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Most seven transmembrane receptors (7TMRs) are G protein-coupled receptors; however, some 7TMRs evoke intracellular signals through β-arrestin as a biased receptor. As several β-arrestin-biased agonists have been reported to be cardioprotective, we examined the role of the chemokine receptor CXCR7 as a β-arrestin-biased receptor in the heart. Among 510 7TMR genes examined, Cxcr7 was the most abundantly expressed in the murine heart. Single-cell RNA-sequencing analysis revealed that Cxcr7 was abundantly expressed in cardiomyocytes and fibroblasts. Cardiomyocyte-specific Cxcr7 null mice showed more prominent cardiac dilatation and dysfunction than control mice 4 weeks after myocardial infarction. In contrast, there was no difference in cardiac phenotypes between fibroblast-specific Cxcr7-knockout mice and control mice even after myocardial infarction. TC14012, a specific agonist of CXCR7, significantly recruited β-arrestin to CXCR7 in CXCR7-expressing cells and activated extracellular signal-regulated kinase (ERK) in neonatal rat cardiomyocytes. Cxcr7 expression was significantly increased and ERK was activated in the border zone of the heart in control, but not Cxcr7 null mice. These results indicate that the abundantly expressed CXCR7 in cardiomyocytes may play a protective role in the heart as a β-arrestin-biased receptor and that CXCR7 may be a novel therapeutic target for myocardial infarction.

Rheumatoid arthritis (RA) is a disease characterized primarily by chronic inflammatory synovitis and is well-known to be associated with significant sex differences in its prevalence and clinical features. Sex steroids have been proposed to be involved in the pathogenesis of RA, but details pertaining to the expression of sex steroid receptors in RA synovial tissue have yet to be fully characterized. In the present study, we examined oestrogen receptor (ER) alpha, ERbeta, progesterone receptor (PR) and androgen receptor (AR) mRNA expression using real-time reverse transcriptase-PCR (RT-PCR) in eight female RA synovial tissues and six female synovial tissues without inflammation, and determined immunolocalization of ERalpha, ERbeta, PR-A, PR-B and AR using immunohistochemistry in synovial tissues obtained from 22 RA patients. Real-time RT-PCR analysis demonstrated the expression of ER, PR and AR mRNAs in both RA and non-inflamed synovial tissues. Relative abundance of ER mRNAs was significantly higher in RA synovial tissue than non-inflamed synovial tissue (P<0.05). In addition, the relative ERalpha/ERbeta mRNA expression ratio was significantly lower in RA than non-inflamed synovial tissue (RA, 2.34 +/- 1.60; and non-inflamed, 20.7 +/- 19.1; P<0.05). There were no significant differences in relative abundance of PR mRNA. Relative abundance of AR mRNA was significantly lower in RA (P<0.05). Immunoreactivity for ERalpha, ERbeta, PR-B and AR was detected in the lining cells, inflammatory cells and fibroblasts in all the patients examined. The labelling indices for ERbeta and PR-B were more abundant in both lining cells (ERbeta, 54.2 +/- 12.2%; PR-B, 73.6 +/- 18.9%) and inflammatory cells (ERbeta, 74.6 +/- 16.2%; PR-B, 75.9 +/- 16.1%) than in fibroblasts (ERbeta, 36.5 +/- 15.6%; PR-B, 49.4 +/- 18.0%). Labelling indices for ERalpha and AR were significantly higher in lining cells (ERalpha, 14.4 +/- 8.6%; AR, 31.2 +/- 11.3%) and fibroblasts (ERalpha, 12.1 +/- 7.5%; AR, 20.1 +/- 9.6%) than those in inflammatory cells (ERalpha, 5.7 +/- 3.3%; AR, 9.2 +/- 4.4%). There were significant differences (P<0.05) in the labelling indices for ERalpha, ERbeta and PR-B between men and women under 50 years of age in fibroblasts of RA synovial tissues. These results indicate that sex steroid receptors are present in RA and non-inflamed synovial tissues, including inflammatory cells in RA, and suggest that sex steroids may play important roles in the regulation of inflammation of RA synovial tissue.

