A sewer process model as planning and management tool--hydrogen sulfide simulation at catchment scale.

The collection system of a major city at the Persian Gulf was simulated for bulk water hydrogen sulfide and the release of sewer gas to the urban atmosphere. Geometry data on 870 km of sanitary sewer and data on dry weather flow entering all nodes in the catchment was exported from a Mike Urban database and imported to the sewer process model WATS. The process model then routed sewage and sewer gas through the system and simulated relevant physical, chemical and biological processes. In its non-calibrated state, the model was used as a planning tool to identify problem areas and to identify locations to install monitoring equipment and make preliminary choices for control strategies in terms of dosing of nitrate and iron salts. The monitoring equipment consisted of flow meters, level gauges, UV-Vis spectroscopes, and H2S gas sensors. Data from the first set of installed monitoring equipment were applied to calibrate and validate the model. It was illustrated how the calibrated model can be applied to assess compliance with quantitative formulated service levels and to design control strategies in terms of dosing of iron and nitrate salts.

Establishment of a quantitative ELISA capable of determining peptide - MHC class I interaction.

Many different assays for measuring peptide-MHC interactions have been suggested over the years. Yet, there is no generally accepted standard method available. We have recently generated preoxidized recombinant MHC class I molecules (MHC-I) which can be purified to homogeneity under denaturing conditions (i.e., in the absence of any contaminating peptides). Such denatured MHC-I molecules are functional equivalents of "empty molecules". When diluted into aqueous buffer containing beta-2 microglobulin (beta2m) and the appropriate peptide, they fold rapidly and efficiently in an entirely peptide dependent manner. Here, we exploit the availability of these molecules to generate a quantitative ELISA-based assay capable of measuring the affinity of the interaction between peptide and MHC-I. This assay is simple and sensitive, and one can easily envisage that the necessary reagents, standards and protocols could be made generally available to the scientific community.

Efficient assembly of recombinant major histocompatibility complex class I molecules with preformed disulfide bonds.

The expression of major histocompatibility class I (MHC-I) crucially depends upon the binding of appropriate peptides. MHC-I from natural sources are therefore always preoccupied with peptides complicating their purification and analysis. Here, we present an efficient solution to this problem. Recombinant MHC-I heavy chains were produced in Escherichia coli and subsequently purified under denaturing conditions. In contrast to common practice, the molecules were not reduced during the purification. The oxidized MHC-I heavy chain isoforms were highly active with respect to peptide binding. This suggests that de novo folding of denatured MHC-I molecules proceed efficiently if directed by preformed disulfide bond(s). Importantly, these molecules express serological epitopes and stain specific T cells; and they bind peptides specifically. Several denatured MHC-I heavy chains were analyzed and shown to be of a quality, which allowed quantitative analysis of peptide binding. The analysis of the specificity of the several hundred human MHC haplotypes, should benefit considerably from the availability of pre-oxidized recombinant MHC-I.