Molecular recognition is an important tool for developing tunable controlled release systems and fabricating biosensors with increased selectivity and sensitivity. The development of polymer-based materials that exploit molecular recognition events such as host-guest complexation, enzyme-substrate and enzyme-inhibitor interactions and nucleic acid hybridization was pursued in this thesis. Using polymers as an anchor for molecular recognition can enhance the affinity, selectivity, and the capacity for immobilization of recognition units, enabling the practical use of affinity-based systems in real applications. To introduce the potential for immobilization while preserving or enhancing the affinity of small molecule recognition units, the affinity of derivatized cyclodextrins for the hydrophobic drug, dexamethasone, was investigated. Cyclodextrins (CDs) are molecules that possess a hydrophilic exterior and a hydrophobic cavity capable of accommodating a wide range of small molecule guests. Analysis of the solubilization capacities, thermodynamic parameters and aggregative potentials of carboxymethyl and hydrazide derivatives of CDs established the dextran-conjugated βCD derivative as an ideal carrier of hydrophobic drugs and the hydrazide βCD derivative as an optimal solubilizer of lipophilic pharmaceuticals, both alone and when incorporated in a polymer-based drug delivery vehicle. To enable non-covalent immobilization and stabilization of biomacromolecular recognition units, a printed layer hydrogel was investigated as a selective diffusion barrier for analyte sensing and enzyme inhibitor recognition. A printable hydrogel platform was developed from an established injectable system composed of aldehyde- and hydrazide-functionalized poly(oligoethylene glycol methacrylate) polymers. The printed layer hydrogel effectively immobilized a wide range of enzymes and protected enzyme activity against time-dependent and protease-induced denaturation, while facilitating the diffusion of small molecules. Furthermore, to demonstrate the potential of the printed film hydrogel immobilization layer to enhance the selectivity of the target, the printable hydrogel platform was used to develop a microarray-based assay for the screening of inhibitors of the model enzyme, β-lactamase. The assay was able to accurately quantify dose-response relationships of a series of established inhibitors, while reducing the required reagent volumes in traditional drug screening campaigns by 95%. Most significantly, the assay demonstrated an ability to discriminate true inhibitors of β-lactamase from a class of non-specific inhibitors called promiscuous aggregating inhibitors. Finally, to enable non-covalent immobilization of DNA recognition units, the printable hydrogel-based microarray was tested for its ability to immobilize DNA recognition sites and promote the detection of DNA hybridization events. A long, concatameric DNA molecule was generated through rolling circle amplification and was used as a sensing material for the detection of a small, fluorophore labeled oligonucleotide. The printable hydrogel was able to effectively entrap the rolling circle amplification product. Properties of the printable hydrogel were investigated for their ability to support the detection of DNA hybridization events. / Thesis / Doctor of Philosophy (PhD) / This thesis describes the development of polymer-based materials that exploit molecular recognition events for drug delivery and biosensing applications. First, cyclodextrins (CDs) are molecules that are capable of binding a wide range of small molecules. A comprehensive analysis of the complexation properties of CD derivatives revealed critical insight regarding their application in polymer-based drug delivery vehicles. Second, a printable hydrogel platform was developed to support the immobilization and activity of biomolecules and establish a biosensing interface that facilitates the diffusion of small molecules but not molecular aggregates. A microarray-based assay was developed by employing the printed hydrogel interface for the screening of inhibitors of the model enzyme, β-lactamase, and the detection of DNA hybridization events.

The global food supply is being stressed by climate change, a growing population, and harmful diseases. One risk to vital cereal crops such as wheat and barley is Fusarium head blight (FHB), caused by the fungal plant pathogen Fusarium graminearum. Ascospores of the fungus are released from perithecia on the residues of corn and small grains and can be transported long distances (>500 m) through the atmosphere. The overall objective of this work was to assess the influence of meteorological conditions and perithecial maturity on ascospore release. The research focuses on F. graminearum because of its damaging impact to staple crops and the global ubiquity of FHB. The first specific objective was to apply state-of-the-science techniques to identify causal meteorological variables of ascospore release. We analyzed field measurements of airborne ascospores against meteorological conditions at Virginia Tech's Kentland Farm, Blacksburg, Virginia, USA and used convergent cross mapping and multivariate state space reconstruction to identify significant causal agents within this complicated natural and dynamic system. We identified relative humidity, solar radiation, wind speed, and air temperature as predictors of ascospore release. Our second research objective was to understand the impact of varying meteorological conditions on ascospore release under controlled environmental conditions. We assessed ascospore release in a chamber with controlled temperature (15°C and 25°C) and relative humidity (60%, 75%, and 95%). Ascospores released from ascospore-producing structures (perithecia) were captured on microscope slides placed inside of 3D-printed ascospore discharge devices. Results showed the sensitivity of ascospore release to relative humidity and temperature, with cool temperature and high relative humidity resulting in greater quantities of ascospores released. Our third research objective was to determine the relationship between the maturity, the number of ascospores, and the hardness of perithecia. A mechanical compression testing instrument was used to investigate the hardness of perithecia at various stages of maturity, producing a mean perithecium compression constant quantifying the uniaxial compression force required to rupture a perithecium. Results indicated that old perithecia contain the greatest amount of ascospores and exhibit increased resiliency, requiring greater forces to rupture, compared to young perithecia. This research has illustrated the complexities of F. graminearum ascospore release by describing the impact of several meteorological conditions and perithecial maturity on the timing and quantity of released ascospores. Collectively, our results may inform wheat growers on the nature and timing of ascospore release, which could help inform FHB management decisions in the future. / Ph. D.

