International nursing advanced competency-based training for intensive care: a europe-wide survey.

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Cytotoxic T-cell precursor frequencies to HER-2 (369 – 377) in patients with HER-2/neu-positive epithelial tumours

HER-2/neu oncoprotein contains several major histocompatibility complex class I-restricted epitopes, which are recognised by cytotoxic T lymphocyte (CTL) on autologous tumours and therefore can be used in immune-based cancer therapies. Of these, the most extensively studied is HER-2(9(369)). In the present report, we used dendritic cells pulsed with HER-2(9(369)) to stimulate, in the presence of IL-7 and IL-12, the production of IFN-gamma by patients' CTL detected by the enzyme-linked immunosorbent spot-assay. Frequencies of peptide-specific precursors were estimated in HLA-A2, HLA-A3 and HLA-A26 patients with HER-2/neu-positive (+) breast, ovarian, lung, colorectal and prostate cancers and healthy individuals. We found increased percentages of such precursors in HLA-A2 (25%) and HLA-A26 (30%) patients, which were significantly higher (60%) in HLA-A3 patients. Our results demonstrate for the first time that pre-existing immunity to HER-2(9(369)) occurs in patients with colorectal, lung and prostate cancer. They also suggest that HER-2(9(369)) can be recognised by CTL, besides HLA-A2, also in the context of HLA-A3 and HLA-A26, thus increasing the applicability of HER-2(9(369))-based vaccinations in a considerably broader patients' population.Journal ArticleResearch Support, Non-U.S. Gov'tinfo:eu-repo/semantics/publishe

Rapid generation of human B-cell lymphomas via combined expression of Myc and Bcl2 and their use as a preclinical model for biological therapies

Although numerous mouse models of B-cell malignancy have been developed via the enforced expression of defined oncogenic lesions, the feasibility of generating lineage-defined human B-cell malignancies using mice reconstituted with modified human hematopoietic stem cells (HSCs) remains unclear. In fact, whether human cells can be transformed as readily as murine cells by simple oncogene combinations is a subject of considerable debate. Here, we describe the development of humanized mouse model of MYC/BCL2-driven ‘double-hit’ lymphoma. By engrafting human HSCs transduced with the oncogene combination into immunodeficient mice, we generate a fatal B malignancy with complete penetrance. This humanized-MYC/BCL2-model (hMB) accurately recapitulates the histopathological and clinical aspects of steroid-, chemotherapy- and rituximab-resistant human ‘double-hit’ lymphomas that involve the MYC and BCL2 loci. Notably, this model can serve as a platform for the evaluation of antibody-based therapeutics. As a proof of principle, we used this model to show that the anti-CD52 antibody alemtuzumab effectively eliminates lymphoma cells from the spleen, liver and peripheral blood, but not from the brain. The hMB humanized mouse model underscores the synergy of MYC and BCL2 in ‘double-hit’ lymphomas in human patients. Additionally, our findings highlight the utility of humanized mouse models in interrogating therapeutic approaches, particularly human-specific monoclonal antibodies.Kathy and Curt Marble Cancer Research FundSingapore-MIT Alliance for Research and TechnologyNational Institutes of Health (U.S.) (Grant R01-CA128803)Virginia and Daniel K. Ludwig Graduate FellowshipNational Institute of General Medical Sciences (U.S.) (Medical Scientist Training Program Grant T32GM007753)MIT School of Science (Cancer Research Fellowship

Membrane-Bound IL-21 Promotes Sustained Ex Vivo Proliferation of Human Natural Killer Cells

NK cells have therapeutic potential for a wide variety of human malignancies. However, because NK cells expand poorly in vitro, have limited life spans in vivo, and represent a small fraction of peripheral white blood cells, obtaining sufficient cell numbers is the major obstacle for NK-cell immunotherapy. Genetically-engineered artificial antigen-presenting cells (aAPCs) expressing membrane-bound IL-15 (mbIL15) have been used to propagate clinical-grade NK cells for human trials of adoptive immunotherapy, but ex vivo proliferation has been limited by telomere shortening. We developed K562-based aAPCs with membrane-bound IL-21 (mbIL21) and assessed their ability to support human NK-cell proliferation. In contrast to mbIL15, mbIL21-expressing aAPCs promoted log-phase NK cell expansion without evidence of senescence for up to 6 weeks of culture. By day 21, parallel expansion of NK cells from 22 donors demonstrated a mean 47,967-fold expansion (median 31,747) when co-cultured with aAPCs expressing mbIL21 compared to 825-fold expansion (median 325) with mbIL15. Despite the significant increase in proliferation, mbIL21-expanded NK cells also showed a significant increase in telomere length compared to freshly obtained NK cells, suggesting a possible mechanism for their sustained proliferation. NK cells expanded with mbIL21 were similar in phenotype and cytotoxicity to those expanded with mbIL15, with retained donor KIR repertoires and high expression of NCRs, CD16, and NKG2D, but had superior cytokine secretion. The mbIL21-expanded NK cells showed increased transcription of the activating receptor CD160, but otherwise had remarkably similar mRNA expression profiles of the 96 genes assessed. mbIL21-expanded NK cells had significant cytotoxicity against all tumor cell lines tested, retained responsiveness to inhibitory KIR ligands, and demonstrated enhanced killing via antibody-dependent cell cytotoxicity. Thus, aAPCs expressing mbIL21 promote improved proliferation of human NK cells with longer telomeres and less senescence, supporting their clinical use in propagating NK cells for adoptive immunotherapy

