Synergistic antitumor effect of TRAIL and adriamycin on the human breast cancer cell line MCF-7

The aim of the present study was to determine the effect of the combination of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and adriamycin (ADM) on the human breast cancer cell line MCF-7 and to identify potential mechanisms of apoptosis. Cell viability was analyzed by the MTT assay and the synergistic effect was assessed by the Webb coefficient. Apoptosis was quantified using the annexin V-FITC and propidium iodide staining flow cytometry. The mRNA expression of TRAIL receptors was measured by RT-PCR. Changes in the quantities of Bax and caspase-9 proteins were determined by Western blot. MCF-7 cells were relatively resistant to TRAIL (IC50 >10 ��g/mL), while MCF-7 cells were sensitive to ADM (IC50 <10 ��g/mL). A subtoxic concentration of ADM (0.5 ��g/mL) combined with 0.1, 1, or 10 ��g/mL TRAIL had a synergistic cytotoxic effect on MCF-7 cells, which was more marked with the combination of TRAIL (0.1 ��g/mL) and ADM (0.5 ��g/mL). In addition, the combined treatment with TRAIL and ADM significantly increased cell apoptosis from 9.8% (TRAIL) or 17% (ADM) to 38.7%, resulting in a synergistic apoptotic effect, which is proposed to be mediated by up-regulation of DR4 and DR5 mRNA expression and increased expression of Bax and caspase-9 proteins. These results suggest that the combination of TRAIL and ADM might be a promising therapy for breast cancer.

Myosin Va is developmentally regulated and expressed in the human cerebellum from birth to old age

Myosin Va functions as a processive, actin-based motor molecule highly enriched in the nervous system, which transports and/or tethers organelles, vesicles, and mRNA and protein translation machinery. Mutation of myosin Va leads to Griscelli disease that is associated with severe neurological deficits and a short life span. Despite playing a critical role in development, the expression of myosin Va in the central nervous system throughout the human life span has not been reported. To address this issue, the cerebellar expression of myosin Va from newborns to elderly humans was studied by immunohistochemistry using an affinity-purified anti-myosin Va antibody. Myosin Va was expressed at all ages from the 10th postnatal day to the 98th year of life, in molecular, Purkinje and granular cerebellar layers. Cerebellar myosin Va expression did not differ essentially in localization or intensity from childhood to old age, except during the postnatal developmental period. Structures resembling granules and climbing fibers in Purkinje cells were deeply stained. In dentate neurons, long processes were deeply stained by anti-myosin Va, as were punctate nuclear structures. During the first postnatal year, myosin Va was differentially expressed in the external granular layer (EGL). In the EGL, proliferating prospective granule cells were not stained by anti-myosin Va antibody. In contrast, premigratory granule cells in the EGL stained moderately. Granule cells exhibiting a migratory profile in the molecular layer were also moderately stained. In conclusion, neuronal myosin Va is developmentally regulated, and appears to be required for cerebellar function from early postnatal life to senescence.

Construction of bovine adenovirus type 3 E1 and E3, substitution plasmids and the sequencing analysis of DNA pol and pTP /

