The spread of clonally related carbapenem-resistant Pseudomonas aeruginosa producing the metallo-␤-lactamase SPM-1 was found in Recife, Brazil. Upstream of bla SPM-1 , a novel common region (CR4) was identified, comprising an open reading frame, orf495, whose product shares significant identity with putative recombinases, such as Orf513. CR4 may be responsible for mobilization and expression of bla SPM-1 .During the last decade, several acquired carbapenem-hydrolyzing enzymes have been identified in Pseudomonas aeruginosa, beginning with IMP-1 and derivatives, which are widespread in Japan (24, 25) and China (4) and are emerging in Europe and Canada (8,13). Then, VIM-1 and VIM-2 enzymes were characterized, first in Italy and France (11,22), respectively, but these enzymes are also highly prevalent in Korea and Greece (7,14,29). Recently a third member of acquired Ambler class B ␤-lactamases, SPM-1, has been identified from a P. aeruginosa isolate in Sao Paulo, Brazil (28).The aim of this study was to analyze carbapenem-resistant P. aeruginosa isolates recovered from different hospitals in Recife, Brazil, from November 2002 to January 2003. Nineteen P. aeruginosa strains resistant to ceftazidime and imipenem were isolated at the Marcelo Magalhaes clinical laboratory in Recife. Ten isolates were from patients of neurological wards and intensive care units of the Hospital Portugues, Recife, whereas nine strains were from patients hospitalized in similar units from several hospitals in Recife. Eleven imipenem-resistant isolates were identified as metallo-␤-lactamase producers by using the double disk-synergy test with EDTA (30), and these were retained for further genetical analysis. They were resistant to all ␤-lactams except aztreonam. Isoelectric focusing analysis performed as described previously (20) showed that the 11 isolates expressed 3 ␤-lactamases, with pI values of 6.5, 7.5, and 8.7, the last likely corresponding to the naturally occurring AmpC enzyme of P. aeruginosa. Under standard PCR conditions (20), a series of primers was used for detection of genes encoding Ambler class B ␤-lactamases of the VIM, IMP, or SPM type as described previously (21). Positive results were obtained using the bla SPM-1 -specific primers (SPM-1A, 5Ј-CTGCTTGGATTCATGGGCGC-3Ј; SPM-1B, 5Ј-CCTTT TCCGCGACCTTGATC-3Ј). The 11 isolates had the same bla SPM-1 gene previously identified in P. aeruginosa (28).A genotypic comparison of the bla SPM-1 -positive P. aeruginosa isolates performed by pulsed-field gel electrophoresis, using restriction enzyme SpeI, revealed that these isolates were clonally related (data not shown) (18, 27). Electroporation of putative plasmid DNA extracts from bla SPM-1 -positive P. aeruginosa isolates performed as described previously (19) failed. Using pulsed-field gel electrophoresis and I-CeuI digestion followed by hybridization with PCR-generated probes as previously reported (9), a strong signal (located at ca. 400 kb) was obtained with the bla SPM-1 -specific probe for SPM-1-producing P. aeruginosa that ...

Objective: To evaluate the impact on clinical recovery and severity of the addition of large doses of vitamin A to the standard treatment for childhood pneumonia. Design: A randomised, double blind, placebo controlled trial. Setting: Study children were recruited at a public hospital in Recife, north east Brazil, an area of marginal vitamin A deficiency. Subjects: 472 children aged 6 to 59 months with clinical diagnosis of pneumonia. Interventions: 200 000 IU (infants) or 400 000 IU (1-4 year olds) of vitamin A in oil or similar capsules of placebo divided into two daily oral doses, in addition to the standard treatment. Main outcome measures: Duration of the episode and incidence of adverse outcomes. Results: The groups were similar with respect to overall duration of pneumonia and incidence of adverse outcomes. Children who received vitamin A, however, were less likely to have fever by day 3 (P = 0.008) and were 29% less likely to fail to respond to the first line antibiotic (P = 0.054). Conclusion: There was little evidence for an effect of vitamin A treatment on the immediate outcome of the pneumonia episode.

Progressive macular hypomelanosis (PMH) is a common skin disorder that causes hypopigmentation in a variety of skin types. Although the underlying aetiology of this condition is unclear, there is circumstantial evidence that links the skin bacterium Propionibacterium acnes to the condition. We now describe the first detailed population genetic analysis of P. acnes isolates recovered from paired lesional and non-lesional skin of PMH patients. Our results demonstrate a strong statistical association between strains from the type III phylogenetic lineage and PMH lesions (P = 0.0019), but not those representing other phylogroups, including those associated with acne (type IA1). We also demonstrate, based on in silico 16S rDNA analysis, that PMH isolates previously recovered from patients in Europe are also consistent with the type III lineage. Using comparative genome analysis, we identified multiple genomic regions that are specific for, or absent from, type III strains compared to other phylogroups. In the former case, these include open reading frames with putative functions in metabolism, transport and transcriptional regulation, as well as predicted proteins of unknown function. Further study of these genomic elements, along with transcriptional and functional analyses, may help to explain why type III strains are associated with PMH.

