Tools and technologies for expert systems: A human factors perspective

It is widely recognized that technologies based on artificial intelligence (AI), especially expert systems, can make significant contributions to the productivity and effectiveness of operations of information and knowledge intensive organizations such as NASA. At the same time, these being relatively new technologies, there is the problem of transfering technology to key personnel of such organizations. The problems of examining the potential of expert systems and of technology transfer is addressed in the context of human factors applications. One of the topics of interest was the investigation of the potential use of expert system building tools, particularly NEXPERT as a technology transfer medium. Two basic conclusions were reached in this regard. First, NEXPERT is an excellent tool for rapid prototyping of experimental expert systems, but not ideal as a delivery vehicle. Therefore, it is not a substitute for general purpose system implementation languages such a LISP or C. This assertion probably holds for nearly all such tools on the market today. Second, an effective technology transfer mechanism is to formulate and implement expert systems for problems which members of the organization in question can relate to. For this purpose, the LIghting EnGineering Expert (LIEGE) was implemented using NEXPERT as the tool for technology transfer and to illustrate the value of expert systems to the activities of the Man-System Division

Directed Evolution of Stabilized Peptides with Bacterial Display

Interactions between proteins govern cellular and the body’s states, including aberrant interactions found in diseases such as in cancers and infections. Small molecule drugs are not ideal in targeting these interactions as their size generally prevents efficient blocking of contacts over large surface areas. Antibodies and related biologics have seen clinical success in the past few decades and can block large surfaces but are typically limited to extracellular targets. Intermediate-size peptides have the potential to bridge this gap, with the ability to target large surface areas inside the cell. Peptide stapling, by chemically linking two or more amino acid residues, can confer affinity improvements, resistance to degradation, and better biological transport properties. As such, stapled peptides show promise as next-generation therapeutics. Unfortunately, existing methods to screen sequence and stapling locations suffer from numerous disadvantages including limited search space, lack of real-time monitoring of selections, and difficulty in incorporating the non-canonical amino acids used for amino acid stapling. In this dissertation, I describe my research on stapled peptide discovery with bacterial incorporation of non-canonical amino acids. To screen stapled peptides of the type desired, we incorporated azidohomoalanine (AHA) into surface displayed peptides, enabling an in situ ‘click’ chemistry reaction to bridge two turns of an alpha helical (i, i+7) amino acid library for directed evolution. Using the p53-MDM2 interaction as a model target, we developed peptides that block MDM2 degradation of the tumor suppressor protein p53, an interaction that is dysregulated in a sizeable fraction of cancers. We generated and displayed a stapled peptide library on the bacterial cell surface with fixed residues for stabilization and binding requirements, while randomizing the remaining amino acids. After multiple rounds of selection, clones were sequenced and characterized. The dissociation constants of the peptide-MDM2 interaction were measured on both the bacterial cell surface by flow cytometry and in solution by bio-layer interferometry. The highest affinity variant, named SPD-M6-V1 with sequence VCDFXCYWNDLXGY (dissociation constant = 1.8 nM; X = azidohomoalanine) was selected for structural characterization by NMR spectroscopy, revealing a bicyclic disulfide and double click-constrained peptide. Sequencing showed that peptides with two cysteines were highly enriched, further suggesting that the MDM2-binding conformation was enforced with a disulfide bond. In addition, SPD-M6-V1 was the most protease-resistant peptide from the library that we tested. Next, we stapled the displayed peptide library with chemically distinct linkers and screened each library separately. We performed deep sequencing to better understand the relationship between amino acid sequence and linker identity in contributing to high affinity MDM2 binding. We found that both linker-specific and linker-agnostic (i.e. MDM2-specific) mutations were enhanced. Finally, we developed a dual-channel, sequential labeling selection strategy to discriminate between high-display, low-affinity peptides and low-display, high-affinity peptides, two categories that would ordinarily overlap in a typical one-color screen in the absence of an independent display marker. In summary, this thesis develops the chemical tools to screen libraries of stabilized peptides on the bacterial cell surface and applies these techniques to select stabilized alpha helices that disrupt the p53-MDM2 interaction.PHDChemical EngineeringUniversity of Michigan, Horace H. Rackham School of Graduate Studieshttp://deepblue.lib.umich.edu/bitstream/2027.42/163094/1/tejasn_1.pd

Synthesis, microstructure and mechanical properties of bulk ultrafine grained Ti-47Al-2Cr (at%) alloy

