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Abstract: In patients with gastroesophageal reflux disease (GERD), esophageal exposure to both acid and duodenogastroesophageal reflux are more common than to acid reflux alone, suggesting that acidic bile acid in duodenal juice may contribute to the pathophysiology and severity of GERD. However, the mechanism whereby esophageal mucosal epithelial cells react to bile acid remains unclear. We visually examined the real-time response of mouse esophageal epithelial cells to bile acids using calcium (Ca2+)-imaging methods. We investigated the effects of seven different bile acids. After stimulation for a few minutes, only Deoxycholate (DCA) under acidic conditions caused a elevation of intracellular Ca2+ concentration ([Ca2+]i)in the cells in dose- and pH-dependent manners. Conjugated bile acids had no effect on the cells. Viability assay of the cells in the presence of DCA was in good agreement with the calcium imaging data. Besides, DCA-induced [Ca2+]i increase in acidic conditions was observed not only in isolated primary cultured cells, but also in cells in the stratified squamous epithelium. This study suggests that DCA can pass through the anatomical barrier of the esophageal epithelium and induce calcium signaling in epithelial cells in a pH-dependent manner. This supports the hypothesis that bile acid reflux together with gastric acid can affect the esophageal mucosa, even under reflux times of a few minutes. [Copyright &y& Elsevier]

Abstract: Transient receptor potential vanilloid 3 (TRPV3), a member of the calcium-permeable thermosensitive TRP (thermoTRP) subfamily of receptors, is an important cutaneous sensor that detects thermal and chemical stimuli. TRPV3 is activated by innocuous warm temperature stimuli (>33° C) and a variety of physiologically active substances. While the corneal epithelium is known to respond to such stimuli, it is unknown whether TRPV3 is involved in this phenomenon. We show here that TRPV3 mRNA and protein are abundantly expressed in the epithelial cells of human and mouse cornea. Carvacrol, an agonist of TRPV3, elevated cytosolic Ca2+ concentration in both primary mouse corneal epithelial cells and cultured human corneal epithelial cells (HCE-T cells). The response to carvacrol was inhibited by ruthenium red, a TRPV channel antagonist. Moreover, repetitive agonist stimulation sensitized the response with gradually increasing amplitude, suggesting that the TRPV3 in the cornea has similar physiological and pharmacological characteristics to that in skin keratinocytes. Finally, a wound healing assay revealed that appropriate calcium ion influx via activated TRPV3 in corneal epithelial cells accelerated their proliferation. Thus, functional TRPV3 is present in corneal epithelial cells and may play a role not only in thermosensation, but also in the regulation of cell proliferation. [Copyright &y& Elsevier]

Abstract: The thermo-transient receptor potential (thermoTRP) subfamily is composed of channels that are important in nociception and thermo-sensing. Here, we show a selective expression of TRPV3 channel in the distal colon throughout the gastrointestinal tract. Expression analyses clearly revealed that TRPV3 mRNA and proteins were expressed in the superficial epithelial cells of the distal colon, but not in those of the stomach, duodenum or proximal colon. In a subset of primary epithelial cells cultured from the distal colon, carvacrol, an agonist for TRPV3, elevated cytosolic Ca2+concentration in a concentration-dependent manner. This response was inhibited by ruthenium red, a TRPV channel antagonist. Organotypic culture supported that the carvacrol-responsive cells were present in superficial epithelial cells. Moreover, application of carvacrol evoked ATP release in primary colonic epithelial cells. We conclude that TRPV3 is present in absorptive cells in the distal colon and may be involved in a variety of cellular functions. [Copyright &y& Elsevier]

Abstract: We report the effects of new N-acylated ambroxol derivatives (TEI-588a, TEI-588b, TEI-589a, TEI-589b, TEI-602a and TEI-602b: a, aromatic amine-acylated derivative; b, aliphatic amine-acylated derivative) induced from ambroxol (a mucolytic agent to treat human lung diseases) on Cl− secretion in human submucosal serous Calu-3 cells under a Na+/K+/2Cl− cotransporter-1 (NKCC1)-mediated hyper-secreting condition. TEI-589a, TEI-589b and TEI-602a diminished hyper-secretion of Cl− by diminishing the activity of NKCC1 without blockade of apical Cl− channel (TEI-589a>TEI-602a>TEI-589b), while any other tested compounds including ambroxol had no effects on Cl− secretion. These indicate that the inhibitory action of an aromatic amine-acylated derivative on Cl− secretion is stronger that that of an aliphatic amine-acylated derivative, and that 3-(2,5-dimethyl)furoyl group has a strong action in inhibition of Cl− secretion than cyclopropanoyl group. We here indicate that TEI-589a, TEI-589b and TEI-602a reduce hyper-secretion to an appropriate level in the airway, providing a possibility that the compound can be an effective drug in airway obstructive diseases including COPD by reducing the airway resistance under a hyper-secreting condition. [Copyright &y& Elsevier]

