Essential Domains of Anaplasma phagocytophilum Invasins Utilized to Infect Mammalian Host Cells

Anaplasma phagocytophilum causes granulocytic anaplasmosis, an emerging disease of humans and domestic animals. The obligate intracellular bacterium uses its invasins OmpA, Asp14, and AipA to infect myeloid and non-phagocytic cells. Identifying the domains of these proteins that mediate binding and entry, and determining the molecular basis of their interactions with host cell receptors would significantly advance understanding of A. phagocytophilum infection. Here, we identified the OmpA binding domain as residues 59 to 74. Polyclonal antibody generated against a peptide spanning OmpA residues 59 to 74 inhibited A. phagocytophilum infection of host cells and binding to its receptor, sialyl Lewis x (sLex-capped P-selectin glycoprotein ligand 1. Molecular docking analyses predicted that OmpA residues G61 and K64 interact with the two sLex sugars that are important for infection, α2,3-sialic acid and α1,3-fucose. Amino acid substitution analyses demonstrated that K64 was necessary, and G61 was contributory, for recombinant OmpA to bind to host cells and competitively inhibit A. phagocytophilum infection. Adherence of OmpA to RF/6A endothelial cells, which express little to no sLex but express the structurally similar glycan, 6-sulfo-sLex, required α2,3-sialic acid and α1,3-fucose and was antagonized by 6-sulfo-sLex antibody. Binding and uptake of OmpA-coated latex beads by myeloid cells was sensitive to sialidase, fucosidase, and sLex antibody. The Asp14 binding domain was also defined, as antibody specific for residues 113 to 124 inhibited infection. Because OmpA, Asp14, and AipA each contribute to the infection process, it was rationalized that the most effective blocking approach would target all three. An antibody cocktail targeting the OmpA, Asp14, and AipA binding domains neutralized A. phagocytophilumbinding and infection of host cells. This study dissects OmpA-receptor interactions and demonstrates the effectiveness of binding domain-specific antibodies for blocking A. phagocytophilum infection

A mathematical model for reactions during top-blowing in the AOD process:validation and results

Abstract In earlier work, a fundamental mathematical model was proposed for side-blowing operation in the argon oxygen decarburization (AOD) process. In the preceding part “Derivation of the Mode,” a new mathematical model was proposed for reactions during top-blowing in the AOD process. In this model it was assumed that reactions occur simultaneously at the surface of the cavity caused by the gas jet and at the surface of the metal droplets ejected from the metal bath. This paper presents validation and preliminary results with twelve industrial heats. In the studied heats, the last combined-blowing stage was altered so that oxygen was introduced from the top lance only. Four heats were conducted using an oxygen–nitrogen mixture (1:1), while eight heats were conducted with pure oxygen. Simultaneously, nitrogen or argon gas was blown via tuye’res in order to provide mixing that is comparable to regular practice. The measured carbon content varied from 0.4 to 0.5 wt pct before the studied stage to 0.1 to 0.2 wt pct after the studied stage. The results suggest that the model is capable of predicting changes in metal bath composition and temperature with a reasonably high degree of accuracy. The calculations indicate that the top slag may supply oxygen for decarburization during top-blowing. Furthermore, it is postulated that the metal droplets generated by the shear stress of top-blowing create a large mass exchange area, which plays an important role in enabling the high decarburization rates observed during top-blowing in the AOD process. The overall rate of decarburization attributable to top-blowing in the last combined-blowing stage was found to be limited by the mass transfer of dissolved carbon