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...

Some strains of the Burkholderia cepacia complex, including the ET12 lineage, have been implicated in epidemic spread amongst cystic fibrosis (CF) patients. Suppression-subtractive hybridisation was used to identify genomic regions within strain J2315 (ET12 lineage; genomovar IIIA) that were absent from a non-transmissible genomovar IIIB strain. Sequence data from 15 subtracted clones were used to interrogate the genome sequence of strain J2315 and identify genomic regions incorporating the subtracted sequences. Many of the genomic regions displayed abnormally low GC content and similarity to sequences implicated in gene transfer. The distribution of three subtracted regions amongst members of the B. cepacia complex varied. A large cluster of genes with strong sequence similarity to capsular production genes from Burkholderia mallei and other bacterial pathogens was identified. This genomic island was detected in some but not all representatives of genomovar IIIA, two out of four genomovar I strains, and one of two strains of Burkholderia multivorans, but was not detected in Burkholderia stabilis, Burkholderia vietnamiensis, genomovar VI or Burkholderia. ambifaria. The polysaccharide production gene cluster of strain J2315 carries an IS 407-like sequence within the gene similar to B. mallei wcbO that is lacking in other ET12 isolates. Genes from this cluster are expressed during exponential growth in broth.

Objective: Following evolution of COVID-19 pandemic, reports pointed on a high prevalence of thyroiditis related thyrotoxicosis. Interpretation of thyroid tests during illness, however, is hampered by changes occurring in the context of non-thyroidal illness syndrome (NTIS). In order to elucidate these findings, we studied thyroid function in carefully selected cohorts of COVID-19 positive and negative patients. Design: Cohort observational study. Methods: We measured TSH, FT4, T3 within 24hours of admission in 196 patients without thyroid disease and/or confounding medications. 102 patients were SARS-CoV-2 positive; 41 admitted in the ICU, 46 in the ward and 15 outpatients. Controls consisted of 94 SARS-CoV-2 negative patients; 39 in the ICU and 55 in the ward. We designated the thyroid hormone patterns as consistent with NTIS, thyrotoxicosis and hypothyroidism. Results: A NTIS pattern was encountered in 60% of ICU and 36% of ward patients, with similar frequencies between SARS-CoV-2 positive and negative patients (46.0% vs 46.8%, p=NS). A thyrotoxicosis pattern was observed in 14.6% SARS-CoV-2 ICU patients vs. 7.7% in ICU negative (p=NS) and, overall in 8.8% of SARS-CoV-2 positive vs. 7.4% of negative patients. In these patients thyroglobulin levels were similar to those with normal thyroid function or NTIS. The hypothyroidism pattern was rare. Conclusions: NTIS pattern is common and relates to the severity of disease rather than SARS-CoV-2 infection. A thyrotoxicosis pattern is less frequently observed with similar frequency between patients with and without COVID-19. It is suggested that thyroid hormone monitoring in COVID-19 should not differ from other critically ill patients.

Background: The cellular transport proteins ABCB1, ABCC1 and ABCG2 have been implicated in the efflux of some antiretroviral drugs, thus decreasing their intracellular concentrations. Decreased drug accumulation in lymphocytes may allow viral replication and the subsequent emergence of viral resistance leading to treatment failure. Expression of HIV co-receptors on the surface of lymphocytes may influence viral tropism and therefore viral pathogenicity and disease progression. Here, we describe the relationship between expression of transport proteins and chemokine receptors in lymphocytes isolated from HIV-infected individuals and also investigate their relationship with demographic, therapeutic and virological factors.Methods: Peripheral blood mononuclear cells (PBMC) isolated from HIV-positive individuals were co-stained for expression of CD4 and ABCB1, ABCC1, ABCG2, CXCR4 and CCR5. The influence of gender, ethnicity, treatment status, viral load and CD4 count was assessed on expression of each protein as well as correlations between expression of the proteins by univariate and multivariate analyses.Results: Expression of ABCB1 was independently associated with gender (n 5 98) and expression of ABCG2 and CXCR4. Gender also correlated with expression of ABCC1 and CXCR4 in univariate analysis with lower expression being detected in females compared with males. Conclusions:Here we confirm that the previously reported correlation between ABCB1 and CXCR4 observed in PBMC isolated from healthy volunteers is also found in HIV-positive individuals. The influence of gender on the expression of drug efflux proteins could be a determinant of intracellular drug concentrations in vivo.