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Summary The gene mutation profiles of gastric neuroendocrine neoplasms are incompletely understood. The purpose of this study was to characterize the molecular pathology of poorly differentiated neuroendocrine carcinoma (NEC) and mixed neuroendocrine‒non-neuroendocrine neoplasm (MiNEN) of the stomach. Surgical cases of gastric NEC (n = 7) and MiNEN (n = 6) were examined by clinical review, immunohistochemistry, microsatellite instability (MSI) analysis and whole-exome sequencing. NEC cases consisted of small- (n = 2) and large-cell types (n = 4). All cases of MiNEN were histologically composed of large-cell type NEC and tubular adenocarcinoma. Whole-exome sequencing analysis detected recurrent mutations in TP53 in 8 cases (62%), and they were more frequently observed in MiNEN than in NEC (100% vs. 29%). Frameshift mutations of APC were observed in two cases of MiNEN. One case of large-cell type NEC had a frameshift mutation with loss of heterozygosity in RB1. The other mutated genes (e.g., ARID1 and KRAS) were detected in a single case each. A high level of MSI was confirmed in one case of MiNEN, which harbored mutations in two well-differentiated neuroendocrine tumor (NET)-related genes (MEN1 and ATRX1). In cases of MiNEN, two histological components shared mutations in TP53, APC and ZNF521, whereas alterations in CTNNB1, KMT2C, PTEN and SPEN were observed in neuroendocrine components only. In conclusion, TP53 is a single, frequently mutated gene in gastric NEC and MiNEN, and alterations in other genes are less common, resembling the mutation profiles of gastric adenocarcinomas. Gene mutations frequently observed in well-differentiated NET were uncommon but not entirely exclusive.

Objectives: Exosomes are potent vehicles for intercellular communication. Rheumatoid arthritis (RA) is a chronic systemic disease of unknown etiology. Local administration of miR-124 precursor to rats with adjuvant-induced arthritis suppresses systemic arthritis and bone destruction. Thus, exosomes may be involved in this disease. We aimed to determine the role of exosomes in the pathology of RA. Methods: Fibroblast-like synoviocytes (FLS) were collected from patients with RA and osteoarthritis (OA). miR-124-3p mimic was transfected into the RA FLS (RA miR-124 FLS). Exosomes were collected from the culture medium by ultracentrifugation. Macrophages were produced from THP-1 cells. MicroRNAs in the exosomes were analyzed using real-time PCR. Proteomics analysis was performed using nanoscale liquid chromatography-tandem mass spectrometry. Macrophage migration was evaluated using a Transwell migration assay. SiRNA was used to knockdown proteins of interest. Results: MicroRNAs in the RA FLS, RA miR-124 FLS, and OA FLS exosomes were similar. Proteomics analysis revealed that pentraxin 3 (PTX3) levels were higher in RA FLS exosomes than in RA miR-124 FLS and OA FLS exosomes, and proteasome 20S subunit beta 5 (PSMB5) levels were lower in RA FLS exosomes than in RA miR-124 FLS and OA FLS exosomes. The RA FLS exosomes promoted and the RA miR-124 FLS exosomes suppressed macrophage migration. PTX3-silenced RA FLS exosomes suppressed and PSMB5-silenced OA FLS exosomes promoted macrophage migration. Conclusions: RA FLS exosomes promote macrophage migration via PTX3 and PSMB5, and miR-124-3p suppresses this migration.