Hot isostatic pressing (HIP) and powder compact forging of Ti/Al/Cr composite powders of composition Ti-47Al-2Cr (at%) have been carried out to synthesize bulk ultrafine grained Ti-47Al-2Cr alloy. The Ti/Al/Cr composite powders were produced using high energy mechanical milling of elemental Ti, Al, Cr powders in a retsch planetary mill. The microstructure and mechanical properties of the bulk consolidated alloy produced using different processing techniques has been investigated. The mechanical properties of the alloy were studied in tension and compression both at room and elevated temperatures especially to know the formability of the material. The bulk alloy samples produced by HIP for 2 hours at 1000 degrees C had porosity of approximately ~ 5%, indicating that the HIP time was not sufficient to close the pores. The microstructure mainly consisted of TiAl as the major phase and Ti(Al) and Ti3Al as minors, the unreacted Ti(Al) phase in the microstructure was mainly due to the initial powder condition, in which a small fraction of powder particles were rich in Ti. Tensile testing of the alloy samples was carried out at different temperatures. At room temperature the alloy was fairly brittle, without any plastic deformation, and had a fracture strength of ~ 100 MPa. At elevated temperatures the samples became ductile, as reflected by considerable amounts of tensile elongations at 800 degrees C and above. The maximum amount of elongation was found to be between 70 - 80% at 900 degrees C. The tensile yield strength at 800 degrees C was in the range of 84-90 MPa and decreased to 55-58 MPa with the testing temperature of the samples to 900 degrees C. In compression the alloy showed plastic yielding and yield strength of ~ 1.4 GPa at room temperature. Compression testing at 900 degrees C revealed that compressive deformations equivalent to a height reduction of 50% could be easily achieved without cracking. Direct powder compact forging using canned powder compacts of the Ti/Al/Cr composite powder was successfully used to produce bulk consolidated Ti-47Al-2Cr alloy samples. It has been observed that the density of the bulk consolidated alloy sample after forging varied from the centre to the periphery. XRD analysis showed that the forged samples, consisted of TiAl (as major phase) along with Ti(Al) and Ti3.3Al phases. Mechanical testing of the samples showed that the samples exhibited brittle type of fracture both in tension and compression at room temperature and the fracture strength of the samples was in the range of 115 - 130 MPa in tension and 1.38-1.4 GPa in compression without any yielding. When being tested at 900 degrees C, the samples became very ductile showing yield strength in the range of 70-90 MPa and elongation to fracture between 80-165% in tension, and a yield strength of ~ 65 MPa and 50% deformation in compression was easily achievable. Nearly fully dense Ti-47-2Cr alloy samples with density of ~98% were produced by using HIP at 1000 degrees C for a duration of 3 hours. TEM observations revealed equiaxed grains with grain sizes in the range of 200-500 nm. The tensile testing of the alloy samples at different temperatures revealed that the brittle to ductile transition temperature of the alloy was in the range of 700 and 750 degrees C, similar to that reported from literatures. The alloy showed significantly higher strengths both at room and at elevated temperatures, due to the low level of porosity in the sample. Elongation of 95 - 117% at 750 degrees C and 70-100% at 800 degrees C was observed. The ultrafine grained Ti-47Al-2Cr alloy produced using a combination of mechanical milling and HIP/powder compact forging has demonstrated good formability at elevated temperatures leaving a large space for secondary processing to improve the quality of the material

The mechanical behaviour of an ultrafine grained Ti-47Al-2Cr (at%) alloy in tension and compression and at different temperatures

A bulk ultrafine grained (UFG) Ti-47Al-2Cr (at%) alloy has been produced using a powder metallurgy process that combines high energy mechanical milling (HEMM) of a mixture of Ti, Al and Cr powders to produce a Ti/Al/Cr composite powder and hot isostatic pressing (HIP) of the composite powder compact. The purpose of the present study is to determine the mechanical behaviour of the alloy in tension and compression at room temperature (RT) and elevated temperatures, and also to compare the compression behaviour of the material with its tensile behaviour. It has been found that due to the residual pores, lack of full level interparticle bonding and high oxygen content (0.87wt%) in the consolidated samples, the UFG TiAl based alloy has a very low room temperature tensile fracture strength of 100 MPa and shows no tensile ductility. However these microstructural defects and high oxygen content have much less significant effect on the room temperature compressive mechanical properties, and the alloy shows a high compressive yield strength of 1410 MPa, and some ductility (plastic strain to fracture 4%). At elevated temperatures of 800oC and above, the alloy shows high tensile and compressive ductility as demonstrated by 75% tensile elongation to fracture and no cracking in upset forging with a height reduction of 50% at 900oC. The yield strength of the alloy at 900oC is 55 MPa in tension and 33 MPa in compression, both of which are lower than those of coarse grained TiAl based alloys with similar compositions at 900oC. This is due to a higher creep rate of the UFG alloy caused by the small grains. The good formability of the UFG TiAl based alloy as reflected by the lower critical temperature above which the alloy becomes highly formable indicates that the material can be used as a suitable precursor for secondary thermomechanical processing and super-plastic forming

Reducing herbicide discharge to sensitive environments using membrane bioreactors

A hybrid MBR/UV/GAC treatment system was researched to remove Ametryn, which is a commonly used herbicide in Australian farmlands, from wastewater. The research revealed that the hybrid system could be successfully used for 100% removal of Ametryn. Two mathematical models were developed to predict the frequency of chemical cleaning of MBR-membrane and the mechanism of fouling of membrane