The purpose of this thesis is to investigate the use of a single reactor to biologically treat wastewater by simultaneously oxidizing ammonia, and reducing nitrate and nitrite. The Environmental Protection Agency (EPA) places strict discharge restrictions on these compounds due to their inherent toxicity to humans, wildlife, and ecosystems. The use of a simultaneous system can assist the conventional wastewater treatment technology that requires separate systems, by creating a system that needs less time and smaller size to reach effluent requirements. To conduct this research, a bench-scale membrane aerated biofilm reactor was built using silicone tubing for aeration. Batch and continuous-flow experiments were conducted to investigate the reactor’s capability to oxidize ammonia using a defined growth media and monitor nitrate production and reduction. Also, wastewater from a local reclamation facility was used to determine the reactor’s ability to nitrify ammonia and denitrify nitrate concentrations within wastewater. The wastewater was taken from different locations within the reclamation facility, and combinations of primary and nitrified effluent were used to monitor ammonia and nitrate concentration changes. The batch experiments showed the greatest changes, and one batch experiment showed a 79% decrease in ammonium concentrations, and followed a first-order kinetics rate constant of -0.0284 hrs-1. The continuous-flow experiments showed much greater fluctuations in results, but one of the experiments showed an ammonia oxidation efficiency of 86%. The wastewater experiments had even greater fluctuations, and the effluent concentrations of ammonia, nitrate and nitrite showed no changes when compared to the influent.

Reduced graphene oxide (rGO) has attracted considerable interest recently as the low cost and chemical stable derivative of pristine graphene with application in many applications such as energy storage, water purification and electronic devices. This dissertation thoroughly investigated stacked rGO membrane fabrication process by vacuum-driven filtration, discovered asymmetry of the two surfaces of the rGO membrane, explored application perspectives of the asymmetric rGO membrane in fog collection and microstructure patterning, and disclosed membrane compaction issue during water filtration and species rejection. In more details, this dissertation revealed that, with suitable pore size, the filtration membrane substrate would leave its physical imprint on the bottom surface of the rGO membrane in the form of surface microstructures, which result in asymmetric dynamic water wettability properties of the two surfaces of the rGO membrane. The asymmetric wettability of the rGO membrane would lead to contrasting fog harvesting behavior of its two surfaces. The physical imprint mechanism was further extended to engineering pre-designed patterns selectively on the bottom surface of the rGO membrane. This dissertation, for the first time, reported the water flux and rejection kinetics, which was related to the compaction of the rGO membrane under pressure in the process of water filtration.

The aim of this work is the preparation of porous polymeric membranes for liquid separations stable in organic solvents, high temperature and/or extreme acid or basic conditions. Polymeric membranes with these properties could replace more traditional and energy-expensive separation processes like distillation, competing with ceramic membranes due to their easy processability and scalability. A limited library of polymers have been successfully used for decades in water-based applications. They are however unstable in organic solvents without an additional treatment, which is usually a crosslinking reaction. In this dissertation different highperformance polymeric membranes and crosslinking strategies are presented and discussed, allowing their use in harsh environments. We present for the first time the preparation of porous membranes using poly(oxindole) derivatives. These polymers were prepared by superacid catalyzed polyhydroxyalkylation, which is a novel one-pot, room-temperature, metal-free polymerization method. The obtained polymers were fully characterized and then manufactured into membranes by the non-solvent induced phase separation method. The crosslinking of these membranes was achieved by different protocols. First, we reacted the oxindole group in the polymer backbone with a variety of dibromides of different chemical structure. Secondly, we incorporated a propargyl side group, followed by a crosslinking in hot glycerol. Moreover, the strategy of crosslinking using propargyl as pendant group was successfully demonstrated in membranes made of poly(benzimidazole) and poly(triazolebisphenol-AF). And thirdly, we prepared membranes from hydroxyl-functionalized poly(oxindole), and conducted a controlled thermal oxidation, which resulted in the crosslinking by phenoxy radicals. In each case, the resulting membranes achieved insolubility in polar aprotic organic solvents, high resistance in acid medium and had high decomposition temperatures. In each case, the resulting membranes achieved not only insolubility in polar aprotic organic solvents and resistance to acid media but also showed high decomposition temperatures. Finally, we demonstrated for the first time the preparation of porous membranes based on recycled poly(ethylene terephthalate) plastic bottles and their potential application for separations in an organic solvent medium.

