70 research outputs found
Production of functional, stable, unmutated recombinant human papillomavirus E6 oncoprotein: Implications for HPV-tumor diagnosis and therapy
No full textBackground: High-risk human papillomaviruses (HR-HPVs) types 16 and 18 are the main etiological agents of cervical cancer, with more than 550,000 new cases each year worldwide. HPVs are also associated with other ano-genital and head-and-neck tumors. The HR-HPV E6 and E7 oncoproteins are responsible for onset and maintenance of the cell transformation state, and they represent appropriate targets for development of diagnostic and therapeutic tools. Methods: The unmutated E6 gene from HPV16 and HPV18 and from low-risk HPV11 was cloned in a prokaryotic expression vector for expression of the Histidine-tagged E6 protein (His6-E6), according to a novel procedure. The structural properties were determined using circular dichroism and fluorescence spectroscopy. His6-E6 oncoprotein immunogenicity was assessed in a mouse model, and its functionality was determined using in vitro GST pull-down and protein degradation assays. Results: The His6-tagged E6 proteins from HPV16, HPV18, and HPV11 E6 genes, without any further modification in the amino-acid sequence, were produced in bacteria as soluble and stable molecules. Structural analyses of HPV16 His6-E6 suggests that it maintains correct folding and conformational properties. C57BL/6 mice immunized with HPV16 His6-E6 developed significant humoral immune responses. The E6 proteins from HPV16, HPV18, and HPV11 were purified according to a new procedure, and investigated for protein-protein interactions. HR-HPV His6-E6 bound p53, the PDZ1 motif from MAGI-1 proteins, the human discs large tumor suppressor, and the human ubiquitin ligase E6-associated protein, thus suggesting that it is biologically active. The purified HR-HPV E6 proteins also targeted the MAGI-3 and p53 proteins for degradation. Conclusions: This new procedure generates a stable, unmutated HPV16 E6 protein, which maintains the E6 properties in in vitro binding assays. This will be useful for basic studies, and for development of diagnostic kits and immunotherapies in preclinical mouse models of HPV-related tumorigenesis. © 2016 The Author(s)
Gender Disparity in Susceptibility to Oxidative Stress and Apoptosis Induced by Autoantibodies Specific to RLIP76 in Vascular Cells
No full textAim: Ral-binding protein 1 (RLIP76) is a cell surface protein that catalyzes the extrusion from the cell of reduced glutathione (GSH) conjugates. We recently demonstrated the presence of serum antibodies to RLIP76 (aaRLIP76) in patients with immune-mediated diseases characterized by vascular dysfunction. The aim of this work was to analyze the possible implication of gender in this issue, investigating the effects of aaRLIP76 in rat vascular smooth muscle cells and human endothelial cells from males and females. Results: We observed that, after aaRLIP76 treatment, vascular cells from females showed a significantly higher susceptibility to the disturbance of intracellular redox balance, in terms of H(2)O(2) and O(2)(center dot) production, 4-hydroxy-t-2,3-nonenal and GSH levels, C-Jun NH2 kinase signaling activation, and apoptosis in comparison with cells from males. Interestingly, under mild oxidative stress (H(2)O(2) 30 mu m for 30 min), these sex-associated differences became significantly more pronounced. Experiments carried out in the presence of sex hormones in the culture medium clearly suggested that estrogens could significantly increase the susceptibility of cells from females to the effects of aaRLIP76, whereas cells from males appeared unaffected. Innovation: These results open a new perspective in the gender-dependent pathogenic mechanisms of autoimmune diseases characterized by vascular dysfunction. Conclusions: Altogether these results suggest that the impairment of RLIP76 by aaRLIP76 can play a role in the damage of vascular cells from females, contributing to the gender-associated pathogenesis of immune-mediated vascular diseases. Antioxid. Redox Signal. 15, 2825-2836
«Un Historia over Annali del Seminario Romano»: gesuiti e celebrazione storiografica nella Roma barocca (1640-47)
No full textThe article intends to explore the theme of historiographical celebration withinthe Society of Jesus through the specific case of the Seminario Romano, foundedby the Pope but managed by the order. The main source analyzed is the unpublishedmanuscript of the Annali del Seminario Romano, written by the JesuitGirolamo Nappi between 1640 and 1647. The work, which provides a verydetailed depiction of the institute, is part of the broader phenomenon of the historiographyof religious orders in the post-tridentine age. Particular attentionhas been paid to the section of the Annali concerning the vite of model formerstudents, through which the author illustrates the values of the Societas Iesu
Structuring Circular Objectives and Design Strategies for the Circular Economy: A Multi-Hierarchical Theoretical Framework
No full textMost frameworks for dealing with the complexity of designing for the circular economy
have limitations in terms of correlating different domains of knowledge, correlating highly complex
design strategies, and facilitating the process of design strategies’ discovery and development.
