The microbial metabolite desaminotyrosine enhances T-cell priming and cancer immunotherapy with immune checkpoint inhibitors

Background Inter-individual differences in response to immune checkpoint inhibitors (ICI) remain a major challenge in cancer treatment. The composition of the gut microbiome has been associated with differential ICI outcome, but the underlying molecular mechanisms remain unclear, and therapeutic modulation challenging. Methods We established an in vivo model to treat C57Bl/6j mice with the type-I interferon (IFN-I)-modulating, bacterial-derived metabolite desaminotyrosine (DAT) to improve ICI therapy. Broad spectrum antibiotics were used to mimic gut microbial dysbiosis and associated ICI resistance. We utilized genetic mouse models to address the role of host IFN-I in DAT-modulated antitumour immunity. Changes in gut microbiota were assessed using 16S-rRNA sequencing analyses. Findings We found that oral supplementation of mice with the microbial metabolite DAT delays tumour growth and promotes ICI immunotherapy with anti-CTLA-4 or anti-PD-1. DAT-enhanced antitumour immunity was associated with more activated T cells and natural killer cells in the tumour microenvironment and was dependent on host IFN-I signalling. Consistent with this, DAT potently enhanced expansion of antigen-specific T cells following vaccination with an IFN-I-inducing adjuvant. DAT supplementation in mice compensated for the negative effects of broad-spectrum antibiotic-induced dysbiosis on anti-CTLA-4-mediated antitumour immunity. Oral administration of DAT altered the gut microbial composition in mice with increased abundance of bacterial taxa that are associated with beneficial response to ICI immunotherapy. Interpretation We introduce the therapeutic use of an IFN-I-modulating bacterial-derived metabolite to overcome resistance to ICI. This approach is a promising strategy particularly for patients with a history of broad-spectrum antibiotic use and associated loss of gut microbial diversity

Cardiomyocyte overexpression of miR-27b induces cardiac hypertrophy and dysfunction in mice

Recent studies have begun to reveal critical roles of microRNAs (miRNAs) in the pathogenesis of cardiac hypertrophy and dysfunction. In this study, we tested whether a transforming growth factor-β (TGF-β)-regulated miRNA played a pivotal role in the development of cardiac hypertrophy and heart failure (HF). We observed that miR-27b was upregulated in hearts of cardiomyocyte-specific Smad4 knockout mice, which developed cardiac hypertrophy. In vitro experiments showed that the miR-27b expression could be inhibited by TGF-β1 and that its overexpression promoted hypertrophic cell growth, while the miR-27b suppression led to inhibition of the hypertrophic cell growth caused by phenylephrine (PE) treatment. Furthermore, the analysis of transgenic mice with cardiomyocyte-specific overexpression of miR-27b revealed that miR-27b overexpression was sufficient to induce cardiac hypertrophy and dysfunction. We validated the peroxisome proliferator-activated receptor-γ (PPAR-γ) as a direct target of miR-27b in cardiomyocyte. Consistently, the miR-27b transgenic mice displayed significantly lower levels of PPAR-γ than the control mice. Furthermore, in vivo silencing of miR-27b using a specific antagomir in a pressure-overload-induced mouse model of HF increased cardiac PPAR-γ expression, attenuated cardiac hypertrophy and dysfunction. The results of our study demonstrate that TGF-β1-regulated miR-27b is involved in the regulation of cardiac hypertrophy, and validate miR-27b as an efficient therapeutic target for cardiac diseases

Highly Expressed miR-375 is not an Intracellular Oncogene in Merkel Cell Polyomavirus-Associated Merkel Cell Carcinoma

miR-375 is a highly abundant miRNA in Merkel cell carcinoma (MCC). In other cancers, it acts as either a tumor suppressor or oncogene. While free-circulating miR-375 serves as a surrogate marker for tumor burden in patients with advanced MCC, its function within MCC cells has not been established. Nearly complete miR-375 knockdown in MCC cell lines was achieved using antagomiRs via nucleofection. The cell viability, growth characteristics, and morphology were not altered by this knockdown. miR-375 target genes and related signaling pathways were determined using Encyclopedia of RNA Interactomes (ENCORI) revealing Hippo signaling and epithelial to mesenchymal transition (EMT)-related genes likely to be regulated. Therefore, their expression was analyzed by multiplexed qRT-PCR after miR-375 knockdown, demonstrating only a limited change in expression. In summary, highly effective miR-375 knockdown in classical MCC cell lines did not significantly change the cell viability, morphology, or oncogenic signaling pathways. These observations render miR-375 an unlikely intracellular oncogene in MCC cells, thus suggesting that likely functions of miR-375 for the intercellular communication of MCC should be addressed

