Uncertainties persist regarding the assessment of the carcinogenic risk associated with galactic cosmic ray (GCR) exposure. The GCR spectrum peaks in the range of 300 MeV/n to 700 MeV/n and is comprised of elemental ions from H to Ni. While Fe ions represent only 0.03% of the GCR spectrum in terms of particle abundance, they are responsible for nearly 30% of the dose equivalent in free space. Because of this, radiation biology studies focusing on understanding the biological effects of GCR exposure generally use Fe ions. Acting as a thin shield, the Martian atmosphere alters the GCR spectrum in a manner that significantly reduces the importance of Fe ions. Additionally, albedo particles emanating from the regolith complicate the radiation environment. The present study uses the Monte Carlo code FLUKA to simulate the response of a tissue-equivalent proportional counter on the surface of Mars to produce dosimetry quantities and microdosimetry distributions. The dose equivalent rate on the surface of Mars was found to be 0.18 Sv/y with an average quality factor of 2.9 and a dose mean lineal energy of 18.4 keV/µm. Albedo neutrons accounted for 25% of the dose equivalent. Additionally, differential energy spectra were generated in order to determine the fractional contribution to frequency, dose, and dose equivalent for each elemental ion from H to Ni on the surface of Mars. Fe ions were found to account for just 1.3% of the dose equivalent while H and He ions were found to account for 32% and 17%, respectively. It is anticipated that these data will provide relevant benchmarks for use in future risk assessment and mission planning studies.

Tissue equivalent proportional counters (TEPC) are used for medical and space activities whenever a combination of high and low LET (lineal energy transfer) radiations are present. With the frequency and duration of space activities increasing, exposure to fast heavy ions from galactic cosmic radiation and solar events is a major concern. The optimum detector geometry is spherical; to obtain an isotropic response, but simple spherical detectors have the disadvantage of a non-uniform electric field. In order to achieve a uniform electric field along the detector axis, spherical tissue equivalent proportional counters have been designed with different structures to modify the electric field. Some detectors use a cylindrical coil that is coaxial with the anode, but they are not reliable because of their sensitivity to microphonic noise and insufficient mechanical strength. In this work a new spherical TEPC was developed. The approach used was to divide the cathode in several rings with different thicknesses, and adjust the potential difference between each ring and the anode to produce an electric field that is nearly constant along the length of the anode. A-150 tissue equivalent plastic is used for the detector walls, the insulator material between the cathode rings is low density polyethylene, and the gas inside the detector is propane. The detector, along with the charge sensitive preamplifier, is encased in a stainless steel vacuum chamber. The gas gain was found to be 497.5 at 782 volts and the response to neutrons as a function of angle was constant ±7%. This spherical tissue equivalent proportional counter detector system will improve the accuracy of dosimetry in space, and as a result improve radiation safety for astronauts.

Tissue-equivalent proportional counters are frequently used to measure dose and dose equivalent in cosmic radiation fields that include high-Z, high-energy (HZE) particles. The fact that particles with different stopping powers can produce the same energy deposition in the same detector means that the measure of lineal energy cannot provide enough information to evaluate the equivalent dose due to HZE particles. To characterize incident particles by mass and velocity, a multiple-detector system composed of three tissue-equivalent proportional counters simulating different size tissue volumes was proposed to be built. This system took advantage of the well-known fact that lineal energy (y) of a HZE particle depends on the site size, as well as the particle mass and energy. Monte Carlo calculations were used to evaluate lineal energy, using GEANT4, in grid-walled (wall-less) proportional counters with simulated unit density site diameter of 0.1, 0.5 and 2.5 micrometers in a uniform HZE particle field. Uniform beams of 1000 MeV/n and 100 MeV/n 56Fe26+, 28Si14+, 16O8+, 12C6+, 4He2+ ions and proton particles bombarding the detectors were simulated. The results of the calculations were used to determine how much additional information about particle charge and velocity could be obtained from such a detector system. Comparison of simulation results with those of walled detectors was included in the study to illustrate the wall effect. The results shows that the detector system is capable of characterizing HZE particles in a mixed unknown field based on the lineal energy spectra as well as the calculated mean lineal energy. This suggests that it may be practical to use such a system to measure the average particle velocity of HZE particles in space. The parameters used in the simulation are also good references for detector construction. There is only limited experimental data for lineal energy resulting from a large uniform field of HZE particles incident on a wall-less detector. However, the Monte Carlo results are consistent with the experimental data available.