MEST is one of the imprinted genes in human. With the assistance of our integration map and the complete sequence in the registry, we mapped a total of 16 genes/transcripts at the 1.5-Mb MEST-flanking region at 7q32. This region has been suggested to form an imprinted gene cluster, because MEST and its three flanking genes/transcripts (MESTIT1, CPA4, and COPG2IT1) were reported to be imprinted, although two (TSGA14 and COPG2) were shown to escape imprinting. In this study, 10 other genes/transcripts were examined for their imprinting status in human fetal tissues. The results indicated that 8 genes/transcripts (NRF1, UBE2H, HSPC216, KIAA0265, FLJ14803, CPA2, CPA1, and DKFZp667F0312) were expressed biallelically. The imprinting status of two (TSGA13 and CPA5) was not conclusive, because of their weak and/or tissue-specific expression and inconstant results. These findings provided evidence that only 4 of the 16 genes/transcripts located to the region show monoallelic expression, while others are not involved in imprinting. Therefore, it is less likely that the MEST-flanking 7q32 region forms a large imprinted domain. [Copyright &y& Elsevier]

The number of patients with heart failure and related deaths is rapidly increasing worldwide, making it a major problem. Cardiac hypertrophy is a crucial preliminary step in heart failure, but its treatment has not yet been fully successful. In this study, we established a system to evaluate cardiomyocyte hypertrophy using a deep learning-based high-throughput screening system and identified drugs that inhibit it. First, primary cultured cardiomyocytes from neonatal rats were stimulated by both angiotensin II and endothelin-1, and cellular images were captured using a phase-contrast microscope. Subsequently, we used a deep learning model for instance segmentation and established a system to automatically and unbiasedly evaluate the cardiomyocyte size and perimeter. Using this system, we screened 100 FDA-approved drugs library and identified 12 drugs that inhibited cardiomyocyte hypertrophy. We focused on ezetimibe, a cholesterol absorption inhibitor, that inhibited cardiomyocyte hypertrophy in a dose-dependent manner in vitro. Additionally, ezetimibe improved the cardiac dysfunction induced by pressure overload in mice. These results suggest that the deep learning-based system is useful for the evaluation of cardiomyocyte hypertrophy and drug screening, leading to the development of new treatments for heart failure. • We developed a system to evaluate hypertrophied cardiomyocytes by deep learning. • We identified 12 drugs which inhibit cardiomyocyte hypertrophy by screening. • Ezetimibe suppressed cardiac hypertrophy and improved cardiac dysfunction in mice. • This system could be useful to develop new treatment options for heart failure. [ABSTRACT FROM AUTHOR]

Abstract: It has been shown that anti-cancer drug induces secretion of serotonin (5-HT) from small intestine which activates serotonin type 3 (5-HT3) receptor to cause nausea and vomiting. In general, antagonist for 5-HT3 receptor is used as anti-emetics during chemotherapy. However, we found that anti-cancer drug irinotecan itself inhibits 5-HT-gated current through the homomeric 5-HT3A and heteromeric 5-HT3AB receptor in a concentration-dependent manner. The inhibitory effect of irinotecan on 5-HT3A receptor was more potent than that on 5-HT3AB receptor. On the other hand, SN-38, a metabolite of irinotecan, had no effect on the responsiveness. Our findings suggest that irinotecan itself could have anti-emetic activities through inhibition of the 5-HT3A and 5-HT3AB receptor. [Copyright &y& Elsevier]

Abstract: Single crystals of iron(II) pyroborate, Fe2B2O5, were prepared at 1000–1050°C under an argon atmosphere. The crystals were transparent, yellowish in color and needle-like or columnar. The crystal structure of Fe2B2O5 was analyzed by single-crystal X-ray diffraction. Refined triclinic unit cell parameters were a=3.2388(2), b=6.1684(5), c=9.3866(8)Å, α=104.613(3)°, β=90.799(2)° and γ=91.731(2)°. The final reliability factors of refinement were R1=0.020 and wR2=0.059 [I > 2σ(I)]. Transmittance over 50% in the visible light region from 500 to 750nm was observed for a single crystal of Fe2B2O5 with a thickness of about 0.3mm. The light absorption edge estimated from a diffuse reflectance spectrum was at around 350nm (3.6eV). Magnetic susceptibility was measured for single crystals at 4–300K. Fe2B2O5 showed antiferromagnetic behavior below the Néel temperature, T N≈70K, and the Weiss temperature was T W=36K. The effective magnetic moment of Fe was 5.3μ B. [Copyright &y& Elsevier]