Research on hadoop greedy scheduler based on the fair

Conference Name:1st International Conference on Engineering and Technology Innovation, ICETI 2011. Conference Address: Kenting, Pingtung, Taiwan. Time:November 11, 2011 - November 15, 2011.Job scheduling technology is one of the Hadoop platform's key technologies, whose main function is to control the execute sequence of job and the distribution of computing resources, which directly relates to the Hadoop platform's overall performance and system resources, usage. However, the existing job scheduling algorithms such as FIFO Scheduler, Fair Scheduler and Capacity Scheduler all have some defects. To overcome theses defects, this paper proposed a new algorithm Hadoop Greedy Scheduler Based on the Fair (HGSF). Firstly, the job pools are sorted by priority from high to low; pools in the same priority are sorted by their minimum requirements. Then their minimum requirements are met in turn. If have spare computing resources, it will be assigned to the job pool which has the highest priority and the minimum difference between the ideal requirement and the minimum requirement. Finally, the algorithm uses delay allocation strategy to improve the localization of the data for computing tasks. The experimental results testified the effectiveness of the proposed algorithm

Highly expressed miR-375 is not an intracellular oncogene in Merkel cell polyomavirus-associated Merkel cell carcinoma

miR-375 is a highly abundant miRNA in Merkel cell carcinoma (MCC). In other cancers, it acts as either a tumor suppressor or oncogene. While free-circulating miR-375 serves as a surrogate marker for tumor burden in patients with advanced MCC, its function within MCC cells has not been established. Nearly complete miR-375 knockdown in MCC cell lines was achieved using antagomiRs via nucleofection. The cell viability, growth characteristics, and morphology were not altered by this knockdown. miR-375 target genes and related signaling pathways were determined using Encyclopedia of RNA Interactomes (ENCORI) revealing Hippo signaling and epithelial to mesenchymal transition (EMT)-related genes likely to be regulated. Therefore, their expression was analyzed by multiplexed qRT-PCR after miR-375 knockdown, demonstrating only a limited change in expression. In summary, highly effective miR-375 knockdown in classical MCC cell lines did not significantly change the cell viability, morphology, or oncogenic signaling pathways. These observations render miR-375 an unlikely intracellular oncogene in MCC cells, thus suggesting that likely functions of miR-375 for the intercellular communication of MCC should be addressed

High-Precision Wafer Bonding Alignment Mark Using Moiré Fringes and Digital Grating

This paper investigates a moiré-based mark for high-precision wafer bonding alignment. During alignment, the mark is combined with digital grating, which has the benefits of high precision and small size. A digital grating is superimposed on the mark to generate moiré fringes. By performing a phase calculation on the moiré fringe images corresponding to the upper and lower wafers, the relative offset of the upper and lower wafers can be accurately calculated. These moiré fringes are exceptionally stable, thereby enhancing the alignment stability. In this study, through practical experiments, we tested the rationality and practicability of the mark

Geft is dispensable for the development of the second heart field

Geft is a guanine nucleotide exchange factor, which can specificallyactivate Rho family of small GTPase by catalyzing theexchange of bound GDP for GTP. Geft is highly expressed inthe excitable tissue as heart and skeletal muscle and plays importantroles in many cellular processes, such as cell proliferation,migration, and cell fate decision. However, the invivo role of Geft remains unknown. Here, we generated a Geftconditional knockout mouse by flanking exons 5-17 of Geftwith loxP sites. Cre-mediated deletion of the Geft gene in heartusing Mef2c-Cre transgenic mice resulted in a dramatic decreaseof Geft expression. Geft knockout mice develop normallyand exhibit no discernable phenotype, suggesting Geft isdispensable for the development of the second heart field inmouse. The Geft conditional knockout mouse will be a valuablegenetic tool for uncovering the in vivo roles of Geft duringdevelopment and in adult homeostasis. (BMB reports2012; 45(3): 153-158

