Efficacy of a 0.5% Propolis -0.9% Pomegranate Buccal Spray Treatment Compared with 2% Miconazole Gel for Denture Stomatitis Treatment in Elderly Patients: a Randomized Clinical Trial.

Statement of the Problem: Natural products have attracted interest as an alternative to synthetic medi-cations for the treatment of oral diseases due to their efficacy and safety. Propolis and pomegranate extracts have both demonstrated efficacy for the treatment of denture stomatitis. However, use of the two compounds together has not been tested for this purpose. Purpose: A comparison was made of the efficacy of a commercially available propolis-pomegranate buccal spray formulation for the treatment of denture stomatitis, compared with miconazole gel, based on stomatitis lesions and Candida spp. concentrations in mouth rinses. Materials and Method: This was an experimental study, characterized as an open-label, parallel two-armed, non-inferiority randomized clinical trial. Forty elderly adults aged < 60 years with denture stoma-titis were randomly allocated to two groups. The patients applied a buccal spray containing 0.5% propo-lis and 0.9% pomegranate extracts or 2% miconazole gel, a standard treatment recommended in Brazil, to the inner surface of their dentures three times a day for 14 days. They were examined at days 1, 7, 14 and stomatitis lesions were categorized according to Newton's score. Mouth rinses were made with saline solution at days 1 and 14 and then assessed for Candida spp. Results: Both treatments reduced the Newton's score, with clinical cure rates of 75 and 40% for the miconazole and propolis-pomegranate groups, respectively. The Candida concentrations in the mouth rinse decreased significantly only in the miconazole group. Conclusion: The propolis-pomegranate spray was less effective than the miconazole treatment. Howev-er, clinical improvement was also observed in patients treated with the propolis-pomegranate buccal spray.

A Fatal Bacteremia Caused by Hypermucousviscous KPC-2 Producing Extensively Drug-Resistant K64-ST11 Klebsiella pneumoniae in Brazil.

We report a fatal bacteremia caused by Klebsiella pneumoniae in a 60-70-year-old patient from Brazil. The genomic analysis of three isolates (from blood culture, nasal and anal swabs) showed that the bacteremia was caused by a KPC-2 producing extensively drug-resistant K64-ST11 hypermucousviscous K. pneumoniae (hmKP) harboring several virulence and antimicrobial resistance genes. Although the isolates did not present virulence markers associated with hypervirulent K. pneumoniae (hvKP), they showed invasion and toxicity to epithelial Hep-2 cells; resistance to cell microbicidal mechanisms; and blood and human serum survival, evidencing their pathogenic potential. This study highlights the risk of infection caused by hmKp strains not characterized as hvKP as well as the clinical implications and difficulty of treatment, especially in elderly or immunocompromised patients.

Ciprofloxacin resistance in uropathogenic Escherichia coli isolates causing community-acquired urinary infections in Brasília, Brazil.

OBJECTIVES: Considering the global concern of ciprofloxacin resistance, the aim of this study was to evaluate the characteristics of ciprofloxacin-resistant (CIP-R) Escherichia coli isolated from patients with community-acquired urinary tract infections (UTIs) in Brasília, Brazil. METHODS: CIP-R E. coli isolated from different outpatients between July 2013 and April 2014 in a tertiary hospital were analysed for antibiotic resistance profile, phylotype, uropathogenic E. coli (UPEC) virulence genes, clonal relationship by enterobacterial repetitive intergenic consensus PCR (ERIC-PCR), and multilocus sequence typing (MLST). RESULTS: Among the 324 UPEC analysed, 263 (81.2%) were ciprofloxacin-sensitive and 61 (18.8%) were CIP-R. Antibiogram analysis of the 61 CIP-R strains showed that 45 (73.8%) were also multidrug-resistant. The most prevalent phylogroups were A and B2 (26/61 and 18/61, respectively). traT (53/61) and aer-traT (24/61) were the most common gene and genotype observed. Dendrogram analysis found that multidrug resistance and virulence genes were distributed among CIP-R strains independently of clonality and phylogroup. Six ERIC clusters (strains sharing ≥85% genetic similarity) were observed. MLST analysis of all strains of each cluster identified sequence types (STs) associated with worldwide antimicrobial resistance dissemination, including B2-ST131 and ST410, as well as STs not yet associated with antimicrobial resistance propagation, such as ST1725 and ST179. CONCLUSIONS: These results demonstrate that ciprofloxacin resistance dissemination by UPEC causing community-acquired UTIs was associated with multidrug resistance and was promoted by pandemic and non-pandemic STs, a concerning scenario for the local population.

Schistosome-Derived Molecules as Modulating Actors of the Immune System and Promising Candidates to Treat Autoimmune and Inflammatory Diseases.

It is long known that some parasite infections are able to modulate specific pathways of host's metabolism and immune responses. This modulation is not only important in order to understand the host-pathogen interactions and to develop treatments against the parasites themselves but also important in the development of treatments against autoimmune and inflammatory diseases. Throughout the life cycle of schistosomes the mammalian hosts are exposed to several biomolecules that are excreted/secreted from the parasite infective stage, named cercariae, from their tegument, present in adult and larval stages, and finally from their eggs. These molecules can induce the activation and modulation of innate and adaptive responses as well as enabling the evasion of the parasite from host defense mechanisms. Immunomodulatory effects of helminth infections and egg molecules are clear, as well as their ability to downregulate proinflammatory cytokines, upregulate anti-inflammatory cytokines, and drive a Th2 type of immune response. We believe that schistosomes can be used as a model to understand the potential applications of helminths and helminth-derived molecules against autoimmune and inflammatory diseases.

