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Abstract Background Polyriboinosinic-polyribocytidylic acid (polyI:C) triggers a strong innate immune response that mimics immune activation by viral infections. Induction of interferon-induced transmembrane protein 3 (Ifitm3) in astrocytes has a crucial role in polyI:C-induced neurodevelopmental abnormalities. Through a quantitative proteomic screen, we previously identified candidate astroglial factors, such as matrix metalloproteinase-3 (Mmp3) and follistatin-like 1 (Fstl1), in polyl:C-induced neurodevelopmental impairment. Here, we characterized the Ifitm3-dependent inflammatory processes focusing on astrocyte-derived Fstl1 following polyI:C treatment to assess the neuropathologic role of Fstl1. Methods Astrocytes were treated with PBS (control) or polyI:C (10 μg/mL). The conditioned medium was collected 24 h after the polyI:C treatment and used as astrocyte condition medium (ACM). The expression of Fstl1 mRNA and extracellular Fstl1 protein levels were analyzed by quantitative PCR and western blotting, respectively. For functional studies, neurons were treated with ACM and the effects of ACM on dendritic elongation were assayed. To examine the role of Fstl1, recombinant Fstl1 protein and siRNA for Fstl1 were used. To investigate the expression of Fstl1 in vivo, neonatal mice were treated with vehicle or polyI:C on postnatal day 2 to 6. Results ACM prepared with polyI:C (polyI:C ACM) contained significantly higher Fstl1 protein than control ACM, but no increase in Fstl1 was observed in polyI:C ACM derived from Ifitm3-deficient astrocytes. We found that the production of Fstl1 involves the inflammatory responsive molecule Ifitm3 in astrocytes and influences neuronal differentiation. In agreement, the levels of Fstl1 increased in the hippocampus of polyI:C-treated neonatal mice. COS7 cells co-transfected with both Fstl1 and Ifitm3 had higher extracellular levels of Fstl1 than the cells transfected with Fstl1 alone. Treatment of primary cultured hippocampal neurons with recombinant Fstl1 impaired dendritic elongation, and the deleterious effect of polyI:C ACM on dendritic elongation was attenuated by knockdown of Fstl1 in astrocytes. Conclusions The extracellular level of Fstl1 is regulated by Ifitm3 in astrocytes, which could be involved in polyI:C-induced neurodevelopmental impairment.

Inhalation therapy using small-interfering RNA (siRNA) is a potentially effective therapeutic strategy for lung cancer because of its high gene-silencing effects and sequence specificity. Previous studies reported that intratracheal administration of siRNA using pressurized metered dose inhalers or nebulizers could suppress tumor growth in murine lung metastatic models. Although dry powder inhalers are promising devices due to their low cost, good portability, and preservability, the anti-tumor effects of siRNA dry powder have not been elucidated. To evaluate the gene-silencing and anti-tumor effects of intratracheally delivered siRNA dry powder, vascular endothelial growth factor-specific siRNA (VEGF-siRNA) dry powder was administered intratracheally to mice with metastatic lung tumors consisting of B16F10 melanoma cells or Lewis lung carcinoma cells. A single intratracheal administration of VEGF-siRNA dry powder reduced VEGF levels in both bronchoalveolar lavage fluid and lung tumor tissue. Furthermore, repeated intratracheal administration of VEGF-siRNA dry powder suppressed the number of visible metastatic foci on the lung surface and tumor area in lung tissues. Taken together, intratracheal administration of siRNA dry powder could be a novel therapeutic strategy for lung cancer through the suppression of specific genes expressed in lung tumor tissue. Keywords: siRNA, dry powder, intratracheal administration, lung cancer, vascular endothelial growth factor

Summary: Dopamine (DA) activates mitogen-activated protein kinase (MAPK) via protein kinase A (PKA)/Rap1 in medium spiny neurons (MSNs) expressing the dopamine D1 receptor (D1R) in the nucleus accumbens (NAc), thereby regulating reward-related behavior. However, how MAPK regulates reward-related learning and memory through gene expression is poorly understood. Here, to identify the relevant transcriptional factors, we perform proteomic analysis using affinity beads coated with cyclic AMP response element binding protein (CREB)-binding protein (CBP), a transcriptional coactivator involved in reward-related behavior. We identify more than 400 CBP-interacting proteins, including Neuronal Per Arnt Sim domain protein 4 (Npas4). We find that MAPK phosphorylates Npas4 downstream of PKA, increasing the Npas4-CBP interaction and the transcriptional activity of Npas4 at the brain-derived neurotrophic factor (BDNF) promoter. The deletion of Npas4 in D1R-expressing MSNs impairs cocaine-induced place preference, which is rescued by Npas4-wild-type (WT), but not by a phospho-deficient Npas4 mutant. These observations suggest that MAPK phosphorylates Npas4 in D1R-MSNs and increases transcriptional activity to enhance reward-related learning and memory. : Funahashi et al. isolate and concentrate a transcriptional factor from the mouse striatum using affinity beads coated with CBP and identify more than 400 CBP-interacting proteins, including Npas4. MAPK phosphorylates Npas4 in D1R-MSNs and increases the interaction between Npas4 and CBP, thereby regulating transcriptional activity to enhance reward-related learning and memory. Keywords: Phosphorylation, Dopamine, Gene expression, Reward, Behavior, Learning, Memory, MAPK, Npas4, CBP