Background One of the main complications in patients with muscular dystrophies is cardiac dysfunction. The literature on cardiac involvement in patients with Fukuyama congenital muscular dystrophy (FCMD) is limited. Aim To compare cardiac involvement between patients with FCMD and Duchenne muscular dystrophy (DMD). Methods We compared cardiac involvement between 30 patients with FCMD and 181 patients with DMD using echocardiography and serum biomarkers. All patients were receiving regular checkups at Kobe University Hospital. We used single regression analysis to compare echocardiographic parameters, age, and serum biomarkers. Results Almost all clinical and echocardiographic parameters were lower in patients with FCMD than DMD. The brain natriuretic peptide concentration in patients with FCMD showed no correlation with age or left ventricular ejection fraction ( r = 0.231, p = 0.22 and r = 0.058, p = 0.76, respectively). A log-rank test revealed that the risk of left ventricular systolic dysfunction was lower in patients with FCMD than DMD ( p = 0.046, hazard ratio = 0.348). Conclusion The clinical progression of cardiac dysfunction is significantly milder in patients with FCMD than DMD, while skeletal muscle involvement is significantly worse in patients with FCMD. These data suggest that the pathophysiological findings of FCMD can be explained by less severe cardiac dysfunction in FCMD than DMD.

Following the Fukushima Dai-ichi Nuclear Power Plant accident in March 2011, extensive radiation monitoring and environmental clean-up activities have been conducted throughout the Fukushima region. Outside air temperatures there reach 40 °C in summer and −20 °C in winter, which are beyond the quoted operational range of many radiation survey instruments. Herein, temperature performance of four types of portable Japanese radiation survey instruments widely used in Fukushima was experimentally investigated using a temperature-controlled chamber. They included two ionization chamber type instruments, Fuji NHA1 and Aloka ICS-323C, and two NaI(Tl) scintillation type ones, Fuji NHC7 and Aloka TCS-172B. Experimental results showed significantly diverse characteristics on the temperature dependences from one type of instrument to another. For example, NHA1 overestimated the ambient dose-equivalent rate by as much as 17% at −30 °C and 10% at 40 °C, whereas the TCS-172B readings underestimated the rate by 30% at −30 °C and 7% at 40 °C.

Purpose To evaluate the incidence of fragility fractures associated with high-dose glucocorticoid therapy in patients with systemic rheumatic disease. Methods: A retrospective study of patients who were treated with high-dose prednisolone (> 0.8 mg/kg) for systemic rheumatic disease at Kobe University Hospital from April 1988 to March 2012. The primary outcome was a major osteoporotic fracture (defined as a clinical vertebral, hip, forearm, or proximal humerus fracture) after high-dose glucocorticoid therapy. For postmenopausal women and men over 40 of age, the patient's fracture risk at the beginning of high-dose glucocorticoid therapy was assessed by the World Health Organization's Fracture Risk Assessment Tool (FRAX®). Results Of 229 patients (median age: 49 years), 57 suffered a fragility fracture during the observation period (median observation period: 1558 days). Of 84 premenopausal patients, 5 suffered a fracture. In contrast, of 86 postmenopausal female, 36 suffered a fracture. Fragility fractures were far more frequent than predicted by the FRAX® score. Patients with FRAX® scores over 8.3% had a particularly high risk of fracture. Conclusions Fragility fractures associated with high-dose glucocorticoid therapy are common among postmenopausal women. Extreme care should be taken especially for postmenopausal women when high-dose glucocorticoid therapy is required., Highlights • We evaluated the incidence of the fragility fractures associated with high-dose glucocorticoid therapy. • Of 86 postmenopausal female, 36 suffered a fracture during the observation period. • Patients with FRAX scores over 8.3% had a particularly high risk of fracture.

After the accident at the Fukushima Daiichi nuclear power plant (NPP), outdoor school swimming pools at Fukushima were decontaminated to curb the redistribution of radioactivity into downstream farmlands. In the process, the radioactivity concentrations of the pool water and sediment substances (residue) were measured to estimate the deposition density of the fallout. At a pool situated 50 km away from the NPP, the average concentrations of radiocesium ( 134+137 Cs) for the water and residue were quantified as 170 Bq L −1 and 3.6 × 10 5 Bq kg −1 , respectively. Taking account of the radioactivity concentrations and of the water balance in and around the pool, the deposition density of radiocesium, as of August 2011, was precisely determined to be 0.32 ± 0.03 MBq m −2 ( k = 1). The density corroborated the previous results obtained by other methods, i.e., airborne surveys, in-situ Ge surveys and soil samplings at neighboring locations. Other than radiocesium, the only gamma-emitting nuclide detected was 110m Ag, with a concentration of 560 Bq kg −1 in the residue. The radioactivity concentrations of 89 Sr, 90 Sr, 238 Pu and 239+240 Pu in the water were all less than the minimum detectable activities – 2, 0.1, 0.002 and 0.002 Bq L −1 , respectively.

