Coregulator Control of Androgen Receptor Action by a Novel Nuclear Receptor-Binding Motif

The androgen receptor (AR) is a ligand-activated transcription factor that is essential for prostate cancer development. It is activated by androgens through its ligand-binding domain (LBD), which consists predominantly of 11 α-helices. Upon ligand binding, the last helix is reorganized to an agonist conformation termed activator function-2 (AF-2) for coactivator binding. Several coactivators bind to the AF-2 pocket through conserved LXXLL or FXXLF sequences to enhance the activity of the receptor. Recently, a small compound-binding surface adjacent to AF-2 has been identified as an allosteric modulator of the AF-2 activity and is termed binding function-3 (BF-3). However, the role of BF-3 in vivo is currently unknown, and little is understood about what proteins can bind to it. Here we demonstrate that a duplicated GARRPR motif at the N terminus of the cochaperone Bag-1L functions through the BF-3 pocket. These findings are supported by the fact that a selective BF-3 inhibitor or mutations within the BF-3 pocket abolish the interaction between the GARRPR motif(s) and the BF-3. Conversely, amino acid exchanges in the two GARRPR motifs of Bag-1L can impair the interaction between Bag-1L and AR without altering the ability of Bag-1L to bind to chromatin. Furthermore, the mutant Bag-1L increases androgen-dependent activation of a subset of AR targets in a genome-wide transcriptome analysis, demonstrating a repressive function of the GARRPR/BF-3 interaction. We have therefore identified GARRPR as a novel BF-3 regulatory sequence important for fine-tuning the activity of the AR.

Transgenic tomato plants expressing the antigen gene PfCP-2.9 of Plasmodium falciparum

The objective of this work was to obtain transgenic tomato plants expressing the PfCP-2.9 protein (a chimera of the antigens MSP1 and AMA1 of Plasmodium falciparum). Cotyledons of seven-day-old tomatoes, cultivar Summers, were transformed via Agrobacterium tumefaciens. Transgenic expression in the T0 plants was verified in the DNA extracted from fruits. PCR analysis was used to test the presence of the gene of interest in the T1 generation. Reverse transcriptase PCR provided evidence of gene expression at the RNA level, and Western blot analysis confirmed the presence of the protein of interest in the T1 plants. This is the first report of successful transformation with the expression of a malaria antigen (PfCP-2.9) in transgenic tomato plants from the T0 and T1 generations.

Absorptive capacity and the effect of social integration mechanisms in knowledge utilization

Tiedosta on tullut määräävä tekijä yrityksensuorituskyvylle. Yritykset hankkivat aktiivisesti uutta tietoa ulkoisesta ympäristöstään ja tallentavat sitä tietokantoihinsa. Uusi tieto on innovaatioiden ja uusien ideoiden peruselementti. Uudet ideat pitää myös kaupallistaa, jotta niiden avulla voidaan hankkia kilpailuetua. Absorptiivisen kapasiteetin malli yhdistää tiedon prosessointiin liittyvät kyvykkyydet, jotka vaikuttavat yrityksen kykyyn hyödyntää tietoa tehokkaasti. Ennen kuin tietoa voidaan käyttää uusien tuotteiden ja palveluiden luomiseen, täytyy sitä jakaa yrityksessä ja muuttaa se yrityksen toimintaa palvelevaksi. Aiemmissa tutkimuksissa innovaatiot ovat vahvasti liitetty yrityksen kykyyn uudistua. Tämä pro gradu -tutkielma tutkii sosiaalisten integraatiomekanismien vaikutusta potentiaalisen absorptiivisen kapasiteetin muuttamiseen toteutuneeksi absorptiiviseksi kapasiteetiksi. Yksilöiden ja osastojen välisen yhteistyön sekä luottamuksen vaikutus tiedon sisäistämiseen tutkittiin. Tutkielma pohjautuu monikansallisessa yrityksessä keväällä 2006 suoritettuun uudistumiskyky-tutkimukseen. Tutkielma keskittyy yrityksen kykyyn uudistua uuden tiedon ja innovaatioiden avulla. Tutkielma on kvantitatiivinen tapaustutkimus. Tutkielmassa tehtyjen havaintojen mukaan sosiaaliset integraatiomekanismit ovat tärkeitä uuden tiedon hyödyntämisessä. Tiedon eksplisiittyyden havaitaan vaikuttavan tiedon muuttamiseen yritykselle hyödylliseksi resurssiksi.

