Imunorreatividade para endotelina-1 no plasma e fluido folicular de eqüinos e bovinos

Endothelin-1, a recently isolated peptide, has been observed in different tissues and is involved in several reproductive functions. The objective of the present study was to determine the concentrations of immunoreactive endothelin-1 in the plasma of horses and cattle and in the follicular fluid of horses using a commercial radioimmunoassay kit. Mean immunoreactivity was 47.5%, the minimum detectable dose was 1 pg/tube and the interassay and intra-assay coefficients of variation were 2.36% and 7.26%, respectively. The parallelism between the standard curves and the curves determined with different volumes of plasma and follicular fluid validates the radioimmunoassay of endothelin-1 for these species. To our knowledge, this is the first report in the literature of detection of endothelin-1 in the plasma and follicular fluid of horses and in the plasma of cattle by radioimmunoassay. The present results suggest the use of the radioimmunoassay method employing commercial kits for the determination of endothelin-1 concentrations in horse and cattle plasma and horse follicular fluid because of its high sensitivity, accuracy and reproducibility, in order to study the multiple functions of endothelin-1 in these animals species, and especially in the endocrine-reproductive system.A endotelina-1, um peptídeo isolado recentemente, tem sido observada em diferentes tecidos e está envolvida em várias funções reprodutivas. Propusemo-nos a determinar as concentrações da endotelina-1 por radioimunoensaio no plasma de eqüinos e bovinos e no fluido folicular de eqüinos. Foram testados 2 tipos de inibidores de degradação protéica no plasma e fluido folicular. A imunorreatividade média foi de 47,5%, a dose mínima detectável foi de 1 pg/tubo e o coeficiente de variação inter e intra-ensaio foi de 2,36% e 7,26%, respectivamente. O paralelismo entre a curva padrão e as curvas determinadas com diferentes volumes de plasma e fluido folicular valida o radioimunoensaio de endotelina-1 para estas espécies. Segundo nosso conhecimento, este é o primeiro relato de literatura que detecta por radioimunoensaio a endotelina-1 no plasma e fluido folicular de eqüinos e plasma de bovinos. Os resultados do presente trabalho sugerem a utilização do método de radioimunoensaio utilizando-se kits comerciais para a determinação das concentrações da endotelina-1 no plasma de eqüinos e bovinos e fluido folicular de eqüinos, pelo fato de este apresentar alta sensibilidade, precisão e reprodutibilidade, podendo assim ser utilizado no estudo de múltiplas funções da endotelina-1 nestas espécies animais e, em especial, no sistema endócrino-reprodutivo

Identification of bacteria in drinking and purified water during the monitoring of a typical water purification system

BACKGROUND: A typical purification system that provides purified water which meets ionic and organic chemical standards, must be protected from microbial proliferation to minimize cross-contamination for use in cleaning and preparations in pharmaceutical industries and in health environments. METHODOLOGY: Samples of water were taken directly from the public distribution water tank at twelve different stages of a typical purification system were analyzed for the identification of isolated bacteria. Two miniature kits were used: (i) identification system (api 20 NE, Bio-Mérieux) for non-enteric and non-fermenting gram-negative rods; and (ii) identification system (BBL crystal, Becton and Dickson) for enteric and non-fermenting gram-negative rods. The efficiency of the chemical sanitizers used in the stages of the system, over the isolated and identified bacteria in the sampling water, was evaluated by the minimum inhibitory concentration (MIC) method. RESULTS: The 78 isolated colonies were identified as the following bacteria genera: Pseudomonas, Flavobacterium and Acinetobacter. According to the miniature kits used in the identification, there was a prevalence of isolation of P. aeruginosa 32.05%, P. picketti (Ralstonia picketti) 23.08%, P. vesiculares 12.82%,P. diminuta 11.54%, F. aureum 6.42%, P. fluorescens 5.13%, A. lwoffi 2.56%, P. putida 2.56%, P. alcaligenes 1.28%, P. paucimobilis 1.28%, and F. multivorum 1.28%. CONCLUSIONS: We found that research was required for the identification of gram-negative non-fermenting bacteria, which were isolated from drinking water and water purification systems, since Pseudomonas genera represents opportunistic pathogens which disperse and adhere easily to surfaces, forming a biofilm which interferes with the cleaning and disinfection procedures in hospital and industrial environments