SS-A/Ro52 (Ro52), an autoantigen in systemic autoimmune diseases such as systemic lupus erythematosus and Sjogren’s syndrome, has E3 ligase activity to ubiquitinate proteins that protect against viral infection. To investigate Ro52’s role during stress, we transiently knocked it down in HeLa cells by siRo52 transfection. We found that Ro52 low HeLa cells were significantly more resistant to apoptosis than wild-type HeLa cells when stimulated by H 2 O 2 - or diamide-induced oxidative stress, IFN-α, IFN-γ and anti-Fas antibody, etoposide, or γ-irradiation. Furthermore, Ro52-mediated apoptosis was not influenced by p53 protein level in HeLa cells. Depleting Ro52 in HeLa cells caused Bcl-2, but not other Bcl-2 family molecules, to be upregulated. Taken together, our data showed that Ro52 is a universal proapoptotic molecule, and that its proapoptotic effect does not depend on p53, but is exerted through negative regulation of the anti-apoptotic protein Bcl-2. These findings shed light on a new physiological role for Ro52 that is important to intracellular immunity.

Galectin-3 is highly expressed in epithelial cells including keratinocytes and is involved in the pathogenesis of inflammatory skin diseases by affecting the functions of immune cells. For example, galectin-3 can contribute to atopic dermatitis (AD) by promoting polarization toward a Th2 immune response by regulating dendritic cell (DC) and T cell functions. In addition, galectin-3 may be involved in the development of contact hypersensitivity by regulating the migratory capacity of antigen presenting cells. Galectin-3 may act as a regulator of epithelial tumor progression and development through various signaling pathways, such as inhibiting keratinocyte apoptosis through regulation of the activation status of extracellular signal-regulated kinase (ERK) and activated protein kinase B (AKT). Galectin-3 is detected at different stages of melanoma development. In contrast, a marked decrease in the expression of galectin-3 is observed in non-melanoma skin cancers, such as squamous cell carcinoma (SCC) and basal cell carcinoma (BCC). Galectin-3 may play an important role in tumor cell growth, apoptosis, cell motility, invasion, and metastasis. Galectin-3 may be a novel therapeutic target for a variety of skin diseases.

Target photons mixed in the 144, 250 and 565 keV mono-energetic neutron calibration fields were measured using a cylindrical NaI(Tl) detector with 7.62 cm both in diameter and in length. The ambient dose equivalent H*(10) of the photons was evaluated by applying the “G(E) function” to the measured pulse height spectrum. Neutrons induce photons by nuclear reactions in the NaI(Tl) detector and affect the pulse height spectrum. In order to eliminate the influence of these neutron events, the time-of-flight technique was applied with operating the accelerator in the pulse mode. The ratios by the ambient dose equivalent H*(10) of the photons to the 144, 250 and 565 keV neutrons were evaluated to be 3.3%, 4.7% and 0.9%, respectively. Although high energy photons ranging from 6 to 7 MeV are emitted by the 19F(p,αγ)16O reactions, the dose of the target photons is low enough to calibrate neutron dosemeters except for ones with high sensitivity to the photons.

