Highlights • Δ FosB mRNA gives rise to ΔFosB and to the minor alternatively translated Δ2ΔFosB. • Overexpression of ΔFosB alone confirms its pro-reward and pro-resilience phenotype. • In contrast, ...Δ2ΔFosB has no effect on cocaine reward or stress vulnerability. • Full-length FosB, encoded by FosB mRNA, also does not affect reward or resilience.
AdpA is a global transcriptional activator triggering morphological differentiation and secondary metabolism in Streptomyces griseus. AdpA influences the expression of >1000 genes; however, the ...overall picture of the AdpA regulon remains obscure. Here, we took snapshots of the distribution of AdpA across the chromosome in living S. griseus cells using chromatin immunoprecipitation/chromatin affinity precipitation-seq analysis. In both liquid and solid cultures, AdpA bound to >1200 similar sites, which were located on not only in putative regulatory regions (65%), but also in regions (35%) that appeared not to affect transcription. Transcriptome analysis indicated that ~40% of the AdpA-binding sites in putative regulatory regions were involved in gene regulation. AdpA was indicated to act as a transcriptional repressor as well as an activator. Expression profiles of AdpA-target genes were very different between liquid and solid cultures, despite their similar AdpA-binding profiles. We concluded that AdpA directly controls >500 genes in cooperation with other regulatory proteins. A comprehensive competitive gel mobility shift assay of AdpA with 304 selected AdpA-binding sites revealed several unique characteristics of the DNA-binding property of AdpA. This study provides the first experimental insight into the extent of the AdpA regulon, indicating that many genes are under the direct control of AdpA.
Objectives: Kawasaki disease (KD) is a systemic vasculitis of early childhood. Intravenous immunoglobulin (IVIG) is the standard treatment for KD. However, IVIG is not effective in approximately 15% ...of children with KD, and the mechanisms for this are unclear. We investigated changes in monocyte and T-cell activation from pre- to post-IVIG in IVIG-effective and IVIG-resistant KD.
Method: We analysed peripheral CD14
+
CD16
+
cells and human leucocyte antigen-DR (HLA-DR) expression on CD4
+
and CD8
+
cells in 46 children with KD who were admitted to Yamaguchi University Hospital between January 2011 and May 2016. We compared the kinetics in the absolute numbers of CD14
+
CD16
+
cells, CD4
+
HLA-DR
+
cells, and CD8
+
HLA-DR
+
cells before and after IVIG treatment between IVIG-effective and IVIG-resistant groups.
Results: Among the 46 subjects, 30 had IVIG-effective KD and 16 had IVIG-resistant KD. The absolute number of CD14
+
CD16
+
cells in the IVIG-effective group decreased significantly after IVIG, while that in the IVIG-resistant group showed no change after IVIG. The absolute number of CD4
+
HLA-DR
+
cells increased significantly after IVIG in both groups. The absolute number of CD8
+
HLA-DR
+
cells before IVIG was low and significantly increased after IVIG in the IVIG-resistant group, while that in the IVIG-effective group showed no change after IVIG.
Conclusions: Our results suggest that insufficient control of monocyte suppression and T-cell activation, especially in terms of the CD8-related immune system, are associated with IVIG resistance. The restoration of T-cell suppression may be important for KD recovery. These findings provide insight into the mechanism of IVIG resistance.
Abstract
In the low energy ring (LER) for positrons in the SuperKEKB, a vertical beam size blow-up was observed when the bunch current was approximately 1 mA. If a beam size blow-up occurs, the ...design luminosity cannot be achieved. Therefore, beam size blow-ups must be pre-vented. According to calculations, the bunch current threshold of the transverse mode coupling instability (TMCI) is 2 mA or more, and the observed value is 50% or smaller. Ordinary TMCI cannot explain this vertical beam size blow-up. This paper shows that the cause of the vertical beam size blow-up can be determined by analyzing factors such as beam oscillation. The study results showed that the vertical beam size blow-up in the LER was caused by a -1 mode instability.
To construct an infrastructure for genome-wide association studies of common diseases or drug sensitivities, we have been systematically exploring common variants by resequencing genomic regions ...containing genes in DNA from 24 Japanese individuals. We have analyzed a total of 154 Mb, corresponding to approximately 5% of the human genome, and so far have identified 174 269 single-nucleotide polymorphisms and 16 293 insertion/deletion polymorphisms within gene regions, i.e., one polymorphism in 807 bp on average. Our data are freely available via our web site (http://snp.ims.u-tokyo.ac.jp) and will facilitate studies to identify genes associated with susceptibility to common diseases and genes involved in sensitivity to therapeutic drugs.