Unsteady pressure distributions around a simplified sedan automobile model were investigated by conducting dynamic wind-tunnel testing using the newly developed forced oscillating apparatus, HEXA-X3, which can produce 6-degrees-of-freedom motion. The effects of heaving and pitching oscillation were investigated as the model simulated a vehicle running on a flat road at approximately 40 m/s and 1 Hz oscillation. The effects of the ground plate on unsteady pressure distributions over the model surfaces were measured while simulating heaving and pitching motion at Strouhal-number conditions similar to those for actual vehicles. The influence of the tubing on the frequency response of the pressure sensor was evaluated to be negligible by conducting a calibration experiment first. In the static case, the overall pressure distribution was consistent with that for a typical sedan, and the influence of the local relative flow velocity changes due to the contraction effect was observed in the underside of the model. In the forced oscillation tests, the effect of heaving and pitching motions on the flow around the underside was investigated. Effects of oscillation parameters, specifically amplitude and frequency, were investigated using the gain and phase-lag normalized by data from the steady model. Results of the test indicate that there is a characteristic distribution in pressure fluctuation, and the phenomena that become dominant in the flow around the underside vary according to location. The dynamic heaving motion was shown to change the pressure distribution, possibly due to changes in the effective angle of attack in addition to the static effect. The pitching test showed that a dynamic camber effect works in addition to those effects.

The data presented here are related to the research article entitled “Loss of Eed leads to lineage instability and increased CD8 expression of mouse CD4+ T cells upon TGFβ signaling” [1]. The cited research article investigates the molecular mechanism of CD8α upregulation observed in Eed-deficient (∆Eed) CD4+ T cells upon activation in the presence of TGFβ. This data report describes the effect of retinoic acid (RA) and/or anti-interferon-gamma (IFNγ) antibody supplementation on up-regulation of CD8α and Foxp3 in ∆Eed CD4+ T cells, the effect of dose or timing of TGFβ treatment on CD4+ T cell identity of ∆Eed, adding further information regarding the conditions that induces CD8α, and mRNA expression changes of genes encoding polycomb repressive complex 2 (PRC2) subunits by TGFβ treatment. Keywords: TGFβ, PRC2, Eed, CD4+ CD8α+ T cells

Background: Development of novel agents targeting the androgen axis has led to improved overall survival in castration-resistant prostate cancer (CRPC). This study aimed to investigate the optimal timing of treatment with one such agent, abiraterone acetate (AA), in Japanese patients. Materials and methods: Between July 2014 and February 2016, 106 CRPC patients were administered AA in Nagoya City University Hospital, Nagoya, Japan and in four affiliated hospitals following failure of primary combined androgen blockade (CAB). Of these, records of 69 patients treated before chemotherapy were retrospectively analyzed. Patients were divided into two AA treatment groups: (1) first- or second-line after diagnosis of CRPC, designated the Early Group, and (2) third-line onwards, designated the Deferred Group. Prostate-specific antigen (PSA) response rate, ≥ 50% PSA decline rate with treatment, progression-free survival (PFS), and overall survival (OS) were compared between the two groups. National Cancer Institute Common Terminology Criteria for Adverse Events, version 4.0 was used to classify adverse events. Results: In 24 patients in the Early Group and 45 patients in the Deferred Group, no significant differences in baseline parameters were observed between groups. PSA response rate, ≥ 50% PSA decline rate and PFS (but not OS) were significantly better in the Early Group than in the Deferred Group. Serum aspartate aminotransferase/alanine aminotransferase elevations were the most common Grade 3 treatment-related toxicities, and were clinically manageable. In subgroup analyses of the Early Group, comparison of first-line AA with second-line AA after flutamide treatment showed no changes in PSA response rate, PFS, or OS. Conclusion: This study suggests improved favorable outcomes of first- or second-line AA treatment in Japanese chemotherapy-naïve CRPC patients after failed CAB; statistical confirmation of such improvement was evident for PFS, but not OS. In addition, early AA treatment exhibited an acceptable safety profile.

Protein phosphorylation plays critical roles in a variety of intracellular signaling pathways and physiological functions that are controlled by neurotransmitters and neuromodulators in the brain. Dysregulation of these signaling pathways has been implicated in neurodevelopmental disorders, including autism spectrum disorder, attention deficit hyperactivity disorder and schizophrenia. While recent advances in mass spectrometry-based proteomics have allowed us to identify approximately 280,000 phosphorylation sites, it remains largely unknown which sites are phosphorylated by which kinases. To overcome this issue, previously, we developed methods for comprehensive screening of the target substrates of given kinases, such as PKA and Rho-kinase, upon stimulation by extracellular signals and identified many candidate substrates for specific kinases and their phosphorylation sites. Here, we developed a novel online database to provide information about the phosphorylation signals identified by our methods, as well as those previously reported in the literature. The “KANPHOS” (Kinase-Associated Neural Phospho-Signaling) database and its web portal were built based on a next-generation XooNIps neuroinformatics tool. To explore the functionality of the KANPHOS database, we obtained phosphoproteomics data for adenosine-A2A-receptor signaling and its downstream MAPK-mediated signaling in the striatum/nucleus accumbens, registered them in KANPHOS, and analyzed the related pathways.