Multidrug resistance dissemination by extended-spectrum ß-lactamase-producing Escherichia coli causing community-acquired urinary tract infection in the Central-Western Region, Brazil.

The aim of this work was to analyse extended-spectrum ß-lactamase (ESBL)-producing Escherichia coli strains isolated from outpatients with signs of cystitis in Hospital Universitário de Brasília (Brasília, Brazil) during the period July 2013 to April 2014. E. coli isolated from urine culture were identified and their antibiotic susceptibility profile was determined by VITEK 2. ESBL-producing strains identified were submitted to PCR for Clermont phylotyping, CTX-M group typing and virulence determinant detection, and clonal relationships were determined by enterobacterial repetitive intergenic consensus (ERIC)-PCR. One strain belonging to each cluster of the dendrogram obtained by ERIC-PCR was selected for multilocus sequence typing (MLST). Among 324 uropathogenic E. coli (UPEC) analysed, 23 (7.1%) were identified as producing ESBL. All ESBL-producing strains were multidrug-resistant (MDR), i.e. presented non-susceptibility to at least one agent in three or more antimicrobial categories. Of the 23 ESBL-producing UPEC strains, 9 were assigned to phylogenetic group B2 and 7 each belonged to phylogenetic groups D and A. Virulence genotyping showed that aer was the most prevalent gene observed among the strains (21/23), followed by traT (18/23), pap (5/23), afa (5/23), PAI (5/23), cnf (3/23) and sfa (1/23). Analysis of the dendrogram showed that multidrug resistance and CTX-M ESBL groups were distributed among all strains, independent of clonality and phylogroup. Sequence types (STs) associated with pandemic resistance clones, such as B2-ST131 and D-ST648, were observed among the isolates. In conclusion, the results showed worrisome evidence of the potential for antibiotic multiresistant dissemination among community-acquired urinary tract infection caused by UPEC.

Mutant Brucella abortus membrane fusogenic protein induces protection against challenge infection in mice.

Brucella species can cause brucellosis, a zoonotic disease that causes serious livestock economic losses and represents a public health threat. The mechanism of virulence of Brucella spp. is not yet fully understood. Therefore, it is crucial to identify new molecules that serve as virulence factors to better understand this host-pathogen interplay. Here, we evaluated the role of the Brucella membrane fusogenic protein (Mfp) and outer membrane protein 19 (Omp19) in bacterial pathogenesis. In this study, we showed that B. abortus Δmfp::kan and Δomp19::kan deletion mutant strains have reduced persistence in vivo in C57BL/6 and interferon regulatory factor 1 (IRF-1) knockout (KO) mice. Additionally, 24 h after macrophage infection with a Δmfp::kan or Δomp19::kan strain expressing green fluorescent protein (GFP) approximately 80% or 65% of Brucella-containing vacuoles (BCVs) retained the late endosomal/lysosomal marker LAMP-1, respectively, whereas around 60% of BCVs containing wild-type S2308 were found in LAMP-1-negative compartments. B. abortus Δomp19::kan was attenuated in vivo but had a residual virulence in C57BL/6 and IRF-1 KO mice, whereas the Δmfp::kan strain had a lower virulence in these same mouse models. Furthermore, Δmfp::kan and Δomp19::kan strains were used as live vaccines. Challenge experiments revealed that in C57BL/6 and IRF-1 KO mice, the Δmfp::kan strain induced greater protection than the vaccine RB51 and protection similar that of vaccine S19. However, a Δomp19::kan strain induced protection similar to that of RB51. Thus, these results demonstrate that Brucella Mfp and Omp19 are critical for full bacterial virulence and that the Δmfp::kan mutant may serve as a potential vaccine candidate in future studies.

Acylation-dependent export of Trypanosoma cruzi phosphoinositide-specific phospholipase C to the outer surface of amastigotes.

Phosphoinositide phospholipase C (PI-PLC) plays an essential role in cell signaling. A unique Trypanosoma cruzi PI-PLC (TcPI-PLC) is lipid-modified in its N terminus and localizes to the plasma membrane of amastigotes. Here, we show that TcPI-PLC is located onto the extracellular phase of the plasma membrane of amastigotes and that its N-terminal 20 amino acids are necessary and sufficient to target the fused GFP to the outer surface of the parasite. Mutagenesis of the predicted acylated residues confirmed that myristoylation of a glycine residue in the 2nd position and acyl modification of a cysteine in the 4th but not in the 8th or 15th position of the coding sequence are required for correct plasma membrane localization in T. cruzi epimastigotes or amastigotes. Interestingly, mutagenesis of the cysteine at the 8th position increased its flagellar localization. When expressed as fusion constructs with GFP, the N-terminal 6 and 10 amino acids fused to GFP are predominantly located in the cytosol and concentrated in a compartment that co-localizes with a Golgi complex marker. The N-terminal 20 amino acids of TcPI-PLC associate with lipid rafts when dually acylated. Taken together, these results indicate that N-terminal acyl modifications serve as a molecular addressing system for sending TcPI-PLC to the outer surface of the cell.