Structural and Functional Analysis of the Caspase –dependent and –independent Domains of the X-linked Inhibitor of Apoptosis Protein in Inflammatory Breast Cancer Tumor Biology

Inflammatory breast cancer (IBC) is an extremely rare but highly aggressive form of breast cancer characterized by the rapid development of therapeutic resistance leading to particularly poor survival. Our previous work focused on the elucidation of factors that mediate therapeutic resistance in IBC and identified increased expression of the anti-apoptotic protein, X-linked inhibitor of apoptosis protein (XIAP), to correlate with the development of resistance to chemotherapeutics. Although XIAP is classically thought of as an inhibitor of caspase activation, multiple studies have revealed that XIAP can also function as a signaling intermediate in numerous pathways. Based on preliminary evidence revealing high expression of XIAP in pre-treatment IBC cells rather than only subsequent to the development of resistance, we hypothesized that XIAP could play an important signaling role in IBC pathobiology outside of its heavily published apoptotic inhibition function. Further, based on our discovery of inhibition of chemotherapeutic efficacy, we postulated that XIAP overexpression might also play a role in resistance to other forms of therapy, such as immunotherapy. Finally, we posited that targeting of specific redox adaptive mechanisms, which are observed to be a significant barrier to successful treatment of IBC, could overcome therapeutic resistance and enhance the efficacy of chemo-, radio-, and immuno- therapies. To address these hypotheses our objectives were: 1. to determine a role for XIAP in IBC pathobiology and to elucidate the upstream regulators and downstream effectors of XIAP; 2. to evaluate and describe a role for XIAP in the inhibition of immunotherapy; and 3. to develop and characterize novel redox modulatory strategies that target identified mechanisms to prevent or reverse therapeutic resistance.

Using various genomic and proteomic approaches, combined with analysis of cellular viability, proliferation, and growth parameters both in vitro and in vivo, we demonstrate that XIAP plays a central role in both IBC pathobiology in a manner mostly independent of its role as a caspase-binding protein. Modulation of XIAP expression in cells derived from patients prior to any therapeutic intervention significantly altered key aspects IBC biology including, but not limited to: IBC-specific gene signatures; the tumorigenic capacity of tumor cells; and the metastatic phenotype of IBC, all of which are revealed to functionally hinge on XIAP-mediated NFκB activation, a robust molecular determinant of IBC. Identification of the mechanism of XIAP-mediated NFκB activation led to the characterization of novel peptide-based antagonist which was further used to identify that increased NFκB activation was responsible for redox adaptation previously observed in therapy-resistant IBC cells. Lastly, we describe the targeting of this XIAP-NFκB-ROS axis using a novel redox modulatory strategy both in vitro and in vivo. Together, the data presented here characterize a novel and crucial role for XIAP both in therapeutic resistance and the pathobiology of IBC; these results confirm our previous work in acquired therapeutic resistance and establish the feasibility of targeting XIAP-NFκB and the redox adaptive phenotype of IBC as a means to enhance survival of patients.

DnaG interacts with a linker region that joins the N- and C-domains of DnaB and induces the formation of 3-fold symmetric rings

Loading of the replicative ring helicase onto the origin of replication (oriC) is the final outcome of a well coordinated series of events that collectively constitute a primosomal cascade. Once the ring helicase is loaded, it recruits the primase and signals the switch to the polymerization mode. The transient nature of the helicase-primase (DnaB-DnaG) interaction in the Escherichia coli system has hindered our efforts to elucidate its structure and function. Taking advantage of the stable DnaB-DnaG complex in Bacillus stearothermophilus, we have reviewed conflicting mutagenic data from other bacterial systems and shown that DnaG interacts with the flexible linker that connects the N- and C-terminal domains of DnaB. Furthermore, atomic force microscopy (AFM) imaging experiments show that binding of the primase to the helicase induces predominantly a 3-fold symmetric morphology to the hexameric ring. Overall, three DnaG molecules appear to interact with the hexameric ring helicase but a small number of complexes with two and even one DnaG molecule bound to DnaB were also detected. The structural/functional significance of these data is discussed and a speculative structural model for this complex is suggested.

