Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

This paper explores the fit of a stochastic volatility model, in which the Box-Cox transformation of the squared volatility follows an autoregressive Gaussian distribution, to the continuously compounded daily returns of the Australian stock index. Estimation was difficult, and over-fitting likely, because more variables are present than data. We developed a revised model that held a couple of these variables fixed and then, further, a model which reduced the number of variables significantly by grouping trading days. A Metropolis-Hastings algorithm was used to simulate the joint density and derive estimated volatilities. Though autocorrelations were higher with a smaller Box-Cox transformation parameter, the fit of the distribution was much better.

Oxidative phosphorylation apparatus (OXPHOS) is responsible for production of majority of ATP in mammalian organisms. This process, occurring in the inner mitochondrial membrane, is partly regulated by nuclear-encoded subunits of cytochrome c oxidase (COX), the terminal enzyme of electron transport chain. Cox4 subunit, participating in OXPHOS regulation, is an early-assembly state subunit, which is necessary for incorporation of Cox2 catalytic subunit, thus for assembly of catalytically functional COX enzyme. Moreover, regulated expression of two isoforms (Cox4i1, Cox4i2) of Cox4 subunit is hypothesized to optimize respiratory chain function according to tissue oxygen supply. However, the functional impact of the isoform switch for mammalian tissues and cells is still only partly understood. In the present thesis, unique HEK293 cell line-based model with complete absence of subunit Cox4 (knock-out, KO) was prepared employing novel CRISPR CAS9-10A paired nickase technology and further characterized. Knock-out of both isoforms Cox4i1 and Cox4i2 (COX4i1/4i2 KO clones) showed general decrease of majority of Cox subunits resulting in total absence of fully assembled COX. Moreover, detected Complex I subunits as well as the content of assembled Complex I were decreased in COX4i1/4i2 KO clones. On the...

Mehrere Studien haben gezeigt, daß die Cyclooxygenase-2 (COX-2) eine bedeutende Rolle sowohl bei Entstehung als auch Progression maligner Tumoren spielt. COX-2-Inhibitoren werden bereits in klinischen Studien zur Krebstherapie getestet. COX-2 ist die induzierbare Isoform der Cyclooxygenase - dem Schlüsselenzym der Synthese von Prostaglandinen und anderen Eicosanoiden. Im Tier- und Zellkulturmodell konnten COX-Hemmer anti-Tumor-Effekte hervorrufen. Es ist jedoch unklar, ob diese Effekte durch Hemmung des COX-Enzyms oder durch COX-unabhängige Mechanismen vermittelt werden. Wir untersuchten daher die Auswirkung der COX-Inhibition zum einen durch den selektiven COX-2-Hemmer NS-398 sowie zum anderen durch COX-Isoform-spezifische RNA-Interferenz (RNAi) in zwei humanen Ovarialkarzinomzellinien (OVCAR-3 und SKOV-3). OVCAR-3 zeigte eine konstitutive COX-1-Expression und eine durch IL-1beta induzierbare COX-2-Expression. SKOV-3 war COX-1- und COX-2-negativ. IL-1beta führte bei OVCAR-3 zu einer vermehrten Produktion von Prostaglandin E2 (PGE2), die durch eine gegen die COX-2 gerichtete siRNA gehemmt werden konnte, wohingegen COX-1-siRNA keinen Effekt hatte. Das deutet darauf hin, daß die COX-2 die Hauptquelle von PGE2 in OVCAR-3 ist. 1mikroM NS-398 waren ausreichend, um die PGE2-Produktion und somit auch die COX-2 in OVCAR-3 zu inhibieren. Höhere Konzentrationen NS-398 (>10mikroM) hatten einen antiproliferativen Effekt. Auch in der COX-2-negativen Zellinie SKOV-3 trat diese Wachstumshemmung auf; sie war nicht durch exogene Zufuhr von PGE2 (10mikroM) reversibel. Durchflußzytometrische Zellzyklusanalyse ergab, daß der Wachstumshemmung in beiden Zellinien ein G0/G1-Zellzyklusarrest zugrunde liegt. Dagegen führten weder COX-1- noch COX-2-Ausschaltung durch RNAi zu ähnlichen Auswirkungen auf Proliferation bzw. Zellzyklus. Diese Ergebnisse zeigen, dass ein COX-2-unabhängiger Mechanismus für den durch NS-398 induzierten G0/G1-Arrest verantwortlich ist. / Several studies have provided evidence that the enzyme Cyclooxygenase-2 (COX-2) plays an important role in tumor development and progression. COX-2-inhibitors are already evaluated in clinical trials as cancer therapeutics. COX-2 is the inducible isoform of cyclooxygenase - the rate-limiting enzyme in the synthesis of prostaglandins and other eicosanoids. COX-inhibitors cause antitumor effects in animal models and in cell culture experiments. However, it is not clear, whether these effects are due to inhibition of the COX-enzyme or mediated via a COX-independent mechanism. We therefore investigated the effects of COX inhibition by the selective COX-2-inhibitor NS-398, as well as by COX-isoform specific RNA interference (RNAi) in the human ovarian carcinoma cell lines OVCAR-3 and SKOV-3. OVCAR-3 cells showed a constitutive expression of COX-1, and an inducible COX-2 expression. COX-2 was induced through stimulation with Interleukin-1beta, leading to production of high levels of Prostaglandin E2 (PGE2). SKOV-3 cells were negative for both COX isoforms. Selective COX-2-suppression by RNAi reduced PGE2 production in OVCAR-3, whereas COX-1-siRNA had no effect on PGE2 synthesis. Thus, COX-2 is the main source of PGE2 in OVCAR-3 cells. In these cells, 1microM NS-398 was sufficient to completely inhibit PGE2-synthesis - and thus the activity of the COX-2 enzyme. Increasing amounts of NS-398 (>10microM) had an antiproliferative effect. This growth inhibition was also observed in the COX-negative cell line SKOV-3, it could not be reverted by exogenous addition of PGE2 (10microM). Flowcytometric analysis of the cell cycle revealed that this growth inhibition was based on a G0/G1-cell-cycle-arrest. In contrast, suppression of COX-1 or COX-2 by RNAi had no effect on proliferation or cell cycle progression. These results suggest that a COX-independent mechanism is responsible for the G0/G1-arrest induced by NS-398.

