RFLP analysis of a PCR-amplified fragment of the 16S rRNA gene as a tool to identify Enterococcus strains

Restriction fragment length polymorphism (RFLP) analysis of a PCR-amplified fragment of the 16S rRNA gene was performed on reference strains belonging to 21 different enterococcal species and on 75 Enterococcus isolates recovered from poultry meat, pasteurised milk and fresh cheese. PCR amplification generated a 275 bp fragment, which was digested with three restriction endonucleases (DdeI, HaeIII, HinfI). The strains were divided into five groups (groups A-E) on the basis of their restriction patterns. Five biochemical tests (arabinose, arginine, manitol, methyl-â-D-glucopyranoside and raffinose) were then performed in addition to RFLP analysis to narrow the identification of enterococcal strains to the species level. PCR-RFLP, in conjunction with the selected biochemical tests, allowed the precise identification of the 21 species of Enterococcus included in the present study. This proposed method is relatively simple and rapid and can be useful as an adjunct tool for accurate identification of Enterococcus.

Influence of pursed-lip breathing on heart rate variability and cardiorespiratory parameters in subjects with chronic obstructive pulmonary disease (COPD) / Influência da respiração freno-labial sobre a variabilidade da frequência cardíaca e parâmetros cardiorrespiratórios em pacientes com doença pulmonar obstrutiva crônica (DPOC)

OBJECTIVES: To assess the effects of pursed-lip breathing (PLB) at rest on the behavior of heart rate (HR) and its variability, and on variations in blood pressure (BP), respiratory rate (RR) and pulse oxygen saturation (SpO2) in subjects with chronic obstructive pulmonary disease (COPD). METHODS: Sixteen subjects with COPD (seven in GOLD stage I, three in GOLD stage II and six in GOLD stage III; mean age 64±11 years; mean FEV1 60±25 percent of predicted value) were assessed at rest, in a seated position, under the following conditions: ten minutes of normal breathing without PLB (R1), eight minutes with PLB (R2) and ten minutes of normal breathing once more (R3). HR was recorded, beat-to-beat, by means of a Polar S810 heart monitor. The RMSSD index (root mean square of the difference between successive R-R intervals) was determined. BP, RR and SpO2 were also assessed during the trials. ANOVA for repeated measures followed by the Tukey test and Kruskal-Wallis test were used for data analysis, with a 5 percent significance level. RESULTS: There was a significant increase in the RMSSD index during R2, in comparison with R1. The HR variation between inspiration and expiration was 8.98 bpm, and the variation between HR at rest and HR with PLB was 8.25 bpm. During R2, RR decreased and SpO2 increased significantly in comparison with R1 and R3. BP values did not show significant changes. CONCLUSIONS: The results showed that PLB produced significant changes in HR, RR and SpO2, and did not alter BP in subjects with COPD. Furthermore, analysis of the RMSSD index showed that PLB promoted increased parasympathetic activity in these subjects, thus indicating that this technique influenced the autonomic cardiac modulation.

OBJETIVOS: Avaliar os efeitos da respiração freno-labial (RFL), em repouso, sobre o comportamento da frequência cardíaca (FC) e sua variabilidade e variações na pressão arterial (PA), frequência respiratória (FR) e saturação parcial de oxigênio (SpO2) em pacientes com doença pulmonar obstrutiva crônica (DPOC). MÉTODOS: Dezesseis pacientes com DPOC (7: estágio GOLD I, 3: estágio GOLD II e 6: estágio GOLD III; média de idade=64±11; média de VEF1=60±25 por cento do predito) foram avaliados em repouso, na posição sentada, nas seguintes condições: 10 minutos respirando normalmente sem RFL (R1), 8 minutos com RFL (R2) e 10 minutos respirando novamente normalmente (R3). A FC foi registrada, batimento a batimento, por meio do frequencímetro Polar S810, e o índice RMSSD (raiz quadrada da média das diferenças entre intervalos RR sucessivos) foi determinado. PA, FR e SpO2 foram também avaliados durante o protocolo. ANOVA para medidas repetidas, seguida pelo teste de Tukey e teste de Kruskal-Wallis foram usados para análise dos dados, com nível de significância de 5 por cento. RESULTADOS: Ocorreu aumento significante no índice RMSSD durante R2 em comparação com R1. A variação na FC inspiração/expiração foi de 8,98 bpm, e a variação na FC em repouso/RFL foi de 8,25 bpm. Durante R2, FR diminuiu e SpO2 aumentou significativamente em comparação a R1 e R3. Os valores de PA não apresentaram modificações significativas. CONCLUSÕES: Os resultados mostraram que a RFL produziu modificações significativas na FC, FR e SpO2 e não alterou a PA em pacientes com DPOC. Além disso, a análise do índice RMSSD mostrou que a RFL promoveu aumento da atividade parassimpática nesses pacientes, indicando que essa técnica influencia a modulação autonômica cardíaca.

Evaluation of the Antimicrobial Activity of Sparfloxacin, Relative to Other Oral Antibiotics Against 1,125 Bacterial Isolates from 10 Medical Centers in Brazil.

A multicenter study was carried out in order to compare the in vitro activity of sparfloxacin to ciprofloxacin, amoxicillin/clavulanic acid, cephalexin, cefuroxine and azithromycin, against 1,125 microorganisms recently isolated from clinical specimens, most of them representative of respiratory tract infections. Sparfloxacin demonstrated potent action and was more active than the beta-lactam agents and azithromycin against most of the bacterial strains tested. Sparfloxacin was more potent (96% and 95% sensitivity, with MIC(90) of 0.19µg/mL and 0.5µg/mL, respectively)than the other antimicrobial agents tested against the Enterobacteriaceae family (Escherichia coli and Elebsiella pneumoniae). It was found to be equivalent in activity to ciprofloxacin (96% and 91% sensitivity and MIC(90) of 0.25 and 0.75µg/mL, respectively). Sparfloxacin was also found to be very active against the most fastidious microorganisms commonly associated to respiratory tract infections such as the penicillin-susceptible and resistant Streptococcus pneumoniae (MIC(90) 0.5µg/mL and 0.38µg/mL, respectively), ampicillin-susceptible and -resistant Haemophilus influenzae (MIC(90) 0.016µg/mL and 0.38µg/mL, respectively), beta-lactamase producing Moraxella catarrhalis (MIC(90) 0.032µg/mL) and non beta-lactamase producing M.catarrhalis (MIC(90) 0.5µg/mL).