Browsing by Author "Cabrita, J"
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- Detection of Helicobacter pylori DNA in human feces by PCRPublication . Monteiro, L; Gras, N; Vidal, R; Cabrita, J; Megraud, FIn this study, the stability of Helicobacter pylori DNA in human feces and the effect of a diet lacking in plant material, the suspected source of PCR inhibitors in human feces, were investigated. in addition, a method to remove these inhibitors was developed. Stools inoculated with H, pylori were used as a model. For this purpose, a H. pylori suspension (10(8) CFU/ml) was used to spike stool samples obtained from four healthy adults known to be H. pylori negative. The evaluation of the stability of H, pylori DNA in frees showed that DNA was degraded after 3 days of contact with fecal material at 37 degreesC. A 2-day diet completely free of plant material was sufficient to eliminate PCR inhibitors from human feces. However, inhibitors were detected 48 h after a normal diet was resumed. A new technique consisting of agarose blocks containing embedded DNA as a template for PCR amplification was used for removal of inhibitors, following DNA extraction by a modified QIAamp tissue method (Qiagen, Hilden, Germany). When this method was applied to inhibiting stool samples known to have an inhibitory effect and spiked with H. pylori (5.10(8) CFU/g), a positive PCR was obtained showing that inhibitors present in the original DNA samples were completely removed. The agarose embedded DNA block method is highly efficient and provides clean, high quality template DNA for PCR purposes avoiding icing and fastidious conventional extraction methods, in conclusion, this study confirms that H, pylori DNA degrades with time in stools. A diet free of plant material or a special DNA preparation can be used to remove inhibitors and to allow the detection of H. pylori. (C) 2001 Elsevier Science B.V. All rights reserved.
- Estilo de vida e obesidade em adolescentes da região de LisboaPublication . Soveral, G; Lopes, F; Ferreira, C; Cabrita, J
- Study of drug utilization among students at Lisbon University in PortugalPublication . Cabrita, J; Ferreira, HS; Iglesias, P; Baptista, TM; Rocha, E; da Silva, AL; Miguel, JP
- Using a pharmacoepidemiological approach to estimate diabetes type 2 prevalence in PortugalPublication . Duarte-Ramos, F; Cabrita, JPurpose To estimate the prevalence of drug-treated diabetes type 2 in Portugal using drug consumption data and accounting for the proportion of patients treated with combinations of oral hypoglycaemic agents (OHAs). Methods The prevalence of diabetes treated by OHAs was calculated on the basis of: LMS-Health data for Portugal (2003) and two measures of daily drug intake-defined daily dose (DDD) and prescribed daily dose (PDD), after correcting for the proportion of patients treated with two or more OHAs. The PDD and proportion of patients in combination therapies have been obtained in a descriptive, cross-sectional national survey, conducted in 2003, by inquiry of 1046 type 2 diabetics countrywide distributed. Results Drug use study: We have studied 1046 type 2 diabetics (539 women, 501 men, 6 sex unkown), with a mean age of 64.5 (SD = 11) years. OHAs were prescribed as monotherapy in 46.8% (489/1046) of the patients and the remaining 557 (53.2%) received from 2 to 4 OHAs concomitantly. Diabetes prevalence: Using DDD as the mean daily intake consumption unit, the prevalence of drug-treated type 2 diabetes was 4.15%, which dropped to 2.52% when the proportion of OHAs associations was taken into account. Using PDD these values were 4.48% and 2.72%, respectively. Conclusions Since a high proportion of patients are treated with combination of OHAs in clinical practice, it is imperative to account for that confounder in order to improve the accuracy of estimate from drug consumption data. The use of this methodology provided a slight under-estimation of diabetes prevalence, compared with the National Health Authorities values (3%-5%). Nevertheless we consider this as an efficient tool to estimate drug-treated diabetes prevalence that should be implemented in a regular way for longitudinal observations, in order to generate signals. Copyright (c) 2005 John Wiley & Sons, Ltd.