The relative ease to concentrate and purify adenoviruses, their well characterized mid-sized genome, and the ability to delete non-essential regions from their genome to accommodate foreign gene, made adenoviruses a suitable candidate for the construction of vectors. The use of adenoviral vectors in gene therapy, vaccination, and as a general vector system for expressing foreign genes have been documented for some time. In this study, the objective was to rescue a BAV3 E1 or E3 recombinant vector carrying the kanamycin resistant gene, a dominant selectable marker with useful applications in studying vectored gene expression in mammalian cells. To accomplish the objective of this study, more information about BAV3 DNA sequences was required in order to make the manipulation of the virus genome accessible. Therefore, sequencing of the BAV3 genome from 1 1 .7% to 30.8% was carried out. Analysis of the determined sequences revealed the primary structure of important viral gene products coded by E2 including BAV3 DNA pol and precursor to terminal protein. Comparative analysis of these proteins with their counterparts from human and non human adenoviruses revealed important insights as to the evolutionary lineage of BAV3. In order to insert the kanamycin resistance gene in either E1 or E3, it was necessary to delete BAV3 sequences to accommodate the foreign gene so as not to exceed the limit of the packaging capacity of the virus. To construct a recombinant BAV3 in which a foreign gene was inserted in the deleted E1 region, an E1 shuttle vector was constructed. This involved the deletion from the viral sequences a region between 1.3% to 9% and inserting the kanamycin resistance gene to replace the deletion. The E1 shuttle vector contained the left (0%- 53.9%) segment of the genome and was expected to generate BAV3 recombinants that can be grown and propagated in cells that can complement the missing E1 functions. To construct a similar shuttle vector for E3 deletion, DNA sequences extending from 78.9% to 82.5% (1281 bp) were deleted from within the E3 region that had been cloned into a plasmid vector. The deleted region corresponds to those that have been shown to be non-essential for viral replication in cell culture. The resulting plasmid was used to construct another recombinant plasmid with BAV3 DNA sequences extending from 37.1% to 100% and with a deletion of E3 sequences that were replaced by kanamycin resistance gene. This shuttle plasmid was used in cotransfections with digested viral DNA in an attempt to rescue a recombinant BAV3 carrying the kanamycin resistance gene to replace the deleted E3. In spite of repeated attempts of transfection, El or E3 recombinant BAV3 were not isolated. It seems that other approaches should be applied to make a final conclusion on BAV3 infectivity.

The backwoods of our backyards : Ontario pioneers and the crisis of ecology /

Early in his landmark ecocritical book The Comedy of Survival, Joseph Meeker develops an intriguing hypothesis about human behaviour. He remarks the species Homo sapiens tend to behave like an invasive or pioneering organism, entering a bio-geographical region and aggressively outcompeting all other species for space and resources. Moreover, he suggests, human cultural traditions, at least in the West, have reinforced such behaviour, continually insisting that the impulses he describes are both necessary and right. While Meeker's work goes on to assess a number of literary works in both the tragic and comic modes, his work never fully explores this hypothesis in the context of human pioneers; that is, there is no ~xploration o( how these themes manifest themselves within our culture and what role they might play in the culture of specific pioneering groups. This project is an attempt at just such an analysis, examining the validity of Meeker's hypothesis through a case study of settler literature in Upper Canada/Ontario between the . years 1800-1867. It explores Meeker's work within three main areas: first, Chapter Two situates his book historically within the field of ecocriticism, showing what came before and the explosion of ecocritical inquiry that followed its release. This chapter also delves into the rift between the natural sciences and humanities, arguing that a move towards deeper interdisciplinarity is r:tecessary for the future. Chapter Three examines the biological and ecological ground on which Meeker rests his hypothesis through exploring evolutionary biology as well as invasive and pioneer species behaviour. Lastly, Chapter Four examines how these ecological principles are manifested in the writings of early Canadian settlers, suggesting that Meeker's hypothesis indeed finds itself on stable footing.

The condition of Niagara Falls, and the measures needed to preserve them : eight letters published in the New York Evening post, the New York tribune, and the Boston daily advertiser, during the summer of 1882 /

Appendix: Extracts from Harper's weekly, New York herald, New York tribune and the Nation.

Report of the Committee appointed to inquire into the Present condition and distribution of the flags, standards and colors, which have been taken by the forces of the United States from their enemies, and whether it would be expedient to make any provision in relation to them.--

Adam Seybet, Chairman.

Beyond the Veil: A Case Study of Context, Culture, Curriculum, and Constructivism at Dubai Women's College

This case study of curriculum at Dubai Women's College (DWC) examines perceptions of international educators who designed and implemented curriculum for female Emirati higher-educational students in the UAE, and sheds light on the complex social, cultural, and religious factors affecting educational practice. Participants were faculty and supervisors, mainly foreign nationals, while students at DWC are exclusively Emirati. Theories prominent in this study are: constructivist learning theory, trans formative curriculum theory, and sociological theory. Change and empowerment theory figure prominently in this study. Findings reveal this unique group of educators understand curriculum theory as a "contextualized" construct and argue that theory and practice must be viewed through an international lens of religious, cultural, and social contexts. As well, the study explores how mandated "standards" in education-in the form of the International English Language Testing System (IEL TS) and integrated, constructivist curriculum, as taught in the Higher Diploma Year 1 program-function as dual curricular emphases in this context. The study found that tensions among these dual emphases existed and were mediated through specific strategies, including the use of authentic texts to mirror the IEL TS examination during in-class activities, and the relevance of curricular tasks.

