Macrophages (monocytes/microglia) could play a critical role in central nervous system repair. We have previously found a synchronism between the regression of spontaneous axonal regeneration and the deactivation of macrophages 34 wk after a compression-injury of rat spinal cord. To explore whether reactivation of endogenous macrophages might be beneficial for spinal cord repair, we have studied the effects of granulocyte-macrophage colony stimulating factor (GM-CSF) in the same paraplegia model and in cell cultures. There is significant, though transient, improvement of locomotor recovery after a single delayed intraperitoneal injection of 2g GM-CSF. This improvement is associated with an increased expression of 5HT at the level of the CPG (T13-L2). At longer survival delays, axonal regeneration is significantly enhanced in GM-CSF-treated rats. We then studied the effects of GM-CSF on brain-derived neurotrophic factor (BDNF)secretion by macrophages/microglia, inflammatory reaction and phagocytosis by macrophages/microglia. In vivo, at short post-treatment delays, we found that GM-CSF increases significantly the expression of Cr3 and BDNF by macrophages at the lesion site. In vitro, BV2 microglial cells expressed higher levels of BDNF in the presence of GM-CSF and neurons cocultured with microglial cells activated by GM-CSF generated more neurites, an effect blocked by a BDNF antibody. In vivo, we showed that GM-CSF treatment (either immediate or delayed) does not increase IL-6 expression by macrophages/microglia or astrocytes. We showed that a delayed GM-CSF treatment down regulates IL-1 expression by astrocytes. In vivo, we showed that a delayed GM-CSF treatment can decrease MAG expression at the lesion site. These experiments suggest that GM-CSF could be an interesting treatment option for spinal cord injury and that its beneficial effects might be mediated by BDNF.

In this thesis, two different vaccines were evaluated as adjuvant therapy for patients with colorectal cancer. The ability of the two candidate vaccines to generate antigen-specific cellular and humoral responses, respectively, was studied. The effectiveness of granulocyte colony stimulating factor (GM-CSF) as a cytokine adjuvant to augment the immune response was also examined. The first vaccination strategy involved immunization with the recombinant tumor-associated protein, carcinoembryonic antigen (CEA). Recombinant CEA was administered at 4 different dose levels 7 times during one year. Peripheral blood samples were regularly analyzed during 36 months. This vaccination regimen induced a strong immunoglobulin 1 (IgG1) and IgG4 response, a moderate IgG2 response and a weak IgG3 response against CEA. GM-CSF markedly augmented the effect on IgG1 and IgG4 as well as the T cell response. In contrast, dose of rCEA had no or modest effect on induced immune responses. The response gradually increased during the 12 months immunization period. Responses of all three IgG subclasses and of T cells were protracted up to 36 months. The anti-CEA IgG titers related significantly to survival. Functional HLA-DR epitopes of CEA could be defined. These major histocompatibility class II epitopes may serve as putative components of a peptide-based vaccination strategy. The other vaccine strategy consisted of the tumor-associated antigen epithelial cell adhesion molecule (Ep-Cam) expressed as a transgene in a viral vector, ALVAC. Patients were immunized subcutaneously/intradermally 3 times over 6 weeks and monitored for immune responses for 46 weeks. No anti-Ep-Cam specific humoral response was induced, but Ep-Cam specific type 1 T cells (interpheron-gamma production) were induced, mainly in the GM-CSF group. The cytotoxic cellular response appeared late, or a few months after the last immunization. Both vaccines were well tolerated. Since GM-CSF was an important component for both regimens, immungenicity of this cytokine was assessed. Multiple immunizations with low dose GM-CSF were associated with a low incidence of GM-CSF antibodies that did not neutralize the biological effect of GM-CSF. In conclusion, both vaccines are promising candidate vaccines. GM-CSF is necessary to induce a strong humoral and cellular immune response. Large clinical trials are urgently warranted to evaluate the clinical efficacy.

The current research is directed towards dissection of host genetic factors involved in host immune response and the malaria disease outcome. A possible association between FcγRIIa polymorphism and anti-malarial antibody (A.M.A) responses were investigated in Sudanese patients in relation to clinical outcome of falciparum malaria. The frequency of the R/R131 genotype was significantly higher in patients with severe malaria as compared with mild malaria. A.M.A IgG3 was shown to be associated with reduced risk of clinical malaria in individuals carrying the H/H131 genotype. Low levels of IgG2 reactive with the Pf332-C231 antigen were associated with lower risk of severe malaria in individuals carrying the H131 allele. Fulani and Masaleit, two sympatric ethnic groups in Sudan, are characterized by marked differences in susceptibility to falciparum malaria. We investigated whether the two populations differ in the frequency of GM/KM allotypes. The distribution of GM/KM phenotypes differed significantly among the two groups, with Gm 6 being significantly lower among the Fulani, and the combined frequency of Km 1,3 and Gm 1,17 5,6,13,14 phenotypes was found to be higher among Masaleit. In interethnic study we investigated whether the two groups differ in the frequency of FcγRIIa and HbAS genotypes. The frequency of the H/H131, R/R13 and HbAS genotypes differed significantly among the two groups. Moreover, the Fulani showed higher levels of A.M.A IgG2 and lower IgG1 and IgG3 when compared to their sympatric non-Fulani neighbours. A tri-allelic SNP (C/T/A) in the CRP gene was investigated for possible ethnic associations. The A allele, which is associated with higher basal CRP levels, was found to be less frequent in the Fulani compared with non-Fulani ethnic groups both in Sudan and Mali. In conclusion, our results suggest possible associations between FcγRIIa, CRP genotypes, GM/KM allotypes, and anti-malarial antibody responses and the clinical outcome of falciparum malaria.

