Experiences of Users with an Online Self-Guided Mental Health Training Program Using Gamification.

Young adulthood is a period of high risk for the development of mental health concerns. Increasing well-being among young adults is important to prevent mental health concerns and their consequences. Self-compassion has been identified as a modifiable trait with the potential to protect against mental health concerns. An online self-guided mental health training program using gamification was developed and the user experience was evaluated in a 6-week experimental design. During this period, 294 participants were allocated to use the online training program via a website. User experience was assessed via self-report questionnaires, and interaction data for the training program were also collected. Results showed that those who completed the intervention (n= 47) visited the website on average 3.2 days a week, with a mean of 45.8 interactions during the 6 weeks. Participants report positive user experiences of the online training, on average a System Usability Scale Brooke (1) score of 79.1 (out of 100) at the end-point. Participants showed positive engagement with story elements of the training, based on an average score of 4.1 (out of 5) in the evaluation of the story at the end-point. This study found the online self-compassion intervention for youth to be acceptable, although some features seem preferred by users as compared to others. Gamification in the form of a guiding story and a reward structure seemed to be a promising element for successfully motivating participants and serving as a guiding metaphor for self-compassion.

Fast isolation of RNA to detect expression of tumor markers.

The expression status of several tumor-related proteins is of great interest in clinical examination and research. As a completion to conventional antibody staining, RT-PCR is often used today. Reliable isolation of RNA from a low number of cells is very often a critical stage of such an examination. We demonstrate here a simple and fast method to isolate RNA from only 10,000 cells and applied it to the detection of CEA, c-ERB-B2, and mdr-1 as often studied models for tumor markers.

Immortalization and transformation of human fibroblasts by regulated expression of polyoma virus T antigens.

We have established conditions for the immortalization of human fibroblasts by the large T antigen of the rodent virus polyoma. This allows the mechanism of immortalization to be studied, without interference by transformation events, in cells with relatively stable chromosomes. Large T antigen could immortalize human fibroblasts if expression was driven by a heterologous promoter like the immediate early promoter/enhancer of cytomegalovirus or the inducible mouse mammary tumour virus (MMTV) promoter. Using the latter promoter and dexamethasone, three clones were obtained, the immortalized phenotype of which was strictly dependent on the induction of T-antigen expression. At least one of these clones became mortal after removal of the inducing agent. The expression of large T antigen was paralleled by PCNA gene expression, as shown by nuclear run-off transcription, whereas none of a number of other known proto-oncogenes was influenced in its activity. Immortalized fibroblasts were readily transformed by polyoma virus middle T antigen expressed from the MMTV promoter or by the activated c-Ha-ras oncogene. The reversibility of immortalization and transformation is considered.

Identification and characterization of human hemopoietic mast cell colonies.

Persisting mast cell colonies from human bone marrow and cord blood cells grown in semisolid agar cultures for over 56 days have been positively identified and characterized using morphology and cytochemistry. Mast cells demonstrated the following features: Cytoplasmic granules frequently contained the specific and characteristic papyrus rolls (transmission electron microscopy); mature cells were positive to the mouse monoclonal antibody YB5.B8 specific for human mast cells (raised against acute myeloid leukemia cells) and RPA-M1 specific for human monocytes but negative to the human basophil monoclonal antibody Bsp-1; morphologically the cells were large (diameter 20-25 micron), deeply basophilic, and contained granules that measured up to 2 micron in diameter (May-Grünwald-Giemsa stain); the presence of heparin by the thrombin clotting time and positive staining with toluidine blue and alcian blue; the presence of histamine by a positive fluorescent o-phthalaldehyde stain; the presence of IgE receptor sites with human IgE and a rabbit anti-human IgE second antibody; and a unique zone of lysis around mast cell colonies occurred when cultured on peripheral blood feeder layers in agar plates that was not present around monocytic, neutrophilic, or eosinophilic colonies under the same culture conditions. Our results identify the cells in persisting colonies as mast cells and describe some specific characteristics that distinguish these cells from basophils.

Neuroimmunological electron microscopy with microwave-accelerated fixation.

Immunoelectron microscopy is an important tool used to determine the precise location of immune complexes. Standard concentrations of glutaraldehyde destroy these complexes. This paper describes a method in which the period of glutaraldehyde fixation is shortened by concomitant microwave treatment. Using 1.25% glutaraldehyde and microwave fixation ideal preservation and demonstration of MHC class I antigen on Schwann cells was obtained by the peroxidase method.

