The mechanism of allosteric coupling between the external ligand-binding domain and the internal signaling domain of bacterial chemoreceptors is poorly understood. Genetic, biochemical, and biophysical evidence suggests that transmembrane helix 2 (TM2) undergoes a piston-like displacement of approximately 1-3 Angstroms toward the cytoplasm upon the binding of aspartate to the Tar receptor. The signal is then transmitted to the cytoplasmic signaling domain via the HAMP domain, a conserved motif found in all methyl-accepting chemotaxis proteins (MCPs) and most histidine protein kinases (HPKs). HAMP forms a parallel four-helix bundle consisting of a dimer of two amphipathic helices (AS1 and AS2) connected by a flexible linker. The MLLT sequence between residues Arg-214, at the end of TM2, and the conserved residue Pro-219, at the beginning of AS1 of the HAMP domain (the TM2-HAMP junction), is predicted to be able to form a helical extension of TM2. We hypothesized that perturbing the native secondary structure and/or the length of the TM2-HAMP junction would disrupt the ability of HAMP to communicate the input signal from TM2 to the kinase-control domain. To test this hypothesis, we designed two experiments. First, constructs were made in which 1 to 3 Gly residues were inserted between T218 and P219. Second, Tar variants were constructed in which 1 to 9 Gly residues were inserted between R214 and P219. The results suggest that increasing the length and flexibility of the TM2-HAMP connection tends to uncouple signal propagation between the TM2 and the HAMP elements and suggests that HAMP alone causes an inhibitory effect on the cytoplasmic signaling domain. To determine whether the predicted helical register of the MLLT sequence is an important component of the propagation of the transmembrane signal from TM2 to the HAMP domain, we added and subtracted helical residues to the MLLT sequence. The results suggest that helical register and length of the TM2-HAMP junction are essential for optimal receptor function.

The nervous system coordinates a sequence of muscle movements to give rise to animal behaviors. In complex invertebrates or lab-studied vertebrates, due to the large number of cells in their nervous systems and the complexities of their behaviors, it is difficult to address how circuits process information to direct each motor output of the behavior. In this dissertation, I used the Caenorhabditis elegans male copulation behavior as a model to address how a compact circuit coordinates different behavioral programs. Insertion of a male copulatory organ into a suitable mate is a conserved and necessary behavioral step for most terrestrial mating. However, the detailed molecular and cellular mechanisms for this distinct social interaction have not been elucidated in any animal. During mating, the C. elegans male cloaca is positioned over the hermaphrodite’s vulva as he attempts to insert his copulatory spicules repetitively. Rhythmic spicule thrusts cease when insertion is sensed. Circuit components consisting of sensory/motor neurons and sex muscles for these steps have been previously identified, but it was unclear how their outputs are integrated to generate a coordinated behavior pattern. Here, I show that contraction of the male oblique muscles is required to sustain genital contact between the sexes. These muscles are innervated by the postcloacal sensilla (p.c.s.) sensory/motor neurons, which secret ACh to activate the levamisole-sensitive AChR and the ACR-16-containing ionotropic AChR on the oblique muscles. For spicules to rhythmically thrust during genital contact, activity of the oblique muscles and the gubernacular muscles is transmitted to the spicule protractor muscles instantaneously via gap junctions between these muscles and causes shallow protractor contractions. The rhythmic protractor contractions eventually switch to sustained contraction, as the SPC sensory-motor neurons integrate information of spicule position at the vulva with inputs from the hook and cloacal sensilla. The ERG-like K+ channel, UNC-103, which decreases the spicule circuit excitability, is likely to set a threshold requirement for integration of these inputs, so that sustained spicule muscle contraction is not stimulated by fewer inputs. In addition, I demonstrate that a cholinergic signaling pathway mediated by a muscarinic acetylcholine receptor, GAR-3, is used to enhance the ionotropic AChRs-mediated fast synaptic transmission in the copulation circuit. GAR-3 is expressed in multiple cells of the copulation circuit, but mainly in the cholinergic p.c.s. neurons and SPC neurons. Activation of GAR-3 is coupled to Gαq to trigger downstream signal transduction events that modulate neurotransmitter release from these neurons. Males with a loss-of-function allele of the gar-3 gene are defective in inserting their spicules into the hermaphrodite’s vulva efficiently. Since the p.c.s. neurons regulate the male’s contact with the hermaphrodite’s vulva, and the SPC neurons are required for spicule insertion during mating, GAR-3 probably facilitates male mating behavior via enhancing synaptic transmission from these neurons to their postsynaptic partners.

