published_or_final_version / Medicine / Master / Doctor of Medicine

published_or_final_version / Medicine / Master / Doctor of Medicine

Absolute quantification of I-131 activity in tumors and normal tissue is essential for internal dose estimates and is one of the greatest challenges in contemporary Nuclear Medicine. Therefore, for obtaining more accurate results, the most suitable and clinical applicable method, for both patients and scientists, for the quantification of I-131 has to be found. In the theoretical part of this project, the two methods for the quantitation are explained and analyzed as well as the factors that affect the obtained clinical image followed by the corrections that must be applied for gaining a more discrete image. For the planar technique, two methods for scatter and septal penetration correction were used applying two different matrix sizes specifically, 256x256 and 512x512 matrix sizes. On the other hand, for the SPECT technique, a 64x64 matrix size was used and a comparison between an auto-ROI and a same pattern of ROI segmentation that was produced manually for the purpose of this project, was performed. Furthermore, for patient specific dosimetry the total absorbed dose for the thyroid gland was calculated. Figures and tables for planar and SPECT technique were produced for each matrix size along with the final curve of the absorbed dose of the patient. For the planar technique it was observed that the method proposed by Anne Larsson, according to statistical analysis, gives results with lower statistical errors than the method proposed by Macey. In addition, 512x512 matrix size for the planar technique leads to lower statistical errors when compared with the results from using the 256x256 matrix size. Moreover, for the SPECT technique, the auto-sum that the software of the camera provides gives exactly the same results compared with the manual-sum. Finally, the total thyroid absorbed dose was 45,0472 Gy a result very close to the ICRU proposed dose value. / Η απόλυτη ποσοτικοποίηση της ενεργότητας του I-131 σε καρκινικούς και σε φυσιολογικούς ιστούς είναι απαραίτητη για την εκτίμηση της εσωτερικής δόσης και αποτελεί μία από τις μεγαλύτερες προκλήσεις της σύγχρονης Πυρηνικής Ιατρικής. Για τον λόγο αυτό, για την απόκτηση ακριβέστερων αποτελεσμάτων, θα πρέπει να βρεθεί η καταλληλότερη μέθοδος ποσοτικοποίησης, που να είναι εύχρηστη τόσο για τον ασθενή όσο και για το επιστημονικό προσωπικό των νοσοκομείων. Στο θεωρητικό μέρος της παρούσας εργασίας, αναλύονται οι δύο μέθοδοι ποσοτικοποίησης καθώς και οι παράγοντες που επηρεάζουν την ποιότητα της εικόνας ακολουθούμενοι από τους διορθωτικούς παράγοντες για την απόκτηση μιας ευκρινέστερης κλινικής εικόνας. Όσον αφορά την Planar τεχνική, περιγράφονται δύο μέθοδοι για διόρθωση της σκεδαζόμενης ακτινοβολίας καθώς επίσης και του septal penetration στην τελική εικόνα, χρησιμοποιώντας δύο διαφορετικά μεγέθη μήτρας, συγκεκριμένα, 256x256 και 512x512. Για την τεχνική SPECT, το μέγεθος μήτρας που χρησιμοποιήθηκε ήταν 64x64 και στην συνέχεια έγινε σύγκριση της περιοχής ενδιαφέροντος (ROI) που έδωσε το λογισμικό της κάμερας με την περιοχή ενδιαφέροντος που σχεδιάστηκε χειροκίνητα για την συγκεκριμένη μελέτη. Επίσης, υπολογίστηκε η συνολική απορροφούμενη δόση στον θυρεοειδή αδένα ενός ασθενούς. Κατασκευάστηκαν διαγράμματα τόσο για την planar όσο και για την SPECT τεχνική και για τα δύο μεγέθη μήτρας καθώς και η καμπύλη απορροφούμενης δόσης του ασθενούς. Παρατηρήθηκε ότι στην planar τεχνική, η μέθοδος που προτείνεται από την Anne Larsson, δίνει καλύτερα αποτελέσματα και μικρότερα σφάλματα συγκρινόμενη με την μέθοδο που προτείνεται από τον Macey. Επιπλέον, παρατηρήθηκε ότι το 512x512 μέγεθος μήτρας δίνει πιο ευκρινή εικόνα απ’ ότι το 256x256 μέγεθος μήτρας. Επίσης, με την SPECT τεχνική, παρατηρείται ότι χρησιμοποιώντας το αυτόματο άθροισμα των κρούσεων που δίνεται από το λογισμικό της κάμερας προκύπτουν τα ίδια ακριβώς αποτελέσματα με την χρήση του αθροίσματος των κρούσεων όταν αυτό γίνεται χειροκίνητα. Τέλος, η τελική απορροφούμενη δόση του ασθενούς υπολογίστηκε 45,0472 Gy, ένα αποτέλεσμα που δεν αποκλείει από τις τιμές που προτείνονται από το ICRU.