This paper discusses how managers and designers can create products that can be circulated for
several life cycles by considering five different circular objectives (i.e., maintenance/longevity, reuse,
refurbishment, remanufacture and/or recycling). To achieve one or more of these objectives, multiple
design strategies can be used at various phases of each product life cycle and throughout the product’s
lifetime. A literature review is used in this article to evaluate how circular objectives and design
strategies are classified in terms of relevance, product life cycle phases, and product life cycles.
The three classifications are merged to create a novel conceptual framework, which is then tested
through the use of four circular case studies to map out life cycles, circular objectives, and design
strategies. The framework may help managers and designers better understand how their businesses
and products interact along the supply chain, allowing them to establish more effective product
lifetime plans
Semantic Service Publication and Retrieval in the MAIS Project
No full textAccording to the Service Oriented Computing paradigm, a lot of services have been recently implemented by several companies. Since the number of services are growing, tools able to aid people and software to look for a service are required. Current solutions, such as UDDI, even if compliant with the Service Oriented Computing paradigm, do not provide an efficient way to search for services, since they mainly rely on keyword-based approaches. In this paper we present an approach based on a semantic service description supporting the service publication and retrieval processes. Such an approach is based on an UDDI extension in which a service ontology is considered
Metabolomic systems biology of trypanosomes
No full textMetabolomics analysis, which aims at the systematic identification and quantification of all metabolites in biological systems, is emerging as a powerful new tool to identify biomarkers of disease, report on cellular responses to environmental perturbation, and to identify the targets of drugs. Here we discuss recent developments in metabolomic analysis, from the perspective of trypanosome research, highlighting remaining challenges and the most promising areas for future research
Para além das fronteiras : uma leitura do 'Menino do Pijama Listrado'
No full textO objetivo desse trabalho é analisar criticamente o romance O Menino do Pijama Listrado do autor irlandês John Boyne. O foco da análise é a relação entre dois meninos, Bruno e Shmuel, que se conhecem através de uma cerca e a relação de Bruno com os demais membros de sua família. Esse trabalho analisa a questão de fronteiras e barreiras existentes na obra, que podem ser visíveis, invisíveis ou simbólicas. O encontro de culturas que acontece no entre-lugar ou terceiro espaço também é analisado, assim como as questões de alteridade. O referencial teórico utilizado abrange autores que tratam de questões sobre fronteiras, como Claude Raffestin e Sandra Jatahy Pesavento, de discurso e de poder, como Michael Foucault e de diferença cultural e alteridade, como Homi Bhabha, Mary Louisi Pratt e Rodolfo Franconi.The aim of this monograph is to critically analyze the novel The Boy in the Striped Pajamas by the Irish author John Boyne. The focus of analysis is the relationship between two boys, Bruno and Shmuel, who get to know each other through a fence, and the relationship between Bruno and his family. The encounter of cultures that happens in an in-between or third space is also analyzed, as well as alterity issues. The theoretical background used in the analyses includes authors who investigate boundary issues, such as Claude Raffestin and Sandra Jatahy Pesavento, discourse and power, such as Michael Foucault and culture and alterity, such as Homi Bhabha, Mary Louisi Pratt and Rodolfo Franconi
Single exposure of human fibroblasts (WI-38) to a sub-cytotoxic dose of UVB induces premature senescence
No full textAbstractIn this work, we present a new model of stress-induced premature senescence obtained by exposing human fibroblasts (WI-38) at early passages (passages 2–4) to a single sub-cytotoxic dose of UVB (200mJ/cm2). We show that this treatment leads to the appearance of several biomarkers of senescence such as enlarged and flattened cell morphology, the presence of nuclear heterochromatic foci and β-galactosidase activity. Furthermore, we demonstrate that a mild ROS production and p53 activation are upstream events required for the induction of premature senescence. Our method represents an alternative in vitro model in photoaging research and could be used to test potential anti-photoaging compounds
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