Efecto determinante de la motivación de viaje sobre la imagen de destino en turistas de ocio a un destino urbano: El caso de Monterrey, México. Una aproximación por medio de mínimos cuadrados parciales (PLS) = Determinant effect of travel motivation on the destination image of leisure tourist in an urban destination: the case of Monterrey, México. An approach through partial least squares (PLS)

Los turistas urbanos se caracterizan por ser uno de los segmentos de mayor crecimiento en los mercados turísticos actuales. Monterrey (México), uno de los principales destinos urbanos del país, ha apostado en la actualidad por mejorar su competitividad. Esta investigación se propuso encontrar evidencia acerca de la relación causal de la motivación de viaje sobre la imagen percibida del destino, dos variables importantes por su influencia en la satisfacción de los visitantes. Una revisión de la literatura permitió proponer constructos teóricos integrados en un instrumento para la recogida de datos vía encuesta a una muestra representativa. Por medio del método de regresión y ecuaciones estructurales por mínimos cuadrados parciales (PLS), se identificaron los componentes principales de ambas variables y se obtuvo un modelo explicativo de la imagen percibida del destino en función de la motivación de viaje. Finalmente, se emiten recomendaciones para la gestión del destino urbano en función de los resultados obtenidos. ABSTRACT: Abstract Urban tourists are recognized as one of the fastest growing segments in today’s tourism markets. Monterrey, Mexico, one of the main urban destinations in the country aims at improving its competitiveness. This research work had the purpose of finding evidence on the causal relationship between travel motivation and destination image, two important variables because of their influence on visitors’ satisfaction. A literature review enabled the proposal of a research instrument with theoretically based constructs to gather data through survey from a representative sample. Using regression and structural equations modelling by partial least squares (pls) a set of main components of both variables were identified thus enabling the obtention of a explanatory model of destination image in terms of travel motivations. Finally based on the results some recommendations of tourism management are given.

Methods for assessing physical activity: a systematic review focused on older adults

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Leisure as a Facilitator of Posttraumatic Growth in Individuals Living with Cancer

Although there is a growing body of literature that shifts the focus of chronic illness and trauma research to personal growth, there is limited literature on the role that leisure has in this process (e.g., Tedeschi & Calhoun, 2004). This qualitative study explored the role of leisure in the phenomenon of posttraumatic growth (PTG) for individuals living with cancer. The findings revealed that leisure influences PTG in four domains: (a) building meaningful relationships, (b) providing experiences to develop and maintain a sense of self, (c) creating opportunities to experience positive emotions, and (d) finding purpose in life. Findings provide insight on how individuals living with cancer perceive the role that leisure has in facilitating positive change after diagnosis. These findings will better enable healthcare and leisure providers to understand the unique needs of individuals living with cancer, and help them to facilitate meaningful leisure programs to encourage PTG.

Health related quality of life is differently associated with leisure-time physical activity intensities according to gender: a cross-sectional approach

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Correlates of sports practice, occupational and leisure-time physical activity in Brazilian adolescents

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

The lens of ludic engagement : evaluating participation in interactive art installations

Designers and artists have integrated recent advances in interactive, tangible and ubiquitous computing technologies to create new forms of interactive environments in the domains of work, recreation, culture and leisure. Many designs of technology systems begin with the workplace in mind, and with function, ease of use, and efficiency high on the list of priorities. [1] These priorities do not fit well with works designed for an interactive art environment, where the aims are many, and where the focus on utility and functionality is to support a playful, ambiguous or even experimental experience for the participants. To evaluate such works requires an integration of art-criticism techniques with more recent Human Computer Interaction (HCI) methods, and an understanding of the different nature of engagement in these environments. This paper begins a process of mapping a set of priorities for amplifying engagement in interactive art installations. I first define the concept of ludic engagement and its usefulness as a lens for both design and evaluation in these settings. I then detail two fieldwork evaluations I conducted within two exhibitions of interactive artworks, and discuss their outcomes and the future directions of this research.

