Supporting the development of scientific enquiry and conceptual understanding in science with deaf and typically hearing preschool children through a home-based science intervention

The study sought to determine the effects of a pilot home-based science intervention on deaf and typically hearing children’s conceptual understanding of science and their abilities to reason about and communicate their understanding (scientific enquiry skills). Data show that by age 5 years a science attainment gap exists between deaf children and those typically hearing yet early interventions with deaf children typically neglect the development of science. We created an intervention to support caregivers (N = 17) to incorporate the language of science into their daily routines. Participants were placed into a control or intervention group; within each group were caregivers of deaf and typically hearing children. Over a period of 13 months, caregivers completed five assessments (each at two different time points); four on their child’s understanding of concepts and one on their developing scientific enquiry skills. All caregivers, irrespective of group, reported positive gains in their child’s conceptual understanding. Caregivers of deaf children reported greater gains in scientific enquiry skills than those of typically hearing children. This offers preliminary evidence to suggest that longer-term interventions may provide an opportunity to narrow the attainment gap between deaf and typically hearing children.</p

Constraining and enabling factors to using long-term climate information in decision-making

We carry out a structured review of the peer-reviewed literature to assess the factors that constrain and enable the uptake of long-term climate information in a wide range of sectoral investment and planning decisions. Common applications of long-term climate information are shown to relate to urban planning and infrastructure, as well as flood and coastal management. Analysis of the identified literature highlights five categories of constraints: disconnection between users and producers of climate information, limitations of climate information, financial and technical constraints, political economy and institutional constraints and finally psycho-social constraints. Five categories of enablers to the uptake of long-term climate information in decision-making are also identified: collaboration and bridge work, increased accessibility of climate information, improvement in the underlying science, institutional reform and windows of opportunity for building trust. Policy relevance Our review suggests that stand-alone interventions aimed at promoting the uptake of climate information into decision-making are unlikely to succeed without genuine and sustained relationships between producers and users. We also highlight that not every decision requires consideration of long-term climate information for successful outcomes to be achieved. This is particularly the case in the context of developing countries, where the immediacy of development challenges means that decision makers often prioritize short-term interventions. Care should therefore be taken to ensure that information is targeted towards investments and planning decisions that are relevant to longer-term timescales.</p

A20 controls maturation and cytokine production of Mф.

<p><b>A</b>. Expression of costimulatory molecules and MHC class II molecule on the adenoviral-transduced BMMф in response to stimulation of LPS. <b>B</b>. Production of inflammatory cytokines by the adenoviral-transduced BMMфs, as tested by ELISA. <b>C</b>. NO production by adenoviral-transduced BMMфs, as tested by Griess assay. Experiments were repeated three times with similar results. *p<0.05, **p<0.01 Ad-shA20- vs. Ad-con-transduced Mф.</p

A20-silenced Mф elicits a cytotoxic CD4⁺ T cell response via activation of IFN-γ signaling and by an MHC-class-II-restricted mechanism.

<p>A. Adenoviral-transduced BMMфs were used to immunize IFNGR^−/− mice or the wildtype littermates (2–3 mice/group) twice. The inguinal LNs were harvested for analyzing expression of granzyme B in CD4⁺ or CD8⁺ T cells by ICS. p<0.01 Ad-shA20-IFNGR KO mice vs. Ad-ShA20 WT mice. B. Adenoviral-transduced BMMфs were used to immunize Stat1^−/− mice or the wild-type littermates twice (2–3 mice/group). The LNs were harvested for analyzing expression of granzyme B in CD4⁺ (p<0.05, Ad-shA20-Stat1 KO mice vs. Ad-shA20-WT mice) or CD8⁺ T cells by ICS. C. BMMфs were prepared from MHCII^−/− mice or the wild-type littermates. The adenoviral-transduced BMMфs were used to immunize wild-type mice (2–3 mice/group) twice. The LNs were harvested for analyzing expression of granzyme B in CD4⁺ (p<0.01, Ad-shA20-MHC-II KO Mф immunization vs. Ad-shA20-WT Mф immunization) or CD8⁺ T cells by ICS. Experiments were repeated with similar results.</p

IFN-γ enhances MФ to prime cytotoxic T cells response <i>in vitro.</i>

<p>BMMфs were transduced with Ad-con and cocultured with CD4⁺ OT-II (<b>A</b>) or CD8⁺ OT-I (<b>B</b>) T cells in the presence of the different doses of IL-6, IL-12 or IFN-γ (2.5 ug/ml or 10 ug/ml ) for 3–5 days. A20-silenced MФ priming OT-II or OT-I T cells was used as positive control. Expression of granzyme B in T cells was assessed by ICS assay. The data is a representative of three independent experiments. p<0.01, OT-II/con-Mф+IFN-γ(10 ug/ml) vs. OT-II/con-Mф or OT-I/con-Mф+IFN-γ (10 ug/ml) vs. OT-I/con-Mф.</p

A20-silenced Mф immunization enhances NK cell-, CD8⁺ T cell- and CD4⁺ T cell-mediated cytotoxicity.