An experimental study was conducted to determine the effect of complex geometries on the burning rate of materials made using additive manufacturing. Controlling heat release rate has applicability in limiting fire hazards as well as for designing fuels for optimal burning rate. The burning rate of a structure is a function of the material properties as well as the airflow through it, which is dictated by the geometry. This burning rate is generally proportional to the porosity for objects in which the flow is limited by the path constriction. The relations between porosity and burning rate are well studied for wood cribs, which are layers of wood sticks. Crib and other objects with various geometric features were constructed of ABS plastic and coal powder using additive manufacturing processes. A cone calorimeter using oxygen calorimetry was used to measure the heat release rate of the crib specimens. Within the flow limited burning regime, the burning rate of an object is proportional to the porosity factor. Porosity factors calculated from a 1-D theoretical burn rate model as well as from two empirical models were found to correlate the heat release rate results for the crib samples. The heat release rate results of the complex geometries generally correlated to the same porosity factor; however, the model was modified to account for differences between regularly shaped cribs and objects with different sized flow areas. Using the empirical models provides good correlation for the crib burning data and gives a clearer delineation between the flow-limited and surface area controlled regimes. / Master of Science

Nitrogen (N) fertilizer is a required mineral nutrient in containerized crop production that is necessary for crop growth and development. Due to production aspects, the N added to crops far exceeds the amount that the plant uses and such inefficiency results in adverse environmental impacts related to N gaseous and aqueous emissions from containers on the production site. Growers are responsible for optimizing nutrient usage in crop production. Three studies were conducted to investigate and better understand the fate of applied N fertilizers, the transformations associated with individual N sources, and the influence of substrate texture on losses of aqueous and gaseous N species. The first study conducted a mass balance looking at the four major avenues of N fate in an open-air container production setting (plant uptake, immobilized or bound N in a pine bark substrate, leached aqueous N, and gaseous emissions of N), the mass balance was speciated to measure applied and intermediary forms of N fertilizer species to provide insight into the overall fate of applied N. Show Off® Forsythia ×intermedia' Mindor' were grown using two control-release fertilizer (CRF) treatments [AN (ammonium-nitrate based) or UAN (urea ammonium-nitrate)] products. This study determined that 97% of the released N from the CRF treatments was lost via aqueous or gaseous pathways. The aqueous losses were inferred to be predominately composed of NO3-N, while the gaseous emissions were inferred to be predominately lost as inert nitrogen gas (N2). During a second experiment, individual N sources treatments [urea (CH4N2O), ammonium (NH4+), and nitrate (NO3-)] were applied to established containers of At LastⓇ Rosa x 'HORCOGJIL' grown in a pine bark substrate in either open wall high tunnel or a glass greenhouse to determine subsequent reaction sequence and fate based on applied N source. By applying an individual form of N it was determined that based on the N source applied, a sequential set of reactions occurs based on the N source. This study determined that the reactive N gaseous species occurred from the hydrolysis of CH4N2O-N to NH4+ and the nitrification of NH4+ to NO3- and then the denitrification of NO3- to N2. Hibiscus moscheutos' Vintage wine' was grown in either a coarse or fine texture substrate utilizing either a water-soluble fertilizer or a CRF to compare the influence of pine bark texture on N leachate losses and RN gaseous emissions. There were few differences between the two substrate texture treatments related to aqueous or gaseous N losses. In both experiments, the Hibiscus grown in the fine texture substrate resulted in higher above and below-ground biomass at experimental termination. Working with growers to develop best management practices will help to improve the use of N fertilizers and impact growers economically, while simultaneously reducing losses leading to less environmental impact on the areas surrounding production sites. / Doctor of Philosophy / Nitrogen (N) fertilizer is a crucial mineral nutrient input to produce container crops, however excessive application can have detrimental effects on the environment including gaseous N emissions and N leaching leading to water pollution. Therefore, three studies were conducted to investigate N losses during production and potential mitigation strategies using common management practices in the production of container crops. During the first study investigating how N fertilizer is lost from production, results showed that a significant portion of the N added to the containers is either emitted from the containers into the atmosphere or leached from the container. Only a small fraction of the applied N was utilized by the plants for growth and development. The second study investigated the reactions and transformations of different N fertilizers sources. When applying single N sources urea (CH4N2O), ammonium (NH4+), or nitrate (NO3-) result in a set of sequential reactions that occur based on the applied N source. Urea is hydrolyzed via CH4N2O hydrolysis leading to the formation of NH4+ which is nitrified via nitrification to NO3- which is denitrified via denitrification leading to the production of N2 gas. In the final study two pine bark substrate classes were compared when using either a water-soluble fertilizer (WSF) or a controlled-release fertilizer (CRF). Surprisingly there were only a few differences between the two substrate treatments in either the WSF or CRF studies. This body of work show the importance of investigating N fertilizer usage in container crop production. Collaboration between researchers and growers is crucial to develop management practices that maximize the associated economic input of N fertilizers and minimize losses of N that are detrimental to the environment.