The incidence of differentiated thyroid cancer (DTC) is rising, but controversy exists in many aspects of its treatment. This study described the change in incidence of DTC in Ontario, variations in management including extent of thyroidectomy and the influence of provider volume, and the impact of these parameters on recurrence and thyroid cancer-specific death (TCSD). A population-based study identified all new cases of DTC between 1992-2007. The incidence of DTC increased dramatically (annual percentage change 7.6%). Linkage to administrative databases revealed that extent of thyroidectomy is influenced by various factors including patient gender, age, year of diagnosis, surgeon specialty, and hospital setting, but not provider volume. Total thyroidectomy is associated with a lower recurrence rate. There is a significant association between provider volume and recurrence, with lower volume surgeons having a higher recurrence risk. Extent of thyroidectomy and provider volume did not influence TCSD. Such variations in management may lead to disparities in health outcomes.

The incidence of differentiated thyroid cancer (DTC) is rising, but controversy exists in many aspects of its treatment. This study described the change in incidence of DTC in Ontario, variations in management including extent of thyroidectomy and the influence of provider volume, and the impact of these parameters on recurrence and thyroid cancer-specific death (TCSD). A population-based study identified all new cases of DTC between 1992-2007. The incidence of DTC increased dramatically (annual percentage change 7.6%). Linkage to administrative databases revealed that extent of thyroidectomy is influenced by various factors including patient gender, age, year of diagnosis, surgeon specialty, and hospital setting, but not provider volume. Total thyroidectomy is associated with a lower recurrence rate. There is a significant association between provider volume and recurrence, with lower volume surgeons having a higher recurrence risk. Extent of thyroidectomy and provider volume did not influence TCSD. Such variations in management may lead to disparities in health outcomes.

In vitro embryo production (IVP) has been successfully used for infertility treatment, farm animal breeding and preservation of endangered species. Embryo culture media and its composition play an important role in the developmental competency of in vitro produced embryos. To optimize IVP it is important to understand the physiology of the in vivo embryo medium. Previously, we confirmed the presence of thyroid hormones (THs) in the bovine reproductive tract and their beneficial effect on embryo quality in vitro. The aim of this study was to further investigate the developmental effect and mechanisms behind THs function in bovine early embryos by supplementing IVP media with 50ng/ml T3 and T4. In vitro embryo culture (IVC) media fortification with THs significantly improved blastocyst and hatching rates, which was observed in both fast and slow cleaving embryos. THs treatment significantly skewed male/female ratio toward femaleness (43% vs. 57%). Only %60 of supplemented THs was bioavailable in the IVC media. Gene microarray and blastocyst genes expression profile analysis showed that THs supplementation altered mRNA expression profile in blastocysts. 1,234 genes were expressed differentially in the treated embryos and these differences were statistically significant (>1.5 fold at p < 0.05); these findings were confirmed by qPCR. TH-related genes, including THs receptor (TRs) mRNA and deiodinases (DIOs), were expressed in the gene array data of both treated and control groups. TR mRNA was expressed in all stages of early embryo development (EED) and the expression was not inducible by THs. This finding was consistent in both gene array and qPCR. Suppression of embryo gene transcription by α-amanitin resulted in a significantly higher level of TR mRNA from 2-cell to 16-cell treated and control embryos. TR protein was expressed in both treated and control blastocysts but, differentially located and distributed in response to THs in the blastocysts. This study highlights the importance of THs in EED and therefore the need to include these factors in culture media. Furthermore, this study has better characterized THs effects and identified possible regulatory roles of these hormones in EED. Further investigation into embryo competency and pregnancy outcome can provide more details. / Canada Research Chair(CRC), EmbryoGene, Ontario Graduate Scholarship(OGS) and Agriculture and Agrifood Canada.