Absorbed dose distributions in lineal energy for neutrons and gamma rays were measured by using both a tissue-equivalent walled counter (TEPC) and a graphite-walled low pressure proportional counter (GPC) in the Am-Be neutron source facility at UOIT. A series of measurements were performed with the counters filled with propane-based TE gas (55.1% C3H8, 39.5% CO2 and 5.4% N2) at operating gas pressures corresponding to tissue spheres 2.0 , 4.0 and 8.0 μm in diameter. The results of these measurements indicated satisfactory performance of counters to measure microdosimetric spectra extending down to event-sizes that cover the gamma component of a mixed field. The spectra and the related mean values ̅y F and ̅y D are compared with other similar work but with monoenergetic neutrons of different energy range, the agreement between them is good. An assessment of the performance of different size TEPC has been done. An excellent agreement between their event size spectra was found and the proton edge appears at the same position on the lineal energy scale and differences in microdosimetric parameters ̅ and ̅ is not exceeding 3%, which is in the region of counting statistics. In Am-Be neutron field, the efficiency of the TEPCs was measured to have an average value of 250 counts per μSv or equivalently about 4.17 counts per minutes per μSv/hr. This efficiency is reasonable for dose equivalent measurements but needs a long integration period. The measurements showed that the dose equivalent which depends on the measurement of energy deposition by the secondary charged particles was originated mainly from elastic collisions of the incident neutrons with hydrogen atoms. Moreover the number of events in the sensitive gas is dominated by proton recoils. A non- negligible fraction of the dose equivalent resulted from gamma interactions, alpha and recoil nuclei. The energy deposition patterns in these micro-scale targets are strongly dependent on radiation quality, so differences of linear energy transfer (LET) of the components in a mixed radiation field are significant. Accordingly, in a radiation field with an unknown gamma ray energy spectrum, absorbed dose for neutrons can be obtained by the separation of neutron induced events from gamma events using their distribution in lineal energy. To separate neutron dose from gamma dose a simple lineal energy threshold technique has been used in addition to a more sophisticated methods using γ-fitting and the graphite-walled counter measurements. The results of this study will establish the degree of error introduced by using a lineal energy threshold, which is likely to be used in any hand-held neutron monitor based on TEPCs. / UOIT

The thymus is the major site for T-cell generation and thus is important for the adaptive immune system. Development of a properly selected, functional T-cell repertoire relies on interactions between developing T cells and a series of functionally distinct thymic stroma cell types including the cortical and medullary thymic epithelial cells (TECs). The thymus is one of the first organs to degenerate in normal healthy ageing. Related to this, there is strong interest in developing protocols for improving thymus function in patients by cell replacement or regenerative therapies. Thymic epithelial progenitor cells (TEPCs) represent a potential source of cells for thymus transplantation. However, the only source of these cells for transplantation is currently fetal thymus tissue. If TEPCs could be generated from pluripotent cells, this could provide an alternative source of cells for transplantation. The work described in this thesis therefore had two central aims (i) to test the stability of thymic epithelial progenitor cells in vivo and (ii) to investigate the possibility of generating TEPCs or TECs from mouse embryonic stem (ES) cells. The forkhead transcription factor, Foxn1, is essential for the development of a functionally mature thymic epithelium, but is not necessary for formation of the thymic primordium or for medullary thymic epithelial sub-lineage specification. By reactivating Foxn1 expression postnatally in mice carrying a revertible hypomorphic allele of Foxn1, Foxn1R, I herein demonstrate that TEPCs that can express only low levels of Foxn1 mRNA can persist postnatally in the thymic rudiment in mice until at least 6 months of age, and retain the potential to give rise to both cortical and medullary thymic epithelial cells (cTECs and mTECs). These data demonstrate that the TEPC-state is remarkably stable in vivo under conditions of low Foxn1 expression. In parallel with this work, I confirmed the possibility of generating Foxn1-expressing cells from mouse ES cells by using a Foxn1 reporter cell line. As the thymic epithelium has a single origin in the third pharyngeal pouch (3pp) endoderm, I then tested whether or not TEPCs and /or TECs were generated during ES cell differentiation via existing protocols for generating anterior definitive endoderm differentiation cells from mouse ES cells. From this work, I showed that genes expressed in the 3pp and/or TEPC,-including Plet-1, Tbx1, Hoxa3 and Pax9, were induced by differentiation of ES cells using these protocols. I further showed that cells expressing both Plet-1, a marker of foregut endoderm and 3pp, and EpCAM, a marker of proliferating epithelial cells, were induced using a novel protocol (2i ADE) for generating ES cells from ADE. However, gene expression analysis and functional testing suggested that the majority of these cells were non-thymus lineage. I subsequently developed a novel protocol which combined this 2i ADE protocol with co-culturing of the differentiating ES cells with fetal thymic lobes, and demonstrated that this further induced 3pp and TEPC related genes. Finally, I modified the culture conditions in this protocol to conditions predicted to better support TEPC/TEC, and showed that in these conditions, the TEPC-specific markers Foxn1 and IL-7 were induced more strongly than in any other conditions tested. The data presented in this thesis therefore represent an advance towards an optimized protocol for successfully generating TEPCs from ES cells in vitro.