Resumo:As neoplasias testiculares caninas são achados ocasionais. São descritas como benignas, mas podem metastatizar ou mostrar características de malignidade com o avanço da idade do animal, o que torna necessário o melhor entendimento do comportamento destas neoplasias no cão. O objetivo deste estudo foi caracterizar a expressão imuno-histoquímica de Ki-67, COX-2, MMP-9 e p53 em neoplasias testiculares de 50 cães, verificar a relação entre o padrão histológico e raça, idade e posicionamento testicular e verificar a expressão imuno-histoquímica dos anticorpos avaliados nos diferentes tipos de neoplasia. A avaliação imuno-histoquímica das proteínas Ki-67, MMP-9 e p53 foi mais intensa nos seminomas, enquanto que os leydigocitomas exibiram maior marcação para COX-2. A avaliação histológica e imuno-histoquímica dos subtipos de seminomas em cães, particularmente o seminoma difuso, podem resultar em diferença significativa que permita a utilização destas proteínas como marcadores de prognóstico. As características histopatológicas em associação ao histórico dos animais e à expressão das proteínas estudadas podem contribuir para a caracterização do comportamento biológico nas neoplasias testiculares caninas / Abstract:testicular neoplasms are sporadic findings. They are described as benign, but they can metastasize or express malignant features with the animal ageing, which makes necessary the better understanding of theses neoplasms behavior in dogs. The aim of this study was to characterize the immunoexpression of Ki-67, COX-2, MMP-9 and p53 in testicular neoplasms of fifty dogs, to verify the relation among histological pattern of the neoplasms and breed, age and testicular localization; and also to verify the immunoexpression of these markers in different types of testicular tumours. Ki-67, MMP-9 and p53 immunostaining were more intense in seminomas, whereas COX-2 presented a more intense staining in Leydig cell tumors. The histological and immunohistochemical analyses of the subtypes of canine seminomas, especially the diffuse seminoma, may result in significant differences which would allow the use of these markers as prognostic factors. The histopathological features associated with the day-history of the animals and with these markers may contribute to the characterization of the biological behavior of canine testicular neoplasms / Orientador:Gisele Fabrino Machado / Banca:Felipe Augusto Ruiz Sueiro / Banca:Rosemary de Oliveira Vasconcelos / Mestre

