Additional file 1 of Exploration of key factors in Gingival Crevicular fluids from patients undergoing Periodontally Accelerated Osteogenic Orthodontics (PAOO) using proteome analysis

Supplementary Material

Analysis of Carotenoids and Gene Expression in Apple Germplasm Resources Reveals the Role of <i>MdCRTISO</i> and <i>MdLCYE</i> in the Accumulation of Carotenoids

Carotenoids play an important role in the coloring and nutritional value of apple (Malus spp.) fruits. Here, six carotenoids, including lutein, zeaxanthin, β-carotene, β-cryptoxanthin, violaxanthin, and neoxanthin, were detected in 105 fruits of apple germplasm resources, which showed a skewed distribution in both the peel and pulp. There were more carotenoids in the peel than in the pulp, and lutein and β-carotene were the primary carotenoids that were present. The expression levels of most carotenoid pathway genes in germplasm fruits during fruit development were higher in the fruits that had an abundance of carotenoids. A linear relationship analysis showed that the expression levels of MdCRTISO and MdLCYE were highly correlated with the content of carotenoids. The leaves accumulated the greatest number of carotenoids, while the roots had the lowest amount. MdCRTISO and MdLCYE were highly expressed in the fruits compared to other tissues. Transgenic calli and transiently transformed fruits confirmed that MdCRTISO and MdLCYE affected the biosynthesis of carotenoids owing to their effects on the expression of other genes for enzymes in the carotenoid pathway. Our findings will extend the understanding of carotenoid biosynthesis in apple and excavate apple germplasm resources with rich carotenoids to breed high-quality apples

Table_1_Effects of Exogenous Dopamine on the Uptake, Transport, and Resorption of Apple Ionome Under Moderate Drought.pdf

<p>The frequency and intensity of water deficits is expected to increase because of global warming. Drought stress is often one of the most limiting factors for plant growth. We conducted greenhouse pot experiments to address how dopamine affects the drought-resistance traits of apple trees at the physiological and molecular levels. Our factorial design consisted of dopamine and no-dopamine applications combined with well-watered and moderate-drought conditions. Seedling biomass, photosynthesis rates, chlorophyll concentrations, and stomatal apertures were markedly reduced under stress but dopamine treatment mitigated the inhibiting effects of drought on plant growth and helped maintain strong photosynthesis, chlorophyll levels, and stomatal functioning. Concentrations of most macro-, micro-, and trace elements decreased in response to drought. This stress also diminished the uptake and transport of elements in the leaves and stems, but increased the partitioning of elements in the roots. Nutrient resorption proficiency decreased while nutrient resorption efficiency increased for most analyzed elements. Exogenous dopamine significantly increased the concentrations, uptake, and transport of nutrients under drought stress, and also altered their distribution within the whole plant. However, this molecule had a negative effect on nutrient resorption. Although transcript levels of a key chlorophyll degradation gene, pheide a oxygenase, and senescence-associate gene 12 were elevated upon drought treatment, dopamine significantly suppressed the upregulation of those genes under such stress conditions. These observations indicate that dopamine has an important anti-senescence effect that might be helpful for regulating nutrient uptake, transport, and resorption, and ultimately influencing overall plant growth. Thus, understanding the role of dopamine in drought tolerance introduces new possibilities to use this compound for agricultural purposes.</p

Tregs Promote the Differentiation of Th17 Cells in Silica-Induced Lung Fibrosis in Mice

<div><h3>Background</h3><p>Silicosis is an occupational lung disease caused by inhalation of silica dust and characterized by lung inflammation and fibrosis. Previous study showed that Tregs regulate the process of silicosis by modulating the maintenance of immune homeostasis in the lung. Th17 cells share reciprocal developmental pathway with Tregs and play a pivotal role in the immunopathogenesis of many lung diseases by recruiting and activating neutrophils, but the regulatory function of Tregs on Th17 response in silica induced lung fibrosis remains to be explored.</p> <h3>Methodology/Principal Findings</h3><p>To evaluate the role of Th17 and IL-17 in the development of silicosis and their interaction with Tregs, Treg-depleted mice model was generated and exposed to silica to establish experimental model of silica-induced lung fibrosis. Here we showed that silica increased Th17 response in lung fibrosis. Tregs depletion enhanced the neutrophils accumulation and attenuated Th17 response in silica induced lung fibrosis. Both mRNA and protein results showed that Tregs exerted its modulatory function on Th17 cells and IL-17 by regulating TGF-β1 and IL-1β.</p> <h3>Conclusion/Significance</h3><p>Our study suggested that Tregs could promote Th17 cells differentiation by regulating TGF-β1 and IL-1β in silica induced lung fibrosis of mice, which further the understanding of the progress of silicosis and provide a new insight in the regulatory mechanism of Th17 by Tregs in lung inflammation.</p> </div

The regulatory function of Tregs on Th17 and IL-17A may depend on IL-1β but not IL-6 and IL-23.

<p>The IL-1β (A), IL-6 (B) and IL-23 (C) mRNA were assayed by realtime RT-PCR by using −ΔΔCt method. Results (n = 5) are shown as mean±SEM (one-way analysis of variance followed by pair-wise comparison with the Student-Newman-Keuls test.*, as compared with the saline control group, P<0.05; Δ, as compared with the silica group, P<0.05; +, as compared with 3day of the same group, P<0.05; #, as compared with 7day of the same group, P<0.05).</p

Th1 type of cytokines took the advantage over Th2 cytokine in the depletion of Tregs.

<p>Typical Th1 IL-2 (A), IFN-γ (B), IL-12 (C) and Th2 IL-4 (D) cytokines (day 3) were assayed by realtime RT-PCR by using −ΔΔCt method. Results (n = 5) are shown as mean±SEM (one-way analysis of variance followed by pair-wise comparison with the Student-Newman-Keuls test. *, as compared with the saline control group, P<0.05; Δ, as compared with the silica group, P<0.05).</p

TGF-β1 and IL-10 decreased in the Tregs depletion group and TGF-β1 may contribute to the Th17 cells differentiation.

<p>The IL-10 (A) and TGF-β1 (B) mRNA (day 3) were assayed by realtime RT-PCR by using −ΔΔCt method. Results (n = 5) are shown as mean±SEM (one-way analysis of variance followed by pair-wise comparison with the Student-Newman-Keuls test. *, as compared with the saline control group, P<0.05; Δ, as compared with the silica group, P<0.05).</p

Histopathology changes in mouse lungs after instillation with HE staining (×200).

<p>The scale on the graph above was 50 µm; date was day3, day7, day28. Lung sections were stained with H&E. The degree of inflammation was assessed by the histological analysis of six random fields per sample (with n = 5 mice per group).</p

Cell infiltration and alveolar change of the mice lungs in each group at day 3, 7, 28 and 56.

<p>The degree of inflammation was assessed by the histological analysis of six random fields per sample (n = 5).</p

Depletion of Tregs decreased Th17 response in silica induced lung fibrosis.

<p>The RORγ-t (A) and IL-17A (B) mRNA were assayed by realtime RT-PCR by using −ΔΔCt method. Results (n = 5) are shown as mean±SEM (one-way analysis of variance followed by pair-wise comparison with the Student-Newman-Keuls test. *, as compare with the saline control group, P<0.05; Δ, as compared with the silica group, P<0.05; + compared with 3day of the same group, P<0.05; #, as compared with 7day of the same group, P<0.05).</p