Effect of wall growth on the kinetic modeling of nitrite oxidation in a CSTR

A simple kinetic model was developed for describing nitrite oxidation by autotrophic aerobic nitrifiers in a continuous stirred tank reactor (CSTR), in which mixed (suspended and attached) growth conditions prevail. The CSTR system was operated under conditions of constant nitrite feed concentration and varying volumetric flow rates. Experimental data from steady-state conditions in the CSTR system and from batch experiments were used for the determination of the model's kinetic parameters. Model predictions were verified against experimental data obtained under transient operating conditions, when volumetric flow rate and nitrite feed concentration disturbances were imposed on the CSTR. The presented kinetic modeling procedure is quite simple and general and therefore can also be applied to other mixed growth biological systems.     

Utilization of cross‐linked laccase aggregates in a perfusion basket reactor for the continuous elimination of endocrine‐disrupting chemicals

A perfusion basket reactor (BR) was developed for the continuous utilization of insolubilized laccase as cross‐linked enzyme aggregates (CLEAs). The BR consisted of an unbaffled basket made of a metallic filtration module filled with CLEAs and continuously agitated by a 3‐blade marine propeller. The agitation conditions influenced both the apparent laccase activity in the reactor and the stability of the biocatalyst. Optimal laccase activity was obtained at a rotational speed of 12.5 rps and the highest stability was reached at speeds of 1.7 rps or lower. The activity and stability of the biocatalyst were affected drastically upon the appearance of vortices in the reaction medium. This reactor was used for the continuous elimination of the endocrine disrupting chemicals (EDCs) nonylphenol (NP), bisphenol A (BPA), and triclosan (TCS). Optimization of EDC elimination by laccase CLEAs as a function of temperature and pH was achieved by response surface methodology using a central composite factorial design. The optimal conditions of pH and temperature were, respectively, 4.8 and 40.3°C for the elimination of p353NP (a branched isomer of NP), 4.7 and 48.0°C for BPA, and 4.9 and 41.2°C for TCS. Finally, the BR was used for the continuous elimination of these EDCs from a 5 mg L^?1 aqueous solution using 1 mg of CLEAs at pH 5 and room temperature. Our results showed that at least 85% of these EDCs could be eliminated with a hydraulic retention time of 325 min. The performances of the BR were quite stable over a 7‐day period of continuous treatment. Furthermore, this system could eliminate the same EDCs from a 100 mg L^?1 solution. Finally, a mathematical model combining the Michaelis–Menten kinetics of the laccase CLEAs and the continuous stirred tank reactor behavior of the BR was developed to predict the elimination of these xenobiotics. Biotechnol. Bioeng. 2009;102: 1582–1592. © 2008 Wiley Periodicals, Inc.     

High‐throughput quantification of selenium in individual serum proteins from a healthy human population using HPLC on‐line with isotope dilution inductively coupled plasma‐MS

In this study, a method, based on dual column affinity chromatography hyphenated to isotope dilution inductively coupled plasma–quadrupole MS, was developed for selenium determination in selenoprotein P, glutathione peroxidase, and selenoalbumin in human serum samples from a group of healthy volunteers (n=399). Method improvement was achieved using methanol‐enhanced isotope dilution which resulted in improved sensitivity and removal of isobaric interferences. Although no human serum reference materials are currently certified for their selenium species levels, method development was conducted using human serum reference material BCR 637 and 639 as their Se species content has been reported in the previous studies, and thus comparisons were possible. The mean selenium concentrations determined for the 399 healthy volunteer serum samples were 23±10 ng Se mL^?1 for glutathione peroxidase, 49±15 ng Se mL^?1 for selenoprotein P and 11±4 ng Se mL^?1 for selenoalbumin. These values are found to be in close agreement with published values for a limited number of healthy volunteer samples, and to establish baseline Se levels in serum proteins for an apparently healthy group of individuals, thus allowing for subsequent comparisons with respective values determined for groups of individuals with selenium related health issues, as well as assist in the discovery of potential selenium biomarkers. Also, the relationship between Se serum protein levels and some anthropometric characteristics of the volunteer population were investigated. Additionally, further development of the analytical method used in this study was achieved by adding a size exclusion chromatography column after the two affinity columns via a switching valve. This allowed for the separation of small selenium‐containing molecules from glutathione peroxidase and thus enhanced the overall confidence in its identification.     

The effects of a twenty-four-week aquatic training program on muscular strength performance in healthy elderly women

The purpose of the present study was to determine the effectiveness of a 24-week aquatic training (AT) program, which included both aerobic and resistance components, on muscle strength (isometric and dynamic), flexibility, and functional mobility in healthy women over 60 years of age. Twenty-two subjects were assigned randomly to either an AT (n = 12) or a control (C, n = 10) group. Volunteers participated in a supervised shallow-water exercise program for 60 minutes a day, 3 days a week; the exercise program consisted of a 10-minute warm-up and stretching, 25 minutes of endurance-type exercise (dancing) at 80% of heart rate (HR)(max), 20 minutes of upper- and lower-body resistance exercises with specialized water-resistance equipment, and a 5-minute cool down. Maximal isometric torque of knee extensors (KEXT) and knee flexors (KFLEX) were evaluated by a Cybex Norm dynamometer, grip strength (HGR) was evaluated using a Jamar hydraulic dynamometer, and dynamic strength was evaluated via the 3 repetition maximum (3RM) test for chest press, knee extension, lat pull down, and leg press. Jumping performance was evaluated using the squat jump (SJ), functional mobility with the timed up-and-go (TUG) test, and trunk flexion with the sit-and-reach test. Body composition was measured using the bioelectrical impedance method. The AT induced significant improvements in KEXT (10.5%) and KFLEX (13.4%) peak torque, HGR strength (13%), 3RM (25.7-29.4%), SJ (24.6%), sit-and-reach (11.6%), and TUG (19.8%) performance. The AT group demonstrated a significant increase in lean body mass (3.4%). No significant changes in these variables were observed in the C group. The results indicate that AT, with both aerobic and resistance components, is an alternative training method for improving neuromuscular and functional fitness performance in healthy elderly women.     