Validación de la versión portuguesa del Cuestionario de Eficacia Clínica y Práctica Basada en Evidencias

OBJETIVOS: descrever o processo de tradução e validação linguística e cultural para o contexto português do Questionário de Eficácia Clínica e Prática Baseada em Evidências (QECPBE). MÉTODO: desenvolveu-se um estudo metodológico e transversal. Foi efetuada tradução e retroversão, de acordo com os padrões usuais. Na determinação das características psicométricas do QECPBE utilizou-se a Análise de Componentes Principais com rotação ortogonal, segundo o método Varimax, seguida de análise fatorial confirmatória. A consistência interna foi determinada pelo valor alfa de Cronbach. A coleta de dados ocorreu entre dezembro de 2013 e fevereiro de 2014. RESULTADOS: participaram 358 enfermeiros que exercem a prática clínica num centro hospitalar do norte de Portugal. O QECPBE apresenta 20 itens e três subescalas: Práticas (α=0,74); Atitudes (α=0,75); Conhecimentos/Habilidades e Competências (α=0,95), apresentando consistência interna global de α=0,74. No modelo testado obteve-se variância explicada de 55,86%. O modelo demonstrou um bom ajuste: χ2(167)=520,009; p=0,0001; χ2df=3,114; CFI=0,908; GFI=0,865; PCFI=0,798; PGFI=0,678; RMSEA=0,077 (IC90%=0,07-0,08). CONCLUSÃO: através da análise fatorial confirmatória realizada demonstrou-se que o questionário é válido e adequado para utilização no contexto estudado.OBJETIVOS: describir el proceso de traducción y validación lingüística y cultural para el contexto portugués del Cuestionario de Eficacia Clínica y Práctica Basada en Evidencias (CECPBE). MÉTODO: se desarrolló un estudio metodológico y transversal. Fue efectuada traducción y retroversión de acuerdo con los estándares usuales. En la determinación de las características psicométricas del CECPBE se utilizó el Análisis de Componentes Principales con rotación ortogonal, según el método Varimax, seguido por análisis factorial confirmatorio. La consistencia interna fue determinada por el valor alfa de Cronbach. La recolección de datos ocurrió entre diciembre de 2013 y febrero de 2014. RESULTADOS: participaron 358 enfermeros que ejercían la práctica clínica en un centro hospitalario en el norte de Portugal. El CECPBE presenta 20 ítems y tres subescalas: Prácticas (α=0,74); Actitudes (α=0,75); Conocimientos/Habilidades y Competencias (α=0,95), presentando consistencia interna global de α=0,74. En el modelo probado se obtuvo variancia explicada de 55,86%. El modelo demostró un buen ajuste: χ2(167)=520,009; p=0,0001; χ2df=3,114; CFI=0,908; GFI=0,865; PCFI=0,798; PGFI = 0,678; RMSEA = 0,077 (IC90%=0,07-0,08). CONCLUSIÓN: a través del análisis factorial confirmatorio se demostró que el cuestionario es válido y adecuado para utilización en el contexto estudiado.OBJECTIVES: to describe the process of translation and linguistic and cultural validation of the Evidence Based Practice Questionnaire for the Portuguese context: Questionário de Eficácia Clínica e Prática Baseada em Evidências (QECPBE). METHOD: a methodological and cross-sectional study was developed. The translation and back translation was performed according to traditional standards. Principal Components Analysis with orthogonal rotation according to the Varimax method was used to verify the QECPBE's psychometric characteristics, followed by confirmatory factor analysis. Internal consistency was determined by Cronbach's alpha. Data were collected between December 2013 and February 2014. RESULTS: 358 nurses delivering care in a hospital facility in North of Portugal participated in the study. QECPBE contains 20 items and three subscales: Practice (α=0.74); Attitudes (α=0.75); Knowledge/Skills and Competencies (α=0.95), presenting an overall internal consistency of α=0.74. The tested model explained 55.86% of the variance and presented good fit: χ2(167)=520.009; p = 0.0001; χ2df=3.114; CFI=0.908; GFI=0.865; PCFI=0.798; PGFI=0.678; RMSEA=0.077 (CI90%=0.07-0.08). CONCLUSION: confirmatory factor analysis revealed the questionnaire is valid and appropriate to be used in the studied context

