Rapid yeast DNA extraction by boiling and freeze-thawing without using chemical reagents and DNA purification.

The purpose of this work was to study a rapid yeast DNA extraction by boiling and freeze-thawing processes without using chemical reagents or any purification procedures, to obtain a high grade PCR-product. A specific DNA fragment of the 18S region of Dekkera bruxellensis and Saccharomyces cerevisiae was chosen. The described boiling and freeze-thawing protocols generated the PCR-grade product preparations and could be used to process many samples. The amplification of the fragments could be observed after 30 and 35 cycles. These processes of extraction without using any kind of chemical reagents, especial water, and purification procedures proved to be efficient, reproducible, simple, fast, and inexpensive

Avaliação do sistema comercial CandiFast para identificação de leveduras isoladas do mosto de uvas brancas e tintas.

A região do Vale dos Vinhedos, localizada em Bento Gonçalves - RS, possui indicação geográfica para elaboração de vinhos no Sul do Brasil, o que torna de extrema importância a seleção e a identificação de linhagens autóctones para elaboração de vinhos de qualidade.Resumo

Oral and anal high-risk human papilloma virus infection in HIV-positive men who have sex with men over a 24-month longitudinal study: Complexity and vaccine implications

BackgroundFew studies focused on longitudinal modifications over time of high-risk HPV (HR-HPV) at anal and oral sites in HIV+ men who have sex with men (MSM).MethodsWe described patterns and longitudinal changes of HR-HPV detection and the prevalence of HR-HPV covered by the nonavalent HPV vaccine (vax-HPV) at oral and anal sites in 165 HIV+ MSM followed in an Italian hospital. The samples were collected at baseline and after 24months (follow-up). The presence of HPV was investigated with Inno-LiPA HPV Genotyping Extra II.ResultsMedian age was 44years (IQR 36-53), median CD4+ cell count at nadir was 312 cells/mm(3) (IQR 187-450). A total of 120 subjects (72.7%) were receiving successful antiretroviral therapy (ART). At baseline and follow-up, the frequency of HR-HPV was significantly higher in the anal site (65.4% vs 9.4 and 62.4% vs 6.8%, respectively). Only 2.9% of subjects were persistently HR-HPV negative at both sites. All oral HR-HPV were single at baseline vs 54.6% at baseline at the anal site (p=0.005), and all oral HR-HPV were single at follow-up vs 54.4% at anal site at follow-up (p=0.002). The lowest rate of concordance between the oral and anal results was found for HR-HPV detection; almost all HR-HPV positive results at both anal and oral sites had different HR-HPV.The most frequent HR-HPV in anal swabs at baseline and follow-up were HPV-16 and HPV-52.At follow-up at anal site, 37.5% of patients had different HR-HPV genotypes respect to baseline, 28.8% of subjects with 1 HR-HPV at baseline had an increased number of HR-HPV, and patients on ART showed a lower frequency of confirmed anal HR-HPV detection than untreated patients (p=0.03) over time. Additionally,54.6 and 50.5% of patients had only HR-vax-HPV at anal site at baseline and follow-up, respectively; 15.2% had only HR-vax-HPV at baseline and follow-up.ConclusionsWe believe that it is important testing multiple sites over time in HIV-positive MSM. ART seems to protect men from anal HR-HPV confirmed detection. Vaccination programmes could reduce the number of HR-HPV genotypes at anal site and the risk of the first HR-HPV acquisition at the oral site

Inflammatory cytokines and biofilm production sustain Staphylococcus aureus outgrowth and persistence: A pivotal interplay in the pathogenesis of Atopic Dermatitis

Individuals with Atopic dermatitis (AD) are highly susceptible to Staphylococcus aureus colonization. However, the mechanisms driving this process as well as the impact of S. aureus in AD pathogenesis are still incompletely understood. In this study, we analysed the role of biofilm in sustaining S. aureus chronic persistence and its impact on AD severity. Further we explored whether key inflammatory cytokines overexpressed in AD might provide a selective advantage to S. aureus. Results show that the strength of biofilm production by S. aureus correlated with the severity of the skin lesion, being significantly higher (P < 0.01) in patients with a more severe form of the disease as compared to those individuals with mild AD. Additionally, interleukin (IL)-β and interferon γ (IFN-γ), but not interleukin (IL)-6, induced a concentration-dependent increase of S. aureus growth. This effect was not observed with coagulase-negative staphylococci isolated from the skin of AD patients. These findings indicate that inflammatory cytokines such as IL1-β and IFN-γ, can selectively promote S. aureus outgrowth, thus subverting the composition of the healthy skin microbiome. Moreover, biofilm production by S. aureus plays a relevant role in further supporting chronic colonization and disease severity, while providing an increased tolerance to antimicrobials

