Calcium kinetics with microgram stable isotope doses and saliva sampling

Studies of calcium kinetics require administration of tracer doses of calcium and subsequent repeated sampling of biological fluids. This study was designed to develop techniques that would allow estimation of calcium kinetics by using small (micrograms) doses of isotopes instead of the more common large (mg) doses to minimize tracer perturbation of the system and reduce cost, and to explore the use of saliva sampling as an alternative to blood sampling. Subjects received an oral dose (133 micrograms) of 43Ca and an i.v. dose (7.7 micrograms) of 46Ca. Isotopic enrichment in blood, urine, saliva and feces was well above thermal ionization mass spectrometry measurement precision up to 170 h after dosing. Fractional calcium absorptions determined from isotopic ratios in blood, urine and saliva were similar. Compartmental modeling revealed that kinetic parameters determined from serum or saliva data were similar, decreasing the necessity for blood samples. It is concluded from these results that calcium kinetics can be assessed with micrograms doses of stable isotopes, thereby reducing tracer costs and with saliva samples, thereby reducing the amount of blood needed.     

A control strategy for reducing aeration costs during low loading periods.

The efficiency of the aeration system in a full-scale activated sludge basin with 3 separately controlled aeration zones was improved for the low loading period in summer. The air flow rate to each aeration zone is currently regulated to hold a preset dissolved oxygen concentration (DO). Four different DO setpoint combinations were tested, each one for a one week period, using dynamic off-gas testing to measure the standardised oxygen transfer efficiency (alphaSOTE). As the DO setpoints were lowered, the total air flow rate to the basin decreased initially. A low DO in the first zones slowed biomass activity and pushed the load towards the end of the aeration basin. The relationship between alphaSOTE and the specific diffuser flow rate qD is different for each zone. In Zone 1 there was a strong decrease in alphaSOTE as qD increased, while Zones 2 and 3 were fairly independent of qD, Zone 2 at a higher level than Zone 3. Aeration costs were reduced by 15% for the most efficient combination. To achieve even more savings, a control strategy adjusting oxygen transfer rates over the aeration basin to the necessary oxygen transfer rates is suggested. It is based on changing the DO setpoints to reach the lowest total air flow rate while meeting the effluent requirements.     

Ozonation of polycyclic aromatic hydrocarbons in oil/water-emulsions: mass transfer and reaction kinetics

The ozonation of highly condensed polycyclic aromatic hydrocarbons (PAH) was studied in oil/water-emulsions, which are comparable to poorly water-soluble PAH in industrial wastewaters and at contaminated sites. As there was a lack of knowledge about the ozonation in oil/water-emulsions, first the ozone mass transfer was studied and optimized from the gas to the water phase and from the water to the oil phase. The ratio of mass transfer and oxidation reaction was determined by the Hatta-number and revealed a slow, quasi homogeneous reaction of ozone with PAH inside the oil droplets. Because the ozone gas concentration had no influence under the optimized conditions, the selective PAH-ozonation could be described microkinetically by a direct ozone reaction of pseudo-first order regarding PAH-concentrations. The determined PAH mean reaction rate constants of 1.02 min(-1) in oil/water-emulsions are in the upper range as found for PAH dissolved in water. These results give a new insight into the ozonation in the three-phase systems and into the treatment of highly condensed, hardly biodegradable PAH.     

Crystal structure of the complex between VEGF and a receptor-blocking peptide

Vascular endothelial growth factor (VEGF) is a specific and potent angiogenic factor and, therefore, a prime therapeutic target for the development of antagonists for the treatment of cancer. As a first step toward this goal, phage display was used to generate peptides that bind to the receptor-binding domain (residues 8-109) of VEGF and compete with receptor [Fairbrother, W. J., Christinger, H. W., Cochran, A. G., Fuh, G., Keenan, C. J., Quan, C., Shriver, S. K., Tom, J. Y. K., Wells, J. A., and Cunningham, B. C. (1999) Biochemistry 38, 17754-17764]. The crystal structure of VEGF in complex with one of these peptides was solved and refined to a resolution of 1.9 A. The 20-mer peptide is unstructured in solution and adopts a largely extended conformation when bound to VEGF. Residues 3-8 form a beta-strand which pairs with strand beta6 of VEGF via six hydrogen bonds. The C-terminal four residues of the peptide point away from the growth factor, consistent with NMR data indicating that these residues are flexible in the complex in solution. In contrast, shortening the N-terminus of the peptide leads to decreased binding affinities. Truncation studies show that the peptide can be reduced to 14 residues with only moderate effect on binding affinity. However, because of the extended conformation and the scarcity of specific side-chain interactions with VEGF, the peptide is not a promising lead for small-molecule development. The interface between the peptide and VEGF contains a subset of the residues recognized by a neutralizing Fab fragment and overlaps partially with the binding site for the Flt-1 receptor. The location of the peptide-binding site and the hydrophilic character of the interactions with VEGF resemble more the binding mode of the Fab fragment than that of the receptor.     

Effect of silica and hydroxyapatite mineralization on the mechanical properties and the biocompatibility of nanocomposite collagen scaffolds

A recently established materials concept of biomimetic composites based on silica, collagen, and calcium phosphates was adapted for the preparation of porous scaffolds suitable for tissue engineering applications. Mineralization was achieved by directed nucleation of silica on the templating organic phase during a sol-gel process with or without addition of hydroxyapatite. Both mineral phases (25 wt %, individually or combined in equal shares) influenced the scaffold's morphology at the nanoscale. Enhancement of apparent density and compressive strength was similar for silica or hydroxyapatite mineralization; however the stiffening effect of hydroxyapatite was much higher. All scaffold modifications provided proper conditions for adhesion, proliferation, and osteogenic differentiation of human bone marrow stromal cells. The open porosity allowed cells to migrate throughout the scaffolds while maintaining their viability, both confirmed by MTT staining and confocal laser scanning microscopy. Initial cell distributions were graduated due to collagen mineralization, but balanced out over the cultivation time of 28 days. RT-PCR analyses revealed higher gene expression of ALP but lower expression of BSP II and osteocalcin because of collagen mineralization. The results demonstrate that both silica and hydroxyapatite offer comparable possibilities to tailor mechanical properties of collagen-based scaffolds without being detrimental to in vitro biocompatibility.