were studied, which accounted for 69.2% of the patients in Progressive Care Units. The mean age was 55.7 years. Males predominated. Sixty four point two percent of patients reported one or more past illnesses. Most admissions to emergency services were due to clinical reasons (51.3%). Sixty nine point two percent of patients received mechanical ventilation for 3 to 21 days. The average stay was 14.7 days. Seventy one point eight percent received a combined antimicrobial treatment and most of them were discharged alive (64.1%). Overall mortality was 35.9%. Conclusions: there was a predominance of males, patients over 60 years of age and clinical cases. The study patients needed mechanical ventilation for a medium length of time and combined antimicrobial treatment. Most patients were discharged alive, and mortality was within the range of that reported in the scientific literature.
RESUMO
We have found a direct relationship between protein production in Pichia pastoris and the number of introduced synthetic genes of miniproinsulin (MPI), fused to the Saccharomyces cerevisiae pre-pro alpha factor used as secretion signal, and inserted between the alcohol oxidase 1 (AOX1) promoter and terminator sequences. Two consecutive approaches were followed to increase the number of integrated cassettes: the head-to-tail expression cassette multimerization procedure and re-transformation with a dominant selection marker. This increased expression from 19 to 250 mg l(-1) when about 11 copies have been integrated. Further, the correct position of one of the disulphide bridges of the purified molecule was verified by digestion with Glu-C endoprotease, followed by mass spectrometry of the isolated fragments.
Assuntos
Biotecnologia/métodos , Técnicas Genéticas , Pichia/metabolismo , Proinsulina/genética , Proinsulina/metabolismo , Oxirredutases do Álcool/genética , Sequência de Aminoácidos , Clonagem Molecular , Dissulfetos/química , Eletroforese em Gel de Poliacrilamida , Fermentação , Dosagem de Genes , Vetores Genéticos , Humanos , Cinética , Espectrometria de Massas , Dados de Sequência Molecular , Mapeamento de Peptídeos , Peptídeos/química , Plasmídeos/metabolismo , Regiões Promotoras Genéticas , Saccharomyces cerevisiae/metabolismo , Serina Endopeptidases/metabolismo , Fatores de TempoRESUMO
Increased expression of recombinant mini-proinsulin in Pichia pastoris in 2.5 l bioreactors was achieved by increasing the cultivation pH from 5.1 to 6.3, by decreasing the temperature from 28 to 22 degrees C, and by periodical addition of ammonium sulfate and EDTA to the culture broth. Using this procedure, mini-proinsulin reached nearly 0.3 g l(-1) in the culture supernatant after 160 h of growth.