Additional file 3 of Perceived environmental barriers and facilitators of refugee children’s physical activity in/around refugee accommodation: a qualitative case study in Berlin

Additional file 3. (a) playable clock poll example for children in stage I; (b) environmental scale paper example for children’s drawing in A3 paper

Adaptive network traffic control with approximate dynamic programming based on a non-homogeneous Poisson demand model

In this study, we develop a stochastic dynamic traffic-flow model subject to practical restrictions under the non-homogeneous Poisson vehicle arrival process. Using the cell transmission strategy, we establish traffic dynamics between two intersections. We also discuss simulating the random demand of source links from an estimated intensity function. Additionally, we propose an algorithm to optimize time interval division for aggregated data, aiming to enhance estimation performance. We explore applying our traffic flow model to the adaptive traffic network management problem, which is formulated as a Markov decision process. Leveraging approximate dynamic programming with recursive least squares-temporal difference learning, we achieve adaptive optimal policies. To validate our approach, we conduct a series of numerical experiments with random demands. The results of non-homogeneous Poisson demand conducted using random numbers and a real-word dataset indicate high efficiency with the piecewise constant, I-SMOOTH, and MNO-PQRS estimators. Compared to the Webster and Max-pressure control systems, our proposed approximate dynamic programming-based model exhibits superior stability and applicability.</p

Additional file 1 of Perceived environmental barriers and facilitators of refugee children’s physical activity in/around refugee accommodation: a qualitative case study in Berlin

Additional file 1. Questionnaire for parents of their children’s daily playing (English version)*

Graft-versus-Host Disease Is Enhanced by Selective CD73 Blockade in Mice

<div><p>CD73 functions as an ecto-5′-nucleotidase to produce extracellular adenosine that has anti-inflammatory and immunosuppressive activity. We here demonstrate that CD73 helps control graft-versus-host disease (GVHD) in mouse models. Survival of wild-type (WT) recipients of either allogeneic donor naïve CD73 knock-out (KO) or WT T cells was similar suggesting that donor naïve T cell CD73 did not contribute to GVHD. By contrast, donor CD73 KO CD4⁺CD25⁺ regulatory T cells (Treg) had significantly impaired ability to mitigate GVHD mortality compared to WT Treg, suggesting that CD73 on Treg is critical for GVHD protection. However, compared to donor CD73, recipient CD73 is more effective in limiting GVHD. Pharmacological blockade of A2A receptor exacerbated GVHD in WT recipients, but not in CD73 KO recipients, suggesting that A2 receptor signaling is primarily implicated in CD73-mediated GVHD protection. Moreover, pharmacological blockade of CD73 enzymatic activity induced stronger alloreactive T cell activity, worsened GVHD and enhanced the graft-versus-leukemia (GVL) effect. These findings suggest that both donor and recipient CD73 protects against GVHD but also limits GVL effects. Thus, either enhancing or blocking CD73 activity has great potential clinical application in allogeneic bone marrow transplants.</p> </div

Variant analysis of MCM2.

<p>(a) Pedigree of Family #51 with hearing loss. This pedigree demonstrates nonsyndromic hearing loss of autosomal dominant inheritance. Open symbols, unaffected; solid symbols, affected. Squares, male; circles, female; slashed, deceased individual. Slanting arrow, the proband. Family members with the C>T symbol denote c.130C>T variant in <i>MCM2</i>, those with the C/C symbol had no variant in <i>MCM2</i>, and those without the symbols were not examined. Segregation of hearing loss with the c.130C>T variant in <i>MCM2</i> is remarkable in this pedigree. (b) Results of pure tone audiometry for eight members of Family #51 with hearing loss. Threshold data were obtained from the worse ears of the eight affected family members. We can clearly deduce from this graph that hearing loss is variable and mild to severe in degree in these cases. (c) A heterozygous c.130C>T variant in exon 2 of the <i>MCM2</i> gene was identified in the affected members in this pedigree. (d) Conservation analysis showed that Arg44 in human MCM2 is conserved across human, bovine, zebra finch, green anole, zebrafish, nile tilapia, acorn worm, sea urchin lineages.</p

MCM2 expression in inner ears.

<p>(a) Western blot of cochlear tissues from guinea pigs. The three lanes were total cochlear protein (Total), cochlear tissues without spiral ligament or stria vasculari (without sl. or sv.), and spiral ligament and stria vascularis parts (sl. and sv.), respectively. GAPDH (glyceraldehyde-3-phosphate dehydrogenase) was used as the reference. (b-f) Whole mounts of the hearing sensory epithelial layer from mature rat were prepared for immunofluorescence using anti-MCM2 antibody. Serial sections of confocal microscopic scanning were taken from superficial to deeper layers with an interval of 1 μm/layer at the site of one line of inner hair cells (double arrows) and three lines of outer hair cells (1, 2, 3 in the panels). MCM2 was stained with Alexa Fluor 488 and nuclei with DAPI. The nuclei of the outer hair cells are not shown as they lie deeper than the level of section G. MCM2 was distributed in the cytoplasm and was highly expressed in the hair skin plate of the two kinds of hair cell. Scale = 10 μm. (g) Cochlear frozen sections from mature rat were prepared for immunofluorescence using anti-MCM2 antibody. MCM2 was stained with Alexa Fluor 488. Three outer hair cells on the left and one inner hair cell on the right were shown in the panel. Scale = 10 μm.</p

CD73 deficient (KO) CD4⁺CD25⁺ Tregs have reduced ability to inhibit GVHD.