Stroke causes balance dysfunction, leading to decreased physical activity and increased falls. Thus, effective balance exercises are needed to improve balance dysfunction. This single-blind, single-center randomized controlled trial evaluated the long-term and continuous effects of balance exercise using a real-time postural feedback system to improve balancing ability safely. Thirty participants were randomized into intervention (n = 15) and control (n = 15) groups; 11 in each group completed the final evaluation. The effect of the intervention was evaluated by muscle strength of knee extension, physical performance (short physical performance battery, the center of pressure trajectory length per second, and Timed Up and Go test [TUG]), and self-reported questionnaires (modified Gait Efficacy Scale [mGES] and the Fall Efficacy Scale) at pre (0 week), post (6-week), and at follow-up (10-week) visits. The TUG and mGES showed a significant interactive (group * time) effect (p = 0.007 and p = 0.038, respectively). The intervention group showed significant decreasing time to perform TUG from pre- to post-intervention (p = 0.015) and pre-intervention to follow-up (p = 0.016); mGES showed a significant change from pre-intervention to follow-up (p = 0.036). Thus, balance exercise using a real-time postural feedback system can confer a positive effect on the walking ability in patients with chronic stroke and increase their self-confidence in gait performance.

Abstract Background Pheochromocytomas are rare neuroendocrine tumors, with a malignancy frequency of approximately 10%. The treatment of malignant pheochromocytoma is palliative, and the traditional management strategy has limited efficacy. Furthermore, no clear criteria exist for the treatment of metastatic pheochromocytoma, especially for unresectable lesions. We report a case of complete remission of metastatic pheochromocytoma in 123I-metaiodobenzylguanidine (MIBG) scintigraphy after a single session of 131I-MIBG therapy. Case presentation A 61-year-old woman had a right adrenal grand tumor and lymph node metastasis on the hilum of the right kidney, both of which incorporated MIBG. After surgery, immunostaining of a tumor specimen showed expression of the tumor makers chromogranin and synaptophysin. One year postoperatively, abdominal computed tomography revealed a local recurrence and retroperitoneal lymph node swelling. The local recurrence was positive for MIBG uptake, whereas the swollen retroperitoneal lymph nodes were negative. She underwent surgery again, but the local recurrence was unresectable because of rigid adhesion to the surrounding tissue. Immunostaining of an intraoperatively extracted swollen retroperitoneal lymph node showed expression of tumor markers. The patient then underwent a single session of 131I-MIBG therapy (7.4 GBq, 200 mCi), after which the residual lesions no longer incorporated MIBG, and a complete response in 123I- metaiodobenzylguanidine (MIBG) scintigraphy was achieved. The 131I-MIBG treatment was repeated 6 months later. None of the lesions were positive for MIBG uptake. Conclusions 131I-MIBG therapy efficaciously treats unresectable lesions that are positive for MIBG uptake.

Abstract The alkaliphilic, halophilic bacterium Halomonas sp. KM-1 can utilize glucose for the intracellular storage of the bioplastic poly-(R)-3-hydroxybutyric acid (PHB) and extracellular secretion of pyruvate under aerobic conditions. In this study, we investigated the effects of sodium chloride concentration on PHB accumulation and pyruvate secretion in the KM-1 strain and, unexpectedly, observed that oxaloacetate, an important intermediate chemical in the TCA cycle, glycogenesis, and aspartic acid biosynthesis, was secreted. We then further analyzed oxaloacetate productivity after changing the sodium chloride additive concentration, additive time-shift, and culture temperature. In 42-h batch-cultivation experiments, we found that wild-type Halomonas sp. KM-1 secreted 39.0 g/L oxaloacetate at a rate of 0.93 g/(L h). The halophilic bacteria Halomonas has already gained attention for industrial chemical-production processes owing to its unique properties, such as contamination-free culture conditions and a tolerance for high substrate concentrations. Moreover, no commercial scale oxaloacetate production was previously reported to result from bacterial fermentation. Oxaloacetate is an important intermediate chemical in biosynthesis and is used as a health food based on its role in energy synthesis. Thus, these data provided important insights into the production of oxaloacetate and other derivative chemicals using this strain.

The underlying histopathology is very important in determining patient management, as the histopathology usually has direct repercussions on the treatment response and clinical course. However, the impact of the method used to assess renal biopsies, i.e., light microscopy (LM), immunofluorescence (IF), and electron microscopy (EM), on the occurrence of a difficult biopsy classification in the native kidneys of pediatric nephrotic patients is unknown. A 12-month-old Japanese boy was diagnosed with nephrotic syndrome (NS); he was administered prednisolone (60 mg/m2/day), and a continuous albumin infusion was started. A renal biopsy using LM revealed minimal change. However, an IF study showed granular staining for immunoglobulin G along the glomerular basement membrane. Therefore, he was diagnosed with membranous nephropathy (MN). As his proteinuria was so severe, we started immunosuppressant therapy and continued the albumin infusion for more than 2 months. However, he did not attain complete remission. A month later, EM examination of his renal biopsy showed extensive foot process fusion without electron-dense deposits. Although the result of the IF study suggested MN, the results of the LM and EM studies indicated minimal change. We finally diagnosed the patient with minimal change NS, in consideration of his clinical condition and course. Because of the failure of previous treatments, pulse steroid therapy was started. After five rounds of therapy the patient attained complete remission. A difficult renal biopsy finding classification, dependent on the diagnostic method used, might occur in the native kidneys of pediatric nephrotic patients. Therefore, a diagnosis should be made after considering all renal biopsy findings and the clinical course.

