Search

Background: In recent years a new concept of health, 'positive health', has been developed, which focusses on a person's resilience instead of merely the absence of disease. A previous survey among a variety of stakeholders in general health care showed that there are differences in how dimensions of positive health are valued. Patients valued the spiritual and societal participation dimension higher than physicians and policymakers., Aim: To investigate how the six dimensions of positive health are valued by patients, health care professionals and policymakers in mental health care in the Netherlands, and to test whether these values differ from such stakeholders in general healthcare., Method: In a cross-sectional survey patients (N= 458), healthcare professionals (N=250) and policy makers (N=47) of two mental health care institutions in the Netherlands filled in an online survey. The results were compared to the results of the study by Huber e.a. (2016) by ANCOVA, paired T-tests and cohens' d., Results: Respondents valued all dimensions equally high. No significant differences between groups were found, except for a significant difference on daily functioning. Patients rated this dimension significantly higher than other stakeholders. The equal significance of the six dimensions is in contrast with the findings of a previous survey among stakeholders in general health care., Conclusion: In contrast to stakeholders in general health care, those in mental health care valued all dimensions of health of equal and high importance. Only daily functioning was rated lower by professionals and policymakers than by patients.

Members of the TNF superfamily, including Fas, Fas ligand, and CD40, have been shown to be expressed on tumor cells. In the studies described in this work, we report that another family member, the ligand for 4-1BB (CD137), is expressed on various human carcinoma cell lines, on cells of solid tumors derived from these cell lines, and cells obtained from human tumors. Expression of 4-1BB ligand (4-1BBL) mRNA was detected by both RT-PCR and Northern blot analysis, and expression of 4-1BBL protein was detected by Western blot analysis of whole cell lysates and by FACS analysis of tumor cells and cell lines. Incubation of tumor cells with a 4-1BB-Ig fusion protein led to the production of IL-8 by the cells, demonstrating that the 4-1BBL is functionally active and signals back into the tumor cells. Furthermore, 4-1BBL expressed on the carcinoma cells functioned as a costimulatory molecule for the production of cytokines (most notably IFN-gamma) in cocultures of T cells and tumor cells. These findings suggest that 4-1BBL expressed on carcinoma cells may significantly influence the outcome of a T cell-tumor cell interaction.

Apoptosis, or programmed cell death, is a physiological form of cell death that plays a critical role in the development and maintenance of multicellular organisms. Apoptosis is characterized based on morphological and biochemical criteria. Morphological characteristics include cell shrinkage, cytoplasmic condensation, chromatin segregation and condensation, membrane blebbing, and the formation of membrane-bound apoptotic bodies, whereas the biochemical hallmark of apoptosis is internucleosomal DNA cleavage into oligonucleosome-length fragments. A great deal of research is aimed at defining the molecular mechanisms that play a role in apoptosis. As one of the common end points of experiments related to apoptosis is in fact the death of the cell, it has become important to develop reliable assays to measure cell death that may be compared among the various systems being investigated. This chapter reviews many of the current methods used to measure apoptotic cell death and points out strengths and weaknesses of each approach with respect to the system being examined and the questions being asked. Traditional cell-based methods, including light and electron microscopy, vital dyes, and nuclear stains, are described. Biochemical methods such as DNA laddering, lactate dehydrogenase enzyme release, and MTT/XTT enzyme activity are described as well. Additionally, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling of DNA fragments (TUNEL) and in situ end labeling (ISEL) techniques are reviewed, which when used in conjunction with standard flow cytometric staining methods may yield informative data relating cell death to various cellular parameters, including cell cycle and cell phenotype. The use of one or more of the methods described in this chapter for measuring cell death should enable investigators to accurately assess apoptosis in the context of the various models being examined and help define causal relationships between the mechanisms that regulate apoptosis and the cell death event itself.

