Cost-Effectiveness of Constructing Minimal Shelter to Store INDOT Equipment (Weather Protection)

Currently vehicles used by INDOT are more likely to be subject to maintenance and repair than to replacement. The extent of wear and tear on the vehicles is likely to be impacted by the lack of covered storage in all districts. There are many different levels of covered storage—from tent-like structures to completely covered garages to temperature-controlled environments—each of which has different associated costs. But the associated reduction in equipment wear, speed of startup of equipment, and perhaps better mileage may all reflect savings to INDOT. This project evaluates INDOT’s expected return on investment to create covered areas for equipment (vehicles, other assets), while also assessing the cost difference between vehicles left in covered versus uncovered areas. This project can be used to benchmark certain lot sizes of equipment in order to decide whether or not implementing an indoor storage facility is efficient given the harsh conditions that the equipment may be facing during certain periods of the year. The information can be used to see the impact of weathering on snowplow trucks, the maintenance costs that can be reduced, and the payback period of building a shelter facility

Isolation of balamuthia mandrillaris-specific antibody fragments from a bacteriophage antibody display library

Balamuthia mandrillaris is a protist pathogen that can cause encephalitis with a mortality rate of more than 95%. Early diagnosis followed by aggressive treatment is a pre-requisite for successful prognosis. Current methods for identifying this organism rely on culture and microscopy, antibody-based methods using animals, or involve the use of molecular tools that are expensive. Here, we describe the isolation of antibody fragments that can be used for the unequivocal identification of B. mandrillaris. B. mandrillaris-specific antibody fragments were isolated from a bacteriophage antibody display library. Individual clones were studied by enzyme-linked immunosorbent assay, and immunofluorescence. Four antibody clones showed specific binding to B. mandrillaris. The usefulness of phage antibody display technology as a diagnostic tool for isolating antibody fragments against B. mandrillaris antigens and studying their biological role(s) is discussed further

Apoptosis in Acanthamoeba castellanii belonging to the T4 genotype

Here we describe features of apoptosis in unicellular Acanthamoeba castellanii belonging to the T4 genotype. When exposed to apoptosis-inducing compounds such as doxorubicin, A. castellanii trophozoites exhibited cell shrinkage and membrane blebbing as observed microscopically, DNA fragmentation using agarose gel electrophoresis, and phosphatidylserine (PS) externalization using annexin V immunostaining. Overall, these findings suggest the existence of apoptosis in A. castellanii possibly mediated by intrinsic apoptotic cascade. Further research in this field could provide avenues to selectively induce apoptosis in A. castellanii by triggering intrinsic apoptotic cascade

\u27Targeting the feast of a sleeping beast\u27: Nutrient and mineral dependencies of encysted acanthamoeba castellanii

Acanthamoeba spp. cause a corneal infection, Acanthamoeba keratitis (AK), and a cerebral infection, granulomatous amoebic encephalitis (GAE). Though aggressive chemotherapy has been able to kill the active trophozoite form of Acanthamoeba, the encysted form of this parasite has remained problematic to resist physiological concentrations of drugs. The emergence of encysted amoeba into active trophozoite form poses a challenge to eradicate this parasite. Acanthamoeba trophozoites have active metabolic machinery that furnishes energy in the form of ATPs by subjecting carbohydrates and lipids to undergo pathways including glycolysis and beta-oxidation of free fatty acids respectively. However, very little is known about the metabolic preferences and dependencies of an encysted trophozoite on minerals or potential nutrients that it consumes to live in an encysted state. Here, we investigate the metabolic and nutrient preferences of the encysted trophozoite of A. castellanii and the possibility to target them by drugs that act on calcium ion dependencies of the encysted amoeba. The experimental assays, immunostaining coupled with bioinformatics tools show that the encysted Acanthamoeba uses diverse nutrient pathways to obtain energy in the quiescent encysted state. These findings highlight potential pathways that can be targeted in eradicating amoebae cysts successfully