Awareness of high school students for a healthy way of life - sources and methods

Having basic knowledge, habits and skills about health is essential for living a healthy life. Basic components, connected with the health of students, are a healthy diet, physical activities, social and psychic health.The goal of this research is to determine and analyze the level of awareness of healthy living among students from grades eight to eleven.A total of one hundred and sixty (160) students from four schools, all located in the Varna, have taken part in the research. The research was conducted from February till March 2015. We handed out individual polls, the results of which were processed with mathematical statistic and graphical analyses.Independently from the discretion, that they are informed (90%), the results show that there are things the students do not know. A big part of them rely on information from friends or the internet, as around half of the students (55%) say that they count on their schools for information about health related questions. The information is mostly given by their form-teacher (68%) and the schools psychologist (56%). Medical specialists and lectors are rarely engaged (26%). A big part of the students say that they find the traditional ways of giving out information very helpful

The JDRF Network for the Pancreatic Organ Donor with Diabetes (nPOD): A novel Resource and Study Approach in Type 1 Diabetes Research

The JDRF nPOD organization has three goals: To establish a biobank from organ donors with T1D. To distribute tissues to nPOD investigators in support of diverse studies of human T1D. To promote collaboration and data sharing among nPOD investigators and achieve a comprehensive understanding of human T1D. The JDRF nPOD supports 71 projects in key research areas. Findings from nPOD investigators include abnormalities of self-antigen expression in the pancreatic lymph node, which could impair self-tolerance, the localization of antigen-specific autoreactive T-cells in the insulitic lesions, evidence of heterogeneity and distinct patterns of beta-cell destruction, lack of insulitis in donors with a single autoantibody, and beta-cell persistence in patients with long standing disease. Programmatic expansions will apply the nPOD model to additional areas of the diabetes spectrum, such as diabetes complications and diabetes in patients after transplantation. The JDRF nPOD is organizing working groups, which apply the concepts of real-time data sharing to accelerate the rate of discovery. The first working group is addressing viruses in diabetes. Ultimately, nPOD provides a scientific venue and a “cloud” environment consisting of shared tissues, data sharing and collaborative, coordinated study design developed with collective input

Transient B-Cell Depletion with Anti-CD20 in Combination with Proinsulin DNA Vaccine or Oral Insulin: Immunologic Effects and Efficacy in NOD Mice

<div><p>A recent type 1 diabetes (T1D) clinical trial of rituximab (a B cell-depleting anti-CD20 antibody) achieved some therapeutic benefit in preserving C-peptide for a period of approximately nine months in patients with recently diagnosed diabetes. Our previous data in the NOD mouse demonstrated that co-administration of antigen (insulin) with anti-CD3 antibody (a T cell-directed immunomodulator) offers better protection than either entity alone, indicating that novel combination therapies that include a T1D-related autoantigen are possible. To accelerate the identification and development of novel combination therapies that can be advanced into the clinic, we have evaluated the combination of a mouse anti-CD20 antibody with either oral insulin or a proinsulin-expressing DNA vaccine. Anti-CD20 alone, given once or on 4 consecutive days, produced transient B cell depletion but did not prevent or reverse T1D in the NOD mouse. Oral insulin alone (twice weekly for 6 weeks) was also ineffective, while proinsulin DNA (weekly for up to 12 weeks) showed a trend toward modest efficacy. Combination of anti-CD20 with oral insulin was ineffective in reversing diabetes in NOD mice whose glycemia was controlled with SC insulin pellets; these experiments were performed in three independent labs. Combination of anti-CD20 with proinsulin DNA was also ineffective in diabetes reversal, but did show modest efficacy in diabetes prevention (p = 0.04). In the prevention studies, anti-CD20 plus proinsulin resulted in modest increases in Tregs in pancreatic lymph nodes and elevated levels of proinsulin-specific CD4+ T-cells that produced IL-4. Thus, combination therapy with anti-CD20 and either oral insulin or proinsulin does not protect hyperglycemic NOD mice, but the combination with proinsulin offers limited efficacy in T1D prevention, potentially by augmentation of proinsulin-specific IL-4 production.</p> </div

Combination therapy increases numbers of CD4+ Foxp3+ cells in pancreas draining lymph node 60-days post treatment.