The O’Brien lab has demonstrated that Heat shock protein 27 (HSP27)shows attenuated expression in human coronary arteries as the degree of atherosclerosis progresses. Moreover, over-expression of HSP27 reduces atherogenesis in mice. The precise mechanism(s) for HSP27-mediated "atheroprotection" are incompletely understood. Nuclear Factor-kappaB (NF-κB) is a key signaling modulator in atherogenesis. Hence, this project sought to determine if recombinant HSP27 (rHSP27) alters NF-κB signaling to affect atheroprotection. Treatment of THP1 macrophages with rHSP27 resulted in degradation of IκBα, coincided with nuclear translocation of the p65 subunit and produced transcriptional evidence of activation of NF-κB signaling. When the transcriptional profile of THP1 macrophages treated with rHSP27 was analyzed using NF-κB-pathway-specific qRT-PCR arrays, among the regulated genes, IL-10 and GM-CSF mRNA levels were markedly increased, as were parallel translational effects observed. These data provide new mechanistic insights into the atheroprotective effects of HSP27.

This thesis contains three essays on topics in agricultural economics. Essays one and two share a focus on international trade and economic development, and essays two and three apply dynamic tools to agricultural economic policy issues.<p>Essay one analyses trade-related implications of a developing country's decision to adopt genetically-modified crop technology. A fixed-proportions model is constructed that evaluates the welfare implications of a range of adoption policies and export market responses. The model in this essay illustrates the importance of the prospective adopter formulating a projection of probable export market effects before making an adoption decision and of the role that high transaction costs may play in a developing country's adoption decision. The model also considers the effects of a new policy tool; a check-off style levy on genetically-modified technology in place of a technology-use agreement. A levy could be useful tool in developing countries, which are characterised by high transaction costs. <p>Essay two models the effects of emergency food aid on a recipient country's agricultural industry. This essay formulates a definition of needed aid in the context of a food emergency and constructs an optimal control model that solves a path of aid shipments that best meets that need. The effects of a range of food aid paths on recipient-country agricultural production are illustrated through numerical simulations. There are two key results. First, a non-optimal amount of aid can hinder a recipient-country's recovery from an exogenous food shock. Second, an exogenous shock can affect farmer revenue and therefore impact planting decisions. This effect must be considered in aid allocation policies. <p>Essay three uses time-series econometric techniques to develop a demand model that assesses the effectiveness of commodity advertising. This essay describes the importance of considering long-run and dynamic effects in demand systems, especially in the case of closely substitutable commodities. A demand system that tests for and accommodates dynamic and time-series properties is developed and applied to US meat data. The results of this model are compared to a traditional static demand system. The dynamic model produces econometrically and theoretically sound results and generates some more intuitively appealing estimates.

The O’Brien lab has demonstrated that Heat shock protein 27 (HSP27)shows attenuated expression in human coronary arteries as the degree of atherosclerosis progresses. Moreover, over-expression of HSP27 reduces atherogenesis in mice. The precise mechanism(s) for HSP27-mediated "atheroprotection" are incompletely understood. Nuclear Factor-kappaB (NF-κB) is a key signaling modulator in atherogenesis. Hence, this project sought to determine if recombinant HSP27 (rHSP27) alters NF-κB signaling to affect atheroprotection. Treatment of THP1 macrophages with rHSP27 resulted in degradation of IκBα, coincided with nuclear translocation of the p65 subunit and produced transcriptional evidence of activation of NF-κB signaling. When the transcriptional profile of THP1 macrophages treated with rHSP27 was analyzed using NF-κB-pathway-specific qRT-PCR arrays, among the regulated genes, IL-10 and GM-CSF mRNA levels were markedly increased, as were parallel translational effects observed. These data provide new mechanistic insights into the atheroprotective effects of HSP27.