Phosphorus (P) recovery and re-use is very important today for sustainable nutrient cycling and water quality protection due to the declining global P reserves and increasingly stringent wastewater treatment regulations. P recovery as struvite (MgNH4PO4·6H2O) is a promising technology because it can be used as a slow-release fertilizer. The objective of this study was to investigate different pretreatment methods to enhance struvite precipitation in dairy manure. Generally there are two challenges that need to be overcome to precipitate struvite in liquid dairy manure. The first is the relatively high calcium (Ca) concentration. The present study investigated the effectiveness of two calcium binding reagents to reduce the calcium inhibitory effects to enhance the struvite precipitation. A chemical equilibrium model (Visual MINTEQ 2.60) was used to determine the pH to acidify manure and to precipitate struvite. Then, bench-scale experiments were conducted to validate the model results using synthetic and untreated manure. First, the manure was acidified (pH 4.5) to liberate the particulate-bounded Mg2+, Ca2+ and inorganic P. Second, EDTA and/or oxalate compounds were added to the manure. Third, pH was increased to 7.5 for struvite precipitation from the liquid filtrate. Results showed that struvite-containing crystals were obtained from the samples treated with calcium removal compounds, with a total suspended solids (TSS) concentration less than 4,000 mg/L. The second challenge is the low dissolved reactive P (DRP, i.e. PO4-P) to total P (TP) ratio. This study investigated different pretreatment methods for P release to enhance struvite precipitation in liquid dairy manure: enhanced biological phosphorus removal (EBPR), microwave heating (MW) and anaerobic digestion (AD). All of the pretreatment methods resulted in P release but struvite crystals were observed only in precipitates obtained in manure pretreated with MW+H2SO4 acidification. Without oxalic acid to reduce the Ca2+ effects, all the other pretreatment methods were not effective in enhancing struvite precipitation in liquid dairy manure. Hardly any struvite or crystalline-like solids were found in the non-centrifuged samples, regardless of any pretreatment, indicating the effects of suspended solids and organic matter on struvite precipitation. A partial economic analysis was conducted to assess the chemical costs of P recovery from liquid dairy manure with different pretreatment methods; P recovery amounts as struvite were modeled by Visual Minteq 2.61.Three pretreatment conditions (untreated manure + oxalic acid, MW + H2SO4 acidification, and AD) were selected based on the precipitation results. MW + H2SO4 acidification produced the highest struvite quantities among the pretreatment methods to enhance struvite recovery, also with the lowest chemical addition costs. / Master of Science

Energy release rates are extracted from non-linear finite element analyses of a thin film bonded to a rigid substrate that is shaft-loaded at its free edge. This geometry is of interest because it simulates a probe test that has proven to be useful in characterizing the adhesion of thin, microelectronic coatings bonded to silicon wafers. Preliminary experimental results indicate that out-of-plane rather than in-plane loading dominates failure in the system. This work therefore focuses on out-of-plane film loading. To validate finite element and energy release rate methodologies, energy release rates from finite element analyses of pressurized and shaft-loaded blister tests are first correlated to theoretical limit cases. Upon validation, mode I, mode II, and mode III energy release rates are extracted from three-dimensional continuum finite element models of the edge-loaded thin film by a three-dimensional modified crack closure method. Having assumed a circular debond as observed experimentally, energy release rates are determined by a step-wise approach around the circumference. The progression of debond is simulated in multiple analyses by altering the boundary conditions associated with increasing the debond radius. Mechanical loading is supplemented with thermal loading, introducing residual stresses in the non-linear analyses. A sensitivity analysis of energy release rates to residual stress is performed. The results indicate that inclusion of residual stress has an important role in both the magnitude and mode-mixity of energy release rates in the thin film. Increasing the length of debond effectively transitions the film from a shearing mode to a bending mode, thereby significantly impacting each mode of energy release rate differently. / Master of Science