The prognostic significance of glandular differentiation in urothelial carcinoma (UC) is controversial, and thus far there is no established treatment strategy against metastasis of glandular component. We describe here a case of metastatic UC with glandular differentiation that had histological disappearance of adenocarcinoma components at autopsy after sequential chemotherapy with S-1 and cisplatin (CDDP) and with mFOLFOX6 (fluorouracil, oxaliplatin, and leucovorin) plus bevacizumab (mFOLFOX6+Bev). A 62-year-old Asian male was diagnosed with invasive UC with glandular differentiation (T2N0M0) by radical cystectomy and ileal conduit, and careful follow-up observation was made. Eight years after radical operation, peritoneal metastases occurred, and a biopsy specimen using colon fiber revealed high-grade adenocarcinomas with an immunohistochemical profile that included positivity for cytokeratin 7 (CK7) and negativity for cytokeratin 20 (CK20) and uroplakin, which was identical to the radical cystectomy specimen. Thus, he received combination chemotherapy consisting of S-1 and CDDP; however, the peritoneal metastasis worsened after 2 cycles. Therefore, second-line mFOLFOX6+Bev chemotherapy was performed for a total of 5 courses. In spite of this, the patient died, and the final diagnosis by autopsy was multiple metastases of infiltrating pure UC to the lung, bone, and peritoneum. Interestingly, there were no pathological findings of adenocarcinoma, and the immunohistochemical profile of the metastatic lesions was identical to that of the previous specimens from the bladder and colon. This suggests that sequential chemotherapy of S-1 and CDDP and second-line mFOLFOX6+Bev might be a feasible option in metastatic UC with glandular differentiation.

Houttuynia cordata Thunb. (HCT) is a well-known Asian medicinal plant with biological activities used in the treatment of many diseases including cancer. This study investigated the effects of HCT extract and its ethyl acetate fraction (EA) on prostate carcinogenesis and castration-resistant prostate cancer (CRPC). HCT and EA induced apoptosis in androgen-sensitive prostate cancer cells (LNCaP) and CRPC cells (PCai1) through activation of caspases, down-regulation of androgen receptor, and inactivation of AKT/ERK/MAPK signaling. Rutin was found to be a major component in HCT (44.00 ± 5.61 mg/g) and EA (81.34 ± 5.21 mg/g) in a previous study. Rutin had similar effects to HCT/EA on LNCaP cells and was considered to be one of the active compounds. Moreover, HCT/EA inhibited cell migration and epithelial-mesenchymal transition phenotypes via STAT3/Snail/Twist pathways in LNCaP cells. The consumption of 1% HCT-mixed diet significantly decreased the incidence of adenocarcinoma in the lateral prostate lobe of the Transgenic rat for adenocarcinoma of prostate model. Similarly, tumor growth of PCai1 xenografts was significantly suppressed by 1% HCT treatment. HCT also induced caspase-dependent apoptosis via AKT inactivation in both in vivo models. Together, the results of in vitro and in vivo studies indicate that HCT has inhibitory effects against prostate carcinogenesis and CRPC. This plant therefore should receive more attention as a source for the future development of non-toxic chemopreventive agents against various cancers.

The benefits of probiotic supplementation to lactating mothers on human milk cytokines are inconclusive. Thus, we performed a comprehensive open-label pilot trial analysis of 27 human milk cytokines in lactating women with allergies (one to three months postpartum) to determine the effect of supplementation with a mixture of new probiotic strains. Participants voluntarily joined the probiotic (n = 41) or no supplementation control (n = 19) groups. The probiotic group took three probiotic tablets (Lactobacillus casei LC5, Bifidobacterium longum BG7, and Bacillus coagulans SANK70258) daily for one to three months postpartum. Milk samples were collected at one, two, and three months postpartum, and cytokine levels were measured using multiplex assays. The effects were analyzed using multivariate regression models. Eleven cytokines showed a positive rate of over 50% in the milk samples throughout testing in both groups. The positive rates of IL-1 receptor antagonist and IL-7 changed significantly with lactation progression in logistic regression models after adjusting for time and supplementation, whereas rates of other cytokines showed no significant differences. The lactational change patterns of IL-10 concentrations differed significantly between the two groups. A short-term supplementation of probiotics affects human milk cytokine levels in lactating women with a possible placebo effect still existing. Future placebo-controlled studies are needed to support these results, based on the estimated sample sizes in this study.

In this study, the propagation time and attenuation rate distributions of each sound source grid point (200 × 200) to a microphone of an arbitrary position across the shear layer, which are required for beamforming, were reconstructed by the reduced-order model with sparse sampling for acceleration of the computation. First, the propagation time and attenuation rate distributions, including the refraction of sound by the shear layer were calculated over 100 patterns of combinations of the wind speed and the microphone position, as training data. The dominant modes and optimum sampling points were discovered from the training data. Subsequently, data-driven sparse sampling for reconstruction was applied and the propagation time and the attenuation rate from each grid point (200 × 200) to a microphone were quickly calculated for the given microphone position and wind speed. The error of the obtained calculation result is 1% or less, and the approximation by data-driven sparse sampling is concluded to be effective.