Identification of Brucella by MALDI-TOF Mass Spectrometry. Fast and Reliable Identification from Agar Plates and Blood Cultures

BACKGROUND: MALDI-TOF mass spectrometry (MS) is a reliable method for bacteria identification. Some databases used for this purpose lack reference profiles for Brucella species, which is still an important pathogen in wide areas around the world. We report the creation of profiles for MALDI-TOF Biotyper 2.0 database (Bruker Daltonics, Germany) and their usefulness for identifying brucellae from culture plates and blood cultures. METHODOLOGY/PRINCIPAL FINDINGS: We created MALDI Biotyper 2.0 profiles for type strains belonging to B. melitensis biotypes 1, 2 and 3; B. abortus biotypes 1, 2, 5 and 9; B. suis, B. canis, B ceti and B. pinnipedialis. Then, 131 clinical isolates grown on plate cultures were used in triplicate to check identification. Identification at genus level was always correct, although in most cases the three replicates reported different identification at species level. Simulated blood cultures were performed with type strains belonging to the main human pathogenic species (B. melitensis, B. abortus, B. suis and B. canis), and studied by MALDI-TOF MS in triplicate. Identification at genus level was always correct. CONCLUSIONS/SIGNIFICANCE: MALDI-TOF MS is reliable for Brucella identification to the genus level from culture plates and directly from blood culture bottles

Design procedures of reinforced concrete framed buildings in Nepal and its impact on seismic safety

The present paper analyses the design procedure and its impact on seismic safety of the structures. For this, a representative reinforced concrete frame building (WDS) is designed and the results are compared with similar buildings detailed with: i) Current Construction Practices (CCP); ii) the Nepal Building Code (NBC) and iii) the Modified Nepal Building Code (NBC+) recommendations. The seismic performance evaluation is done with global strength, inter-storey drift and displacement of the structures. Likewise, the sensitivity of the structural and geometrical parameters of the RC frame building is studied through nonlinear analysis. The study parameters considered for parametric analysis are column size, beam size, inter-storey height, bay length, bay width, and compressive strength of concrete. The results show that the influence on the structural behaviour, particularly by variation in column size and inter-storey height. Additionally, the influence of the seismic zone factor on reinforcement demand of the structure is studied. The result shows that structures designed for high to medium seismic hazard demands double the reinforcement in beams compared to structures in low seismic zone

Accumulation of heavy metals by aquatic macrophytes in the Marano and Grado lagoon (Northern Adriatic sea, Italy)

Approximately 3% of the Italian territory is included in the National Priority List of Polluted sites. An example is represented by the \u201cLaguna di Grado e Marano\u201d which lies in Friuli Venezia Giulia (NE Italy). It includes about 4,200 ha of dry land and ca. 6,800 ha of wetlands and lagoons. The main pollutant, occurring particularly in wetlands and lagoon, is represented by mercury. However, heavy metals and organic pollutants were recorded as well. During Summer 2010, a survey was performed with the objective to observe, if any, the bioaccumulation of heavy metals in the tissues of Cymodocea nodosa, Zostera marina, Zostera nolti, aquatic macrophytes commonly found in the lagoon. Samples of sediments and specimens of macrophytes, were collected and analyzed for the heavy metal content (Cd, Co, Cd, Cu, Hg, Mn, Ni, Pb and Zn). The in concentrations, with the exception of Hg, were quite low. The Hg levels found in the sediments were comprised within the range 610-12,997 \u3bcg kg-1. In Cymodocea nodosa and Zostera marina, Hg was mostly accumulated in the leaves; the average Hg concentrations were, respectively, 44.5 and 61.8 \u3bcg kg-1. The opposite was found in Zostera noltii, being the leaf concentration of Hg (183 \u3bcg kg-1) more than doubled when compared to the root concentration (71 \u3bcg kg-1). For all the species, the bioconcentration factor was very low (<<1)