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The possibility has been indicated that polymers with helical chirality, such as poly-γ-benzyl-L-glutamate (PBLG) and poly-L-lactic acid (PLLA), exhibit a large shear piezoelectric constant. To attempt the realization of a PBLG membrane with a large piezoelectric constant, we fabricated the PBLG membrane oriented by magnetic field force. Concretely, the PBLG membranes were casted from 1,2-dichloroethane solution with various PBLG concentrations under the magnetic field generation equipment incorporating a superconducting magnet. First, the orientation of the chain molecules of the PBLG membranes obtained was observed macroscopically by means of a polarizing microscope (POM). The orientation of the chain molecules of the PBLG membranes was recognized for the case of casting from the PBLG 1,2-dichloroethane solution in the liquid crystal state. Also, from X-ray photograph measurements, it was found that the orientation direction of the chain molecules of PBLG was perpendicular to the magnetic field direction. We then measured the shear piezoelectric constant d 14 of the oriented PBLG membranes. With increasing the strength of the applied magnetic field in the casting process for the film preparation, d 14 * of the PBLG membranes obtained increases. Finally, a large piezoelectric constant of 26 pC/N was found in the PBLG membrane. It is assumed that d 14 * is not saturated even at the magnetic field of 10 T.

In-beam γ spectroscopy of the stable nucleus 127 I has been studied experimentally using the 124 Sn( 7 Li, 4nγ ) 127 I reaction at a beam energy of 32 MeV. The high-spin level scheme of 127 I is extended significantly. Negative-parity levels built on the 11/2 − , πh 11/2 particle state are observed up to (35/2 − ) and described as a decoupled band, extending our knowledge of decoupled structures to the most neutron-rich stable iodine isotope. Two I = 2 yrast positive-parity sequences are proposed to be associated with the πg 7/2 configuration due to observations of several strong interband transitions, and two weakly populated I = 2 positive-parity bands are newly identified and interpreted as arising mainly from the πd 5/2 configuration. Three-quasiparticle configurations are assigned to the I π = 15/2 + and 23/2 + states according to the existing knowledge in neighboring nuclei; irregular noncollective and regular collective excitations built on these two (15/2 + and 23/2 + ) states are observed to coexist at similar energies. The observed three-quasiparticle band structures are further interpreted with the aid of configuration-constrained potential energy surface calculations.

A R T I C L E I N F O Keywords:Heart failure Single-cell spatial analysis Immunostaining Single-molecule fluorescence in situ hybridization A B S T R A C T Background: The heart responds to hemodynamic overload through cardiac hypertrophy and activation of the fetal gene program. However, these changes have not been thoroughly examined in individual cardiomyocytes, and the relation between cardiomyocyte size and fetal gene expression remains elusive. We established a method of high-throughput single-molecule RNA imaging analysis of in vivo cardiomyocytes and determined spatial and temporal changes during the development of heart failure. Methods and results: We applied three novel single-cell analysis methods, namely, single-cell quantitative PCR (sc-qPCR), single-cell RNA sequencing (scRNA-seq), and single-molecule fluorescence in situ hybridization (smFISH). Isolated cardiomyocytes and cross sections from pressure overloaded murine hearts after transverse aortic constriction (TAC) were analyzed at an early hypertrophy stage (2 weeks, TAC2W) and at a late heart failure stage (8 weeks, TAC8W). Expression of myosin heavy chain β (Myh7), a representative fetal gene, was induced in some cardiomyocytes in TAC2W hearts and in more cardiomyocytes in TAC8W hearts. Expression levels of Myh7 varied considerably among cardiomyocytes. Myh7-expressing cardiomyocytes were significantly more abundant in the middle layer, compared with the inner or outer layers of TAC2W hearts, while such spatial differences were not observed in TAC8W hearts. Expression levels of Myh7 were inversely correlated with cardiomyocyte size and expression levels of mitochondria-related genes. Conclusions: We developed a new image-analysis pipeline to allow automated and unbiased quantification of gene expression at the single-cell level and determined the spatial and temporal regulation of heterogenous Myh7 expression in cardiomyocytes after pressure overload.

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