TBN1 is a nuclease with antitumor activity. The main goal of this work was to estimate how TBN1 and its modificated variants are stable in the ?leaf factory? system used for its production and whether it can be enhanced or influenced by chosen potential ?modificators? i. e. silencing supressors, transcription factors, glycosyltransferases and kinases. Nicotiana benthamina plants were infiltrated with the mixture of Agrobacterium tumefaciens strains bearing the nuclease plant expression vectors and co-infiltrated with the ?modifying? vectors. The nuclease and protein analyses revealed that nuclease TBN1 wt and its modificated variants are stable in the used ?leaf factory? system as to their molecular mass, only quantitative changes were detected. Expreximents showed that activity and production of the nucleases increased upon coinfiltration with silencing supressor and decreased upon coexpression with chosen transcription factor. Glycosyltransferases and kinases influenced activity and production only insignificantly. The experiments also revealed that modificated variants of TBN1 have different molecular weight suggesting that different N-glycosylation domains have different length of sugar chain and influence on nuclear activity. Our data show that this expression in planta seems to be suitable for production for study of antitumor activity of these nucleases.

Résumé : Au repos, l'activité du cerveau d'un humain sain est caractérisée par de larges fluctuations dans la bande de fréquences de 8-13 Hz d'un électroencéphalogramme (EEG), connue sous le nom de bande alpha. Bien qu'il soit établi que son activité varie d'un individu à l'autre, peu d'études se sont intéressées à la façon dont elle peut être reliée aux variations morphologiques des structures du cerveau. Entre autres, on pense que le corps géniculé latéral (CGL) et ses fibres efférentes (la radiation optique) jouent un rôle clé sur l'activité alpha, bien qu'il n'est pas certain que leur forme ou leur grosseur contribuent à sa variabilité inter-individuelle. Considérant l'utilisation courante d'EEG dans la recherche fondamentale ou clinique, ce sujet est important, mais difficile à traiter vu les problèmes associés à une bonne segmentation du CGL et de la radiation optique. Pour cette raison, nous avons utilisé la résonance magnétique de diffusion (IRMd), la résonance magnétique fonctionnelle (IRMf) et l'EEG sur 20 sujets sains pour mesurer la structure et la fonction, respectivement. L'analyse de la structure a nécessité une nouvelle approche semi-automatique pour segmenter le CGL et la radiation optique, qui nous a permis de mesurer plusieurs variables, telles que le volume et la position. Ces mesures correspondent bien aux connaissances sur la morphologie de ces structures basées sur des études post-mortem, et pourtant, nous avons trouvé que leur variabilité inter-sujet n'influençait pas la puissance des ondes alpha ou leur fréquence-type (p>0.05). Ces résultats suggèrent que la variabilité alpha soit médiée par d'autres sources structurelles. Notre méthodologie pourra servir pour de futures recherches sur l'influence de l'anatomie sur la fonction en IRMf, tomographie par émission de positron (TEP), EEG, etc., ou pour améliorer les recherches cliniques sur la radiation optique. / Abstract : At rest, healthy human brain activity is characterized by large electroencephalography (EEG) fluctuations in the 8-13 Hz range, commonly referred to as the alpha band. Although it is well known that EEG alpha activity varies across individuals, few studies have investigated how this may be related to underlying morphological variations in brain structure. Specifically, it is generally believed that the lateral geniculate nucleus (LGN) and its efferent fibres (optic radiation, OR) play a key role in alpha activity, yet it is unclear whether their shape or size variations contribute to its inter-subject variability. Given the widespread use of EEG alpha in basic and clinical research, addressing this is important, though difficult given the problems associated with reliably segmenting the LGN and OR. For this, we employed a multi-modal approach and combined diffusion magnetic resonance imaging (dMRI), functional magnetic resonance imaging (fMRI) and EEG in 20 healthy subjects to measure structure and function, respectively. For the former, we developed a new, semi-automated approach for segmenting the OR and LGN, from which we extracted several structural metrics such as volume, position and diffusivity. Although these measures corresponded well with known morphology based on previous post-mortem studies, we nonetheless found that their inter-subject variability was not significantly correlated to alpha power or peak frequency (p > 0.05). Our results therefore suggest that alpha variability may be mediated by an alternative structural source and our proposed methodology may in general help in better understanding the influence of anatomy on function.

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Medium Mn TRIP steels are amongst the most widely researched third generation advanced high strength steels (3G-AHSSs) as they are ideal candidates for automotive light-weighting applications owing to their superior strength and ductility balance. However, the thermal processing cycles of these steels need to be compatible with the industrial continuous galvanizing line (CGL) in order to successfully employ them in the automotive manufacturing industry. The main objective of the present research was to develop a CGL compatible thermal processing cycle for a prototype medium Mn steel that would produce significant volume fractions of chemically stable retained austenite and exhibit mechanical properties consistent with established 3G-AHSS targets. In that regard, the effects of intercritical annealing (IA) time and temperature and starting microstructure were determined in the first part of this research. The as-received tempered martensite (S-TM) and heat treated martensite (S-M) were the two different starting microstructures studied in this research. In this case, the overaging temperature (OT) treatment (460°C for 20s) was kept constant. It was found that high volume fractions (≥ 0.30) of retained austenite were achieved for S-M samples intercritically annealed at 675°C for shorter times (i.e. 60 to 120s) compared to S-TM samples. TEM analysis of the S-M samples showed that most of the retained austenite was present in a film type morphology, which is known to be more stable chemically and mechanically compared to the block type morphology. The tensile test results showed that although both the S-TM and S-M samples exhibited a high strength/ductility balance, the S-M samples, particularly the S-M 675°C + 120s samples, showed more potential in terms of CGL compatibility and achieving 3G-AHSS target mechanical properties. The effect of OT holding time was determined in the second part of this research. In that regard, the OT holding time was varied form 20s to 120s for selected S-TM and S-M samples. The S-TM 710°C samples with increased OT holding times (60s and 120s) had a significant increase in retained austenite volume fraction compared to the baseline 20s OT samples. However, the retained austenite volume fractions did not change for S-M samples regardless of OT holding time. It was also found that the mechanical properties of the annealed S-TM and S-M steels depended on the OT holding time. For the S-TM samples with > 120s IA holding times, longer OT holding times (60s and 120s) produced chemically unstable retained austenite which transformed rapidly at low strain resulting in low UTS × TE products. However, although longer OT holding times significantly increased the yield strength of the annealed S-M samples, the UTS × TE product decreased significantly owing to decreased retained austenite stability. Finally, based on the results of this research, it was concluded that the prototype medium Mn TRIP steel can achieve 3G-AHSS target mechanical properties using CGL-compatible thermal processing cycles. Moreover, depending on successful reactive wetting, it may be possible to perform both thermal processing and galvanizing of this steel in the industrial CGL. / Thesis / Master of Applied Science (MASc)