Functions of the human papillomavirus type 16 E4 protein

1. Abstract Cervical cancer is thought to be the consequence of infection by human papillomaviruses (HPV). In the majority of cases, DNA from HPV type 16 (HPV16) is found in malignant cervical lesions. The initial steps leading to transformation of an infected cell are not clearly understood but in most cases, disruption and integration of the episomal viral DNA must take place. As a consequence, the E2 and E4 genes are usually not expressed whereas the E6 and E7 oncogenes are highly expressed. However, in a normal infection in which the viral DNA is maintained as an episome, all viral genes are expressed. The pattern according to which the viral proteins are made, and therefore the life cycle of the virus, is tightly linked to the differentiation process of the host keratinocyte. The study of the viral oncogenes E6 and E7 has revealed crucial functions in the process of malignant transformation such as degradation of the p53 tumor suppressor protein, deregulation of the Retinoblastoma protein pathway and activation of the telomerase ribonucleoprotein. All these steps are necessary for cancerous lesions to develop. However, the loss of the E2 gene product seems to be necessary for sufficient expression of E6 and E7 in order to achieve such effects. In normal infections, the E4 protein is made abundantly in the later stages of the viral life cycle. Though extensive amounts of work have been carried out to define the function of E4, it still remains unclear. In this study, several approaches have been used to try and determine the functions of E4. First, a cell-penetrating fusion protein was designed and produced in order to circumvent the chronic difficulties of expressing E4 in mammalian cells. Unfortunately, this approach was not successful due to precipitation of the purified fusion protein. Second, the observation that E4 accumulates in cells having modified their adhesion properties led to the hypothesis that E4 might be involved in the differentiation process of keratinocytes. Preliminary results suggest that E4 triggers differentiation. Last, as E4 has been reported to collapse the cytokeratin network of keratinocytes, a direct approach using atomic force microscopy has allowed us to test the potential modification of mechanical properties of cells harboring reorganized cytokeratin networks. If so, a potential role for E4 in viral particle release could be hypothesized. 2. Résumé Il a été établi que le cancer du col de l'utérus se développe essentiellement à la suite d'une infection par le virus du papillome humain (HPV). Dans la majorité des cas analysés, de l'ADN du HPV de type 16 (HPV16) est détecté. Les étapes initiales de la transformation d'une cellule infectée sont mal connues mais il semble qu'une rupture du génome viral, normalement épisomal, suivi d'une intégration dans le génome de la cellule hôte soient des étapes nécessaires dans la plupart des cas. Or il semble qu'il y ait une sélection pour les cas où l'expression des oncogènes viraux E6 et E7 soit favorisée alors que l'expression des gènes E2 et E4 est en général impossible. Par contre, dans une infection dite normale où le génome viral n'est pas rompu, il n'y pas développement de cancer et tous les gènes viraux sont exprimés. L'ordre dans lequel les protéines virales sont produites, et donc le cycle de réplication du virus, est intimement lié au processus de différentiation de la cellule hôte. L'étude des protéines oncogènes E6 et E7 a révélé des fonctions clés dans le processus de transformation des cellules infectées telles que la dégradation du suppresseur de tumeur p53, la dérégulation de la voie de signalisation Rb ainsi que l'activation de la télomérase. Toutes ces activités sont nécessaires au développement de lésions cancéreuses. Toutefois, il semble que l'expression du gène E2 doit être empêchée afin que suffisamment des protéines E6 et E7 soient produites. Lorsque le gène E2 est exprimé, et donc lorsque le génome viral n'est pas rompu, les protéines E6 et E7 n'entraînent pas de telles conséquences. Le gène E4, qui se trouve dans la séquence codante de E2, a aussi besoin d'un génome viral intact pour être exprimé. Dans une infection normale, le gène E4 est exprimé abondamment dans les dernières étapes de la réplication du virus. Bien que de nombreuses études aient été menées afin de déterminer la fonction virale à E4, aucun résultat n'apparaît évident. Dans ce travail, plusieurs approches ont été utilisées afin d'adresser cette question. Premièrement, une protéine de fusion TAT-E4 a été produite et purifiée. Cette protéine, pouvant entrer dans les cellules vivantes par diffusion au travers de la membrane plasmique, aurait permis d'éviter ainsi les problèmes chroniques rencontrés lors de l'expression de E4 dans les cellules mammifères. Malheureusement, cette stratégie n'a pas pu être utilisée à cause de la précipitation de la protéine purifiée. Ensuite, l'observation que E4 s'accumule dans les cellules ayant modifié leurs propriétés d'adhésion a suggéré que E4 pourrait être impliqué dans le procédé de différentiation des kératinocytes. Des résultats préliminaires supportent cette possibilité. Enfin, il a été montré que E4 pouvait induire une réorganisation du réseau des cytokératines. Une approche directe utilisant le microscope à force atomique nous a ainsi permis de tester une potentielle modification des propriétés mécaniques de cellules ayant modifié leur réseau de cytokératines en présence de E4. Si tel est le cas, un rôle dans la libération de particules virales peut être proposé pour E4.