Abstract: New compounds CaY2Ge3O10 and CaY2Ge4O12 were prepared by heating mixtures of CaCO3, Y2O3 and GeO2 at 1200°C. CaY2Ge3O10 is stable at 1300°C, while CaY2Ge4O12 decomposes into a melt and CaY2Ge3O10 at approximately 1250°C. We obtained single crystals of CaY2Ge3O10 by cooling a sample with an initial composition of Ca:Y:Ge=1:2:8 from 1300°C with a rate of −6°C/h. The crystal structure of CaY2Ge3O10 was determined by single crystal X-ray diffraction. CaY2Ge3O10 crystallizes in the monoclinic space group with , , and , , and for . In the structure of CaY2Ge3O10, Ca and Y atoms are situated disorderly in three 7-fold coordination sites between isolated germanate groups of triple GeO4 tetrahedra, Ge3O10. The structural formula of CaY2Ge3O10 is expressed as (Ca0.45Y0.55)(Ca0.46Y0.54)(Ca0.09Y0.91)Ge3O10. The crystal structure of CaY2Ge4O12 was analyzed by the Rietveld method for the X-ray powder diffraction pattern. CaY2Ge4O12 is isotypic with SrNa2P4O12, crystallizing in the orthorhombic space group P4/nbm, , , , , . CaY2Ge4O12 contains four-membered GeO4-tetrahedra rings, Ge4O12. Eight-fold coordinated square-anitiprism sites and 6-fold octahedral sites between the layers of the Ge4O12 rings are occupied by Y atom and Ca/Y atoms, respectively The structural formula is Y(Ca0.5Y0.5)2Ge4O12. [Copyright &y& Elsevier]

Small fibre neuropathy (SFN) is an initial pathology of diabetic polyneuropathy (DPN). Serum lipopolysaccharide binding protein levels are positively correlated with the pain threshold in the foot, suggesting that the abundance of gut Gram-negative bacilli, which are a source of lipopolysaccharides, may be involved in the development of DPN. Furthermore, the abundance of the gut and oral microbiota is assumed to be involved in the pathogenesis of diabetes. Nevertheless, the association between SFN and the microbiota has not been clarified. A total of 1056 individuals were recruited in the 2018 Iwaki Health Promotion Project. Pain sensation was evaluated based on the pain threshold from intraepidermal electrical stimulation (PINT). Patients with PINT scores <0.15 mA were categorized into the low-PINT group (n = 718); otherwise, they were categorized into the high-PINT group (n = 283). Furthermore, each group was divided into the subjects with or without glucose tolerance based on HbA1c levels, fasting blood glucose levels and diabetic history. Principal coordinate analysis and α- and β-diversity of the microbiota were evaluated. The correlation between clinical and microbiota data was examined. Oral microbiota diversity showed no structural differences according to PINT scores, whereas principal coordinate analysis and α- and β-diversity revealed significant structural differences in gut microbiota (p < 0.01, p < 0.05 and p < 0.05, respectively), even after the participants with glucose intolerance were excluded (p < 0.01, p < 0.05 and p < 0.05, respectively). The relative abundance of the genus Bacteroides was significantly lower in high-PINT participants compared with low-PINT participants (10 ± 6.7% vs. 11.3 ± 7.0%, p < 0.01), even after the exclusion of subjects with diabetes and impaired fasting glucose (10.0 ± 6.5% vs. 11.2 ± 6.9%, p < 0.05). In univariate linear regression analyses, PINT was significantly correlated with metabolic syndrome parameters, eGFR, uric acid level and the abundance of Bacteroides. The correlation between Bacteroides and PINT scores remained significant after adjustment for multiple factors (β = −0.07181, p < 0.05). Changes of bacterial diversity and a low abundance of gut Bacteroides were correlated with elevated PINT scores in the Japanese population. This correlation may represent a new therapeutic option for SFN. • Small fibre neuropathy may be correlated with changes in the bacterial floor. • PINT is a method to evaluate pain threshold as a symptom of SFN. • Structure of the microbiota in the gut changed according to the PINT score. • Abundance of Bacteroides in the gut was significantly reduced in high-PINT subjects. • Changes in the gut microbiota may be a pathophysiology related to pain threshold. [ABSTRACT FROM AUTHOR]

