Wild Lactobacillus strains: Technological characterisation and design of Coalho cheese lactic culture

To design a specific lactic culture for the controlled manufacture of coalho cheese, 13 Lactobacillus rhamnosus, two Lactobacillus fermentum and one Lactobacillus plantarum strains isolated from artisanal coalho cheeses were identified and characterised. Two Lb. rhamnosus, one Lb. plantarum and one Lb. fermentum were selected and grouped in pairs designing four different culture formulations that demonstrated a good performance in cheesemaking experiments at pilot scale. Further studies to adjust the balance of strains used are necessary to attain adequate sensorial and technological attributes as expected for artisanal cheeses.     

Contents of barbaloin-related compounds in aloe drinks and their change during storage

The contents of barbaloin (BA), isoBA, aloin-dimers A, B, C, D and aloe-emodin (AE) in aloe drinks were investigated. BA and isoBA were detected in 30 of the 31 samples at the levels of 120-570 micrograms/mL and 120-580 micrograms/mL, respectively. Aloin-dimers A, B, C and D were detected in 8 of the 31 samples at the levels of 12-38 micrograms/mL, 13-39 micrograms/mL, 11-36 micrograms/mL and 16-69 micrograms/mL, respectively. AE was detected in all of the 31 samples at the levels of 0.03-1.3 micrograms/mL. When aloe drinks were stored for 4 weeks at 5 degrees C after opening the bottle, decrease of BA and isoBA, and increase of AE and aloin-dimers A, B, C and D were observed in most cases. However, in a few aloe drinks, all of BA, isoBA, aloin-dimers A, B, C, D and AE decreased. In these drinks, the existence of aloin-trimer was elucidated by LC/MS analyses. These data suggested that BA in aloe drinks is converted to the dimer and then to the trimer during storage.     

Determination of sucralose in foods by HPLC using pre-column derivatization

The development of a sensitive pre-column derivatization high-performance liquid chromatography (HPLC) method for determination of sucralose is reported. Sucralose is converted into a strongly ultraviolet (UV)-absorbing derivative, possessing strong absorption at 260 nm, by treatment with p-nitrobenzoyl chloride (PNBCl). Homogenized samples were dialyzed and washed with a Bond Elut ENV cartridge, then the eluate was evaporated to dryness and the residue was derivatized. Subsequently, the sucralose derivative was purified with hexane-ethyl actate (9:1) in a silica cartridge, and then the sucralose derivative was eluted with acetone. HPLC was performed on a phenyl column, using acetonitrile-water (73:27) as a mobile phase with UV detection (260 nm). The calibration curve was linear in the range of 1 microgram/mL to 50 micrograms/mL of sucralose. The recoveries of sucralose from eight kinds of foods spiked at the levels of 0.20 and 0.05 g/kg of sucralose were more than 76.2% with SD values in the range from 0.90% to 4.31%. The quantitative limit of the developed method was 0.005 g/kg for sucralose in samples.     

Antimicrobial activity of nutmeg against Escherichia coli O157

We examined the difference between Escherichia coli O157 and non-pathogenic E. coli in their tolerance to spices. Various spices (5 g each) were homogenized at 25 degrees C for 10 min with 5 ml of 70% ethyl alcohol, and the supernatant solutions obtained by centrifugation were used as spice extracts. When the E. coli strains were incubated with each spice extract at concentrations of 0.01% and 0.1%, a noteworthy difference was observed between the O157 and non-pathogenic strains in their tolerance to nutmeg. The populations of the non-pathogenic strains could not be reduced, but those of the O157 strains were remarkably reduced. Antibacterial activity by the nutmeg extract was also found against the enteropathogenic E. coli O111, but not against enterotoxigenic (O6 and O148) and enteroinvasive (O29 and O124) E. coli. When we examined the antibacterial effect of volatile oils in nutmeg on the O157 and non-pathogenic E. coli strains, all O157 strains tested were found to be more sensitive to beta-pinene than non-pathogenic E. coli strains.     

Different patterns of biofilm formation in Staphylococcus aureus under food-related stress conditions

Staphylococcus aureus and its biofilm formation are recognized as a serious clinical problem. S. aureus is also a food borne pathogen, and little is known regarding biofilm formation of food-related strains. We have studied biofilm formation of both food-related and clinical S. aureus strains grown under different stress conditions (temperature, sodium chloride, glucose and ethanol) relevant for food processing. Strong biofilm formers were identified among food-related S. aureus strains, and biofilm formation was affected by environmental conditions relevant for the food industry. The results showed that temperatures suboptimal for growth increased the production of biofilm. The combined presence of sodium chloride and glucose enhanced the biofilm formation. Both temperature and osmolarity affected the expression of several biofilm associated genes (e.g. icaA and rbf). Variations in gene expression (e.g. icaA, agrA and sigB) between strains were also observed. Our results support the existence of both ica-dependent and ica-independent mechanisms of biofilm production in S. aureus. The phenotypic and genotypic results showed highly diverse and complex patterns of biofilm formation in S. aureus. This clearly demonstrates that caution must be exercised before drawing general conclusions about gene expression in S. aureus in relation to regulation of biofilm formation. The results are relevant for food safety as they indicate that food processing conditions could promote biofilm formation by S. aureus.     