The political ecology of weeds : a scalar approach to landscape transformations

How do plants that move and spread across landscapes become branded as weeds and thereby objects of contention and control? We outline a political ecology approach that builds on a Lefebvrian understanding of the production of space, identifying three scalar moments that make plants into 'weeds' in different spatial contexts and landscapes. The three moments are: the operational scale, which relates to empirical phenomena in nature and society; the observational scale, which defines formal concepts of these phenomena and their implicit or explicit 'biopower' across institutional and spatial categories; and the interpretive scale, which is communicated through stories and actions expressing human feelings or concerns regarding the phenomena and processes of socio-spatial change. Together, these three scalar moments interact to produce a political ecology of landscape transformation, where biophysical and socio-cultural processes of daily life encounter formal categories and modes of control as well as emotive and normative expectations in shaping landscapes. Using three exemplar 'weeds' - acacia, lantana and ambrosia - our political ecology approach to landscape transformations shows that weeds do not act alone and that invasives are not inherently bad organisms. Humans and weeds go together; plants take advantage of spaces and opportunities that we create. Human desires for preserving certain social values in landscapes in contradiction to actual transformations is often at the heart of definitions of and conflicts over weeds or invasives.

Cysteine cathepsin proteases: regulators of cancer progression and therapeutic response.

Cysteine cathepsin protease activity is frequently dysregulated in the context of neoplastic transformation. Increased activity and aberrant localization of proteases within the tumour microenvironment have a potent role in driving cancer progression, proliferation, invasion and metastasis. Recent studies have also uncovered functions for cathepsins in the suppression of the response to therapeutic intervention in various malignancies. However, cathepsins can be either tumour promoting or tumour suppressive depending on the context, which emphasizes the importance of rigorous in vivo analyses to ascertain function. Here, we review the basic research and clinical findings that underlie the roles of cathepsins in cancer, and provide a roadmap for the rational integration of cathepsin-targeting agents into clinical treatment.