The efficacy of chemical agents in cleaning and disinfection programs

BACKGROUND: Due to the growing number of outbreaks of infection in hospital nurseries, it becomes essential to set up a sanitation program that indicates that the appropriate chemical agent was chosen for application in the most effective way. METHOD: For the purpose of evaluating the efficacy of a chemical agent, the minimum inhibitory concentration (MIC) was reached by the classic method of successive broth dilutions. The reference bacteria utilized were Bacillus subtilis var. globigii ATCC 9372, Bacillus stearothermophilus ATCC 7953, Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 25923. The strains of Enterobacter cloacae IAL 1976 (Adolfo Lutz Institute), Serratia marcescens IAL 1478 and Acinetobactev calcoaceticus IAL 124 (ATCC 19606), were isolated from material collected from babies involved in outbreaks of infection in hospital nurseries. RESULTS: The MIC intervals, which reduced bacteria populations over 08 log(10), were: 59 to 156 mg/L of quaternarium ammonium compounds (QACs); 63 to 10000 mg/L of chlorhexidine digluconate; 1375 to 3250 mg/L of glutaraldehyde; 39 to 246 mg/L of formaldehyde; 43750 to 87500 mg/L of isopropanol or ethanol; 1250 to 6250 mg/L of iodine in polyvinyl-pyrolidone complexes, 150 to 4491 mg/L of chlorine-releasing-agents (CRAs); 469 to 2500 mg/L of hydrogen peroxide; and, 2310 to 18500 mg/L of peracetic acid. CONCLUSIONS: Chlorhexidine showed non inhibitory activity over germinating spores. A. calcoaceticus, was observed to show resistance to the majority of the agents tested, followed by E. cloacae and S. marcescens

Post-intervention Status in Patients With Refractory Myasthenia Gravis Treated With Eculizumab During REGAIN and Its Open-Label Extension

OBJECTIVE: To evaluate whether eculizumab helps patients with anti-acetylcholine receptor-positive (AChR+) refractory generalized myasthenia gravis (gMG) achieve the Myasthenia Gravis Foundation of America (MGFA) post-intervention status of minimal manifestations (MM), we assessed patients' status throughout REGAIN (Safety and Efficacy of Eculizumab in AChR+ Refractory Generalized Myasthenia Gravis) and its open-label extension. METHODS: Patients who completed the REGAIN randomized controlled trial and continued into the open-label extension were included in this tertiary endpoint analysis. Patients were assessed for the MGFA post-intervention status of improved, unchanged, worse, MM, and pharmacologic remission at defined time points during REGAIN and through week 130 of the open-label study. RESULTS: A total of 117 patients completed REGAIN and continued into the open-label study (eculizumab/eculizumab: 56; placebo/eculizumab: 61). At week 26 of REGAIN, more eculizumab-treated patients than placebo-treated patients achieved a status of improved (60.7% vs 41.7%) or MM (25.0% vs 13.3%; common OR: 2.3; 95% CI: 1.1-4.5). After 130 weeks of eculizumab treatment, 88.0% of patients achieved improved status and 57.3% of patients achieved MM status. The safety profile of eculizumab was consistent with its known profile and no new safety signals were detected. CONCLUSION: Eculizumab led to rapid and sustained achievement of MM in patients with AChR+ refractory gMG. These findings support the use of eculizumab in this previously difficult-to-treat patient population. CLINICALTRIALSGOV IDENTIFIER: REGAIN, NCT01997229; REGAIN open-label extension, NCT02301624. CLASSIFICATION OF EVIDENCE: This study provides Class II evidence that, after 26 weeks of eculizumab treatment, 25.0% of adults with AChR+ refractory gMG achieved MM, compared with 13.3% who received placebo