This thesis focuses on two fundamental topics, specifically in medical statistics: the modelling of correlated survival datasets and the variable selection of the significant covariates and random effects. In particular, two types of survival data are considered: the classical survival datasets, where subjects are likely to experience only one type of event and the competing risks datasets, where subjects are likely to experience one of several types of event. In Chapter 2, among other topics, we highlight the importance of adding frailty terms on the proposed models in order to account for the association between the survival time and characteristics of subjects/groups. The main novelty of this thesis is to simultaneously select fixed effects and frailty terms through the proposed statistical models for each survival dataset. Chapter 3 covers the analysis of the classical survival dataset through the proposed Cox Proportional Hazard (PH) model. Utilizing a Cox PH frailty model, may increase the dimension of variable components and estimation of the unknown coefficients becomes very challenging. The method proposed for the analysis of classical survival datasets involves simultaneous variable selection on both fixed effects and frailty terms through penalty functions. The benefit of penalty functions is that they identify the non-significant parameters and set them to have a zero effect in the model. Hence, the idea is to 'doubly-penalize' the partial likelihood of the Cox PH frailty model; one penalty for each term. Estimation and selection implemented through Newton-Raphson algorithms, whereas closed iterative forms for the estimation and selection of fixed effects and prediction of frailty terms were obtained. For the selection of frailty terms, penalties imposed on their variances since frailties are random effects. Based on the same idea, we further extend the simultaneous variable selection in the competing risks datasets in Chapter 4, using extended cause-specific frailty models. Two different scenarios are considered for frailty terms; in the first case we consider that frailty terms vary among different types of events (similar to the fixed effects) whereas in the second case we consider shared frailties over all the types of events. Moreover, our 'individual penalization' approach allows for one covariate to be significant for some types of events, in contrast to the frequently used 'group-penalization' where a covariate is entirely removed when it is not significant over all the events. For both proposed methods, simulation studies were conduced and showed that the proposed procedure followed for each analysis works well in simultaneously selecting and estimating significant fixed effects and frailty terms. The proposed methods are also applied to real datasets analysis; Kidney catheter infections, Diabetes Type 2 and Breast Cancer datasets. Association of the survival times and unmeasured characteristics of the subjects was studied as well as a variable selection for fixed effects and frailties implemented successfully.

No description available.

Lung cancer is the most frequent fatal cancer in the United States. Following the notion in actuarial math analysis, we assume an exponential form for the baseline hazard function and combine Cox proportional hazard regression for the survival study of a group of lung cancer patients. The covariates in the hazard function are estimated by maximum likelihood estimation following the proportional hazards regression analysis. Although the proportional hazards model does not give an explicit baseline hazard function, the baseline hazard function can be estimated by fitting the data with a non-linear least square technique. The survival model is then examined by a neural network simulation. The neural network learns the survival pattern from available hospital data and gives survival prediction for random covariate combinations. The simulation results support the covariate estimation in the survival model.

The abuse of methylenedioxymethamphetamine (MDMA) during pregnancy is of concern. MDMA treatment of rats during a period of brain growth analogous to late human gestation leads to neurochemical and behavioral changes. MDMA from postnatal day (P)11–20 in rats produces reductions in serotonin and deficits in spatial and route-based navigation. In this experiment we examined the impact of MDMA from P11 to P20 (20 mg/kg twice daily, 8 h apart) on neuronal architecture. Golgi impregnated sections showed significant changes. In the nucleus accumbens, the dendrites were shorter with fewer spines, whereas in the dentate gyrus the dendritic length was decreased but with more spines, and for the entorhinal cortex, reductions in basilar and apical dendritic lengths in MDMA animals compared with saline animals were seen. The data show that neuronal cytoarchitectural changes are long-lasting following developmental MDMA exposure and are in regions consistent with the learning and memory deficits observed in such animals.

To increase diversity of enzymes and proteins, cells mix and match exonic and intronic regions retained in mature mRNAs by alternative splicing. An estimated 94% of all multi-exon genes express one or more alternatively spliced transcripts generating proteins with similar or modified functions. Cyclooxygenase is a signaling enzyme that catalyzes the rate-limiting step in the synthesis of diverse bioactive lipids termed prostaglandins. Prostaglandins are involved in myriad physiological and pathopysiological processes including vasoregulation, stomach mucosal maintenance, parturition, pain, fever, inflammation, neoplasia and angiogenesis and are inhibited by aspirin-like drugs known as NSAIDs. In 2002 an alternatively spliced, intron-1 retaining variant of COX-1 was cloned from canine brain tissue. This new variant, termed COX-3 or COX-1b, is an enzymatically active prostaglandin synthase expressed at relatively high levels in a tissue and cell type dependant manner in all species examined. In humans and most rodent species intron-1 is 94 and 98 nucleotides long respectively. Retention of the intron in these species introduces a frameshift and is predicted to result in translation of a very small 8-16kD protein with little similarity to either 72kD COX-1 or COX-2, calling into question the role of this variant. In this dissertation, I present my results from cloning and ectopically expressing a complete and accurate COX-3 cDNA from both rat and human. I confirmed that COX-3 mRNA encodes multiple large molecular weight cyclooxygenase-like proteins in the same reading frame as COX-1. Translation of these proteins relies on several recoding mechanisms including cap-independent translation initiation, alternative start site selection, and ribosomal frameshifting. Using siRNA and Western blotting I have identified some of these proteins in tissues and cells. Two COX-3 encoded proteins are active prostaglandin synthase enzymes with activities similar to COX-1 and represent novel targets of NSAIDs. Other COX-3 proteins have unknown function, but their size and cellular location suggest potential roles as diverse as cytosolic enzymes and nuclear factors.