Bactericidal and anti-inflammatory effects of Moquilea tomentosa Benth. flavonoid-rich leaf extract.

BACKGROUND: Natural products are an important source of bioproducts with pharmacological properties. Here we investigate the components of leaves from M. tomentosa Benth. (Fritsch) (Chrysobalanaceae) and its effects on bacterial cell growth, biofilm production and macrophage activity. METHODS: The effect of the different leaf extracts against bacterial cell growth was performed using the microdilution method. The most active extract was analyzed by mass spectrometry, and its effect on bacterial biofilm production was evaluated on polystyrene plates. The extract effect on macrophage activity was tested in the RAW264.7 cell line, which was stimulated with different concentrations of the extract in the presence or absence of LPS. RESULTS: We show that the ethyl acetate (EtOAc) extract was the most effective against bacterial cell growth. EtOAc extract DI-ESI (-)MSn analysis showed the presence of a glycosylated flavonoid tentatively assigned as myricetin 3-O-xylosyl-rhamnoside (MW 596). Also, the EtOAc extract increased biofilm formation by S. aureus and inhibited cytokine and NO production induced by LPS in RAW macrophages. CONCLUSION: M. tomentosa flavonoid-enriched EtOAc extract presented a bactericidal and anti-inflammatory pharmacological potential.

Panton-Valentine leukocidin (PVL) isoforms among Staphylococcus aureus lineages isolated from hospitals in Rio de Janeiro, Brazil.

Panton-Valentine leukocidin (PVL) is a Staphylococcus aureus virulence factor codified by lukSF-PV genes. Single-nucleotide polymorphisms (SNPs) at lukSF-PV genes can lead to two PVL sequence variants (R and H) generating different PVL isoforms. This study analyzed lukSF-PV genes SNPs among four different clonal lineages (STs/CC 1, 5, 8, and 30) of nine S. aureus isolated at Brazilian hospitals. The sequenced products showed SNPs at seven sites (positions 121, 470, 527, 663, 856, 1396, and 1729), leading to non-synonymous substitutions in all isolates investigated. Our findings showed new R and H isoforms variants in S. aureus isolated in Brazil and suggest a possible relationship between H2b isoform and the ST30/CC30 lineage.

Resistance to Antiseptics and Disinfectants of Planktonic and Biofilm-Associated Forms of Corynebacterium striatum.

Disinfection and antisepsis are of primary importance in controlling nosocomial infections and outbreaks by pathogens expressing multiple resistance to antimicrobial agents (multidrug-resistant [MDR]) used in therapy. Nowadays, infections related to health services (HAIs) due to MDR and multidrug-susceptible (MDS) Corynebacterium striatum should not be underestimated, including patients using invasive medical devices. The virulence potential of C. striatum needs further investigation. Currently, susceptibility profiles of planktonic and/or sessile forms of four C. striatum strains of different pulsed-field gel electrophoresis types were examined as biocides based on the manufacturer's recommendations: 2% glutaraldehyde (GA), 2% peracetic acid (PA), 1% potassium monopersulfate (Virkon®; VK), 1% sodium hypochlorite (SH), and 70% ethyl alcohol (ET). Time-kill assays using 2% bovine serum albumin (BSA) were performed for evaluation of influence of organic matter on biocides effects. Planktonic forms expressed GA resistance at different levels. C. striatum viability was observed until 2, 4, 20, and 30 min for MDR 2369/II, MDS 1954/IV, MDR 1987/I, and MDS 1961/III strains, respectively. In contrast to GA, the biocides PA, VK24h, SH, and ET had higher effective bacterial mortality. However, storage of VK (48 hr) reduced their biocide activities. Moreover, mature biofilms were produced on abiotic substrates, including steel surfaces. Post-treatment with GA (30 min), survival of sessile forms was ≥100% than planktonic forms of all C. striatum tested strains. Independent of biocides tested, BSA increased the survival of planktonic and sessile forms (p ≤ 0.005). Present data indicated that hospital staff should be aware of dissemination and eradication of HAIs by C. striatum presenting resistance to biocides, including high-level disinfectants, such as GA.

Corynebacterium diphtheriae 67-72p hemagglutinin, characterized as the protein DIP0733, contributes to invasion and induction of apoptosis in HEp-2 cells.

Although Corynebacterium diphtheriae has been classically described as an exclusively extracellular pathogen, there is growing evidence that it may be internalized by epithelial cells. The aim of the present report was to investigate the nature and involvement of the surface-exposed non-fimbrial 67-72 kDa proteins (67-72p), previously characterized as adhesin/hemagglutinin, in C. diphtheriae internalization by HEp-2 cells. Transmission electron microscopy and bacterial internalization inhibition assays indicated the role of 67-72p as invasin for strains of varied sources. Cytoskeletal changes with accumulation of polymerized actin in HEp-2 cells beneath adherent 67-72p-adsorbed microspheres were observed by the Fluorescent actin staining test. Trypan blue staining method and Methylthiazole tetrazolium reduction assay showed a significant decrease in viability of HEp-2 cells treated with 67-72p. Morphological changes in HEp-2 cells observed after treatment with 67-72p included vacuolization, nuclear fragmentation and the formation of apoptotic bodies. Flow cytometry revealed an apoptotic volume decrease in HEp-2 cells treated with 67-72p. Moreover, a double-staining assay using Propidium Iodide/Annexin V gave information about the numbers of vital vs. early apoptotic cells and late apoptotic or secondary necrotic cells. The comparative analysis of MALDI-TOF MS experiments with the probes provided for 67-72p CDC-E8392 with an in silico proteome deduced from the complete genome sequence of C. diphtheriae identified with significant scores 67-72p as the protein DIP0733. In conclusion, DIP0733 (67-72p) may be directly implicated in bacterial invasion and apoptosis of epithelial cells in the early stages of diphtheria and C. diphtheriae invasive infection.