The characteristics of fast-response pressure-sensitive paints (PSPs) in low-pressure conditions were evaluated. Three representative porous binders were investigated: polymer-ceramic PSP (PC-PSP), anodized-aluminum PSP (AA-PSP), and thin-layer chromatography PSP (TLC-PSP). For each PSP, two types of luminophores, Pt(II) meso-tetra (pentafluorophenyl) porphine (PtTFPP) and tris(bathophenanthroline) ruthenium dichloride (Ru(dpp)3), were used as sensor molecules. Pressure sensitivities, temperature sensitivities, and photodegradation rates were measured and evaluated using a pressure chamber. The effect of ambient pressure on the frequency response was investigated using an acoustic resonance tube. The diffusivity coefficients of PSPs were estimated from the measured frequency response and luminescent lifetime, and the governing factor of the frequency response under low-pressure conditions was identified. The results of static calibration show that PC-PSP/PtTFPP, AA-PSP/Ru(dpp)3, and TLC-PSP/PtTFPP have high pressure sensitivities that exceed 4%/kPa under low-pressure conditions and that temperature sensitivity and photodegradation rates become lower as the ambient pressure decreases. Dynamic calibration results show that the dynamic characteristics of PSPs with PtTFPP are dependent on the ambient pressure, whereas those of PSPs with Ru(dpp)3 are not influenced by the ambient pressure. This observation indicates that the governing factor in the frequency response under low-pressure conditions is the lifetime for PC-PSP and TLC-PSP, whereas the governing factor for AA-PSP is diffusion.

Abstract In this paper, a new model for electroadhesion between two surface-insulated plates under DC electric field is presented and control of dynamic responses of electroadhesion force is discussed. Under DC electric field, even if the voltage difference between the plates is constant, electroadhesion force increases or decreases over time depending on the insulating materials. The increase had been explained by Johnsen–Rahbek (JR) model, but the decrease had not been focused or modeled by physically meaningful way. In addition, the previous models did not explicitly consider the mechanical behaviors of the electrodes, although the mechanical behaviors considerably affect the response. In this work, we introduced a new model that combines both electrical and mechanical behaviors. The electrical part, which is based on JR model, explained the force decrease under DC field, in addition to the force increase that had been explained using JR model. The mechanical part was represented by a combination of a spring and a damper. Numerical simulations using the model successfully reproduced characteristics behaviors of electroadhesion force, which include force decay under constant voltages and relatively smaller initial force. Using the inverse model, we carried out experiments to control dynamic responses of electroadhesion force, which successfully controlled force responses against pulse voltages. Through the experiments, we also showed the importance of the neutralization of surface charges for obtaining reproducible responses.

Urothelial carcinoma of the bladder in childhood is extremely rare. We report on a 12-year-old girl with urothelial carcinoma of the bladder who presented with gross hematuria. Immunohistochemical staining showed normal levels of cytokeratin 20 expression and decreased levels of Ki67 and p53 expression. Human papillomavirus, which can be a cause of urothelial carcinoma of the bladder, was not detected by polymerase chain reaction. We reviewed the literature focusing on diagnostic tools for urothelial carcinoma of the bladder and determined that abdominal ultrasonography, with an extremely high sensitivity of 98%, could be a useful tool for postoperative follow-up in childhood.

Background: Dietary probiotics supplementation in lactating mothers may help prevent allergic disease in infants. However, owing to a lack of consistency in nutritional and safety outcomes associated with probiotics, this topic remains controversial.Methods: In this open-label pilot trial conducted between April 2013 and December 2013, we evaluated the safety of probiotic supplementation with 5 × 109 CFU of Lactobacillus casei LC5, 5 × 109 CFU of Bifidobacterium longum BG7, and 2 × 108 CFU of Bacillus coagulans SANK70258 in lactating women who exhibited allergies for 2 months (1–3 months postpartum); we also evaluated the effects of probiotic supplementation on transforming growth factor-β (TGF-β) and immunoglobulin A (IgA) levels in human milk. Participants self-selected to join the probiotic (n = 41; age [median (interquartile range [IQR]), y] 33 [27–39], body mass index [BMI] [median (IQR), kg/m2] 21.8 [19.5–22.8]) or no supplementation control group (n = 19; age [median (IQR), y] 33 [23–43], BMI [median (IQR), kg/m2) 19.6 [18.4–22.1]). Probiotics (three tablets) received were taken as daily supplements. Milk samples were collected at 1, 2, and 3 months postpartum, and TGF-β1, TGF-β2, and IgA levels were measured.Results: No adverse effects were observed in the probiotic group, according to the self-recorded diary during the study period. Milk IgA decreased with increasing postpartum months in both groups. In contrast, TGF- β1 and β2 were not affected by lactation periods, and showed different patterns over time between the two groups. TGF-β1, TGF-β1, and IgA levels were significantly correlated at baseline (respectively p < 0.05). However, the correlation between TGF-β1 and IgA became non-significant by the end of the intervention (p = 0.063).Conclusion: Altogether, probiotic supplementation was tolerated with respect to no dropout and 91.5% adherence. Although probiotic supplementation might affect human milk TGF-β levels, a positive effect of probiotic supplementation was not entirely supported. Future placebo-controlled studies are needed to further support the efficacy and safety of probiotic supplementation.Clinical Trial Registration:www.umin.ac.jp/ctr/, identifier: UMIN000036059.

