Molecular and functional characterization of Lactobacillus sanfranciscensis strains isolated from sourdoughs

Fifty isolates of Lactobacillus sanfranciscensis from Italian sourdoughs were identified and typed by a polyphasic approach which included genotypic and phenotypic criteria. Genotypic diversity was characterized by Ribosomal Intergenic Spacer Analysis (RISA) of PCR amplified 16S-23S rDNA spacer region, denaturing gradient gel electrophoresis (DGGE) of PCR amplified rpoB (beta subunit of RNA polymerase) gene, and rep-PCR (PCR amplification of repetitive bacterial DNA elements) analyses. The RISA analysis produced a unique electrophoretical profile of four bands (ranging from 300 to 600 bp) for all L. sanfranciscensis isolates. The DGGE analysis of rpoB gene allowed the subdivision of isolates in four clusters. The resolution found by using rep-PCR with primers BOXA1R and REP1R-I/REP2-I allowed the widening of the level of isolates heterogeneity. Phenotypic diversity was evaluated by Biolog System and characterization of several technological traits (e.g., acidification kinetics, proteinase and peptidase activities). L. sanfranciscensis isolates used a large varieties of carbon sources such as dextrin, D-fructose, L-fucose, alpha-D-glucose, maltose, palatinose, L-rhanmose, L- and D,L-lactic acids and L-methionine. The acidification activity and related quotient of fermentation, and the peptidase (PepN, PepV, PepT, PepI, PepX, PepQ and PepR) activities markedly varied among strains. The same was found concerning the capacity to liberate amino acids during sourdough fermentation. This study could be considered as an example of a computerized analysis of the genotypic and phenotypic traits to reliably and rapidly differentiate sourdough isolates. Although some L. sanfranciscensis isolates combined several technological traits, the association of more selected strains seemed to be a requisite to get optimal sourdough characteristics.     

Reduced E-cadherin expression correlates with unfavorable prognosis in adenoid cystic carcinoma of salivary glands of the oral cavity

OBJECTIVE: To describe the epidemiology, diagnosis, risk factors, patient impact, and treatment strategies for recurrent Clostridium difficile-associated disease (CDAD). DESIGN: Data were collected as part of a blinded, placebo-controlled clinical trial testing a new combination treatment for recurrent CDAD. Retrospective data regarding prior CDAD episodes were collected from interviews and medical-chart review. Prospective data on the current CDAD episode, risk factors, and recurrence rates were collected during a 2-month follow-up. SETTINGS: National referral study. PARTICIPANTS: Patients with recurrent CDAD. INTERVENTIONS: Treatment with a 10-day course of low-dose (500 mg/d) or high-dose (2 g/d) vancomycin or metronidazole (1 g/d). RESULTS: Recurrent CDAD was found to have a lengthy course involving multiple episodes of diarrhea, abdominal cramping, nausea, and fever. CDAD may recur over several years despite frequent treatment with antibiotics. Recurrence rates were similar regardless of the choice or dose of antibiotic. Recurrent CDAD is not a trivial disease: patients may have multiple episodes (as many as 14), may require hospitalization, and the mean lifetime cost of direct medical care was $10,970 per patient. Fortunately, the disease does not become progressively more severe as the number of episodes increase. Two risk factors predictive for recurrent CDAD were found: increasing age and a decreased quality-of-life score at enrollment. CONCLUSIONS: Recurrent CDAD is a persistent disease that may result in prolonged hospital stays, additional medical costs, and rare serious complications.     