Catalyzed and non-catalyzed synthesis of bioactive monastrol

The bioactive 3,4-dihydropyrimidin-2(1H)-thione derivative known as Monastrol was synthesized under catalyzed and non-catalyzed conditions through the Biginelli multicomponent reaction under solvent-free conditions. The use of two Lewis acids (FeCl3 and CuCl2) and two Brønsted acids (HCl and CF3COOH) as catalysts improved the reaction yields of the transformation compared with the non-catalyzed reaction. The experiments investigated catalysis and its role, the importance of multicomponent reactions and their green features, and the application of these concepts to the synthesis of a biologically important structure.

Magnetic sputtering of complex oxides

HTSC materials are relevant in modern microelectronics, because of their transformation from the normal state to the superconducting. That is why the idea of producing HTSC in industrial amounts is actual nowadays. To decrease cost of their production it is important to use magnetron sputtering systems which give the best results for essential parameters. Modeling is the simplest and the fastest way to determine optimum sputtering condition. This thesis concentrates on determination the phases of the whole sputtering process and to find out basic factors of each phase using the modeling. It was find out, that the main factors which influence on the mode of occurrence of the initial stages are the current density of the magnetron discharge and the pressure of sputtering gas. With the modeling also velocity dependences were obtained for YBCO and SmFeAsO. These were compared and difference between them was examined. To support represented model comparison was made with experimental results. This showed that the model gives good results, very similar to the experimental ones. The results of this work were published in annual conference of the finnish physical society.

Two variants of HIV-1 B serotype are transmitted heterosexually in São Paulo, Brazil

HIV-1 variability may have an important impact on transmission and pathogenicity. Better characterization of the HIV epidemic in Brazil is necessary for the development of vaccine trials in this country. We analyzed sera from 108 HIV-1-infected volunteers from São Paulo City to determine serotype and reactivity for V3 motifs of HIV in this population, and the relationship to transmission mode. We concluded that the HIV-1 B serotype is frequent among heterosexually infected women, even in the absence of anal sex, and that two major V3 motifs, GPGR and GWGR, had similar prevalence among women (48% and 52%, respectively) and men (56% and 44%, respectively). We also observed an equal distribution of these strains regardless of their CD4+ T cell counts, clinical status, and mode of transmission. Even though V3 serology for HIV-1 subtyping is an inexpensive tool for use in developing countries, additional methods, such as heteroduplex mobility assay and direct DNA sequencing, should be included to determine HIV-1 genetic diversity.

Cytogenetic description of breast fibroadenomas: alterations related solely to proliferation?

Twelve breast fibroadenomas were analyzed cytogenetically and only four were found to have clonal alterations. The presence of chromosomal alterations in fibroadenomas must be the consequence of the proliferating process and must not be related to the etiology of this type of lesion. In contrast, the few fibroadenomas that exhibit chromosomal alterations are likely to be those presenting a risk of neoplastic transformation. Clonal numerical alterations involved chromosomes 8, 18, 19, and 21. Of the chromosomal alterations found in the present study, only monosomy of chromosomes 19 and 21 has been reported in breast fibroadenomas. The loss of chromosome 21 was the most frequent alteration found in our sample. The study of benign proliferations and their comparison with chromosome alterations in their malignant counterparts ought to result in a better understanding of the genes acting on cell proliferation alone, and of the genes that cause these cells to exhibit varied behaviors such as recurrences, spontaneous regression and fast growth.

The transforming network of a foreign company operating in Mongolia. Case: A Finnish firm in Mongolia