Section I: This section is an introduction to the field of nuclear receptors. A general overview of nuclear receptor-mediated transcriptional regulation is followed by a review of literature on retinoid and thyroid receptor-mediated signaling. Section II: An introduction to NPDC-1 (neural proliferation, differentiation, and control), its discovery and characterization with regards to developmental expression and cellular localization. In addition NPDC-1 has been found to associate with a number of cell cycle regulatory proteins. NPDC-1 is characterized as a regulator of nuclear receptor-mediated transcriptional regulation. NPDC-1 was also demonstrated to be regulated post-transcriptionally through the ubiquitin/proteosome degradation pathway. Section III: Axolotl NPDC-1 (aNPDC-1) was cloned from axolotl brain and analyzed for homology to NPDC-1 from higher vertebrates. The tissue distribution and developmental expression of axolotl NPDC-1 were also examined. Section IV: The axolotl homolog for RAR (aRAR) was isolated from axolotl brain. Axolotl NPDC-1 and aRAR were then examined in a series of assays for interactions. Axolotl NPDC-1 was found to repress transcription mediated by aRAR to a smaller extent than the repression observed in higher vertebrates. The DNA binding of aRAR-RXR was increased in the presence of aNPDC-1 and complex mobility was also observed. The domain of interaction between aNPDC-1, aRAR and hRXR was localized in the amino terminus of aNPDC-1. Axolotl NPDC-1 was also demonstrated to repress proliferation as measured by reduced [3H] thymidine incorporation. Section V: The axolotl homologs of TR and TR (aTR) genes were isolated and utilized in a series of experiments to demonstrate an interaction between aTRs and aNPDC-1. As observed for RE, aNPDC-1 increases the binding of aTR-RXR heterodimer to xDR4, but no change in the mobility of the complex was observed. Interaction between aNPDC-1, aTR and aTR was localized to the amino terminus of aNPDC-1. In contrast to previous observations for other nuclear receptors, aNPDC-1 was found to stimulate transcription mediated by axolotl TRs, suggesting a role for aNPDC-1 in axolotl metamorphosis. Section VI: A summary of data presented in the previous sections as well as a presentation of future directions and a proposed model for NPDC-1 actions in retinoid and thyroid-receptor mediated signaling in axolotl.

Microarray technology allows researchers to monitor the mRNA transcription levels of thousands of genes in parallel which opens the door for more advanced cancer research. This thesis focuses on a case study of papillary thyroid carcinoma data. Fourteen publicly available Affymetrix microarray data sets were used where seven samples were collected from normal thyroid tissue and the remaining seven were collected from papillary thyroid carcinoma tissue. The present study compared the results obtained from three different normalization processes: MAS5.0, RMA and GCRMA in detecting differentially expressed genes under two conditions. Internal consistencies within the methods as well as the results across three methods were compared. Statistical packages 82.5.1 and Bioconductor 2.08 are used to perform the data analysis. Each step of normalization with MAS5.0 and RMA is described. Statistical package Limma is used to fit a linear model. Finally an empirical Bayes method is used to detect the significantly differentially expressed genes. First, considering all genes a comparison is made among the three normalization methods where RMA and GCRMA showed the maximum agreement in detecting differentially expressed genes. Then using unspecified filtering process a set of genes was selected and the whole process was replicated where the top fifty differentially expressed genes did not show any overlap with each other. / Department of Mathematical Sciences