Antioxidant activity of virgin olive oil phenolic compounds in a micellar system†

The antioxidant activity of two virgin olive oils, obtained from the same olive (Olea europaea sativa) batch but processed with different hammer crushing conditions, was evaluated by measuring their protective action towards linoleic acid peroxidation in a micellar system. The antioxidant efficiency (AE) of the oil phenolic fraction was higher when the olives were processed with a higher hammer crusher rotation rate. HPLC analysis demonstrated that one of the main derivatives of oleuropein, indicated as 3,4-DHPEA-EDA (the dialdehydic form of elenolic acid linked with 3,4-dihydroxyphenylethanol), is present only in the oil with higher AE. 3,4-DHPEA-EDA showed the greatest antioxidant ability among virgin olive oil phenols. Its greater efficiency in the micellar system in comparison with 3,4-dihydroxyphenylethanol (3,4-DHPEA) is related to its greater lipophilicity. It is suggested that the behaviour in the water–micellar environment is representative of that in a real system such as tomato-based sauce with added virgin olive oil. © 1999 Society of Chemical Industry     

Familial study of patients with hereditary hemorrhagic telangiectasia

Rendu-Osler-Weber's disease or Hereditary Hemorrhagic Telangiectasia (HTT) is an autosomal dominant hereditary clinical entity characterized by the presence of telangiectasias on skin, mucous membranes and internal organs. The incidence of hepatic, pulmonary or cerebral complications justifies an early diagnostic. A familiar study of patients previously diagnosed of HTT in our hospital was made, establishing a protocol for the identification of asymptomatic or pauci-symptomatic cases. Fourteen patients were studied: 6 with a previous diagnostic, and 8 familiar contacts. Penetrance was of 85%. Epistaxis (80%) and telangiectasias on skin were the most frequent clinical findings. The visceral lesions found were gastrointestinal (28%), hepatic (15%), urological (15%) and pulmonary (7%). All patients were included in an hepatitis B virus vaccination program. It was also made a morphological study with Werhoeff's staining of the elastic layer, that allowed to distinguish both arterial and venous alterations.     

Hydrogen production by the thermophilic eubacterium Thermotoga neapolitana from storage polysaccharides of the CO2-fixing diatom Thalassiosira weissflogii

Planktic diatoms are the largest primary producers in marine and freshwater habitats. Their dry biomass accumulates up to 50% of lipids and contains water-soluble β-1,3-glucans as major storage products. Because of the world-wide abundance of these photosynthetic protists, β-1,3-glucans may rival cellulose as the polysaccharide with the highest annual production on Earth. Here we show the feasibility of a simple and efficient process leading to bio-hydrogen by dark fermentation of microalgal biomass with the thermophilic bacterium Thermotoga neapolitana. Production of the biogas on minimum medium supplemented only with the extract of the diatom Thalassiosira weissflogii proved that algal biomass per se can serve as substrate for sustaining the biotechnological process with no requirement of any pretreatment and external integration of other nutrients. At the same time, lipids unused for the anaerobic production of the biogas, can be employed for production of bio-diesel, thus considerably increasing the economic potential of these renewable feedstocks.     

Surface treatments with perfluoropolyether derivatives for the hydrophobization of gas diffusion layers for PEM fuel cells

In the present work, preliminary results of different hydrophobic surface treatments for gas diffusion layer (GDL) for PEM fuel cells are presented. This hydrophobic coating consists of new perfluoropolyether (PFPE) derivatives, in comparison to standard polytetrafluoroethylene (PTFE) dispersions. Experimental conditions for an efficient coating of fluoropolymers onto carbon clothes were explored by wet chemical methods.The GDLs obtained were tested in a single fuel cell at the lab scale. The cell testing was run at two temperatures (60 °C and 80 °C) with a relative humidity (RH) of the feeding gases of 80/100%, hydrogen/air respectively.The new PFPE coatings measurably improve the cell performances, and this effect is more evident at 60 °C with respect to 80 °C.     