Association between disruption of CD4 receptor dimerization and increased human immunodeficiency virus type 1 entry

Affiliation: D��partement de microbiologie et immunologie, Facult�� de m��decine, Universit�� de Montr��al & Institut de Recherches Cliniques de Montr��al

Who is to protect whom from what? The human security dilemmas of Iraq, North Korea and Darfur

Brian Job, Professor, Department of Political Science, University of British Columbia, Director, Center of International Relations. Pr��sentation dans le cadre du cycle de conf��rences organis�� par le CRDP intitul�� �� Le droit �� la s��curit�� ... la s��curit�� par le droit ��.

Cloning and Characterisation of the human gene gremlin promoter

L���ost��oarthrose (OA) est une maladie articulaire invalidante caract��ris��e par la perte de l���int��grit�� du cartilage articulaire. Les recherches tentent de comprendre les m��canismes mol��culaires de la maladie afin de trouver des inhibiteurs efficaces pouvant pr��venir la d��gradation du cartilage articulaire. Les BMPs (bone morphogenic proteins) jouent un r��le dans le processus pathophysiologique de cette maladie. Cette ��tude cible le r��le d���un antagoniste des BMPs, le gremlin. Nous avons ��tudi�� la r��gulation de l���expression de gremlin par le clonage et la caract��risation de son promoteur et en d��terminant si gremlin pouvait jouer un r��le autre qu���antagoniste des BMP, en affectant l���expression d���autres g��nes par l���activation d���une cascade de signalisation dans la cellule. Les r��sultats ont identifi�� une r��gion importante dans le promoteur de gremlin qui affecte son activit�� basale et induite, et ont montr�� que le gremlin ne pouvait pas affecter l���expression g��nique et l���activation de signalisation intracellulaire ind��pendamment des BMPs. Cette ��tude d��montre que le r��le de gremlin dans l���OA en est un essentiellement d���antagoniste des BMPs.

The human right to water - normative foundations and ethical implications

La pre��sente contribution examine les fondements normatifs ainsi que les implications e��thiques du droit a�� l���eau, tel qu���il fut reconnu en 2002 par le comite�� onusien des droits e��conomiques, sociaux et culturels. Il sera de��fendu que le droit a�� l���eau potable peut e��tre justifie�� en tant que droit moral fondamental, de par son caracte��re indispensable en vue de la garantie des conditions basiques de survie. Cet e��tat de fait, cependant, s���ave��re moins e��vident au vue d���un droit a�� l���eau d���usage non-domestique. Ici, la discussion se rapproche des de��bats accompagnant le concept beaucoup plus complexe des droits sociaux et e��conomiques. Par rapport a�� ce groupe de droits, la question de l���allocation est des plus controverse��es: a�� qui incombe-t-il de garantir leur respect? Dans le but d���e��viter cette proble��matique d���allocation, le pre��sent essai soule��vera la question de savoir, si la limitation de l���acce��s a�� l���eau peut e��tre conc��ue comme une violation d���autres droits moraux: bien qu���il y ait des cas ou�� des entreprises transnationales de��ploient des activite��s nuisibles a�� l���e��gard des populations pauvres en polluant sciemment leurs ressources en eau ou en initiant et en exe��cutant des strate��gies de privatisation les privant de leurs droits, la crise globale de l���eau ne saura e��tre rattache��e uniquement aux effets de la mondialisation. Pluto��t, l���on reconnai��tra la ne��cessite�� d���efforts positifs et soutenus de la part des pays de��veloppe��s en vue de la re��alisation d���un approvisionnement suffisant en eau pour tous.