CD39 delineates chimeric antigen receptor regulatory T cell subsets with distinct cytotoxic & regulatory functions against human islets

Human regulatory T cells (Treg) suppress other immune cells. Their dysfunction contributes to the pathophysiology of autoimmune diseases, including type 1 diabetes (T1D). Infusion of Tregs is being clinically evaluated as a novel way to prevent or treat T1D. Genetic modification of Tregs, most notably through the introduction of a chimeric antigen receptor (CAR) targeting Tregs to pancreatic islets, may improve their efficacy. We evaluated CAR targeting of human Tregs to monocytes, a human β cell line and human islet β cells in vitro. Targeting of HLA-A2-CAR (A2-CAR) bulk Tregs to HLA-A2+ cells resulted in dichotomous cytotoxic killing of human monocytes and islet β cells. In exploring subsets and mechanisms that may explain this pattern, we found that CD39 expression segregated CAR Treg cytotoxicity. CAR Tregs from individuals with more CD39low/- Tregs and from individuals with genetic polymorphism associated with lower CD39 expression (rs10748643) had more cytotoxicity. Isolated CD39− CAR Tregs had elevated granzyme B expression and cytotoxicity compared to the CD39+ CAR Treg subset. Genetic overexpression of CD39 in CD39low CAR Tregs reduced their cytotoxicity. Importantly, β cells upregulated protein surface expression of PD-L1 and PD-L2 in response to A2-CAR Tregs. Blockade of PD-L1/PD-L2 increased β cell death in A2-CAR Treg co-cultures suggesting that the PD-1/PD-L1 pathway is important in protecting islet β cells in the setting of CAR immunotherapy. In summary, introduction of CAR can enhance biological differences in subsets of Tregs. CD39+ Tregs represent a safer choice for CAR Treg therapies targeting tissues for tolerance induction

Can young students think systemically about the environment? The case of pollution

This paper reports on the thoughts 6-year-olds and 9-year-olds have related to the serious issue of air and sea pollution. More specifically, twenty seven 6-year-olds and thirty 9-year-olds attending two state schools in Volos, a small provincial town in Greece participated in the research which assessed the students’ competence to think systemically about dealing with pollution. The study sheds light on whether students are able to identify interrelations between the components of pollution, and clarifies the mental models they hold. Data was collected through drawings and interviews, subsequent analysis of which indicates that students appeared to hold two mental models related to pollution; pollution is connected both spatially and temporally with visible pollutants; and, pollution is connected indirectly with invisible pollutants. The students seemed to exhibit a kind of systemic thinking, which was done unconsciously to a certain degree. Thus it is a challenge for education to enhance students’ systemic thinking in an attempt to bring it to a more conscious level, which will assist them to reconstruct their mental models of pollution. © 2016 ASPE

Children’s thinking about environmental issues

Background: The conceptions that children develop about environmental issues are likely to form the basis of how they comprehend such issues, influence their behaviour towards the environment and, ultimately, determine the way they address these issues. Being aware of children’s ways of thinking about the environment is, therefore, of paramount importance if educators are to provide effective and efficient teaching in this field of increasing global significance. Purpose: This paper presents an explorative study which sets out to identify the dominant conceptions held by a sample of 9-year-old students in relation to addressing, both collectively and individually, two environmental issues, namely forest destruction and waste. Sample: The sample comprised 30 9-year-old students (15 boys and 15 girls), from 2 state primary schools with diverse socio-economic student populations, in a provincial city in central Greece. Methods: Semi-structured interviews were used to collect the data. The interviews were analysed qualitatively, using a thematic approach. Results: Findings suggested that children’s conceptions about how to address environmental problems could be grouped into two broad conceptions: the first concerned proposing ‘direct actions’ for the short-term addressing of the problems, whereas the second related to a proactive approach for addressing the problem in the longer term, with an emphasis on sustainability. Furthermore, there seemed to be connections evident between the children’s conceptions in terms of their personal contributions and the thoughts they held about collective actions. Conclusions: This study offers insights into children’s approaches to addressing the problems of forest destruction and waste. It was evident that the children believed that they could address the problems by applying whatever methods they are aware of. We argue that this suggests that an action-oriented and problem-solving strategy towards education would be beneficial. A transformative approach is suggested as an effective technique, drawing on hands-on, experiential and project-based activities, so that children can be educated with the aim of becoming competent at making decisions themselves and taking action as active participants and change agents for sustainability. © 2018 NFER

How young children think they can act for the environment: the case of forest and waste

Thirty 6-year-olds from two public schools located in a small provincial town in Greece were used as a sample in an explorative study into the mental models young children hold concerning forest destruction and waste. The study also explored how these mental models might be interrelated either within each issue or between issues. The analysis of data collected in interviews indicates that the mental models held by young children appear to be oriented towards two different trains of thought: one that relates to direct actions for the short-term addressing of problems and one that relates to preventive actions and caring for sustainability. Furthermore, children’s own thoughts about their individual contributions to the two environmental problems under study, appear to interrelate with those of collective contributions. More specifically, children seem to share common ideas on the environmental issues discussed as they voiced the opinion, ‘I can help if I do something that addresses the issue'. Implications for education are also discussed in this paper. © 2016 ASPE