Hydroxyapatite growth inhibition by osteopontin hexapeptide sequences.

The effects of three acidic hexapeptides on in vitro hydroxyapatite growth were characterized by pH-stat kinetic studies, adsorption isotherms, and molecular modeling. The three peptides, pSDEpSDE, SDESDE, and DDDDDD, are equal-length model compounds for the acidic sequences in osteopontin, a protein that inhibits mineral formation in both calcified and noncalcified tissues. Growth rates from 1.67 mM calcium and 1.00 mM phosphate solution were measured at pH 7.4 and 37 degrees C in 150 mM NaCl. pSDEpSDE was a strong growth inhibitor when preadsorbed onto hydroxyapatite (HA) seeds from > or = 0.67 mM solutions, concentrations where adsorption isotherms showed relatively complete surface coverage. The nonphosphorylated SDESDE control showed no growth inhibition. Although it adsorbed to almost the same extent as pSDEpSDE, it rapidly desorbed under the pH-stat growth conditions while pSDEpSDE did not. DDDDDD exhibited weak inhibition as its concentration was increased and similar adsorption/desorption behavior to pSDEpSDE. Molecular modeling yielded binding energy trends based on simple adsorption of peptides on the [100] surface that were consistent with observed inhibition, but not for the [001] surface. The relatively unfavorable binding energies for peptides on the [001] surface suggest that their absorption will be primarily on the [100] face. The kinetic and adsorption data are consistent with phosphorylation of osteopontin acting to control mineral formation.

Confirmation of ethanol compressed gas standard concentrations by an NIST-traceable, absolute chemical method and comparison with wet breath alcohol simulators.

This study compares compressed-gas Ethanol Breath Standards (EBS) with wet simulators as calibration standards for breath alcohol analyzers. After reviewing the technical basis and traceability for the two calibration sources, new analytical data verifying the ethanol content of EBS gas are presented. Based on collection of ethanol in an impinger and titration using a modified California Department of Health method, the data confirm the alcohol content of EBS compressed gas standards by an absolute, wet chemical method. These results establish the concentration accuracy and the NIST traceability of the EBS gas by two independent methods, gas chromatographic analysis traceable to NIST standards and the new wet chemical method. Effluent gas from a wet simulator standard is then analyzed by the wet method to show its equivalence with EBS gas.