Bonds between fibronectin and fibronectin-binding proteins on Staphylococcus aureus and Lactococcus lactis.

Bacterial cell-wall-associated fibronectin binding proteins A and B (FnBPA and FnBPB) form bonds with host fibronectin. This binding reaction is often the initial step in prosthetic device infections. Atomic force microscopy was used to evaluate binding interactions between a fibronectin-coated probe and laboratory-derived Staphylococcus aureus that are (i) defective in both FnBPA and FnBPB (fnbA fnbB double mutant, DU5883), (ii) capable of expressing only FnBPA (fnbA fnbB double mutant complemented with pFNBA4), or (iii) capable of expressing only FnBPB (fnbA fnbB double mutant complemented with pFNBB4). These experiments were repeated using Lactococcus lactis constructs expressing fnbA and fnbB genes from S. aureus. A distinct force signature was observed for those bacteria that expressed FnBPA or FnBPB. Analysis of this force signature with the biomechanical wormlike chain model suggests that parallel bonds form between fibronectin and FnBPs on a bacterium. The strength and covalence of bonds were evaluated via nonlinear regression of force profiles. Binding events were more frequent (p < 0.01) for S. aureus expressing FnBPA or FnBPB than for the S. aureus double mutant. The binding force, frequency, and profile were similar between the FnBPA and FnBPB expressing strains of S. aureus. The absence of both FnBPs from the surface of S. aureus removed its ability to form a detectable bond with fibronectin. By contrast, ectopic expression of FnBPA or FnBPB on the surface of L. lactis conferred fibronectin binding characteristics similar to those of S. aureus. These measurements demonstrate that fibronectin-binding adhesins FnBPA and FnBPB are necessary and sufficient for the binding of S. aureus to prosthetic devices that are coated with host fibronectin.

Investigating the Tradespace between Increased Automation and Optimal Manning on Aircraft Carrier Decks

With increasing prevalence and capabilities of autonomous systems as part of complex heterogeneous manned-unmanned environments (HMUEs), an important consideration is the impact of the introduction of automation on the optimal assignment of human personnel. The US Navy has implemented optimal staffing techniques before in the 1990's and 2000's with a "minimal staffing" approach. The results were poor, leading to the degradation of Naval preparedness. Clearly, another approach to determining optimal staffing is necessary. To this end, the goal of this research is to develop human performance models for use in determining optimal manning of HMUEs. The human performance models are developed using an agent-based simulation of the aircraft carrier flight deck, a representative safety-critical HMUE. The Personnel Multi-Agent Safety and Control Simulation (PMASCS) simulates and analyzes the effects of introducing generalized maintenance crew skill sets and accelerated failure repair times on the overall performance and safety of the carrier flight deck. A behavioral model of four operator types (ordnance officers, chocks and chains, fueling officers, plane captains, and maintenance operators) is presented here along with an aircraft failure model. The main focus of this work is on the maintenance operators and aircraft failure modeling, since they have a direct impact on total launch time, a primary metric for carrier deck performance. With PMASCS I explore the effects of two variables on total launch time of 22 aircraft: 1) skill level of maintenance operators and 2) aircraft failure repair times while on the catapult (referred to as Phase 4 repair times). It is found that neither introducing a generic skill set to maintenance crews nor introducing a technology to accelerate Phase 4 aircraft repair times improves the average total launch time of 22 aircraft. An optimal manning level of 3 maintenance crews is found under all conditions, the point at which any additional maintenance crews does not reduce the total launch time. An additional discussion is included about how these results change if the operations are relieved of the bottleneck of installing the holdback bar at launch time.