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PCR tests were used to assign genomovar status to 39 non-cystic fibrosis (non-CF) and 11 CF Burkholderia cepacia complex isolates from patients in hospitals in Recife, Brazil. Non-CF isolates were assigned to genomovar IIIA (71.8%), genomovar I (15.4%), B. vietnamiensis (7.7%), and B. multivorans (5.1%). CF isolates were assigned to genomovar IIIA (18.2%), B. vietnamiensis (18.2%), and genomovar I (9.1%). Six CF isolates sharing recA PCR-restriction fragment length polymorphism (RFLP) and randomly amplified polymorphic DNA (RAPD) patterns could not be assigned to a genomovar. 16S rDNA sequence obtained from these isolates indicated a closest relationship to B. anthina, but the recA sequence was equally divergent from several genomovars. PCR screening indicated the presence of cblA in only two isolates, whereas the B. cepacia epidemic strain marker was found in 22 of 28 genomovar IIIA isolates. A type III secretion gene was detected in all but genomovar I isolates. RAPD and PCR-RFLP assays, targeting both recA and fliC, indicated a large amount of genetic variability among the isolates, with many novel patterns being observed. Nine genomovar IIIA isolates from different non-CF patients and clinical sources had identical genotypes, indicating the presence of a common clone.Burkholderia cepacia is an important opportunistic respiratory pathogen, particularly in patients with cystic fibrosis (CF) (7) and chronic granulomatous disease (23). In addition, B. cepacia causes catheter-associated urinary tract infections, wound infections, intravenous catheter-associated bacteremia, and endocarditis (25,26).What is now known as the B. cepacia complex was subdivided by DNA-DNA hybridization, whole-cell protein pattern similarity, and phenotypic markers into five genomic species or genomovars (28) B. multivorans, genomovars IIIA and IIIB, B. stabilis, B. vietnamiensis, and B. ambifaria (15). Recently, a test for B. anthina has also been published (27).There have been a number of studies aimed at determining the distribution of genomovars among isolates from CF and non-CF patients (1, 2, 13, 24). In this paper, we report the application of PCR tests to the identification of 39 non-CF and 11 CF B. cepacia isolates from patients in hospitals in Brazil. We further report the results of PCR-based fingerprinting to determine genetic variability among the isolates and data concerning the distribution of genetic markers associated with cable pili, type III secretion (TTS), the B. cepacia epidemic strain marker (BCESM), and a putative polysaccharide export gene cluster. MATERIALS AND METHODSBacterial strains. Thirty-nine non-CF isolates came from inpatients at neurological wards or intensive care units of Hospital Português, a large hospital facility in Recife, Brazil. Most patients were old debilitated individuals with respiratory problems and were undergoing ventilation. The CF isolates were recovered from 11 outpatients younger than 14 years, who were attending the CF unit of Instituto Materno Infantil de Pernambuco. All isolates...

Cholesterol mediates its proliferative and metastatic effects via the metabolite 27-hydroxycholesterol (27-HC), at least in breast and endometrial cancer. We determined the serum lipoprotein profile, intratumoral cholesterol and 27-HC levels in a cohort of patients with well-differentiated papillary thyroid carcinoma (PTC; low/intermediate and high risk), advanced thyroid cancers (poorly differentiated, PDTC and anaplastic thyroid carcinoma, ATC) and benign thyroid tumors, as well as the expression of genes involved in cholesterol metabolism. We investigated the gene expression profile, cellular proliferation, and migration in Nthy-ori 3.1 and CAL-62 cell lines loaded with human low-density lipoprotein (LDL). Patients with more aggressive tumors (high-risk PTC and PDTC/ATC) showed a decrease in blood LDL cholesterol and apolipoprotein B. These changes were associated with an increase in the expression of the thyroid’s LDL receptor, whereas 3-hydroxy-3-methylglutaryl-CoA reductase and 25-hydroxycholesterol 7-alpha-hydroxylase were downregulated, with an intratumoral increase of the 27-HC metabolite. Furthermore, LDL promoted proliferation in both the Nthy-ori 3.1 and CAL-62 thyroid cellular models, but only in ATC cells was its cellular migration increased significantly. We conclude that cholesterol and intratumoral accumulation of 27-HC promote the aggressive behavior process of PTC. Targeting cholesterol metabolism could be a new therapeutic strategy in thyroid tumors with poor prognosis.

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Out of 24 nosocomial strains of Pseudomonas aeruginosa from Recife, Brazil, 15 (62%) were metallo-β-lactamase producers. Such isolates were resistant to main antipseudomonas drugs, except polymixyn B and aztreonam. The enzyme responsible for the carbapenem-resistance belongs to SPM-1 class, and the gene involved, blaspm-1, is likely plasmid located.