The mesopredator release hypothesis suggests that small carnivore populations are negatively affected by competition with larger carnivore species; this could hinder the conservation of mesopredators in areas where large top-carnivores are prioritized. I investigated the ecology of ocelots in Belize and examined the role of mesopredator release in the neotropical carnivore guild to determine if ocelots are limited by competition with jaguars and pumas. I conducted remote camera surveys and sampled habitat within four protected areas: three broadleaf sites and one pine forest site. I measured ocelot activity and habitat-use with respect to the activity of jaguars and pumas at one broadleaf site and one pine forest site; additionally, I calculated estimates of ocelot and jaguar densities within those two sites. Ocelot presence was positively related to jaguar activity in the pine forest and to both jaguar and puma activity in the broadleaf forest. There were few relationships with habitat characteristics; however, in the broadleaf site, ocelot activity was positively related to road width and large avian prey activity. Both jaguar and ocelot densities were low in the pine forest and higher in the broadleaf site. Preliminary findings from the remaining two sites suggest that future results may be similar to those from the first two sites analyzed. I conclude that ocelot populations are not negatively affected by jaguars and that a negative effect of pumas is unlikely. Results imply that mesopredator release does not limit ocelot populations in these areas. / Master of Science

The effects of two forms of mid-rotation release on thinned, fertilized loblolly pine (Pinus taeda L.) plantations in Virginia were examined: aerial imazapyr and basal triclopyr application. Imazapyr measurement plots were installed in nine Piedmont and twelve Coastal Plain plantations operationally released with imazapyr, and triclopyr measurement plots were installed within a controlled fertilization/release study spanning both regions. No differences in volume were detected following triclopyr release. All release dates combined, Piedmont released areas averaged 0.06 m3/tree (18%) greater than the control and Coastal Plain released areas averaged 0.05 m3/tree (14%) greater than the control. Reductions in hardwood basal area, stem density, and shrub stratum cover were observed for both forms of release. Reductions in shrub stratum richness and diversity were also documented for imazapyr release; however, trends indicate that richness and diversity, as well as stem density and shrub stratum cover, may recover to pre-treatment levels. Herbaceous vegetation was increased on triclopyr sites, which was reflected in an elevated turkey (Meleagris gallopavo sylvestris L.) food/brood index. Following imazapyr release, habitat suitability index (HSI) values for pine warblers (Dendroica pinus L.) and black-capped chickadees (Parus atricapillus L.) increased due to reductions in canopy hardwoods and increases in snags. Reduced shrub stratum density resulted in a lower bobwhite quail (Colinus virginianus L.) cover index on imazapyr-released areas. / Master of Science

No / The present work explores inner structuration of in situ gelling system consisting of glyceryl monooleate (GMO) and oleic acid (OA). The system under study involves investigation of microstructural changes which are believed to govern the pharmaceutical performance of final formulation. The changes which are often termed mesophasic transformation were analysed by small angle X-ray scattering (SAXS), differential scanning calorimetry (DSC), rheology and plane polarised light (PPL) microscopy. The current work revealed transformation of blank system from W/O emulsion to reverse hexagonal structure upon addition of structural analogues of ibuprofen. Such transformations are believed to occur due to increased hydrophobic volume within system as probed by SAXS analysis. The findings of SAXS studies were well supported by DSC, rheology and PPL microscopy. The study established inverse relationship between log P value of structural analogues of ibuprofen and the degree of binding of water molecules to surfactant chains. Such relationship had pronounced effect on sol-gel transformation process. The prepared in situ gelling system showed sustained drug release which followed Higuchi model.

Yes / A 16-electron iridium organometallic is reacted with carbon monoxide to form an 18-electron CO-adduct. This CO-adduct is stable for weeks in the solid state, but quickly reverts to its parent 16-e complex in tetrahydrofuran solution, releasing CO(g). Using a simple methodology, we show that this gas can subsequently be used to perform a carbonylation reaction on another molecule. / Royal Society; Academy of Medical Sciences/the Wellcome Trust/the Government Department of Business, Energy and Industrial Strategy/the British Heart Foundation Springboard Award