Performance characterization of nanofiltration membranes based on rigid star amphiphiles

The objective of this study was to develop new nanofiltration (NF) membranes capable of providing significantly greater water permeability and higher rejection of water contaminants compared to state-of-the-art NF membranes. The active layer of the new NF membranes is prepared with rigid star amphiphiles (RSAs) synthesized as part of this study. Performance characterization for a first generation of RSA membranes in a bench-scale apparatus reveals that most of the new membranes provide water permeability of 1.3-3.1 times that of two commercial NF membranes with polyamide active layers while providing comparable rejection of the organic contaminant surrogate Rhodamine WT. However, the rejection of arsenious acid (H3AsO3) by most new NF membranes was found to be lower than that by the two commercial NF membranes tested. Future research efforts of this study will focus on exploring if H3AsO3 rejection could be significantly increased, without negatively affecting water permeability and organic contaminant rejection, by addition of various chemical groups to RSA hydrophobic cores and hydrophilic branches, and by RSA cross-linking.     

Purification and molecular cloning of an intracellular 3-hydroxybutyrate-oligomer hydrolase from Paucimonas lemoignei

An intracellular 3-hydroxybutyrate-oligomer hydrolase was purified from a poly(3-hydroxybutyrate)-degrading bacterium, Paucimonas lemoignei. It hydrolyzed the 3-hydroxybutyrate dimer with the highest specific activity of any of the enzymes reported so far. The gene was cloned and sequenced. The deduced amino acid sequence showed that the enzyme is a homolog of the PhaZc of Ralstonia eutropha H16.     

Role of protein kinase Cbeta in rhythmic contractions of human pregnant myometrium

To assess the role of protein kinase Cbeta (PKCbeta) in human myometrial contractions during pregnancy, we evaluated the effect of a PKCbeta inhibitor (LY333531) on the pregnant and nonpregnant myometrial contractions and compared the level of PKCbeta in the pregnant myometrium with that in the nonpregnant myometrium. The effects of LY333531 on the myometrial contractions were examined by measuring contractile activity (frequency and amplitude). PKCbeta in human myometrium was assessed at mRNA level using real-time PCR method. The characteristics of contractile activity were different between the pregnant and the nonpregnant myometrium. The amplitude of rhythmic contractions in the preterm and term myometrium was increased 2- to 2.5-fold when compared with that in the nonpregnant myometrium, but the frequency of rhythmic contractions was decreased by about half. LY333531 (10(-6) M) reduced the increased amplitude in the preterm and term myometrium by about 50%, and the inhibitory effects of LY333531 in the pregnant myometrium were significantly greater than that in the nonpregnant myometrium (about 50 vs 25%). However, the frequency in the pregnant and nonpregnant myometrium was not influenced by LY333531. Real-time PCR revealed a significant, five- to sevenfold increase in the expression of PKCbeta mRNA in the preterm and term myometrium when compared with the nonpregnant myometrium. These findings suggest that the increased amplitude of human myometrial contractions during pregnancy is related to the increased level of PKCbeta. A PKCbeta inhibitor may reduce preterm uterine contractions and prevent preterm delivery.     

Bromamine decomposition kinetics in aqueous solutions

The objectives of this study are to investigate the kinetics of bromamine decomposition and to identify the corresponding relevant reactions. Experiments were performed with a stopped-flow spectrophotometer system. Experimental variables investigated included pH (6.5-9.5), bromamines concentration (0.15-0.50 mM), ammonia to bromine ratio (5-100), and phosphate and carbonate buffers concentration (5-40 mM). The experimental results were consistent with a reaction scheme that involved the reversible disproportionation of monobromamine into dibromamine and ammonia (2NH2Br (k1)<=>(k(-1)) NHBr2 + NH3), followed by irreversible decomposition of monobromamine and dibromamine into products (2NHBr2 (k2) --> products and NH2Br + NHBr2 (k3) --> products). The monobromamine disproportionation reaction was found to undergo general acid catalysis, and the two subsequent decomposition reactions were found to experience base catalysis. Experimental results were analyzed for the determination of catalysis terms corresponding to H+, NH4+, H2PO4-, HCO3-, and H2O for rate constants k1 and k(-1); HPO4(2-) and H2O for k2; and OH-, CO3(2-), and H2O for k3. These constants were fitted with the Br?nsted relationship, and the resulting fitting expressions were used to calculate any relevant catalysis rate constants that could not be determined at the range of experimental conditions used.     

Metabolic disturbance of tryptophan-nicotinamide conversion pathway by putative endocrine disruptors, bisphenol A and styrene monomer

Bisphenol A, a monomer of polycarbonate plastics, disturbed the conversion pathway of the amino acid tryptophan to the vitamin nicotinamide. The conversion ratio of tryptophan to nicotinamide was reduced to 1/15 by feeding a diet containing 1% bisphenol A. A putative disturbing reaction is kynurenine-->3-hydroxykynurenine, which is catalyzed by kynurenine monohydroxylase. This is an FAD-enzyme and requires NADPH as a coenzyme. Styrene monomer (1% addition to a normal diet) did not affect the food intake or the body weight, but slightly reduced the conversion ratio of tryptophan-nicotinamide.