Dendritic cells (DCs) function mainly as antigen presenting cells (APCs) and as such they play a significant role in activating the adaptive immune system. Dendritic cells express toll-like receptors (TLR), and when these receptors are engaged by their cognate agonists, they promote DC maturation, which is critical in the induction of potent T helper (Th) cell -1 responses. Due to the multifunctional abilities of DCs, they have been explored as vaccine carriers, largely in cancer immunotherapy and some infectious diseases including hepatitis C. Previous studies showed that DCs loaded with mRNA of hepatitis C virus (HCV) antigen(s) induced strong immune responses but immune protection was not complete. Therefore, I expected that adoptive transfer of DCs transfected with HCV NS3/4A and/or NS5A mRNA and further treated with TLR agonist(s) ex vivo would induce HCV-specific immunity in vivo. Bone marrow-derived DCs generated with Flt3L (FL-DCs) or GM-CSF (GM-DCs), and loaded with HCV NS3/4A and/or NS5A mRNA showed maturation characteristics and produced substantial amounts of IL-12 after ex vivo activation with CpG ODN or CpG ODN plus Poly I:C, when compared to their untreated counterparts. Treatment with a combination of CpG ODN and Poly I:C synergized to augment IL-12 production in comparison with stimulation with CpG ODN alone. IL-12 secretion by DCs is pivotal in directing immune responses towards a Th1-bias response, which is needed to eliminate HCV. However, the ex vivo responses of stimulated DCs bearing HCV antigen(s) were not efficiently translated in mice to potentiate vigorous antigen-specific T cell responses. This resulted in a lack of protection after challenge with recombinant vaccinia virus expressing HCV NS3/NS4/NS5 in immunized mice. In contrast, both antigen-specific humoral and cell-mediated immune responses were induced in mice vaccinated with HCV recombinant NS3 or NS5A protein co-formulated with CpG ODN, host defense peptide and polyphosphazene. These responses, however, did not mediate viral clearance, as vaccinated mice remained unprotected from infection with recombinant vaccinia virus expressing HCV antigens. Taken together, these results suggest HCV recombinant protein co-formulated with triple adjuvant to be a better vaccine candidate than the DC-based vaccine.

This thesis contains three essays on topics in agricultural economics. Essays one and two share a focus on international trade and economic development, and essays two and three apply dynamic tools to agricultural economic policy issues.<p>Essay one analyses trade-related implications of a developing country's decision to adopt genetically-modified crop technology. A fixed-proportions model is constructed that evaluates the welfare implications of a range of adoption policies and export market responses. The model in this essay illustrates the importance of the prospective adopter formulating a projection of probable export market effects before making an adoption decision and of the role that high transaction costs may play in a developing country's adoption decision. The model also considers the effects of a new policy tool; a check-off style levy on genetically-modified technology in place of a technology-use agreement. A levy could be useful tool in developing countries, which are characterised by high transaction costs. <p>Essay two models the effects of emergency food aid on a recipient country's agricultural industry. This essay formulates a definition of needed aid in the context of a food emergency and constructs an optimal control model that solves a path of aid shipments that best meets that need. The effects of a range of food aid paths on recipient-country agricultural production are illustrated through numerical simulations. There are two key results. First, a non-optimal amount of aid can hinder a recipient-country's recovery from an exogenous food shock. Second, an exogenous shock can affect farmer revenue and therefore impact planting decisions. This effect must be considered in aid allocation policies. <p>Essay three uses time-series econometric techniques to develop a demand model that assesses the effectiveness of commodity advertising. This essay describes the importance of considering long-run and dynamic effects in demand systems, especially in the case of closely substitutable commodities. A demand system that tests for and accommodates dynamic and time-series properties is developed and applied to US meat data. The results of this model are compared to a traditional static demand system. The dynamic model produces econometrically and theoretically sound results and generates some more intuitively appealing estimates.

High linearity transmitters and receivers should be used to efficiently utilize the available channel bandwidth. Power consumption is also a critical factor that determines the battery life of portable devices and wireless sensors. Three base-band and RF building blocks are designed with the focus of high linearity and low power consumption. An architectural attenuation-predistortion linearization scheme for a wide range of operational transconductance amplifiers (OTAs) is proposed and demonstrated with a transconductance-capacitor (Gm-C) filter. The linearization technique utilizes two matched OTAs to cancel output harmonics, creating a robust architecture. Compensation for process variations and frequency-dependent distortion based on Volterra series analysis is achieved by employing a delay equalization scheme with on-chip programmable resistors. The distortion-cancellation technique enables an IM3 improvement of up to 22dB compared to a commensurate OTA without linearization. A proof-of-concept lowpass filter with the linearized OTAs has a measured IM3 < -70dB and 54.5dB dynamic range over its 195MHz bandwidth. Design methodology for high efficiency class D power amplifier is presented. The high efficiency is achieved by using higher current harmonic to achieve zero voltage switching (ZVS) in class D power amplifier. The matching network is used as a part of the output filter to remove the high order harmonics. Optimum values for passive circuit elements and transistor sizes have been derived in order to achieve the highest possible efficiency. The proposed power amplifier achieves efficiency close to 60 percent at 400 MHz for -2dBm of output power. High efficiency class A power amplifier using dynamic biasing technique is presented. The power consumption of the power amplifier changes dynamically according to the output signal level. Effect of dynamic bias on class A power amplifier linearity is analyzed and the results were verified using simulations. The linearity of the dynamically biased amplifier is improved by adjusting the preamplifier gain to guarantee constant overall gain for different input signal levels.

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