This study sought to determine whether NaCl altered the plasma membrane proteins in alfalfa exhibiting differential NaCl concentrations, and whether caso4 modified the responses. Two alfalfa cultivars, Centurion and Condor, were grown in 0.5 strength Hoagland solution in a greenhouse. The cultivars were exposed to 0, 88, and 132 mM of NaCl alone and mixed with caso4 .H20 at 7 and 14 Mm caso4 for 3, 9, and 60 days. In experiment 1, roots were dried to determine their Na, Ca, K, and Mg concentration. The results were similar to previous reports in which CaS04 alleviated the salt stress by increasing K and Mg levels and reducing Na. In experiment 2, after proteins of the plasma membrane were isolated and their purity was determined by vanadate, ATPase activity showed a significant increase in the presence of calcium. In addition, total plasma membrane protein was analyzed by sodium dodecylsulfate-polyacrylamide gel electrophoresis. Salt treatments induced both quantitative and qualitative changes in proteins. These changes were affected by the length of exposure to treatment solution or the ability of the plants to adapt to the salt stress.

Indiana University-Purdue University Indianapolis (IUPUI) / Insulin resistance, a hallmark of type 2 diabetes (T2D), is associated with accompanying derangements such as hyperinsulinemia that promote the progression of insulin resistance, yet a mechanism(s) is imperfectly understood. Data have demonstrated that hyperinsulinemia promotes insulin resistance as evidenced by diminished ability of insulin to mobilize glucose transporter GLUT4 to the plasma membrane (PM). We found that loss of PM phosphatidylinositol 4,5-bisphosphate (PIP2)-regulated filamentous actin (F-actin) structure contributes to hyperinsulinemia-induced insulin resistance. We tested if increased glucose flux through hexosamine biosynthesis pathway (HBP) causes dysregulation of PM components necessary for GLUT4 translocation. Increased HBP activity was detected in 3T3-L1 adipocytes cultured in hyperinsulinemia (5 nM Ins; 12 h) and also 2 mM glucosamine (GlcN), a distal HBP activator, inducing losses of PM PIP2 and F-actin. In accordance with HBP flux directly weakening PIP2/F-actin structure, inhibition of the rate-limiting HBP enzyme (glutamine:fructose-6-phosphate amidotransferase) restored F-actin and insulin responsiveness. Furthermore, less invasive challenges with glucose led to PIP2/F-actin dysregulation. New findings support a negative correlation between PM cholesterol accrual, PIP2/F-actin structure and GLUT4 regulation. These data stemmed from parallel study aimed at understanding the antidiabetic mechanism of the nutrient chromium (Cr3+). We found that chromium picolinate (CrPic) enhanced insulin-stimulated GLUT4 trafficking via reduction in PM cholesterol. In line with glucose/cholesterol toxicity findings, we demonstrated that therapeutic effects of CrPic occurred solely in adipocytes with increased HBP activity and a concomitant elevation in PM cholesterol. Mechanistically, data are consistent with a role of AMP-activated protein kinase (AMPK) in CrPic action. These data show that CrPic increases AMPK activity and perhaps suppresses cholesterol synthesis via distal phosphorylation and inactivation of 3-hydroxy-3-methylglutaryl CoA reductase (HMGR), a rate-limiting enzyme in cholesterol synthesis. Continued study of the consequence of increased HBP activity revealed alterations in cholesterogenic transcription factors – Sp1, SREBP-1, and NFY – with Sp1 showing a significant increase in O-linked glycosylation. Consistent with Sp1 modification eliciting maximal transcriptional activation of SREBP-1, Hmgr mRNA was significantly enhanced. In conclusion, these data are consistent with a central role of PM cholesterol in glucose transport and suggest perturbations in this lipid have a contributory role in developing insulin resistance.