<p>Splenocytes pooled from 2–3 immunized mice were cultured overnight for NK-mediated cytotoxicity assay or 5–6 days in the presence of OT-I or OT-II peptide for T cells-mediated cytotoxicity assay. The splenocytes cultured with OT-II peptide were selected using anti-CD4 beads prior to cytotoxicity assay. Cytotoxic activities were analyzed by LDH release assay as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048930#s4" target="_blank">Material and Methods</a>. Experiments were repeated three times with similar results. *p<0.05, Ad-shA20-Mф immunization vs. Ad-con-Mф immunization for specific killing.</p

A20-silenced Mф immunization induces enhanced immune protection.

<p><b>A & B.</b> C57BL/6 mice (5–6 mice/group) were immunized twice. The mice were s.c. injected with 5×10⁵ EG-7 (<b>A</b>) or M05 (<b>B</b>). Tumor growth was monitored on the indicated days. * p<0.05, Ad-shA20-Mф immunization vs. Ad-con-Mф immunization. <b>C</b>. CD4^−/− C57BL/6 or the wildtype littermates (5–6 mice/group) were immunized with OT-II-peptide-pulsed, Ad-shA20-transduced BMMфs twice followed by s.c. injection of 5×10⁵ M05 tumor cells. Tumor occurrence and growth were monitored on the indicated days. **p<0.01, wild-type mice vs. CD4^−/− mice. <b>D</b>. Transferred OT-II-specific immune pretection. In vitro primed OT-II T cells (5×10⁶) were transplanted into naïve RAG^−/−C57BL/6 mice (5 mice/group) by retro-orbital injection following s.c injection of OVA-expressed B6SJ1003 tumor cells (6×10⁵). The transplantation of OT-II T cells was repeated one week later. One group of mice were transplanted with CMA-treated, Ad-shA20-transduced Mф-primed OT-II T cells. Tumor growth was monitored on the indicated days. *p<0.05, Ad-shA20-Mф-primed OT-II T cell transfer vs. Ad-con-Mф-primed OT-II T cell transfer, or Ad-shA20-Mф-primed OT-II T cell transfer vs. Ad-shA20-Mф-primed OT-II T cell+ CMT transfer. All the experiments were repeated with similar results.</p

A20-silenced Mф enhances expression of granzyme B in CD4⁺ T cells, CD8⁺T cells or NK cells.

<p><b>A,</b> adenoviral-transduced BMMфs were cocultured with freshly isolated OT-I (<b>uppe</b>r) or OT-II cells (<b>lower</b>) at a raito of 1∶10. 3–5 days later, the cocultured T cells were harvested for analyzing expression of granzyme B by ICS. The data is shown as a representative of 3 independent experiments. (p<0.05, OT-I/shA20-Mф vs. OT-I/con- Mф; p<0.01, OT-II/shA20-Mф vs. OT-II/con-Mф). <b>B,</b> C57BL/6 mice (5–6 mice/group) were immunized (<i>i.p</i>) twice with different adenoviral-transduced Mфs or PBS. Lymphocytes were isolated from the inguinal LNs to analyze expression of granzyme B in NK cells, CD8<b>⁺</b> or CD4<b>⁺</b> T cells by ICS. <b>C.</b> C57BL/6 mice were immunized (<i>i.p</i>) twice with OT-II-pulsed, different adenoviral-transduced BMMфs or PBS. Splenocytes were harvested and in vitro restimulated with OT-II peptide for 48 hrs. CD4⁺ T cells were isolated for analysis of granzyme B expression by qPCR. The data is shown as a representation of three independent experiments. (* p<0.01, shA20- Mф-mice vs. con- Mф-mice).</p

Supplementary figure-2 from Mutant IDH1 Cooperates with ATRX Loss to Drive the Alternative Lengthening of Telomere Phenotype in Glioma

Mutant IDH1 expression is associated with down- regulation of RAP1 and XRCC1</p

Neutralization of IFN-γ reduces A20-silenced MФ to prime cytotoxic T cell response <i>in vitro.</i>

<p>BMMфs were transduced with Ad-shA20 and cocultured with CD4⁺ OT-II (<b>A</b>) or CD8⁺ OT-I (<b>B</b>) T cells in the presence of the different doses of anti-IL-6, anti-IL-12 or anti-IFN-γ (2.5 ug/ml, 10 ug/ml, or 20 ug/ml ) for 3–5 days. Expression of granzyme B in T cells was assessed by ICS assay. The data is a representative of three independent experiments. p<0.01, OT-II/AdshA20-Mф vs. OT-II/AdshA20-Mф+anti-IFN-γ(20 ug/ml).</p