Precise calibration of monitors and dosimeters for use with high energy neutrons necessitates reliable and accurate neutron fluences being evaluated with use of a reference point. A highly efficient Proton Recoil counter Telescope (PRT) to make absolute measurements with use of a reference point was developed to evaluate neutron fluences in quasi-monoenergetic neutron fields. The relatively large design of the PRT componentry and relatively thick, approximately 2 mm, polyethylene converter contributed to high detection efficiency at the reference point over a large irradiation area at a long distance from the target. The polyethylene converter thickness was adjusted to maintain the same carbon density per unit area as the graphite converter for easy background subtraction. The high detection efficiency and thickness adjustment resulted in efficient absolute measurements being made of the neutron fluences of sufficient statistical precision over a short period of time. The neutron detection efficiencies of the PRT were evaluated using MCNPX code at 2.61×10 −6 , 2.16×10 −6 and 1.14×10 −6 for the respective neutron peak energies of 45, 60 and 75 MeV. The neutron fluences were determined to have been evaluated at an uncertainty of within 6.5% using analysis of measured data and the detection efficiencies. The PRT was also designed so as to be capable of simultaneously obtaining TOF data. The TOF data also increased the reliability of neutron fluence measurements and provided useful information for use in interpreting the source of proton events.

Galectin-3 belongs to a family of beta-galactoside-binding animal lectins expressed in several cell types, including epithelial and immune cells. To establish the role of galectin-3 in the development of allergic skin inflammation, we compared inflammatory skin responses of galectin-3-deficient (gal3(-/-)) and wild-type (gal3(+/+)) mice to epicutaneous sensitization with ovalbumin (OVA). OVA-treated gal3(-/-) mice exhibited markedly reduced epidermal thickening, lower eosinophil infiltration, and lower serum IgE levels compared with gal3(+/+) mice. The former evoked lower interleukin-4, but higher interferon-gamma, mRNA expression at OVA-treated skin sites. Moreover, gal3(-/-) splenocytes from OVA-sensitized mice secreted more interleukin-12 compared with gal3(+/+) splenocytes. In addition, antigen presentation by gal3(-/-) dendritic cells to T cells in vitro were T helper cell (Th1)-polarized relative to presentation by gal3(+/+) dendritic cells. When exposed to OVA, recipients engrafted with T cells from gal3(-/-) OVA-specific T cell receptor transgenic mice developed significantly reduced dermatitis and a markedly lower Th2 response compared with recipients of comparable gal3(+/+) T cells. We conclude that galectin-3 is critical for the development of inflammatory Th2 responses to epicutaneously administered antigens; in its absence, mice develop a Th1-polarized response. This regulatory effect of galectin-3 on Th development is exerted at both the dendritic cell and T cell levels. Our studies suggest that galectin-3 may play an important role in the acute phase of human atopic dermatitis.

Secretory phospholipase A2 group IIA (sPLA2-IIA) plays an important role in the pathogenesis of inflammatory diseases. Catalytic activity of this enzyme that generates arachidonic acid is a major target for development of anti-inflammatory agents. Independent of its catalytic activity, sPLA2-IIA induces pro-inflammatory signals in a receptor-mediated mechanism (e.g. through the M-type receptor). However, the M-type receptor is species-specific: sPLA2-IIA binds to the M-type receptor in rodents and rabbits, but not in human. Thus sPLA2-IIA receptors in human have not been established. Here we demonstrated that sPLA2-IIA bound to integrin alphavbeta3 at a high affinity (K(D)=2 x 10(-7) M). We identified amino acid residues in sPLA2-IIA (Arg-74 and Arg-100) that are critical for integrin binding using docking simulation and mutagenesis. The integrin-binding site did not include the catalytic center or the M-type receptor-binding site. sPLA2-IIA also bound to alpha4beta1. We showed that sPLA2-IIA competed with VCAM-1 for binding to alpha4beta1, and bound to a site close to those for VCAM-1 and CS-1 in the alpha4 subunit. Wild type and the catalytically inactive H47Q mutant of sPLA2-IIA induced cell proliferation and ERK1/2 activation in monocytic cells, but the integrin binding-defective R74E/R100E mutant did not. This indicates that integrin binding is required, but catalytic activity is not required, for sPLA2-IIA-induced proliferative signaling. These results suggest that integrins alphavbeta3 and alpha4beta1 may serve as receptors for sPLA2-IIA and mediate pro-inflammatory action of sPLA2-IIA, and that integrin-sPLA2-IIA interaction is a novel therapeutic target.