Functional outcome and health-related quality of life after traumatic brain injury in the framework of the International Classification of Functioning, Disability and Health (ICF)

Traumatic brain injury (TBI) affects people of all ages and is a cause of long-term disability. In recent years, the epidemiological patterns of TBI have been changing. TBI is a heterogeneous disorder with different forms of presentation and highly individual outcome regarding functioning and health-related quality of life (HRQoL). The meaning of disability differs from person to person based on the individual s personality, value system, past experience, and the purpose he or she sees in life. Understanding of all these viewpoints is needed in comprehensive rehabilitation. This study examines the epidemiology of TBI in Finland as well as functioning and HRQoL after TBI, and compares the subjective and objective assessments of outcome. The frame of reference is the International Classification of Functioning, Disability and Health (ICF). The subjects of Study I represent the population of Finnish TBI patients who experienced their first TBI between 1991 and 2005. The 55 Finnish subjects of Studies II and IV participated in the first wave of the international Quality of life after brain injury (QOLIBRI) validation study. The 795 subjects from six language areas of Study III formed the second wave of the QOLIBRI validation study. The average annual incidence of Finnish hospitalised TBI patients during the years 1991-2005 was 101:100 000 in patients who had TBI as the primary diagnosis and did not have a previous TBI in their medical history. Males (59.2%) were at considerably higher risk of getting a TBI than females. The most common external cause of the injury was falls in all age groups. The number of TBI patients ≥ 70 years of age increased by 59.4% while the number of inhabitants older than 70 years increased by 30.3% in the population of Finland during the same time period. The functioning of a sample of 55 persons with TBI was assessed by extracting information from the patients medical documents using the ICF checklist. The most common problems were found in the ICF components of Body Functions (b) and Activities and Participation (d). HRQoL was assessed with the QOLIBRI which showed the highest level of satisfaction on the Emotions, Physical Problems and Daily Life and Autonomy scales. The highest scores were obtained by the youngest participants and participants living independently without the help of other people, and by people who were working. The relationship between the functional outcome and HRQoL was not straightforward. The procedure of linking the QOLIBRI and the GOSE to the ICF showed that these two outcome measures cover the relevant domains of TBI patients functioning. The QOLIBRI provides the patients subjective view, while the GOSE summarises the objective elements of functioning. Our study indicates that there are certain domains of functioning that are not traditionally sufficiently documented but are important for the HRQoL of persons with TBI. This was the finding especially in the domains of interpersonal relationships, social and leisure activities, self, and the environment. Rehabilitation aims to optimize functioning and to minimize the experience of disability among people with health conditions, and it needs to be based on a comprehensive understanding of human functioning. As an integrative model, the ICF may serve as a frame of reference in achieving such an understanding.

Some Aspects of the Kobayashi and Carath,odory Metrics on Pseudoconvex Domains

The purpose of this article is to consider two themes, both of which emanate from and involve the Kobayashi and the Carath,odory metric. First, we study the biholomorphic invariant introduced by B. Fridman on strongly pseudoconvex domains, on weakly pseudoconvex domains of finite type in C (2), and on convex finite type domains in C (n) using the scaling method. Applications include an alternate proof of the Wong-Rosay theorem, a characterization of analytic polyhedra with noncompact automorphism group when the orbit accumulates at a singular boundary point, and a description of the Kobayashi balls on weakly pseudoconvex domains of finite type in C (2) and convex finite type domains in C (n) in terms of Euclidean parameters. Second, a version of Vitushkin's theorem about the uniform extendability of a compact subgroup of automorphisms of a real analytic strongly pseudoconvex domain is proved for C (1)-isometries of the Kobayashi and Carath,odory metrics on a smoothly bounded strongly pseudoconvex domain.