This work has employed the Micro-wave Plasma enhanced Chemical Vapor Deposition (MPCVD) method to fabricate diamond PN junction device. The n+ <111> orientation single-crystal silicon has used as substrates. P-type diamond layer is doped with B(OCH3)3 and the N-type diamond layer is doped with ammonia. The surface structure of diamond film has been observed by scanning electron microscope; and the device rectification property of a PN junction has measured by current-voltage characteristic. The carrier density and mobility of diamond films have been analyzed by Hall measurement. Furthermore, the Cathodoluminescence (CL) spectroscopy showed the defect spectra in diamond PN junction. The N-type diamond film and P-type diamond film have deposited at temperature of 800 ¢J, for 30 minutes and 90 minutes, respectively. The process CVD has performed in the same chamber continually. A I-V curve of sample showed the set on positive voltage 0.5 V and the reverse breakdown voltage of 6 V. Further, CL results revealed a peak at 285 nm (4.4 eV), which represents the CVD diamond band and the other one is at 500 nm (2.5 eV), which stands for donor-acceptor recombination from defect in these diamond films.

There are many intensive researches for zinc compounds due to their wide band gaps and potential applications in visible and UV optoelectronic technologies. Zinc nitride is a n-type semiconductor material having a direct band gap, and is not widely studied. Previous papers reported that zinc nitride is a n-type semiconductor having low resistivity and high electron mobility. Its band gap varies from 1.23 eV to 3.2 eV depending on the process condition. In this work, we successfully fabricated zinc nitride p-n junction by heat treatment on zinc nitride films. The threshold voltage of p-n junction is about 1 V. The Zinc nitride films were prepared by reactive RF magnetron sputtering. The as-grown zinc nitride thin film is a n-type material. It is found that the film treated at 300¢J for 3 hours can be changed to a p-type material. The zinc nitride has a very low resistance (2.2¡Ñ10-2 £[-cm) and high carrier concentration (3.88¡Ñ1019 cm-3) after the heat treatment. The optical band gap of zinc nitride was determined as a direct band gap varying from 1.1 eV to 1.6 eV according to the temperature of heat treatment. The zinc nitride was successfully prepared with various electrical characteristics and band gaps by controlling the temperature of heat treatment.

Thesis (M.S.)--Mississippi State University. Department of Electrical and Computer Engineering. / Title from title screen. Includes bibliographical references.

Peripheral nerve injury (PNI) is common and despite modern microsurgical techniques of repair, functional restoration is always incomplete. This results in impaired sensation and reduced motor function alongside pain and cold intolerance. Traumatic PNI are often associated with loss of nerve tissue, creating a gap, and direct repair of the two damaged nerve stumps is not possible. These types of injuries are reconstructed using autologous nerve grafts but this is far from ideal since it necessitates the sacrifice of a functional nerve from elsewhere in the body. Chronic muscle atrophy because of the prolonged delay in nerve regeneration across gaps is a significant impediment to an optimal functional recovery.   Tissue engineering and regenerative medicine approaches to nerve repair might one day replace the need for autologous nerve grafts. This thesis investigates the effects of adipose derived stem cells (ASC) on nerve regeneration and muscle recovery by using the stem cells for intramuscular injection and combined with a biomaterial, poly-3-hydroxybutyrate (PHB), to create a bioengineered artificial nerve repair construct.  The mechanisms of interaction between the stem cells and neuromuscular system cells were investigated and with a view to translating this work into clinical practice, an optimal source of cells was investigated from human donors.   It was hypothesized that injecting regenerative cells into muscle would reduce nerve injury induced muscle atrophy. A rat sciatic nerve lesion was performed and three different types of cells were injected into the denervated gastrocnemius muscle; either (1) undifferentiated ASC, (2) ASC induced to a ‘Schwann cell-like’ phenotype (dASC) or (3) primary Schwann cells. Nerves were either repaired by direct end-end suture or capped to prevent muscle reinnervation. One month later, functionality was measured using a walking track test, and muscle atrophy was assessed by examining muscle weight and histology. The Schwann cells and dASC groups showed significantly better scores on functional tests when compared with control injections of growth medium alone. Muscle weight and histology were also significantly improved in the cell groups in comparison with the control group.   PHB strips seeded with either primary Schwann cells or dASC suspended in a fibrin glue matrix were used to bridge a 10mm rat sciatic nerve gap. After 12 weeks, functional and morphological analysis (walking track test, electromyography, muscle weight and muscle and nerve histology) was performed. The results showed significantly better functional results for the PHB strips seeded with cells versus the control group with fibrin matrix only. This correlated with less muscle atrophy and greater distal axon myelination in the cell groups.   To further optimize the nerve regeneration and muscle recovery, the nerve gap lesion was repaired by treatment with the bioengineered constructs seeded with dASC or nerve autograft in combination with stem cell injection in the muscle. After 6 weeks, the best results were obtained in the nerve graft group combined with intramuscular dASC injection which showed significantly less atrophy than the other groups. The results also showed that using the stem cells in a matrix on a PHB strip in combination with intramuscular injections could significantly reduce muscle atrophy.   In vitro experiments showed that dASC expressed a wide range of neurotrophic and myogenic factors including BDNF, VEGF-A, IGF-1 and HGF. Stem cell conditioned medium enhanced the proliferation of myoblast cell lines and primary Schwann cells. Various signaling pathways (PKA, MAP kinase) were involved in these effects dependent on the cell type investigated. Furthermore, in direct co-culture with myoblast cells, a small population of the cells fused together to form myotube-like structures and expressed myogenic markers.   Human ASC were isolated from the deep and superficial layers of abdominal fat tissue obtained during abdominoplasty procedures.  Cells from the superficial layer proliferated significantly faster than those from the deep layer. Superficial layer ASC induced significantly enhanced neurite outgrowth from neuronal cell lines when compared with the deep layer cells.  However, RT-PCR and ELISA analysis showed that ASC isolated from both layers expressed similar levels of the neurotrophic factors NGF, BDNF and GDNF.   In summary, these results show that stem cell therapy at both levels (the nerve lesion site and in the target denervated muscle) offers a promising approach for clinical application for treatment of peripheral nerve lesions. The bioengineered artificial nerve construct, combining PHB strip with cells, also provides a beneficial environment for nerve regeneration. Many of the benefits of the ASC are likely to be mediated through their secretome, a rich source of neurotrophic and myogenic factors. Thus adipose tissue contains a pool of regenerative stem cells which have significant potential application to tissue engineering and regenerative medicine for nerve repair.