Diphtheria remains a threat to health in the developing world: an overview

Changes in the epidemiology of diphtheria are occurring worldwide. A large proportion of adults in many industrialized and developing countries are now susceptible to diphtheria. Vaccine-induced immunity wanes over time unless periodic booster is given or exposure to toxigenic Corynebacterium diphtheriae occurs. Immunity gap in adults coupled with large numbers of susceptible children creates the potential for new extensive epidemics. Epidemic emergencies may not be long in coming in countries experiencing rapid industrialization or undergoing sociopolitical instability where many of the factors thought to be important in producing epidemic such as mass population movements and difficult hygienic and economic conditions are present. The continuous circulation of toxigenic C. diphtheriae emphasizes the need to be aware of epidemiological features, clinical signs, and symptoms of diphtheria in vaccine era so that cases can be promptly diagnosed and treated, and further public health measures can be taken to contain this serious disease. This overview focused on worldwide data obtained from diphtheria with particular emphasis to main factors leading to recent epidemics, new clinical forms of C. diphtheriae infections, expression of virulence factors, other than toxin production, control strategies, and laboratory diagnosis procedures.

Influence of stannous chloride on the adhesive properties of Corynebacterium diphtheriae strains.

Stannous ion, as a chloride salt, influenced on the survival and adhesive properties of two toxigenic Corynebacterium diphtheriae of the sucrose-fermenting (241 strain) and non-sucrose-fermenting (CDC-E8392 strain) biotypes. Differences in survival fractions suggested differences in susceptibility of strains to bactericidal effect of stannous chloride (SnCl2). A number of 0.3% bacterial cells of 241 strain and 0.02% of CDC-E8392 strain survived after 220 micro l ml(-1) SnCl2 treatment. Results of polystyrene and spontaneous autoaggregation tests showed an increase in hydrophobicity of SnCl2 treated-bacteria. Spontaneous bacterial autoaggregation was induced in the presence of SnCl2. Stannous chloride also induced adherence to glass and totally inhibited the haemagglutinating activity of the non-sucrose-fermenting CDC-E8392 strain (original titer 32). Decrease in haemagglutination was dependent on SnCl2 concentration used. The presence of SnCl2 exerted differences in the expression of diphtheria bacilli surface carbohydrates possibly related with differences in degrees of haemagglutination and adherence to glass. Lectin-binding assays showed increase in the expression of cell surface receptors to the lectin Canavalia ensiformis (Con A) with affinity for mannose-like residues. The occurrence of cell filamentation suggests genotoxicity of SnCl2 to diphtheria bacilli. SnCl2 treatment was capable of modifying cell morphology, hydrophobins and adhesin expression, suggesting ability of C. diphtheriae to withstand oxidative stressing environment. Therefore, the SnCl2, widely used in nuclear medicine as reducing agent in the 99mTc-labelling process, may influence the outcome of bacterial infections.

Diphtheria remains a threat to health in the developing world--an overview.

Changes in the epidemiology of diphtheria are occurring worldwide. A large proportion of adults in many industrialized and developing countries are now susceptible to diphtheria. Vaccine-induced immunity wanes over time unless periodic booster is given or exposure to toxigenic Corynebacterium diphtheriae occurs. Immunity gap in adults coupled with large numbers of susceptible children creates the potential for new extensive epidemics. Epidemic emergencies may not be long in coming in countries experiencing rapid industrialization or undergoing sociopolitical instability where many of the factors thought to be important in producing epidemic such as mass population movements and difficult hygienic and economic conditions are present. The continuous circulation of toxigenic C. diphtheriae emphasizes the need to be aware of epidemiological features, clinical signs, and symptoms of diphtheria in vaccine era so that cases can be promptly diagnosed and treated, and further public health measures can be taken to contain this serious disease. This overview focused on worldwide data obtained from diphtheria with particular emphasis to main factors leading to recent epidemics, new clinical forms of C. diphtheriae infections, expression of virulence factors, other than toxin production, control strategies, and laboratory diagnosis procedures.

Diphetria in a vaccinated adult in Rio de Janeiro, Brazil

In 1999, a case of diphtheria in a 32-year-old woman was reported. The patient developed a sore throat immediately after participating of a five-day meeting with European workers in Rio de Janeiro. Her history included complete pediatric immunization (DTP) and three doses of adult formulation tetanus and diphtheria toxoid (dT) two years earlier. Clinical diagnosis of diphtheria was not made until microbiologic examination of specimens confirmed toxigenicity of Corynebacterium diphtheriae var. gravis, a biotype currently found circulating within Europe where diphtheria remains epidemic. This case reinforces the potential susceptibility of Brazilian adults to epidemic diphtheria in the vaccine era.