A new protein structure of P-II class snake venom metalloproteinases: it comprises metalloproteinase and disintegrin domains

A new metalloproteinase-disintegrin, named Jerdonitin, was purified from Trimeresurus jerdonii venom with a molecular weight of 36 kDa on SDS-PAGE. It dose-dependently inhibited ADP-induced human platelet aggregation with IC50 of 120 nM. cDNA cloning and sequencing revealed that Jerdonitin belonged to the class II of snake venom metalloproteinases (SVMPs) (P-II class). Different from other P-II class SVMPs, metalloproteinase and disintegrin domains of its natural protein were not separated, confirmed by internal peptide sequencing. Compared to other P-II class SVMPs, Jerdonitin has two additional cysteines (Cys219 and Cys238) located in the spacer domain and disintegrin domain, respectively. They probably form a disulfide bond and therefore the metalloproteinase and disintegrin domains cannot be separated by posttranslationally processing. In summary, comparison of the amino acid sequences of Jerdonitin with those of other P-II class SVMPs by sequence alignment and phylogenetic analysis, in conjunction with natural protein structure data, suggested that it was a new type of P-II class SVMPs. (C) 2003 Elsevier Inc. All rights reserved.

Construction of cytoplasmic molecular markers distinguishing Danio rerio from Gobiocypris rarus at high identity domains based on MP-PCR strategy and Sybr Green I detection

To distinguish the cytoplasm of Danio rerio from that of Gobiocypris rarus, we cloned G. rarus COXI and constructed cytoplasmic molecular markers at the high identity domains of COXI by mutated primer PCR (MP-PCR for short). Then Sybr Green I was used to detect the single amplicon. As a result, we succeeded in getting the cytoplasmic molecular markers, G.M COXI and Z.M COXI, by MP-PCR strategy. They were used to detect the sperm-derived mtDNA in the sexual hybrid embryos (D. rerio female x G. rarus male) before the sphere stage. In the present study, all results demonstrate that MP-PCR approach and Sybr Green I detection are feasible to construct the molecular markers to identify genes that shared high identity.

Leisure and surveillance in Irish childhoods: A cross-class study of Cork city

This research provides an interpretive cross-class analysis of the leisure experience of children, aged between six and ten years, living in Cork city. This study focuses on the cultural dispositions underpinning parental decisions in relation to children’s leisure activities, with a particular emphasis on their child-surveillance practices. In this research, child-surveillance is defined as the adult monitoring of children by technological means, physical supervision, community supervision, or adult supervised activities (Nelson, 2010; Lareau, 2003; Fotel and Thomsen, 2004). This research adds significantly to understandings of Irish childhood by providing the first in-depth qualitative analysis of the surveillance of children’s leisure-time. Since the 1990s, international research on children has highlighted the increasingly structured nature of children’s leisure-time (Lareau, 2011; Valentine & McKendrick, 1997). Furthermore, research on child-surveillance has found an increase in the intensive supervision of children during their unstructured leisure-time (Nelson, 2010; Furedi, 2008; Fotel and Thomsen, 2004). This research bridges the gap between these two key bodies of literature, providing a more integrated overview of children’s experience of leisure in Ireland. Using Bourdieu’s (1992) model of habitus, field and capital, the dispositions that shape parents’ decisions about their children’s leisure time are interrogated. The holistic view of childhood adopted in this research echoes the ‘Whole Child Approach’ by analysing the child’s experience within a wider set of social relationships including family, school, and community. Underpinned by James and Prout’s (1990) paradigm on childhood, this study considers Irish children’s agency in negotiating with parents’ decisions regarding leisure-time. The data collated in this study enhances our understanding of the micro-interactions between parents and children and, the ability of the child to shape their own experience. Moreover, this is the first Irish sociological research to identify and discuss class distinctions in children’s agentic potential during leisure-time.