Hedgehog signalling in allograft vasculopathy: a new therapeutic target?

Allograft vasculopathy (AV) leads to chronic rejection of organ transplants, but its causes are obscure. New research from the Jane-Wit laboratory showed that Sonic Hedgehog (SHH) signalling from damaged graft endothelium drives vasculopathy by promoting proinflammatory cytokine production and NLRP3-inflammasome activation in alloreactive CD4+PTCH1hiPD-1hiT memory cells, offering new diagnostic and therapeutic strategies

C11orf95-RELA reprograms 3D epigenome in supratentorial ependymoma.

Supratentorial ependymoma (ST-EPN) is a type of malignant brain tumor mainly seen in children. Since 2014, it has been known that an intrachromosomal fusion C11orf95-RELA is an oncogenic driver in ST-EPN [Parker et al. Nature 506:451-455 (2014); Pietsch et al. Acta Neuropathol 127:609-611 (2014)] but the molecular mechanisms of oncogenesis are unclear. Here we show that the C11orf95 component of the fusion protein dictates DNA binding activity while the RELA component is required for driving the expression of ependymoma-associated genes. Epigenomic characterizations using ChIP-seq and HiChIP approaches reveal that C11orf95-RELA modulates chromatin states and mediates chromatin interactions, leading to transcriptional reprogramming in ependymoma cells. Our findings provide important characterization of the molecular underpinning of C11orf95-RELA fusion and shed light on potential therapeutic targets for C11orf95-RELA subtype ependymoma

Optimising the Use of TRIzol-extracted Proteins in Surface Enhanced Laser Desorption/ Ionization (SELDI) Analysis

BACKGROUND: Research with clinical specimens is always hampered by the limited availability of relevant samples, necessitating the use of a single sample for multiple assays. TRIzol is a common reagent for RNA extraction, but DNA and protein fractions can also be used for other studies. However, little is known about using TRIzol-extracted proteins in proteomic research, partly because proteins extracted from TRIzol are very resistant to solubilization. RESULTS: To facilitate the use of TRIzol-extracted proteins, we first compared the ability of four different common solubilizing reagents to solubilize the TRIzol-extracted proteins from an osteosarcoma cell line, U2-OS. Then we analyzed the solubilized proteins by Surface Enhanced Laser Desorption/ Ionization technique (SELDI). The results showed that solubilization of TRIzol-extracted proteins with 9.5 M Urea and 2% CHAPS ([3-[(3-cholamidopropyl)-dimethylammonio]propanesulfonate]) (UREA-CHAPS) was significantly better than the standard 1% SDS in terms of solubilization efficiency and the number of detectable ion peaks. Using three different types of SELDI arrays (CM10, H50, and IMAC-Cu), we demonstrated that peak detection with proteins solubilized by UREA-CHAPS was reproducible (r > 0.9). Further SELDI analysis indicated that the number of ion peaks detected in TRIzol-extracted proteins was comparable to a direct extraction method, suggesting many proteins still remain in the TRIzol protein fraction. CONCLUSION: Our results suggest that UREA-CHAPS performed very well in solubilizing TRIzol-extracted proteins for SELDI applications. Protein fractions left over after TRIzol RNA extraction could be a valuable but neglected source for proteomic or biochemical analysis when additional samples are not available

Overexpression of ZFX confers self-renewal and chemoresistance properties in hepatocellular carcinoma

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Hedgehog signalling promotes Th2-differentiation in naive human CD4 T-cells

Original journal article Abstract: Here we show that differentiation of human naïve CD4 T-cells to Th2 is promoted by Hedgehog signaling and attenuated by SMO-inhibition. As Hedgehog proteins are produced by epithelial tissues this finding is important to understanding atopic disease

Systemic Pharmacological Smoothened Inhibition Reduces Lung T-Cell Infiltration and Ameliorates Th2 Inflammation in a Mouse Model of Allergic Airway Disease