Indiana University-Purdue University Indianapolis (IUPUI) / Elaborate vascularization of the retina is crucial for the development and functioning of the eye. The proper patterning of astrocytes is a key event preceding retinal angiogenesis by providing guidance cues for endothelial cells, yet how this is regulated still remains obscure. The dual function of proteoglycans in both extracellular matrix (ECM) composition and cell signal transduction suggests their potential in the regulation of astrocyte migration. The current study demonstrated that non-cell-autonomous regulation by neuroretina cell surface proteoglycan is crucial for PDGF-A regulated astrocyte migration. Ablation of glycosaminoglycan side chains of proteoglycans in neuroretina led to impaired astrocyte migration, incomplete retinal angiogenesis, and hyaloid vessel persistence. This is followed by severe photoreceptor degeneration as a result of reactive gliosis, which cannot be rescued by constitutively activated Kras signaling. Notably, inner limiting membrane (ILM), the basement membrane of the retina, was breached in proteoglycan-deficient retinae prior to the formation of astrocytic network. Herein we propose that cell surface proteoglycans are essential for the initial assembly of ILM, and this cannot be compensated by secreted ECM proteoglycans. In support of this, after removal of ILM in retinal explant by Collagenase digestion, establishment of a new ILM can be achieved by incubation with exogenous laminin-supplemented Matrigel. This basement membrane reconstitution failed, however, in proteoglycan-deficient retinae or in wild type samples digested with a combination of Heparinase and ChABC in addition to Collagenase. Taken together, our study reveals a novel function of neuroretinal cell surface proteoglycans in the initial assembly of basement membrane which subsequently serves as a permissive substratum necessary for astrocyte migration.

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Hemicelluloses (HC) are the second most abundant plant polysaccharides after cellulose, constituting 25-30% of plant materials. In spite of their abundance, HC are not effectively utilized. Recently, considerable interest has been directed to HC-based biomaterials because of their high oxygen barrier properties, which has potential in food packaging applications. In this study, HC were extracted from sugarcane bagasse and southern yellow pine using a hydrothermal technique which utilizes hot compressed water without catalyst. The parameters affecting the yield of extracted HC such as temperature, time and pressure, were tested and optimized. Eighty four percent of xylose was extracted from sugarcane bagasse at the optimum condition, 180 °C 30 min and 1 MPa pressure. In the case of southern yellow pine, 79% of the mannose was extracted at 190 °C for 10 min and 2 MPa pressure. Concentration and isolation of HC from bagasse and southern yellow pine HC extract were performed by membrane filtration and freeze drying systems. Isolated HC were characterized by FT-IR and 13C NMR techniques and used as a starting material for film preparation. Films were prepared in 0/100, 50/50, 60/40, 70/30 and 80/20% ratios of HC and sodium carboxymethylcellulose (CMC). Thirty five percent of sorbitol (w/w of HC and CMC weight) was also added as a plasticizer. Films were evaluated by measuring water absorption, water vapor permeability (WVP), tensile property and oxygen barrier capability. At 55% relative humidity (RH) and 25 °C the water absorption of both sugarcane bagasse and southern yellow pine HC-based films tended to increase as HC content increased. The lowest WVP of sugarcane bagasse (3.84e-12 g/Pa h m) and southern yellow pine HC films (2.18e-12 g/Pa h m) were determined in 60/40 HC/CMC films. Tensile test results showed that as HC content increases the Young’s modulus decreases, deflection at maximum load and percentage of strain at break increase. It implies that the film properties are changing from stiff to elastic. The oxygen permeability for 60/40 bagasse HC/CMC film was 0.005265 cc μm / (m2 day kPa) and for 70/30 pine HC/CMC film was 0.007570 cc μm /(m2 day kPa).