Molecular investigation of ready-to-eat salads for Giardia duodenalis and Cryptosporidium spp. in Portugal.

Foodborne outbreaks are often associated with the consumption of salads. However, published studies on the detection of foodborne pathogens in ready-to-eat salads are scarce. The aim of this study was to detect Giardia duodenalis and Cryptosporidium DNA in ready-to-eat salads, by applying techniques of molecular biology to study the frequency of contamination in salads. A total of 100 packages of ready-to-eat salads containing assorted leafy green vegetables were randomly purchased from hypermarkets located in central regions of Portugal (Coimbra and Viseu). Nested-PCR and qPCR methods were used to detect G. duodenalis and Cryptosporidium DNA. Species and assemblages of the parasites were identified by sequence analysis and PCR. Eighteen of the 100 samples (18%) were positive for G. duodenalis and twelve were sequenced and identified as assemblage A. Cryptosporidium spp. were not detected in any salads. Overall, pre-harvest and post-harvest preventive measures may be need for G. duodenalis control throughout the food production industry, from the field to consumers.

High penetrance of pheochromocytoma associated with the novel C634Y/Y791F double germline mutation in the RET protooncogene.

CONTEXT: Previous studies have shown that double RET mutations may be associated with unusual multiple endocrine neoplasia type 2 (MEN 2) phenotypes. OBJECTIVE: Our objective was to report the clinical features of patients harboring a previously unreported double mutation of the RET gene and to characterize this mutation in vitro. PATIENTS: Sixteen patients from four unrelated families and harboring the C634Y/Y791F double RET germline mutation were included in the study. RESULTS: Large pheochromocytomas measuring 6.0-14 cm and weighing up to 640 g were identified in the four index cases. Three of the four tumors were bilateral. High penetrance of pheochromocytoma was also seen in the C634Y/Y791F-mutation-positive relatives (seven of nine, 77.7%). Of these, two cases had bilateral tumors, one presented with multifocal tumors, two cases had large tumors (>5 cm), and one case, which was diagnosed with a large (5.5 x 4.5 x 4.0 cm) pheochromocytoma, reported early onset of symptoms of the disease (14 yr old). The overall penetrance of pheochromocytoma was 84.6% (11 of 13). Development of medullary thyroid carcinoma in our patients seemed similar to that observed in patients with codon 634 mutations. Haplotype analysis demonstrated that the mutation did not arise from a common ancestor. In vitro studies showed the double C634Y/Y791F RET receptor was significantly more phosphorylated than either activated wild-type receptor or single C634Y and Y791F RET mutants. CONCLUSIONS: Our data suggest that the natural history of the novel C634Y/Y791F double mutation carries a codon 634-like pattern of medullary thyroid carcinoma development, is associated with increased susceptibility to unusually large bilateral pheochromocytomas, and is likely more biologically active than each individual mutation.

Estimated BVDV-prevalence, -contact and -vaccine use in dairy herds in Northern Portugal.

A study to evaluate BVDV-prevalence, recent -contact and -vaccine use in dairy herds in the "Entre Douro e Minho" (EDM) region in North Portugal was carried out in 124 dairy herds in 2003. Herds were visited to ascertain BVDV-vaccine use and to collect a bulk tank milk (BTM) sample and serum from 1268 cattle to analyse BVDV-antibodies using an NS2-3 ELISA. Fifty-three percent of farmers used inactivated BVDV-vaccines whilst the remaining farmers were not presently using BVDV-vaccines. BMT-antibody results included 35% positives, 25% negative and 39% inconclusive, and were similar in vaccinated and non-vaccinated herds (p>0.05) and allowed estimating a 10% BVDV herd-prevalence from prior knowledge of the relationship between BMT-antibody results and probability of PI cattle in the herd. Overall individual seroprevalence was 27% and was 23% in non-vaccinated and 36% in vaccinated cattle (p<0.05). Contact of the herd with BVDV was assessed according to seroprevalence in young and adult cattle in the herd and it was estimated that 35% of herds were infected or had recent contact with BVDV, 40% were not infected and did not have recent contact with BVDV and the BVDV-infection and -contact status of remaining herds was undetermined. The results from this study indicate BVDV is endemic and BVDV-vaccines are widespread in the dairy-cattle population in EDM region in Portugal.

Hypernatremic syndrome due to hypodipsia and diabetes insipidus

We present two patients with tumoral lesions of the hypothalamic area, which developed hypernatremia, polyuria and polydipsia due to hypodipsia and ADH deficiency related to the simultaneous compromising of the thirst regulating center and of ADH production.