Allergic asthma is a common inflammatory airway disease in which Th2 immune response and inflammation are thought to be triggered by inhalation of environmental allergens. Many studies using mouse models and human tissues and genome-wide association have indicated that Sonic Hedgehog (Shh) and the Hedgehog (Hh) signaling pathway are involved in allergic asthma and that Shh is upregulated in the lung on disease induction. We used a papain-induced mouse model of allergic airway inflammation to investigate the impact of systemic pharmacological inhibition of the Hh signal transduction molecule smoothened on allergic airway disease induction and severity. Smoothened-inhibitor treatment reduced the induction of Shh, IL-4, and IL-13 in the lung and decreased serum IgE, as well as the expression of Smo, Il4, Il13, and the mucin gene Muc5ac in lung tissue. Smoothened inhibitor treatment reduced cellular infiltration of eosinophils, mast cells, basophils, and CD4+ T-cells to the lung, and eosinophils and CD4+ T-cells in the bronchoalveolar lavage. In the mediastinal lymph nodes, smoothened inhibitor treatment reduced the number of CD4+ T-cells, and the cell surface expression of Th2 markers ST2 and IL-4rα and expression of Th2 cytokines. Thus, overall pharmacological smoothened inhibition attenuated T-cell infiltration to the lung and Th2 function and reduced disease severity and inflammation in the airway

Microwave Intensified Synthesis: Batch and Flow Chemistry

Many studies have been conducted on organic and inorganic synthesis by microwave heating owing to its special heating mechanism, leading to improved reaction rate, higher purity and yields. We specifically demonstrated microwave heating in the fabrication of nanoparticles and polyester. By fine-tuning the microwave and experimental parameters, the materials prepared have shown excellent physical and bio-properties, e. g. narrow particle size distribution, controlled morphology, varied molecular structure and so forth. We further highlight the recent procedure of using fluidic reactors on preparing both metals and metal oxides nanoparticles. The experimental design strategies and fundamentals of the microwave interaction with chemicals are presented. Furthermore, the key factors and issues facing in this area are also discussed

The Pioneer Transcription Factor Foxa2 Modulates T Helper Differentiation to Reduce Mouse Allergic Airway Disease

Foxa2, a member of the Forkhead box (Fox) family of transcription factors, plays an important role in the regulation of lung function and lung tissue homeostasis. FOXA2 expression is reduced in the lung and airways epithelium of asthmatic patients and in mice absence of Foxa2 from the lung epithelium contributes to airway inflammation and goblet cell hyperplasia. Here we demonstrate a novel role for Foxa2 in the regulation of T helper differentiation and investigate its impact on lung inflammation. Conditional deletion of Foxa2 from T-cells led to increased Th2 cytokine secretion and differentiation, but decreased Th1 differentiation and IFN-γ expression in vitro. Induction of mouse allergic airway inflammation resulted in more severe disease in the conditional Foxa2 knockout than in control mice, with increased cellular infiltration to the lung, characterized by the recruitment of eosinophils and basophils, increased mucus production and increased production of Th2 cytokines and serum IgE. Thus, these experiments suggest that Foxa2 expression in T-cells is required to protect against the Th2 inflammatory response in allergic airway inflammation and that Foxa2 is important in T-cells to maintain the balance of effector cell differentiation and function in the lung

The role of silicon-based nanofillers and polymer crystallization on the breakdown behaviors of polyethylene blend nanocomposites

Good breakdown strength is an important feature for the selection of dielectric materials, especially in high-voltage engineering. Although nanocomposites have been shown to possess many promising dielectric properties, the breakdown strength of nanocomposites is often found to be negatively affected. Recently, imposing nonisothermal crystallization processes on polyethylene blends has been demonstrated to be favorable for breakdown strength improvements of dielectric materials. In an attempt to increase nanocomposites' voltage rating, this work reports on the effects of nonisothermal crystallization (fast, moderate and slow crystallizations) on the structure and dielectric properties of a polyethylene blend (PE) composed of 80% low density polyethylene and 20% high density polyethylene, added with silicon dioxide (SiO2) and silicon nitride (Si3N4) nanofillers. Through breakdown testing, the breakdown performance of Si3N4-based nanocomposites was better than SiO2-based nanocomposites. Since nanofiller dispersion within both nanocomposite systems was comparable, the enhanced breakdown performance of Si3N4-based nanocomposites is attributed to the surface chemistry of Si3N4 containing less hydroxyl groups than SiO2. Furthermore, the breakdown strength of SiO2-based nanocomposites and Si3N4-based nanocomposites improved, with the DC breakdown strength increasing by at least 12% when both the nanocomposites were subjected to moderate crystallization rather than fast and slow crystallizations. This is attributed to changes in the underlying molecular conformation of PE in addition to water-related effects. These results suggest that apart from changes in the nanofiller surface chemistry, changes in the underlying molecular conformation of polymers are also important to improve the breakdown performance of nanocomposites