DEVELOPMENT OF A MECHANISM AND PROCESS TO CONTINUOUSLY PRODUCE SPHERICAL, TORREFIED BIOMASS PELLETS

A recently developed novel biomass fuel pellet, the Q’ Pellet, offers significant improvements over conventional white pellets, with characteristics comparable to those of coal. The Q’ Pellet was initially created at bench scale using a proprietary die and punch design, in which the biomass was torrefied in-situ¬ and then compressed. To bring the benefits of the Q’ Pellet to a commercial level, it must be capable of being produced in a continuous process at a competitive cost. A prototype machine was previously constructed in a first effort to assess continuous processing of the Q’ Pellet. The prototype torrefied biomass in a separate, ex-situ reactor and transported it into a rotary compression stage. Upon evaluation, parts of the prototype were found to be unsuccessful and required a redesign of the material transport method as well as the compression mechanism. A process was developed in which material was torrefied ex-situ and extruded in a pre-compression stage. The extruded biomass overcame multiple handling issues that had been experienced with un-densified biomass, facilitating efficient material transport. Biomass was extruded directly into a novel re-designed pelletizing die, which incorporated a removable cap, ejection pin and a die spring to accommodate a repeatable continuous process. Although after several uses the die required manual intervention due to minor design and manufacturing quality limitations, the system clearly demonstrated the capability of producing the Q’ Pellet in a continuous process. Q’ Pellets produced by the pre-compression method and pelletized in the re-designed die had an average dry basis gross calorific value of 22.04 MJ/kg, pellet durability index of 99.86% and dried to 6.2% of its initial mass following 24 hours submerged in water. This compares well with literature results of 21.29 MJ/kg, 100% pellet durability index and <5% mass increase in a water submersion test. These results indicate that the methods developed herein are capable of producing Q’ Pellets in a continuous process with fuel properties competitive with coal.

The role of a kinesin, KLP-20, on neural development and epidermal morphogenesis in Caenorhabditis elegans

The heterotrimeric kinesin-II motor in Caenorhabditis elegans consists of KLP-20, KLP-11, and KAP-1 subunits and broadly functions in cellular transport for the development of biological structures including cilia and axons. The results of this paper support the ubiquitous and necessary role kinesin-II motors have in development, particularly the KLP-20 microtubule-associating subunit. Mutations in klp-20 result in a variable abnormal (vab) phenotype characterized by observable epidermal defects, although the role of this gene in development and the mechanism by which the vab phenotype is produced is largely unknown. The vab phenotype is highly penetrant in the first larval stage (L1) of C. elegans, which supports that klp-20 functions in early development. Ciliated amphid sensory neurons can be stained with a fluorescent dye, DiI, to simultaneously test cilia structure and function, as well as the morphology of the amphid sensory organ. Reduced dye uptake in klp-20 mutant L1s suggests that the microtubule-based cilia are under-developed as a result of defective kinesin-II function. Consistent observations of the PLM mechanosensory neuron using the zdIs5 reporter suggest that klp-20 has an essential role in neuron development, as mutations to klp-20 result in under-developed PLM axons. Qualitative observations suggest there may be an interaction between the development of the overlying epidermis and the underlying nervous system, as a more severe vab phenotype is observed simultaneously with reduced dye uptake, and hence amphid sensory cilia under-development. Furthermore, a more severe vab phenotype manifested as large bumps on the posterior epidermis appears to be spatially correlated with PLM defects. The results presented and discussed in this paper suggest that KLP-20 has a necessary role in neurodevelopment and epidermal morphogenesis in C. elegans during embryogenesis.

INVESTIGATING THE MECHANISM OF AUTOIMMUNE DISEASE ASSOCIATED-LQTS

The human ether-a-go-go-related gene (hERG) protein passes the rapidly activating delayed rectifier potassium channel (IKr), and malfunction of hERG protein/IKr is the primary cause of acquired long QT syndrome (LQTS). Autoimmune diseases are significantly correlated with prolonged QT intervals, for which autoantibodies have been implicated. The anti-Ro52 autoantibody is the most frequently evaluated, and importantly has been correlated with prolonged QT intervals. Pathological anti-Ro52-hERG interactions have been discussed as a mechanism for autoimmune disease-related LQTS. However, the mechanism is unclear, and it does not explain LQTS in autoimmune diseases which do not commonly express anti-Ro52. In this thesis, I investigated the effects of anti-Ro52 on hERG/IKr function. Through Western blot analysis, whole-cell patch-clamp, and immunofluorescence, I show that anti-Ro52 chronically (12 h) reduced hERG protein expression and hERG current by over 50%, but did not acutely block the channel. My work revealed a novel mechanism in which the Fc portion of anti-Ro52 interacts with the extracellular S5-pore linker of the channel to induce internalization through a tyrosine phosphorylation dependent pathway. This phenomenon extends beyond anti-Ro52 IgG, as other IgG, regardless of their antigen binding specificity, have the potential to reduce hERG expression/current. Rather, the ability of IgG to reduce hERG expression and current is dependent on the IgG subclass, as we show mouse IgG2A was the only mouse IgG subclass which reduced hERG expression. These results provide a novel explanation for autoimmune disease associated LQTS. It also has implications in the development of safe monoclonal antibody drugs.