Demonstration of transforming growth factor activity in mammary epithelial tissues.

Transforming growth factor (TGF) activity has been demonstrated in acid-ethanol extracts of bovine mammary gland, of Ehrlich Ascites Mammary Carcinoma Cells, and of the ascites fluid. The extracts differ in their activity in the soft agar test when using either human or rat fibroblasts. The most active extract obtained from mammary gland tissue was chromatographed and the TGF activity shown to be coeluting with EGF receptor-competing activity. The present data and our previous reports show that TGFs and a growth inhibitor for mammary epithelial cells coexist in bovine mammary gland as separate growth factors.

Secretion of phosphoproteins associated with neoplastic transformation and with the action of transforming growth factors.

Incubation of conditioned media derived from mammary epithelial cells and from fibroblasts with [gamma-32P]ATP revealed much higher intensities of labeled polypeptides in transformed cells compared to normal cells. Conditioned media from human mammary carcinoma cells have in common the presence of characteristic phosphoproteins with molecular masses of about 37,22 and 19.5 kDa. Ehrlich Ascites Mammary Carcinoma Cells secrete a dominant 32 kDa phosphoprotein. Normal fibroblasts secrete elevated levels of phosphoproteins after treatment with transforming growth factors. A phosphoprotein with molecular mass of about 37 kDa becomes secreted preferentially if cell-conditioned media were labeled in vivo. The results indicate that the phosphoproteins comprise a family of secretory polypeptides associated with early steps of transformation.

Immortalization of human Lesch-Nyhan-fibroblasts following infection with Simian virus 40.

Skin fibroblasts of three Lesch-Nyhan patients were successfully transformed to unrestricted growth by Simian virus 40. The transformed character of isolated clones was confirmed by their epitheloid growth in medium containing low concentration of serum, by hyperdiploid karyotype patterns, by detection of SV40 T-antigen, by their growth in soft agar and by the proliferation of cells in vitro up to 200 passages hitherto.

The DNA tumor virus SV 40 induces gene mutations in human cells. Reversion of HPRT deficiency.

The mutagenic effect of papovavirus SV40 on human cells could be demonstrated by reversion of HPRT deficiency in Lesch-Nyhan fibroblasts transformed by the virus. SV40 seems to induce different gene mutations in individually selected cell clones, as was clearly shown by the respective HPRT enzyme properties. The consequences of our results for use of SV40-derived vectors in gene substitution experiments are discussed.

HGPRT structural gene mutation in Lesch-Nyhan-syndrome as indicated by antigenic activity and reversion of the enzyme deficiency.

For three patients with the Lesch-Nyhan syndrome the existence of normal amounts of catalytically inactive hypoxanthine-guanine phosphoribosyltransferase (HGPRT) protein was demonstrated by using antibodies against the normal enzyme subunits. The lack of enzyme activity is reverted in virus transformed cells. Individual revertant cell clones contain different HGPRT enzymes as demonstrated here by isoelectric focusing. The data strongly support the idea of a structural gene mutation as the cause of enzyme deficiency in the Lesch-Nyhan syndrome.

Isolation of a SV40-like Papovavirus from a human glioblastoma.

A human glioblastoma multiforme (M27) tested in early cell cultures by indirect immunofluorescence staining showed SV40-related tumor (T)-antigen, 95% of the cells being positive. SV40-related viral capsid (V)-antigen was absent in all cells tested. Experiments to rescue this virus were performed by fusing M27 cells with CV-I monkey cells, which were permissive for SV40, using polyethylene glycol (PEG) as fusion factor. We succeeded in isolating virus particles SV40-GBM which electron microscopy showed to correspond in size and morphology to papovaviruses. Serological tests (hemagglutination, neutralization, fluorescent antibody) revealed that the virus is indistinguishable from SV40. Despite this apparent antigenic identity SV40-GBM differs slightly from SV40 wild type. This virus can propagate and produce CPE in both CV-I cells and primary fetal human kidney cells. Furthermore digestion of SV40-GBM DNA with the HindII/III restriction endonucleases revealed minor differences compared with the SV40 DNA. Therefore the virus SV40-GBM obtained from glioblastoma cells seems to be closely related to the SV40-PML viruses described earlier.