Carbon source-induced modifications in the mycolic acid content and cell wall permeability of Rhodococcus erythropolis E1

The influence of the carbon source on cell wall properties was analyzed in an efficient alkane-degrading strain of Rhodococcus erythropolis (strain E1), with particular focus on the mycolic acid content. A clear correlation was observed between the carbon source and the mycolic acid profiles as estimated by high-performance liquid chromatography and mass spectrometry. Two types of mycolic acid patterns were observed after growth either on saturated linear alkanes or on short-chain alkanoates. One type of pattern was characterized by the lack of odd-numbered carbon chains and resulted from growth on linear alkanes with even numbers of carbon atoms. The second type of pattern was characterized by mycolic acids with both even- and odd-numbered carbon chains and resulted from growth on compounds with odd-numbered carbon chains, on branched alkanes, or on mixtures of different compounds. Cellular short-chain fatty acids were twice as abundant during growth on a branched alkane (pristane) as during growth on acetate, while equal amounts of mycolic acids were found under both conditions. More hydrocarbon-like compounds and less polysaccharide were exposed at the cell wall surface during growth on alkanes. Whatever the substrate, the cells had the same affinity for aqueous-nonaqueous solvent interfaces. By contrast, bacteria displayed completely opposite susceptibilities to hydrophilic and hydrophobic antibiotics and were found to be strongly stained by hydrophobic dyes after growth on pristane but not after growth on acetate. Taken together, these data show that the cell wall composition of R. erythropolis E1 is influenced by the nutritional regimen and that the most marked effect is a radical change in cell wall permeability.     

Chromium is the proximate clastogenic species for lead chromate-induced clastogenicity in human bronchial cells

Hexavalent chromium (Cr(VI)) is a well-established human lung carcinogen with potentially widespread exposure. Solubility is a key factor in the carcinogenicity of Cr(VI), with the water-insoluble or 'particulate' compounds being the more potent carcinogens. Studies have indicated that the component ions are responsible for their clastogenicity, but it is uncertain whether chromium (Cr), lead (Pb) or some combination of the two is responsible for the clastogenic effects. Accordingly, we compared the clastogenicity of lead chromate (LC) with soluble sodium chromate (SC) and lead glutamate (LG) in WTHBF-6 human lung cells. We found that 1436microM was the maximal intracellular level of Pb after exposure to clastogenic concentrations of LC. However, clastogenesis was not observed after exposure to LG, even when intracellular Pb concentrations reached 13,347microM, indicating that intracellular Pb levels did not reach clastogenic levels in WTHBF-6 cells after LC treatment. By contrast, SC was clastogenic damaging 16 and 44% of metaphase cells at intracellular Cr doses of 312 and 1262microM respectively, which was comparable to the clastogenesis observed after LC treatment. LC damaged 10, 27 and 37% of metaphases at intracellular Cr doses of 288, 926 and 1644microM, respectively. These data indicate that with respect to LC-induced clastogenicity, Cr and not Pb is the proximate clastogenic species in human lung cells.     

A prion protein epitope selective for the pathologically misfolded conformation

Conformational conversion of proteins in disease is likely to be accompanied by molecular surface exposure of previously sequestered amino-acid side chains. We found that induction of beta-sheet structures in recombinant prion proteins is associated with increased solvent accessibility of tyrosine. Antibodies directed against the prion protein repeat motif, tyrosine-tyrosine-arginine, recognize the pathological isoform of the prion protein but not the normal cellular isoform, as assessed by immunoprecipitation, plate capture immunoassay and flow cytometry. Antibody binding to the pathological epitope is saturable and specific, and can be created in vitro by partial denaturation of normal brain prion protein. Conformation-selective exposure of Tyr-Tyr-Arg provides a probe for the distribution and structure of pathologically misfolded prion protein, and may lead to new diagnostics and therapeutics for prion diseases.     

Structure-based design of imidazo[1,2-a]pyrazine derivatives as selective inhibitors of Aurora-A kinase in cells

Co-crystallisation of the imidazo[1,2-a]pyrazine derivative 15 (3-chloro-N-(4-morpholinophenyl)-6-(pyridin-3-yl)imidazo[1,2-a]pyrazin-8-amine) with Aurora-A provided an insight into the interactions of this class of compound with Aurora kinases. This led to the design and synthesis of potent Aurora-A inhibitors demonstrating up to 70-fold selectivity in cell-based Aurora kinase pharmacodynamic biomarker assays.