Amyloidogenesis via interfacial shear in a containerless biochemical reactor aboard the International Space Station.

Fluid interfaces significantly influence the dynamics of protein solutions, effects that can be isolated by performing experiments in microgravity, greatly reducing the amount of solid boundaries present, allowing air-liquid interfaces to become dominant. This investigation examined the effects of protein concentration on interfacial shear-induced fibrillization of insulin in microgravity within a containerless biochemical reactor, the ring-sheared drop (RSD), aboard the international space station (ISS). Human insulin was used as a model amyloidogenic protein for studying protein kinetics with applications to in situ pharmaceutical production, tissue engineering, and diseases such as Alzheimer's, Parkinson's, infectious prions, and type 2 diabetes. Experiments investigated three main stages of amyloidogenesis: nucleation studied by seeding native solutions with fibril aggregates, fibrillization quantified using intrinsic fibrillization rate after fitting measured solution intensity to a sigmoidal function, and gelation observed by detection of solidification fronts. Results demonstrated that in surface-dominated amyloidogenic protein solutions: seeding with fibrils induces fibrillization of native protein, intrinsic fibrillization rate is independent of concentration, and that there is a minimum fibril concentration for gelation with gelation rate and rapidity of onset increasing monotonically with increasing protein concentration. These findings matched well with results of previous studies within ground-based analogs.

Effects of Shear Rate and Protein Concentration on Amyloidogenesis via Interfacial Shear.

The influence of hydrodynamics on protein fibrillization kinetics is relevant to biophysics, biochemical reactors, medicine, and disease. This investigation focused on the effects of interfacial shear on the fibrillization kinetics of insulin. Human insulin served as a model protein for studying shear-induced fibrillization with relevance to amyloid diseases such as Alzheimer's, Parkinson's, prions, and type 2 diabetes. Insulin solutions at different protein concentrations were subjected to shear flows with prescribed interfacial angular velocities using a knife-edge (surface) viscometer (KEV) operating in a laminar axisymmetric flow regime where inertia is significant. Fibrillization kinetics were quantified using intrinsic fibrillization rate and times (onset, half, and end) determined through spectroscopic measurement of monomer extinction curves and fitting to a sigmoidal function. Additionally, the occurrence of gelation was determined through macroscopic imaging and transient fibril microstructure was captured using fluorescence microscopy. The results showed that increasing interfacial shear rate produced a monotonic increase in intrinsic fibrillization rate and a monotonic decrease in fibrillization time. Protein concentration did not significantly impact the intrinsic fibrillization rate or times; however, a minimum fibril concentration for gelation was found. Protein microstructure showed increasing aggregation and plaque/cluster formation with time.

Growth of microorganisms in an interfacially driven space bioreactor analog.

Fluid bioreactors in microgravity environments may utilize alternative methods of containment and mixing. The ring-sheared drop (RSD) is a containerless mixing device which functions in microgravity using surface tension for containment and mixes through interfacially-driven flow. To assess the feasibility of using interfacially driven flow devices, such as the RSD, as bioreactors, Escherichia coli growth and recombinant protein expression were analyzed in a ground-based analog of the RSD called the knife edge surface viscometer (KEV). Results demonstrated that the KEV can facilitate the growth of E. coli and that growth rate increases logarithmically with increasing knife edge rotation rate, similar to the standard growth method on Earth (orbital shaker). Furthermore, the KEV was shown to be viable for supporting recombinant protein expression in E. coli at levels comparable to those achieved using standard growth methods.

JEDI - an executive dashboard and decision support system for lean global military medical resource and logistics management.

Each individual U.S. Air Force, Army, and Navy Surgeon General has integrated oversight of global medical supplies and resources using the Joint Medical Asset Repository (JMAR). A Business Intelligence system called the JMAR Executive Dashboard Initiative (JEDI) was developed over a three-year period to add real-time interactive data-mining tools and executive dashboards. Medical resources can now be efficiently reallocated to military, veteran, family, or civilian purposes and inventories can be maintained at lean levels with peaks managed by interactive dashboards that reduce workload and errors.

Clinical engineering department strategic graphical dashboard to enhance maintenance planning and asset management.

The Clinical Engineering (a.k.a. Biomedical Engineering) Department has heretofore lagged in adoption of some of the leading-edge information system tools used in other industries. This present application is part of a DOD-funded SBIR grant to improve the overall management of medical technology, and describes the capabilities that Strategic Graphical Dashboards (SGDs) can afford. This SGD is built on top of an Oracle database, and uses custom-written graphic objects like gauges, fuel tanks, and Geographic Information System (GIS) maps to improve and accelerate decision making.