In recent years, the rates of heat-related morbidity and mortality have begun to increase with the increase in global warming; in this context, it is noteworthy that the number of patients transported by ambulance in heat-related cases in Japan reached 95,137 in 2018. The estimation of heat-related morbidity forms a key factor in proposing and implementing suitable intervention strategies and ambulance availability and arrangements. Heat-related morbidity is known to be fairly correlated to metrics related to ambient conditions, thus necessitating the exploration of new metrics to more accurately estimate morbidity. In this study, we use an integrated computational technique relating to thermodynamics and thermoregulation to estimate daily peak core temperature elevation and daily water loss, which are linked to heat-related illnesses, from weather data of three different prefectures in Japan (Tokyo, Osaka, and Aichi). The correlations of the computed core temperature elevation and water loss as well as conventional ambient conditions are investigated in terms of number of patients suffering from heat-related illnesses transported by ambulance from 2013 to 2018. The estimated water loss per the proposed computation yields better correlation with the number of patients transported by ambulance. In particular, the weight-sum daily water loss for two to three successive days is found to be an important metric for predicting the number of patients transported by ambulance. For the same ambient conditions, morbidity is found to decrease to 0.4 owing to heat adaption at the end of summer (60 days) as compared with that at the end of the rainy season. Thus, the weighted sum of water loss and daily average ambient temperature for successive days can be used as better metrics than conventional weather data for the application of intervention strategies and planning of ambulance arrangements for heat-related morbidity. Keywords: Heat-related illness, Computational physical modeling, Heat adaptation, Ambulance dispatches

A new dynamic mode decomposition (DMD) method is introduced for simultaneous system identification and denoising in conjunction with the adoption of an extended Kalman filter algorithm. The present paper explains the extended-Kalman-filter-based DMD (EKFDMD) algorithm which is an online algorithm for dataset for a small number of degree of freedom (DoF). It also illustrates that EKFDMD requires significant numerical resources for many-degree-of-freedom (many-DoF) problems and that the combination with truncated proper orthogonal decomposition (trPOD) helps us to apply the EKFDMD algorithm to many-DoF problems, though it prevents the algorithm from being fully online. The numerical experiments of a noisy dataset with a small number of DoFs illustrate that EKFDMD can estimate eigenvalues better than or as well as the existing algorithms, whereas EKFDMD can also denoise the original dataset online. In particular, EKFDMD performs better than existing algorithms for the case in which system noise is present. The EKFDMD with trPOD, which unfortunately is not fully online, can be successfully applied to many-DoF problems, including a fluid-problem example, and the results reveal the superior performance of system identification and denoising.

This paper analyzes stability and tracking performance of displayed-marker-based position tracking on visual displays, such as an LCD (Liquid Crystal Display), which have been utilized in several studies about computer-human interaction systems. The tracking system dealt in this paper consists of an LCD and a target object with photo sensors that is placed on the display surface. A fiducial marker image is displayed beneath the object and the photo sensors detect the relative displacement between the marker and the object. The detected displacement is fed back to the tracking program and the program updates the marker position such that it will track the object. In general, this kind of tracking methods can suffer from display latency that arises from programs, graphic engines, and internal signal processing circuits of LCDs. This paper investigates the latency characteristics to reveal that the lag is not constant; it fluctuates with time. The paper, then, formulates the tracking system and analyzes how the lag affects the tracking stability. Then, the paper analyzes the tracking performance of two different classes of stable tracking algorithms, which are PD control and lag compensation. Based on the analyses, the paper provides a guideline on the selection of tracking algorithms, as well as tracking parameters. The analyzed results, as well as the guideline, are verified by experiments in 1-DOF horizontal motions.

The effects of the oxygen mole fraction on the static properties of pressure-sensitive paint (PSP) were investigated. Sample coupon tests using a calibration chamber were conducted for poly(hexafluoroisopropyl methacrylate)-based PSP (PHFIPM-PSP), polymer/ceramic PSP (PC-PSP), and anodized aluminum PSP (AA-PSP). The oxygen mole fraction was set to 0.1–100%, and the ambient pressure (Pref) was set to 0.5–140 kPa. Localized Stern–Volmer coefficient Blocal increased and then decreased with increasing oxygen mole fraction. Although Blocal depends on both ambient pressure and the oxygen mole fraction, its effect can be characterized as a function of the partial pressure of oxygen. For AA-PSP and PHFIPM-PSP, which are low-pressure- and relatively low-pressure-type PSPs, respectively, Blocal peaks at PO2ref<12 kPa. In contrast, for PC-PSP, which is an atmospheric-pressure-type PSP in the investigated range, Blocal does not have a peak. Blocal has a peak at a relatively high partial pressure of oxygen due to the oxygen permeability of the polymer used in the binder. The peak of SPR, which is the emission intensity change with respect to normalized pressure fluctuation, appears at a lower partial pressure of oxygen than that of Blocal. This is because the intensity of PSP becomes quite low at a high partial pressure of oxygen even if Blocal is high. Hence, the optimal oxygen mole fraction depends on the type of PSP and the ambient pressure range of the experiment. This optimal value can be found on the basis of the partial pressure of oxygen.

An inflammatory myofibroblastic tumor (IMT) is a distinctive neoplasm composed of myofibroblastic and fibroblastic spindle cells, accompanied by inflammatory infiltration of plasma cells, lymphocytes, and eosinophils. IMTs rarely occur in the urinary bladder. It is important to distinguish this tumor from other malignant spindle cell tumors. Herein, we report a patient with an IMT showing muscle invasion, who underwent a transurethral resection of the bladder tumor and, at a later date, partial cystectomy. The resected tumor specimen revealed a proliferation of spindle-shaped cells on a background of plasma cells and lymphocytes. Immunohistochemical staining showed the tumor to be positive for anaplastic lymphoma kinase (ALK), smooth muscle actin, and vascular endothelial growth factor (VEGF). Such histopathological findings were indicative of an IMT, suggesting the use of inhibitors of ALK and VEGF as pharmacotherapy.