Integrins play a role in fibroblast growth factor (FGF) signaling through cross-talk with FGF receptors (FGFRs), but the mechanism underlying the cross-talk is unknown. We discovered that FGF1 directly bound to soluble and cell-surface integrin alphavbeta3 (K(D) about 1 microm). Antagonists to alphavbeta3 (monoclonal antibody 7E3 and cyclic RGDfV) blocked this interaction. alphavbeta3 was the predominant, if not the only, integrin that bound to FGF1, because FGF1 bound only weakly to several beta1 integrins tested. We presented evidence that the CYDMKTTC sequence (the specificity loop) within the ligand-binding site of beta3 plays a role in FGF1 binding. We found that the integrin-binding site of FGF1 overlaps with the heparin-binding site but is distinct from the FGFR-binding site using docking simulation and mutagenesis. We identified an FGF1 mutant (R50E) that was defective in integrin binding but still bound to heparin and FGFR. R50E was defective in inducing DNA synthesis, cell proliferation, cell migration, and chemotaxis, suggesting that the direct integrin binding to FGF1 is critical for FGF signaling. Nevertheless, R50E induced phosphorylation of FGFR1 and FRS2alpha and activation of AKT and ERK1/2. These results suggest that the defect in R50E in FGF signaling is not in the initial activation of FGF signaling pathway components, but in the later steps in FGF signaling. We propose that R50E is a useful tool to identify the role of integrins in FGF signaling.

An efficiency calibration method has been developed to measure radioactivity of volume samples with HPGe detectors. The method has two main procedures. One is a point-source calibration at a representative point near the detector. The position of the representative point is determined by Monte Carlo simulations. The other is correction for absorption by a sample matrix and a sample container. This paper presents application of the method for obtaining efficiency curves for volume samples in the energy range from 22 to 1836 keV. In addition to the standard volume sources of cement and water matrices, measurement was made with some kinds of environmental samples that are often used in environmental monitoring for radiation control works. The composition of the environmental sample was determined by elemental analysis. For these samples, measured absorption correction factors were compared with those by the Monte Carlo simulations. As a result, detection efficiency curves were obtained with good accuracy for variety types of volume samples.

Temporal and spatial distribution of 222 Rn and 220 Rn and their progeny concentrations in the open atmosphere, 222 Rn and 220 Rn exhalation rates, radioactivities in soil and underground water, including some meteorological factors, were measured at a spot of high 222 Rn and 220 Rn concentrations to investigate the source and occurrence mechanism. Hourly mean concentrations of 222 Rn and 220 Rn during August 1993 were about 230 Bq m −3 and 130 Bq m −3 at 1 m above ground surface. Diurnal variations were high at daytime and low at nighttime, which gives an inverse variation compared with the typical variation in the normal environment. 222 Rn concentrations varied quickly in a few minutes and reached a maximum of about 14 kBq m −3 . The maximum daily mean 222 Rn concentration at 5 cm above ground surface was 11 kBq m −3 . Concentrations of 218 Po, 214 Pb, 214 Bi, and 220 Rn progeny were 8∼61 Bqm −3 , 1∼7 Bqm −3 , 0.5∼5 Bq m −3 , and 0.02∼0.14 Bq m −3 , respectively. The equilibrium factor for 222 Rn was estimated to be 0.01∼0.2. The main source of the mysterious spot of high 222 Rn and 220 Rn concentrations was a small but deep channel between the basement of the building and the surrounding soil, and the extremely localized weather condition.