Minimal Symptom Expression' in Patients With Acetylcholine Receptor Antibody-Positive Refractory Generalized Myasthenia Gravis Treated With Eculizumab

The efficacy and tolerability of eculizumab were assessed in REGAIN, a 26-week, phase 3, randomized, double-blind, placebo-controlled study in anti-acetylcholine receptor antibody-positive (AChR+) refractory generalized myasthenia gravis (gMG), and its open-label extension

Reprocessing and reuse of perfusion cannulas

Os procedimentos utilizados para reprocessamento e reuso de artigos médicos de uso único não estão normalizados nem avaliados quanto a segurança. Os objetivos deste estudo foram: (i) determinar o tempo de esterilização e aeração de cânulas de perfusão em ambiente industrial; (ii) avaliar os níveis de resíduos de óxido de etileno, concentração de endotoxinas e resistência a tração de cânulas de perfusão após reprocessamento e reuso em ambiente hospitalar. No ambiente industrial as cânulas foram submetidas á esterilização com 450mg/L de 10% de óxido de etileno e 90% de CO2 por um período de 2 horas a uma temperatura de 45 a 55° C, umidade entre 30 e 90% aeração com 26 trocas de ar por hora com temperatura de 35°C. No ambiente hospitalar a cânula aramada TF 36460 foi utilizada em procedimento cirúrgico com circulação extracorpórea; lavada com água potável, desinfecção com detergente enzimático, enxague com água potável, embalada em papel grau cirúrgico e reprocessada por óxido de etileno com concentração de 500mg/L de 12% de óxido de etileno e 88% de Freon com tempo de exposição de 240 minutos, temperatura de 53°C, umidade entre 40-60% e aeração em temperatura ambiente. No ambiente industrial o tempo necessário para redução de 12 ciclos logarítmicos (SAL 10-6), foi de 120 minutos, no ambiente hospitalar foi de 240 minutos. O tempo de aeração variou de 19 a 28 horas na área industrial; e no ambiente hospitalar foi de 28 horas. No segundo reuso a cânula apresentou residual de óxido de etileno de 83,09 ppm; acima do limite especificado (≤_25ppm). Os testes de resistência a tração e determinação da concentração de endotoxinas foram realizados até o quarto reuso não apresentando resultados fora dos limites especificados. O procedimento de reuso nas condições atuais realizado em ambiente hospitalar foi reprovado, pois não atendeu aos requisitos de residual de óxido de etileno descritos na portaria Brasileira Interministerial n° 482.The procedures used during reprocessing of disposables have not been normalized, nor assessed for safety. The objective of this study is to assess sterility, residual rate of ethylene oxide, pyrogenicity and resistance to traction results observed in perfusion cannulas after the sterilization process in industrial environments and after reprocessing and reuse in the hospital environment. In the industrial environment, the cannulas were submitted to sterilization with 450mg/L of EtO and 90% of CO2 during a period of 2 hours and at a temperature of 45-55°C, humidity ranging from 30 to 90% and aeration of 26 air changes per hour at a temperature of 35°C. For the hospital environment, we selected the TF 36460 wired cannula, and after using it on a surgical procedure with extracorporeal circulation, it was washed with clean water, disinfected with enzymatic detergent, rinsed with clean water, wrapped in surgical paper and reprocessed in 500mg/L ethylene oxide of 12% EtO and 88% Freon and exposed for 240 minutes at a temperature of 53°C, humidity ranging from 40 to 60% and aeration at room temperature. In the industrial environment, the time needed for decreasing 12 logarithm cycles (10-6) was 120 minutes, while in the hospital environment it was 240 minutes. The aeration time ranged between 19 and 28 hours in the industrial area, while in the hospital environment it took 28 hours. In the second reuse the cannula presented residual of ethylene oxide of 83,09 ppm; above of the specified limit (≤25_ppm). The physical test and endotoxins concentration had been carried through until for reuse not presented results out of specified. The currently procedure of reuse in the hospital is disapproved therefore does not take care of the limits of ethylene oxide residues, recommended by Interministerial Brazilian regulation 482