In the globalising business environment ever fewer market areas remain unknown. Mongolia is yet only considered as an isolated strip between two power states. The purpose of this study is to put Mongolia on the map of academic business research. This is done by describing the transforming network of a foreign company operating in Mongolia. The objective of the study is approached through a case study, which presents the transformation of a Finnish company operating in Mongolia. This study aims at providing understanding on how the foreign case company observes the transformations of its network. The transformation within the case company is reflected to the transformations that occur in the Mongolian business environment. This study was conducted through a qualitative, intrinsic case study approach. The empirical data was gathered by using the method of network pictures. The network pictures were completed with the assistance of themed interviews. In order to be able to analyse the transformation within a network, three different time periods were observed: the past period around 2000, the present around 2014, and the estimated future around 2020. The data was collected from four executives positioned either in Finland, Russia or Mongolia. The respondents have a long experience within the case company, they hold managerial position, and therefore were able to offer valuable data for this study. The analytical framework used to analyse the collected data was built on the industrial network model, the ARA (actors-resources-activities)-model. The study shows that the changing business environment of Mongolia was utilised by the case company. In order to better meet the transforming customer wishes, the case company transformed from being a retailer to being a manufacturer. The case company was able to become a pioneer in the market. Thus, the case company has undergone similar kind of rapid transformation as the economy of Mongolia in entirety. This study shows that the general nature of the ARA-model makes it usable for new research contexts. The initial ARA-model offers a way to identify the dimensions of a network and a mean to understand these dimensions. The ARA-model can be applied to different contexts and to all time dimensions, past, present and future. The managerial recommendations offered in this study are directed towards the managers that plan to start operations in Mongolia. While this study is the first of its kind, it offers a good starting point for the future research on the change of Mongolian business networks. Valuable information could, for example, be obtained from a comparative study between the case company of this study and a multinational mining company operating in Mongolia.

Words on wisdom: perspective transformation in a "learning in retirement institute" /

In this study, 7 men and women with an average age of 77 were interviewed regarding their experience of attending courses at a Learning in Retirement Institute (LRI) in southern Ontario. The purpose was to explore the role of wisdom in the learning of these retirees. Explicit theories of wisdom developed by selected philosophers, psychologists, and religious thinkers were compared to the implicit theories of wisdom that respondents expressed. Further comparisons were drawn between these implicit theories of wisdom and the act of perspective transformation in transformative learning. Some evidence was found that the development of wisdom compares favourably to perspective transformation, especially with regards to the behavioural changes associated with critical self-reflection. Among all the respondents, those 3 LRI stude.its who had also moderated courses indicated that they had experienced the most opportunities for critical self-reflection. These 3 also expressed deep satisfaction in having been able to put their learnings to use as teachers. A recommendation of this study is that opportunities for sharing and acting upon the results of discourse within Learning in Retirement Institutes should be implemented. In the absence of evaluation, opportunities for praxis (such as coop placements) must be developed so that students can measure their success against objective criteria and hence attach meaning to their studies.

The establishment of two novel bovine cell lines by transfection with the putative transforming region of bovine adenovirus type 3

Human adenoviruses (Ads), members of the family adenoviridae, are medium-sized DNA viruses which have been used as valuable research tools for the study of RNA processing, oncogenic transformation, and for the development of viral vectors for use in gene delivery and immunization technology. The left 12% of the linear Ad genollle codes for products which are necessary for the efficient replication of the virus, as well as being responsible for the forlllation of tumors in animallllodels. The establishlllent of the 293 cell line, by immortalization of human embryonic kidney cells with th~ E1 region of Ad type S (AdS), has facilitated extensive manipulation of the Ads and the development of recombinant Ad vectors. The study of bovine adenoviruses (BAVs), which cause mild respiratory and gastrointestinal infections in cattle has, on the other hand, been limited primarily to that of infectivity, immunology and clinicallllanifestations. As a result, any potential as gene delivery vehicles has not yet been realized. Continued research into the molecular biolo~gy of BAVs and the development of recolllbinant vectors would benefit from the development of a cell line analogous to that of the 293 cells. In an attelllpt to establish such a cell line, the recombinant plaslllid pKC-neo was constructed, containing the left 0-19.7% of the BAV type 3 (BAV3) genome, and the selectable marker for resistance to the aminoglycoside G418, a neomycin derivative. The plasmid construct was then used to transfect both the Madin-Darby bovine kidney (MDBK) -iicell line and primary bovine lung cells, after which G418-resistant foci were selected for analysis. Two cell lines, E61 (MDBK) and E24 (primary lung), were subsequently selected and analysed for DNA content, revealing the presence of the pKC-neo sequences in their respective genomes. In addition, BAV3 RNA transcripts were detected in the E61 cells. Although the presence of E1 products has yet to be confirmed in both cell lines, the E24 cells exhibit a phenotype characteristic of partial transformation by E1. The apparent immortalization of the primary lung cells will permit exploitation of their ability to take up exogenous DNA at high efficiency.