Implementation and optimization of the HySyLab DMFC single cell test station

In this study, the construction and optimization of a low cost home-assembled DMFC single cell test station is described. The station was built using two flow control systems, for gas (air/oxygen) and liquid (methanol), a temperature controller, an electronic load, a milliohm-meter for resistance measurements and a computerized data acquisition system. The experiments were carried out on a 6.25 cm² single cell, built using two graphite plates with triple serpentine channel geometry for both anodic and cathodic flow fields. The cell was equipped with an MEA composed of Nafion 117 as electrolyte, Pt/C based catalyst as cathode and PtRu (1:1)/C as anode. In order to develop a HySyLab internal standard procedure, MEA activation was carefully studied. The activated MEA was used to determine the optimum values of cell operating parameters, such as oxidant flow, methanol flow and temperature.     

Computational fluid dynamic simulations of image-based stented coronary bifurcation models

One of the relevant phenomenon associated with in-stent restenosis in coronary arteries is an altered haemodynamics in the stented region. Computational fluid dynamics (CFD) offers the possibility to investigate the haemodynamics at a level of detail not always accessible within experimental techniques. CFD can quantify and correlate the local haemodynamics structures which might lead to in-stent restenosis. The aim of this work is to study the fluid dynamics of realistic stented coronary artery models which replicate the complete clinical procedure of stent implantation. Two cases of pathologic left anterior descending coronary arteries with their bifurcations are reconstructed from computed tomography angiography and conventional coronary angiography images. Results of wall shear stress and relative residence time show that the wall regions more prone to the risk of restenosis are located next to stent struts, to the bifurcations and to the stent overlapping zone for both investigated cases. Considering a bulk flow analysis, helical flow structures are generated by the curvature of the zone upstream from the stent and by the bifurcation regions. Helical recirculating microstructures are also visible downstream from the stent struts. This study demonstrates the feasibility to virtually investigate the haemodynamics of patient-specific coronary bifurcation geometries.     

Bacteriological evaluation of goat milk and cheese distributed in the Metropolitan Area of San José, Costa Rica

In the last years, there has been an increase in the production, industrialization and consumption of goat's milk and derivate products, including cheese, worldwide. Nevertheless, in Costa Rica there is no study of these products, reason why the objective of this work was to determine the microbiological characteristics of goat's milk and fresh cheese distributed in the Metropolitan Area of San José, Costa Rica, in order to evaluate its impact in the economical field and as a potential risk for Public Health. A total of 25 raw goat's milk samples, obtained by manual milking from 5 different producers tested in five different dates and 15 cheese samples, elaborated with pasteurized milk, commercially available and coming from three different producers were analyzed. The study included the analysis of spoilage bacteria (total aeobic count and lactic bacteria count), indicators of hygiene (total coliforms), fecal contamination (fecal coliforms), manipulation (Staphylococcus aureus) and pathogens (Listeria monocytogenes and Salmonella spp). High results were obtained for the total aerobic count and lactic bacteria count of the milk and cheese samples, showing a reduced shelf life. Total coliforms, in limits beyond the established ones by the Costa Rican legislation for human consumption raw milk, were found in 100% of milk samples, as well as for fecal coliforms in 76% of them. All cheese samples, except one, were negative for these indicators, suggesting good manufacturing practices. S. aureus counts were low and both Salmonella spp. and L. monocytogenes were not isolated from samples analyzed.     

The corrin moiety of coenzyme B12 is the determinant for switching the btuB riboswitch of E. coli

Riboswitches are regulatory elements in the 5'-untranslated region (5'-UTR) of bacterial mRNAs that bind certain metabolites with high specificity and affinity. The 202 nucleotide (nt)-long btuB riboswitch RNA of E. coli interacts specifically with coenzyme B12 and its derivatives thereby leading to changes in the RNA structure and hence to an altered expression of the downstream btuB gene. We report the investigations of the rearrangement of the three-dimensional structure of the btuB riboswitch upon binding to four different B12 derivatives: coenzyme B12, vitamin B12, adenosyl factor A and adenosyl-cobinamide. In-line probing experiments have shown that the corrin ring plays the crucial role in switching the three-dimensional riboswitch structure. Instead, the apical ligands influence only the binding affinity of the B12 derivative to the btuB riboswitch.