Lymphoepithelioma-like carcinoma of the urinary bladder (LELCB) is a rare variant of infiltrating urothelial carcinoma. We report a case of LELCB in a 43-year-old man. Ultrasonography and cystoscopy revealed two bladder tumors, one on the left side of the trigone and the other on the right side of the trigone. Transurethral resection of the bladder tumors was performed and pathological analysis revealed undifferentiated carcinoma. We therefore performed radical cystectomy and urinary diversion. Immunohistochemically the tumor cells were positive for cytokeratin, but negative for Epstein-Barr virus-encoded small RNA in situ hybridization as found for previous cases of LELCB. The final pathological diagnosis was a lymphoepithelioma-like variant of urothelial carcinoma with perivesical soft tissue invasion. For adjuvant systemic chemotherapy, three courses of cisplatin were administered. The patient subsequently became free of cancer 72 months postoperatively. Based on the literature, pure or predominant LELCB types show favorable prognoses due to their sensitivity to chemotherapy or radiotherapy. An analysis of the apparent diffusion coefficient (ADC) values of bladder tumors examined in our institution revealed that the ADC value measured for this LELCB was relatively low compared to conventional urothelial carcinomas. This suggests that measuring the ADC value of a lymphoepithelioma-like carcinoma prior to operation may be helpful in predicting LELCB.

Pemetrexed is an antifolate agent that is regarded as an alternative second-line chemotherapy against advanced or metastatic urothelial carcinoma (UC). However, there is limited information on pemetrexed in a third-line setting. We report a case of metastatic UC treated with pemetrexed as third-line chemotherapy following gemcitabine and cisplatin (GC) and gemcitabine and docetaxel (GD) therapies. A 73-year-old man with a history of transurethral resection of bladder carcinoma presented with pollakiuria. CT revealed a mass in the left renal pelvis that had invaded into the parenchyma of the left kidney, as well as para-aortic and mediastinum lymph node enlargement. Urinary cytology of the lesion in the left renal pelvis revealed UC. Thus, the patient was diagnosed with left renal pelvic carcinoma (cT3N2M0). After having received 4 courses of GC therapy, another mediastinum lymph node was enlarged. He subsequently received 3 courses of GD therapy as second-line chemotherapy, which showed little efficacy against the metastatic lesions. The patient was administered 3 courses of pemetrexed as third-line chemotherapy; however, its effect on tumor reduction was not sufficient. Finally, metastasis to the liver was observed, and he died 21 months after initiation of chemotherapy. For pathological confirmation, needle biopsy of a metastatic lymph node performed after death revealed high-grade UC and a high positivity of programmed death ligand 1 (PD-L1) in the tumor, which suggested that he could have benefited from anti-PD-L1 antibody immunotherapy. This report describes the outcome of pemetrexed treatment and proposes another possible candidate as third-line chemotherapy against metastatic UC.

There is no established chemotherapy regimen in metastatic primary urethral cancer (mPUC). The efficacy of a cisplatin (CDDP)-based regimen has been reported, however, when the syndrome of inappropriate secretion of antidiuretic hormone (SIADH) occurs, the chemotherapy regimen should be changed to another platinum compound. In this report, we describe a 66-year-old woman who was diagnosed as mPUC with, CDDP-induced SIADH. After switching her to CBDCA and careful managing her sodium balance, three courses of the chemotherapy regimen were completed.

Delayed radiation myelopathy (DRM) is a rare disorder that rapidly leads to disabilities, and the median incubation period was reported to be about 2 years (from 6 months to a few years). In this report, we describe a 61-year-old woman who presented with rapid progressive numbness and weakness in both legs 22 months after palliative radiation therapy with 39 Gy in 3 Gy fractions. She was diagnosed with DRM of the thoracic vertebrae and was treated sequentially with corticosteroids, heparin, and hyperbaric oxygen therapy. However, they were not effective, and complete paralysis of the legs occurred in 3 months.

To probe the effects of pendant side-chain structures on the properties of porous thermoresponsive polymer gels, oligo(ethylene glycol) alkyl ether acrylates were polymerised in an aqueous medium under radical-mediated phase-separation conditions. The monomer structures varied according to the lengths and termini of their ethylene glycol side chains. The porous poly(oligo(ethylene glycol) alkyl ether acrylate) (POEGA) gels exhibited variable lower critical solution temperatures (LCSTs) but similar and rapid swelling–deswelling behaviours. Although the LCST of the poly(tri(ethylene glycol) monomethyl ether acrylate) (PTEGA) gel decreased with increasing aqueous NaCl or CaCl2 concentration, PTEGA showed excellent thermosensitivity in highly concentrated salt solutions, recommending its application in saline environments. Examination of PTEGA adsorption performance in an oil–water emulsion showed that n-tridecane adsorption increased with temperature. Although n-tridecane was effectively adsorbed at 70 °C, its release from the fully adsorbed PTEGA gel was difficult despite a temperature reduction from 70 to 20 °C.