The 4-1BB receptor is an inducible type I membrane protein and member of the tumor necrosis factor receptor (TNFR) superfamily that is rapidly expressed on the surface of CD4+ and CD8+ T cells after antigen- or mitogen-induced activation. Cross-linking of 4-1BB and the T cell receptor (TCR) on activated T cells has been shown to deliver a costimulatory signal to T cells. Here, we expand upon previously published studies by demonstrating that CD8+ T cells when compared with CD4+ T cells are preferentially responsive to both early activation events and proliferative signals provided via the TCR and 4-1BB. In comparison, CD28-mediated costimulatory signals appear to function in a reciprocal manner to those induced through 4-1BB costimulation. In vivo examination of the effects of anti-4-1BB monoclonal antibodies (mAbs) on antigen-induced T cell activation have shown that the administration of epitope-specific anti-4-1BB mAbs amplified the generation of H-2d-specific cytotoxic T cells in a murine model of acute graft versus host disease (GVHD) and enhanced the rapidity of cardiac allograft or skin transplant rejection in mice. Cytokine analysis of in vitro activated CD4+ and CD8+ T cells revealed that anti-4-1BB costimulation markedly enhanced interferon-gamma production by CD8+ T cells and that anti-4-1BB mediated proliferation of CD8+ T cells appears to be IL-2 independent. The results of these studies suggest that regulatory signals delivered by the 4-1BB receptor play an important role in the regulation of cytotoxic T cells in cellular immune responses to antigen.

A variety of neurodegenerative disease states have been associated with oxidative damage or stress. Such stress is thought to be mediated by excessive exposure of cells to reactive oxygen species such as free radicals, which can be generated following cell lysis, oxidative burst (as part of the immune response) or by the presence of an excess of free transition metals. Since the neuronal death observed in neurodegenerative diseases may be related to free radical damage, we were interested in developing a model system to investigate the mechanisms by which reactive oxygen species may damage or kill neurons. To this end, we have recently reported that brief exposure of cultured cortical neurons to H2O2 can induce neuronal death that proceeds via an apoptotic cell suicide pathway. The studies reported here investigate H2O2-induced cell death in more detail. Our data suggest that exposure of cultured cortical neurons to H2O2 can induce apoptotic cell death within 3 h, as assessed by cell viability, morphological and ultrastructural measures. In addition, experiments presented show that exposure to high concentrations of H2O2 (100 microM) causes increases in intracellular free calcium within 3 h, suggesting that increased intracellular calcium may be associated with some aspects of H2O2-induced cell death. However, at intermediate concentrations of H2O2 (30 microM), intracellular calcium remained stable during a 3 h exposure, during which time membrane blebbing was observed in ultrastructural studies. This suggests that some aspects of apoptotic cell death induced by H2O2 may not be associated with increased intracellular free calcium. Thus, this model appears valuable for studies of the mechanism(s) by which oxidative injury may induce apoptotic cell death and damage to neurons in the CNS.

The serum-free mouse embryo (SFME) cell line, derived from 16-day-old mouse embryos in medium in which serum was replaced by growth factors and other supplements, has been cultured for more than 200 generations. SFME cells are nontumorigenic, lack gross chromosomal abnormalities, and display characteristics of CNS progenitor cells. SFME cells show reversible induction of the astrocyte-specific marker glial fibrillary acidic protein when cultured in the presence of TGF-beta or serum. In order to determine if SFME cells exhibit further characteristics of CNS progenitor cells we investigated the expression of the gene encoding nestin, an intermediate filament protein expressed by neuroepithelial stem cells of the CNS. SFME cells express nestin in serum-free medium, and nestin expression is reversibly down-regulated by TGF-beta or serum. These results demonstrate that nestin expression is regulated by factors present in serum and support the hypothesis that SFME cells represent a CNS progenitor cell.

To investigate whether direct oxidative damage may induce neuronal death via an apoptotic cell suicide pathway, we exposed cultured cortical neurons to the prooxidant hydrogen peroxide. Our experiments demonstrate that brief exposures to low concentrations of hydrogen peroxide can induce neuronal cell death via apoptosis as demonstrated by toxicity assays, specific DNA staining, and the appearance of DNA laddering on agarose gels. These results suggest that direct oxidative injury may serve as a general trigger for apoptosis in the CNS, and that other apoptotic stimuli, such as exposure to amyloid-beta peptides, may also involve oxidative injury. Peroxide-induced apoptosis may serve as a valuable model for further studies of apoptosis in neurons.

Mouse embryo cells cultured in vitro in serum-supplemented media undergo growth crisis, resulting in the loss of genomically normal cells prior to the appearance of established, aneuploid cell lines. Mouse embryo cells established and maintained for multiple passages in the absence of serum did not exhibit growth crisis or gross chromosomal aberration. Cells cultured under these conditions were dependent on epidermal growth factor for survival. Proliferation was reversibly inhibited by serum or platelet-free plasma, suggesting that mouse embryo cultures maintained by conventional procedures are under the influence of inhibitory factors.