Reversal of epigenetic silencing of MHC class I chain-related protein A and B improves immune recognition of Merkel cell carcinoma

Merkel cell carcinoma (MCC) is a virally associated cancer characterized by its aggressive behavior and strong immunogenicity. Both viral infection and malignant transformation induce expression of MHC class I chain-related protein (MIC) A and B, which signal stress to cells of the immune system via Natural Killer group 2D (NKG2D) resulting in elimination of target cells. However, despite transformation and the continued presence of virally-encoded proteins, MICs are only expressed in a minority of MCC tumors in situ and are completely absent on MCC cell lines in vitro. This lack of MIC expression was due to epigenetic silencing via MIC promoter hypo-acetylation; indeed, MIC expression was re-induced by pharmacological inhibition of histone deacetylases (HDACs) both in vitro and in vivo. This re-induction of MICs rendered MCC cells more sensitive to immune-mediated lysis. Thus, epigenetic silencing of MICs is an important immune escape mechanism of MCCs

Altered resting-state intra- and inter- network functional connectivity in patients with persistent somatoform pain disorder.

Patients with persistent somatoform pain disorder (PSPD) usually experience various functional impairments in pain, emotion, and cognition, which cannot be fully explained by a physiological process or a physical disorder. However, it is still not clear for the mechanism underlying the pathogenesis of PSPD. The present study aimed to explore the intra- and inter-network functional connectivity (FC) differences between PSPD patients and healthy controls (HCs). Functional magnetic resonance imaging (fMRI) was performed in 13 PSPD patients and 23 age- and gender-matched HCs. We used independent component analysis on resting-state fMRI data to calculate intra- and inter-network FCs, and we used the two-sample t-test to detect the FC differences between groups. Spearman correlation analysis was employed to evaluate the correlations between FCs and clinical assessments. As compared to HCs, PSPD patients showed decreased coactivations in the right superior temporal gyrus within the anterior default-mode network and the anterior cingulate cortex within the salience network, and increased coactivations in the bilateral supplementary motor areas within the sensorimotor network and both the left posterior cingulate cortex and the medial prefrontal cortex within the anterior default-mode network. In addition, we found that the PSPD patients showed decreased FNCs between sensorimotor network and audio network as well as visual network, between default-mode network and executive control network as well as audio network and between salience network and executive control network as well as right frontoparietal network, and increased FNCs between sensorimotor network and left frontoparietal network, salience network as well as cerebellum network, which were negatively correlated with the clinical assessments in PSPD patients. Our findings suggest that PSPD patients experience large-scale reorganization at the level of the functional networks, which suggests a possible mechanism underlying the pathogenesis of PSPD

MCPyV Large T antigen induced atonal homolog 1 (ATOH1) is a lineage-dependency oncogene in Merkel cell carcinoma.

Despite the fact that the transcription factor ATOH1 is a master regulator of Merkel cell development, its role in Merkel cell carcinoma (MCC) carcinogenesis remains controversial. Here, we provide several lines of evidence that ATOH1 is a lineage-dependent oncogene in MCC. Luciferase assays revealed binding of ATOH1 and subsequent activation to the promoter of miR-375, which is one of the most abundant microRNAs in MCCs. Overexpression of ATOH1 in variant MCC cell lines and fibroblasts induced miR-375 expression, whereas ATOH1 knockdown in classical MCC cell lines reduced miR-375 expression. Moreover, ATOH1 overexpression in these cells changed their growth characteristics from adherent to suspension and/orspheroidal growth, that is, resembling the neuroendocrine growth pattern of classical MCC cell lines. Notably, ectopic expression of different Merkel cell polyomavirus (MCPyV)-derived truncated large T antigens induced ATOH1 expression in fibroblasts, which was paralleled by miR-375 expression and similar morphologic changes. In summary, MCPyV-associated carcinogenesis is likely to induce the characteristic neuroendocrine features of MCC via induction of ATOH1; thus, ATOH1 can be regarded as a lineage-dependent oncogene in MCC