Abstract: Eu3+-doped Ca2SnO4 (solid solutions of Ca2− x Eu2 x Sn1− x O4, 0⩽x⩽0.3) and Eu3+ and Y3+-codoped Ca2SnO4 (Ca1.8Y0.2Eu0.2Sn0.8O4) were prepared by solid-state reaction at 1400°C in air. Rietveld analysis of the X-ray powder diffraction patterns revealed that Eu3+ replaced Ca2+ and Sn4+ in Eu3+-doped Ca2SnO4, and that Eu3+ replaced Ca2+ and Y3+ replaced Sn4+ in Ca1.8Y0.2Eu0.2Sn0.8O4. Red luminescence at 616nm due to the electric dipole transition 5 D o→7 F 2 was observed in the photoluminescence (PL) spectra of Ca2− x Eu2 x Sn1− x O4 and Ca1.8Y0.2Eu0.2Sn0.8O4 at room temperature. The maximum PL intensity in the solid solutions of Ca2− x Eu2 x Sn1− x O4 was obtained for x=0.1. The PL intensity of Ca1.8Y0.2Eu0.2Sn0.8O4 was 1.26 times greater than that of Ca2− x Eu2 x Sn1− x O4 with x=0.1. [Copyright &y& Elsevier]

Mutations of the WFS1 gene are responsible for two hereditary diseases, Wolfram syndrome and low frequency sensorineural hearing loss. The WFS1 protein is a glycoprotein located in the endoplasmic reticulum (ER) membrane but its function is poorly understood. Herein we show WFS1 mRNA and protein levels in pancreatic islets to be increased with ER-stress inducers, thapsigargin and dithiothreitol. Another ER-stress inducer, the N-glycosylation inhibitor tunicamycin, also raised WFS1 mRNA but not protein levels. Site-directed mutagenesis showed both Asn-663 and Asn-748 to be N-glycosylated in mouse WFS1 protein. The glycosylation-defective WFS1 protein, in which Asn-663 and Asn-748 had been substituted with aspartate, exhibited an increased protein turnover rate. Consistent with this, the WFS1 protein was more rapidly degraded in the presence of tunicamycin. These data indicate that ER-stress and N-glycosylation play important roles in WFS1 expression and stability, and also suggest regulatory roles for this protein in ER-stress induced cell death. [Copyright &y& Elsevier]

The mouse Atp10a gene is located at the border of an imprinted domain distal to the p-locus on mouse chromosome 7. The localization of Atp10a neighboring the maternally expressed gene Ube3a in the imprinted domain and an unusual inheritance pattern of the obesity phenotype with a p-locus deletion have suggested that Atp10a might be imprinted and associated with body fat. Recently, its human ortholog, ATP10A, was identified as the second imprinted gene with maternal expression in the human chromosome 15q11–q13 imprinted domain. To elucidate the imprinting status of Atp10a, we performed expression analysis in various tissues from reciprocal crosses between C57BL/6 and PWK (divergent strains of Mus musculus) mice. The results revealed that Atp10a was biallelically expressed in all tissues examined. Furthermore, there was no differential methylation in the CpG island and no antisense transcripts of the gene. These findings suggest that the mouse Atp10a gene escapes genomic imprinting. [Copyright &y& Elsevier]

Polyamines produced by both prokaryotes and eukaryotes are bioactive substances with pleiotropic effects. Accumulating evidence has demonstrated that polyamines contribute to anti-inflammatory responses by suppressing the expression of proinflammatory cytokines in mononuclear cells and macrophages. However, the effects of polyamines on CD4+ T cell responses remain to be elucidated. Here, we investigated the effect of polyamines on cell fate decisions of naïve CD4+ T cells in vitro. We found that endogenously generated polyamines are essential for the development of T helper 2 (Th2) cells. Treatment with DL-2-difluoromethylornithine (DFMO), an inhibitor of polyamine biosynthesis, diminished GATA3 expression in CD4+ T cells under Th2-skewed conditions. Supplementation of exogenous polyamines rescued GATA3 downregulation caused by DFMO treatment in CD4+ T cells. Transcriptome analysis revealed that deprivation of endogenous polyamines resulted in upregulated Th9-related genes, such as Il9 , Irf4 , and Batf3 , even under the Th2-skewing conditions. Depletion of intracellular polyamines reduced GATA3 expression but increased IL-9-producing CD4+ T cells under both Th2 and Th9-skewing conditions. Furthermore, oral administration of DFMO increased IL-9-producing CD4+ T cells in small intestine in mice. Thus, our data indicate that polyamines play a critical role in the regulation of the Th2/Th9 balance. • Endogenous polyamine synthesis is essential for the GATA3 expression during T helper 2 (Th2) cell differentiation. • Polyamine synthesis inhibition upregulated the expression of Th9-related genes such as IL-9 under Th2-skewing conditions. • Polyamine synthesis inhibition augmented IL-9 production under Th2 and Th9-skewing conditions. • Deprivation of polyamines increased IL-9-producing CD4+ T cells in vivo. [ABSTRACT FROM AUTHOR]