One of the most difficult issues to be solved in genome-wide association studies is to reduce the amount of genomic DNA required for genotyping. Currently available technologies require too large a ...quantity of genomic DNA to genotype with hundreds or thousands of single-nucleotide polymorphisms (SNPs). To overcome this problem, we combined the Invader assay with multiplex polymerase chain reaction (PCR), carried out in the presence of antibody to Taq polymerase, as well as using a novel 384-well card system that can reduce the required reaction volume. We amplified 100 genomic DNA fragments, each containing one SNP, in a single tube, and analyzed each SNP with the Invader assay. This procedure correctly genotyped 98 of the 100 SNP loci examined in PCR-amplified samples from ten individuals: the genotypes were confirmed by direct sequencing. The reproducibility and universality of the method were confirmed with two additional sets of 100 SNPs. Because we used 40 ng of genomic DNA as a template for multiplex PCR, the amount needed to assay one SNP was only 0.4 ng; therefore, theoretically, more than 200,000 SNPs could be genotyped at once when 100 microg of genomic DNA is available. Our results indicate the feasibility of undertaking genome-wide association studies using blood samples of only 5-10 ml.
In this paper, the mechanical behavior of inclined jointed rock masses during tunneling is considered. Such rock masses can be considered as an assembly of discrete blocks with the discontinuities ...having a significant influence on the mechanical behavior. To simulate this situation, a discrete numerical analysis method, Discontinuous Deformation Analysis (DDA), is applied. The DDA results show the existence of stress arching in the rock masses during tunneling. This stress arching is the primary influence on the stress distribution and surface subsidence. In addition, the stress arching is affected by the dip angle of the jointed rock masses. Moreover, the DDA results are in good agreement with experiments, explaining the reason for the asymmetrical vertical stress and surface subsidence obtained in laboratory tests. These results suggest that DDA can be applied to model the tunneling behavior of complicated discontinuous rock masses.
Inosine triphosphate pyrophosphatase (ITPase), the enzyme that hydrolyzes ITP and other deaminated purine nucleoside triphosphates to the corresponding purine nucleoside monophosphate and ...pyrophosphate, is encoded by the Itpa gene. In this study, we established Itpa knockout (KO) mice and used them to show that ITPase is required for the normal organization of sarcomeres in the heart. Itpa(-/-) mice died about 2 weeks after birth with features of growth retardation and cardiac myofiber disarray, similar to the phenotype of the cardiac alpha-actin KO mouse. Inosine nucleotides were found to accumulate in both the nucleotide pool and RNA of Itpa(-/-) mice. These data suggest that the role of ITPase in mice is to exclude ITP from the ATP pool, and the main target substrate of this enzyme is rITP. Our data also suggest that cardiomyopathy, which is mainly caused by mutations in sarcomeric protein-encoding genes, is also caused by a defect in maintaining the quality of the ATP pool, which is an essential requirement for sarcomere function.
•Topotecan was more myelotoxic to human than murine granulocytes in humanized mice.•Oxaliplatin was more myelotoxic to murine than human granulocytes in humanized mice.•The myelotoxicity of ...Paclitaxel was comparable between human and mouse cells in humanized mice.•Human myelotoxicity of anti-cancer drugs can be predicted in vivo by using the humanized mice.
Evaluating myelotoxicity is essential for ensuring the safety of novel drugs before they are approved for clinical applications. Although in vivo prediction of the maximum tolerated doses (MTDs) of anticancer drugs is usually performed in rodents, the results are not always applicable to clinical treatment because drugs may have different effects in human and rodent cells. Previously, we generated a human IL-3 and GM-CSF transgenic humanized mouse (hu-IL-3/GM Tg), in which human granulocytes effectively differentiated after hematopoietic stem cell transplantation. In this study, we established a novel in vivo preclinical evaluation model for predicting human myelotoxicity of anticancer drugs using these hu-IL-3/GM Tg mice. The myelotoxicity was investigated by kinetic flow cytometry of human or murine granulocytes and by colony-forming unit granulocyte/macrophage (CFU-GM) assays. In both in vivo and in vitro analyses, topotecan was more myelotoxic to human than murine granulocytes. In contrast, oxaliplatin was more myelotoxic to murine granulocytes. The level of myelotoxicity of paclitaxel treatment was comparable between human and mouse cells. These results demonstrate that our humanized mouse model can simultaneously evaluate myelotoxicity against human and mouse cells in vivo, and provides an effective preclinical tool for predicting appropriate doses of anticancer agents for clinical treatment.
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