<p>Eight to ten week old prediabetic NOD mice were given 50 µg of anti-CD20 either alone or in combination with weekly administration of 50 µg proinsulin plasmid for four weeks. Splenocytes and PDLN cells from mono- or combination therapy treated mice were stained with anti-IgM, -B220 (B-cells), -CD4, -CD8, -CD25 and –Foxp3. B-cell frequencies were determined by gating on IgM+ B220+ cells. Frequency of Foxp3+ cells among CD4+ T-cells was determined by gating on CD4+ cells. Cumulative frequencies of B-cells (left panel), CD4+ T-cells (middle panel), and CD4+Foxp3+ cells (right panel) in PDLN (upper panels) or spleen (lower panels) are shown at 30 days and 60 days post combination therapy. Representative data from three independent experiments with similar results is shown. Statistical analysis was performed using unpaired students <i>t-</i>test using Graphpad Prism software. Each dot represents one mouse, with mean value indicated on the graph.</p

Combination therapy with anti-CD20 and proinsulin plasmid prevents T1D more efficiently.

<p>Eight to ten week old prediabetic NOD mice were treated with (<b>A</b>) weekly administration of 50 µg proinsulin plasmid only or, (<b>B</b>) one-time administration of anti-CD20 alone or, (<b>C</b>) anti-CD20 administered in combination with four weekly administrations of proinsulin plasmid. Statistical tests were performed by Kaplan-Meier analysis using Graphpad Prism software.</p

T-cells from combination therapy treated mice produce IL-4 in response to proinsulin peptide stimulation.

<p>At 60-days post anti-CD20 mono- (MT, open bars) or combination therapy (CT, filled black bars), CD4+ T-cells were purified from treated non-diabetic NOD mice. Simultaneously, T-depleted splenocytes (TDS) were obtained from 8–10 week old NOD mice and used as APCs. 250,000 purified CD4+ T-cells were incubated with proinsulin peptide or a mutated insulin B:9–23 (B16:A) peptide in the presence of 5×10⁴ APCs. Following 3-day incubation, IL-4 production was determined by ELISpot assay as previously described <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0054712#pone.0054712-Fousteri1" target="_blank">[34]</a>. Background (media) subtracted spot numbers are shown on the Y-axis. Increased IL-4 production in response to proinsulin peptide was seen in combination therapy treated mice. Representative means ± SEM data from one of two independent experiments with similar results are shown. Statistical analysis was performed using unpaired students <i>t-</i>test using Graphpad Prism software.</p

Novel murine anti-CD20 administration preferentially depletes follicular but not marginal zone B-cells.

<p>Eight week old female NOD mice were treated with 50 µg anti-CD20 and 7 days later various B-cell subsets in the spleen were analyzed. Gating strategy is shown in the top panel. Lymphocytes were gated on (<b>A</b>) B220+ (total B-cells) and further classified as FO B-cells (B220+ IgM^hi, CD21^int), immature B-cells (B220+ IgM^hi CD21⁻), and MZ B-cells (B220+ IgM^hi CD21^hi). Total B220+ B-cell numbers were reduced in the spleen with significant reductions observed in number of follicular and immature B-cells but not MZ B-cells (<b>B</b>). Eight week old female NOD mice were given anti-CD20 i.v. Seven days post anti-CD20 treatment, frequencies of B- and T-cells in peripheral blood were determined by staining with anti-IgM/B220 for B-cells or anti-CD4/CD8 for T-cells; loss of circulating B-cells was associated with compensatory increases in CD4+ and CD8+ T-cell frequencies in NOD mice (<b>C</b>). Frequencies of circulating B-cells returned to similar levels as in untreated NOD mice by 60 days post-treatment (<b>D</b>). Data are means ± SEM. Representative data from 3 independent experiments with similar results are shown. Statistical analysis was performed using unpaired students <i>t-</i>test using Graphpad Prism software.</p

Combination therapy with anti-CD20 and proinsulin plasmid does not offer protection after T1D onset.

<p>NOD mice with hyperglycemia (2 consecutive readings of BGV>180 mg/dl), were treated with either (<b>A</b>) weekly administration of 50 µg proinsulin plasmid or (<b>B</b>) 100 (upper panel) or 250 µg of anti-CD20 alone or (<b>C</b>) 50 (upper panel) or 250 µg of anti-CD20 antibody in combination with weekly administration 50 µg of proinsulin plasmid. Each line represents one individual mouse.</p