Religiosité moderne et transformation personnelle : le cas des pratiquants de l'Ashtanga yoga à Montréal

Dans le contexte de l’expression moderne de la religiosité, l’étude explore comment les méthodes de l’Ashtanga yoga produisent l’expérience de la transformation chez les individus qui les pratiquent et qui atteint, dans une certaine mesure, leur entourage. Ethnographiquement, le mémoire se concentre sur le rituel principal de l’Ashtanga Vinyasa Yoga (AVY), les séances de style Mysore, telle qu’exercé par la communauté de pratiquants de la Sattva Yoga Shala à Montréal. En tant que performance, le rituel met en branle le potentiel de transformation. La pratique de ce yoga implique une discipline et un apprentissage draconien sur une longue échelle temporelle qui amènent, au fur et à mesure, des changements se répercutant dans/et influençant le vécu quotidien des adeptes. Le mémoire résume d’abord les bases littéraires du yoga, et le contexte historique de l’avènement de l’AVY ainsi que celui de sa venue à Montréal. Pour solidifier l’analyse, le travail poursuit par l’idéologie et la praxis du groupe d’étude. Le coeur de l’analyse suit. Nous examinons premièrement les modalités potentielles à la transformation du soi, c’est-à-dire les éléments rituels généraux et ceux spécifiques à l’AVY; puis, nous nous penchons sur ce que ces modalités développent, permettent, et changent chez les répondants, donc leurs impacts. Dans cette recherche, l’Ashtanga Vinyasa Yoga est à la fois une réalité empirique et une catégorie analytique servant à approfondir les connaissances anthropologiques sur le phénomène de la transformation personnelle selon le cadre expérientiel de la religiosité moderne.

Cyberjustice in Brazil : the use of technology to enhance access to justice and procedural celerity

Dans une société mondialisée, où les relations sont intégrées à une vitesse différente avec l'utilisation des technologies de l'information et des communications, l'accès à la justice gagne de nouveaux concepts, mais elle est encore confrontée à de vieux obstacles. La crise mondiale de l'accès à la justice dans le système judiciaire provoque des débats concernant l'égalité en vertu de la loi, la capacité des individus, la connaissance des droits, l'aide juridique, les coûts et les délais. Les deux derniers ont été les facteurs les plus importants du mécontentement des individus avec le système judiciaire. La présente étude a pour objet d'analyser l'incidence de l'utilisation de la technologie dans l’appareil judiciaire, avec l'accent sur la réalité brésilienne, la voie législative et des expériences antérieures dans le développement de logiciels de cyberjustice. La mise en œuvre de ces instruments innovants exige des investissements et de la planification, avec une attention particulière sur l'incidence qu'ils peuvent avoir sur les routines traditionnelles des tribunaux. De nouveaux défis sont sur la voie de ce processus de transformation et doivent être traités avec professionnalisme afin d'éviter l'échec de projets de qualité. En outre, si la technologie peut faire partie des différents aspects de notre quotidien et l'utilisation de modes alternatifs de résolution des conflits en ligne sont considérés comme un succès, pourquoi serait-il difficile de faire ce changement dans la prestation de la justice par le système judiciaire? Des solutions technologiques adoptées dans d'autres pays ne sont pas facilement transférables à un environnement culturel différent, mais il y a toujours la possibilité d'apprendre des expériences des autres et d’éviter de mauvaises voies qui pourraient compromettre la définition globale de l'accès à la justice.