published_or_final_version / abstract / toc / Electrical and Electronic Engineering / Master / Master of Philosophy

Notwithstanding several years of robust growth, solar energy still only accounts for<1% of total electrical generation in the US. Before solar energy can substantially replace fossil fuels subsidy-free at utility scale, further cost reductions and efficiency improvements are needed in complete generating systems. Flat panel silicon PV modules are by far the most dominant solar technology today, but have little room for improvement in efficiency and are limited by balance of system costs. Concentrated PV (CPV) is an alternate approach with long-term potential for much higher efficiency in sunny climates. In CPV modules, large area optics collect and concentrate direct sunlight onto small multi-junction cells with>40% conversion efficiency. Concentrated Solar Power (CSP) uses mirrors to concentrate sunlight onto thermally absorbing receivers, which generate electricity with convention thermal cycles. In this dissertation, four new optical approaches to CPV and CSP with potential for lower cost are analyzed. Common to each approach is the use of large square glass reflectors, which have very low areal cost (~$35/m^2) and field-proven reliability in the CSP industry. Chapter 2 describes a freeform toroidal lens array used to intercept the low concentration line focus of a parabolic trough to produce multiple high concentration foci (>800X) for multi-junction cells. In Chapter 3, three embodiments of dish mirrors and freeform lenslet arrays are explored, including an off-axis system. In each case, a dish mirror illuminates a freeform lenslet array, which divides sunlight equally to a sparse matrix of multi-junction cells. The off-axis optical system achieves +/-0.45° acceptance angle and averages 1215X geometric concentration over 400 multi-junction cells. Chapter 4 proposes a new architecture for CSP central receivers that achieves extremely high collection efficiency (>70%) with unconventional heliostat field tracking. In Chapter 5, the design and preliminary testing of a spectrum-splitting hybrid PV/thermal generator is discussed. This system has the advantage of 'drop-in' capability in existing CSP trough plants and allows for thermal storage, an important mitigation to the intermittency of the solar resource.

The fps proto-oncogene encodes a 92 kDa cytoplasmic tyrosine kinase. Previous studies have shown that Fps expression in the mammary gland changes with development, and Fps has a suppressor function in mammary tumorigenesis. The aim of my thesis was to elucidate the role of the Fps tyrosine kinase in regulating mammary gland development and function. We have shown that the expression of the Fps kinase in the mammary gland increased during pregnancy and reached its maximum during lactation. The level of Fps tyrosine phosphorylation paralleled the expression pattern. Pups reared by fps-null females gained weight more slowly than those reared by wild-type females. Epithelial cells were the primary source of Fps expression. Milk protein and fat content were not affected by the absence of Fps. Similarly, no differences in mammary gland structure were observed with whole mount or histological analysis. Fps was shown to be in a multi-protein complex with E-cadherin, β-catenin and p120-catenin. A strong co-localization signal was observed for Fps and E-cadherin. Immunofluorescence analysis indicated that the localization of E-cadherin and β-catenin was disorganized and less concentrated at sites of cell-cell contacts in the fps-null glands. The interactions between the different adherens junction components were altered in the fps-null tissue. Specifically, less E-cadherin and β-catenin was associated with p120-catenin in the fps-null glands. Suprisingly, no phosphotyrosine differences were detected for the adherens junction components. Conditions were established to grow primary murine epithelial cell cultures that could be used to investigate the function of Fps. Fps expression was up-regulated in these cells in response to lactogenic hormones. A lentiviral system encoding a murine p53 shRNA sequence was used to increase the growth potential of the primary cells. Continual growth of the infected and uninfected primary epithelial cell mixture resulted in the establishment of an immortalized cell line. Immunofluorescent and immunoblot analyses revealed that the cells have undergone an epithelial-to-mesenchymal transition. With the transduction of a myc-epitope tagged Fps into the cells, we have generated cell lines with the appropriate genetic backgrounds to study the function of the Fps kinase in the mammary gland, specifically as it relates to tumorigenesis. / Thesis (Ph.D, Pathology & Molecular Medicine) -- Queen's University, 2008-07-03 11:53:01.135

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