<p>(<b>A</b>) Lethally irradiated BALB/c mice were given i.v. injections of 5×10⁶ T cell depleted BM cells from B6 mice donors alone (n = 5) or with 1×10⁶ (1 M) or 0.5×10⁶ (0.5 M) splenic naïve T cells (CD25⁻CD62L⁺) from WT (n = 10) or CD73 KO (n = 10) B6 donors. (<b>B</b>) Lethally irradiated BALB/c mice were given i.v. injections of 5×10⁶ T cell depleted BM cells from B6 mice donors alone (n = 5) or with 2×10⁷ total splenocytes or CD25 negative (CD25⁻) splenocytes from WT (n = 10) or CD73 KO (n = 10) B6 donors. BM+WT-SP versus BM+CD73 KO-SP, p = 0.0399; BM+WT-SP versus BM+WT-CD25⁻, p = 0.0305. (<b>C</b>) Suppression of the alloresponse of B6 CD4⁺CD25⁻ T responder cells (Tres) to BABL/c cells by B6 WT or CD73 KO Tregs (ratio of Tres/Treg; 1∶1) in the MLR. WT Tregs were incubated with or without APCP at 100 µM before co-culture with indicated cells. NECA at 10 µM was added to the co-cultures with CD73 KO Tregs for the duration of the culture. Data are given as means ± SD of triplicates. (<b>D</b>) Lethally irradiated BALB/c mice were given i.v. injections of 5×10⁶ T cell depleted BM cells from B6 mice donors alone (n = 5) or plus 5×10⁵ purified B6 WT (n = 10) or CD73 KO (n = 10) CD4⁺CD25⁻ T cells ±5×10⁵ CD4⁺CD25⁺ Tregs. BM+WT-T versus BM+WT-T+Treg, p<0.0001; BM+WT-T+WT-Treg versus BM+WT-T+CD73 KO-Treg, p = 0.024.</p

Recipient CD73 limits GVHD development.

<p>(<b>A</b>) Lethally irradiated WT (n = 10) or CD73 KO (n = 10) BALB/c mice were given i.v. injections of 5×10⁶ T cell depleted BM cells from B6 mice donors alone (n = 5) or with 2×10⁷ splenocytes from WT B6 donors. Lethally irradiated WT (n = 10) or CD73 KO (n = 10) B6 mice were given i.v. injections of 5×10⁶ T cell depleted BM cells from BALB/c mice donors alone (n = 5) or with 2×10⁷ splenocytes from WT BALB/c donors (<b>B</b>) or CD73 KO BALB/c donors (<b>C</b>). (<b>A,B</b>) P values indicate the differences between WT versus CD73 KO recipients. (<b>C</b>) BALB WT→B6 CD73KO versus BALB CD73KO→B6 CD73KO, p = 0.0103; BALB WT→B6 CD73KO versus BALB CD73KO→B6 WT, p = 0.0029; BALB CD73KO→B6 WT versus BALB CD73KO→B6 CD73KO, p<0.0001; BALB WT→B6 WT versus BALB CD73KO→B6 WT; p = 0.0284. Results are representative of 2 independently performed experiments with similar results.</p

Changes in HEK293 cells expressing p.R44C variant MCM2.

<p>(a-b) HEK293 cells were treated with liposome transfection reagent only (lipo), empty vector transfection (empty vector), wild-type MCM2 plamid transfection (WT), and variant MCM2 plasmid transfection (M) respectively. After transfection for 48 h, apoptotic cell ratios were determined with annexin-V-PE-staining. Data represent the mean and SD of 3 experiments. The asterisks (**) indicate significant differences between the control and experimental groups or differences between the WT group and the M group (**p<0.001). (c-d) The western blot analysis of whole cell lysates from HEK293 cells showed a level of endogenic expression of MCM2 (as shown in lipo and empty vector groups). The upper bands in lane 3 and 4 (from the left) represent the transfected exogenous MCM2 with a GFP tag. The endogenous level of MCM2 is shown in the lower band. The ratio of cleaved caspase3 densitometry to exogenous MCM2 densitometry is compared. In the histogram, whole cell lysates from the variant MCM2 (M) transfected cells showed an upper regulated expression of cleaved caspase-3 compared with wild-type MCM2 (WT) transfected cells. Data represent the mean and SD of 3 experiments. The asterisks (*) indicate significant differences between the WT group and the M group (*p<0.01). (e) Cell cycle phases of HEK293 cells transiently transfected with wild-type or c.130C>T variant MCM2 cDNA plasmid for 48 h. Cell cycle phases were measured by flow cytometry using a cell cycle assay kit. In cells transfected with wild-type MCM2 cDNA plasmid, G1, S, and G2 phases were 27.08±4.99, 50.69±2.14, 22.21±6.04, respectively; N = 3. In cells transfected with the variant MCM2 cDNA plasmid, G1, S, and G2 phases were 28.61±1.29, 44.80±0.64, 26.57±0.80, respectively; N = 3.</p

CD73 affects GVHD development.

<p>Lethally irradiated BALB/c CD73 KO (<b>A</b>) and WT <b>(C</b>) or B6D2F1 (BDF1) (<b>D</b>) mice were given i.v. injections of 5×10⁶ T cell depleted bone marrow (BM) cells from C57BL/6 (B6) mice donors alone (n = 5) or with 2×10⁷ splenocytes from WT (n = 10) or CD73 KO (n = 10) B6 donors. P values indicate the differences between recipients receiving WT versus CD73 KO cells. Results are representative of 2 independently performed experiments with similar results. (<b>B</b>) Ten days later, pathological analyses of lung, liver, skin, and colon of recipient mice (<b>A</b>) receiving WT or CD73 KO SP were performed by H&E staining. Combined results of pathology scores for 5 mice in each group. Error bars indicate the standard errors of the mean in the group. Representative micrographs from recipient livers (magnification 200X) are shown.</p