There is a growing interest in assessing the neurotoxic potential and endocrine disrupting properties of perfluoroalkyl compounds (PFCs). Several studies have reported in vitro and in vivo effects related to neuronal development, neural cell differentiation, pre- and post- natal development and behaviour. PFC exposure altered hormone levels (e.g. thyroid hormone, estrogen, and testosterone) and the expression of hormone-responsive genes in mammalian and aquatic species. Hormone-mediated events are critical in central nervous system development and function, especially those controlled by thyroid hormones (THs). The studies presented in this thesis are the first to assess the effects of PFCs on primary cultures of neuronal cells in two avian species; the domestic chicken (Gallus domesticus) and herring gull (Larus argentatus). The following TH-responsive genes were examined using real-time RT-PCR: type II iodothyronine 5’-deiodinase (D2), D3, transthyretin (TTR), neurogranin (RC3), octamer motif binding factor (Oct-1), and myelin basic protein (MBP). Several PFCs were shown to alter mRNA expression levels of genes associated with the TH pathway in avian neuronal cells. It was determined that short-chained PFCs (<8 carbons) altered the expression of TH-responsive genes to a greater extent than long-chained PFCs (≥8 carbons). Although several significant changes in mRNA expression were observed in TH-responsive genes following PFC exposure in chicken embryonic neuronal (CEN) cells (Chapter 2), there were fewer changes in herring gull embryonic neuronal (HGEN) cells (Chapter 3). The mRNA levels of D2, D3, TTR, and RC3 were altered following treatment with several short-chained PFCs in CEN cells. Oct-1 and RC3 expression were induced following treatment with several short-chained PFCs in HGEN cells. These studies are the first to report that PFC exposure alters mRNA expression in primary cultures of avian neuronal cells and provide insight into the possible mechanisms of action of PFCs in the avian brain.

Current municipal wastewater treatment plants (MWWTP) technologies are insufficiently removing emerging contaminants of concern. These emerging contaminants are an issue as many are known endocrine disrupting compounds (EDCs). EDCs are contaminants that can have severe and irreversible impacts on highly conserved endocrine systems that are critical during developmental periods in vertebrates as well as during adult life. Many EDCs have non-monotonic dose-response curves yet they are not often tested at low, environmentally relevant concentrations. EDC research to date has focused heavily on xenoestrogenic compounds whereas thyroid hormone (TH) disruption has been largely overlooked. TH is conserved in all vertebrates and plays crucial roles in neural development, basal metabolism, and thermoregulation. TH is comprised of thyroxine (T4), often known as the transport form of TH, and triiodothyronine (T3), the more bioactive form of TH. A TH spike occurs in the perinatal period of humans, and when disrupted, this spike can cause severe developmental defects. An analogous, but perhaps more overt, TH spike occurs in amphibians. TH is the sole hormone that drives amphibian metamorphosis, thus providing an excellent model for TH action. Our lab has previously developed the cultured tailfin (C-fin) assay, which uses biopsies from premetamorphic Rana catesbeiana tadpole tailfins cultured in the presence of an exogenous chemical of concern to assess perturbations to TH- and stress-responsive gene transcript levels by QPCR. This thesis uses the C-fin assay to assess the efficacy of removal of biological TH- and stress-altering activity in conventional municipal wastewater treatment systems. We first assess the successive levels of a full-scale conventional activated sludge (CAS) MWWTP in its ability to reduce perturbations of mRNA transcript levels of the critical TH receptors alpha (thra) and beta (thrb), and stress responsive gene transcripts superoxide dismutase (sod), catalase (cat) and heat shock protein 30 (hsp30). Secondary treatment of wastewater effluents removes cellular stress perturbations when compared to influents, but thr disruptions remain after conventional secondary wastewater treatment. We then assess three pilot-sized conventional secondary MWWTP configurations run at two operational conditions. The C-fin assay results suggest that the current understanding of operational conditions and the efficiency of complex MWWTP configurations is not clear-cut when assessed by biological endpoints such as the transcript abundance perturbations in the C-fin assay. Finally, the C-fin assay is used to investigate transcript profiles of genes of interest when the tissues are treated with the endogenous hormones T3, T4, and estradiol (E2). Our results indicate that T4 acts as more than solely a T3-prohormone and that gene expression levels in response to the two different forms of TH can be T3 or T4 specific. E2 effects, although implicated in altering TH-mediated responses in other contexts, do not affect TH-responsive gene transcripts in the C-fin. The data presented use the novel C-fin assay to challenge and advance the currently accepted views of TH-action, as well as develop necessary yet practical biological knowledge for management of emerging contaminant release from MWWTPs. / Graduate / 0383 / 0768 / 0307 / polaw@uvic.ca