A temperature-sensitive paint (TSP) using a chameleon luminophore [ Tb 0.99 Eu 0.01 ( hfa ) 3 ( dpbp ) ] n is proposed. The chameleon luminophore was dispersed in isobutyl methacrylate polymer in a toluene solvent to fix it on a sample coupon. Temperature and pressure sensitivities of the chameleon luminophore-based TSP were measured using a spectrofluorophotometer. The emission for each wavelength was confirmed to be dependent on the temperature and pressure. The temperature and pressure sensitivities of the TSP were 0.81–2.8%/K and 0.08–0.12%/kPa, respectively. Higher temperature sensitivity can be obtained using the ratio of emissions from the two lanthanide ions, Tb III and Eu III . The temperature sensitivity when using the ratio of the emission intensities at 616 nm derived from Eu III and at 545 nm derived from Tb III was 3.2%/K, which was the highest value in the present study. In addition, the pressure sensitivity for the case using the ratio of the emission intensities at 616 and 545 nm was 4.8 × 10 − 2 % /kPa. Higher temperature sensitivity and lower pressure sensitivity than that with a single wavelength can be achieved using the ratio of the emission intensities at the two peak wavelengths derived from Tb III and Eu III .

The flow control effects of a nanosecond-pulse-driven dielectric barrier discharge plasma actuator (ns-DBDPA) in dynamic stall flow were experimentally investigated. The ns-DBDPA was installed on the leading edge of an airfoil model designed in the form of a helicopter blade. The model was oscillated periodically around 25% of the chord length. Aerodynamic coefficients were calculated using the pressure distribution, which was obtained by the measurement of the unsteady pressure by sensors inside the model. The flow control effect and its sensitivity to pitching oscillation and ns-DBDPA control parameters are discussed using the aerodynamic coefficients. The freestream velocity, the mean of the angle of attack, and the reduced frequency were employed as the oscillation parameters. Moreover, the nondimensional frequency of the pulse voltage, the peak pulse voltage, and the type and position of the ns-DBDPA were adopted as the control parameters. The result shows that the ns-DBDPA can decrease the hysteresis of the aerodynamic coefficients and a flow control effect is obtained in all cases. The flow control effect can be maximized by adopting the low nondimensional frequency of the pulse voltage.

The flow control over the blades of a small horizontal-axis wind turbine (HAWT) model using a dielectric barrier discharge plasma actuator (DBD-PA) was studied based on large-eddy simulations. The numerical simulations were performed with a high-resolution computational method, and the effects of the DBD-PA on the flow fields around the blades were modeled as a spatial body force distribution. The DBD-PA was installed at the leading edge of the blades, and its impacts on the flow fields and axial torque generation were discussed. The increase in the ratios of the computed, cycle-averaged axial torque reasonably agreed with that of the available experimental data. In addition, the computed results presented a maximum of 19% increase in the cycle-averaged axial torque generation by modulating the operating parameters of the DBD-PA because of the suppression of the leading edge separation when the blade’s effective angles of attack were relatively high. Thus, the suppression of the leading edge separation by flow control can lead to a delay in the breakdown of the tip vortex as a secondary effect.

Prostate cancer and castration-resistant prostate cancer (CRPC) remain major health challenges in men. In this study, the inhibitory effects of a hexane insoluble fraction from a purple rice ethanolic extract (PRE-HIF) on prostate carcinogenesis and CRPC were investigated both in vivo and in vitro. In the Transgenic Rat for Adenocarcinoma of Prostate (TRAP) model, 1% PRE-HIF mixed diet-fed rats showed a significantly higher percentage of low-grade prostatic intraepithelial neoplasia and obvious reduction in the incidence of adenocarcinoma in the lateral lobes of the prostate. Additionally, 1% PRE-HIF supplied diet significantly suppressed the tumor growth in a rat CRPC xenograft model of PCai1 cells. In LNCaP and PCai1 cells, PRE-HIF treatment suppressed cell proliferation and induced G0/G1 cell-cycle arrest. Furthermore, androgen receptor (AR), cyclin D1, cdk4, and fatty acid synthase expression were down-regulated while attenuation of p38 mitogen-activated protein kinase, and AMP-activated protein kinase α activation occurred in PRE-HIF treated prostate cancer cells, rat prostate tissues, and CRPC tumors. Due to consistent results with PRE-HIF in PCai1 cells, cyanidin-3-glucoside was characterized as the active compound. Altogether, we surmise that PRE-HIF blocks the development of prostate cancer and CRPC through the inhibition of cell proliferation and metabolic pathways.

A novel dynamic mode decomposition (DMD) method based on a Kalman filter is proposed. This paper explains the fast algorithm of the proposed Kalman filter DMD (KFDMD) in combination with truncated proper orthogonal decomposition for many-degree-of-freedom problems. Numerical experiments reveal that KFDMD can estimate eigenmodes more precisely compared with standard DMD or total least-squares DMD (tlsDMD) methods for the severe noise condition if the nature of the observation noise is known, though tlsDMD works better than KFDMD in the low and medium noise level. Moreover, KFDMD can track the eigenmodes precisely even when the system matrix varies with time similar to online DMD, and this extension is naturally conducted owing to the characteristics of the Kalman filter. In summary, the KFDMD is a promising tool with strong antinoise characteristics for analyzing sequential datasets.

Primitive neuroectodermal tumor (PNET) is a member of the Ewing's sarcoma family of tumors (ESFT). We report a case of PNET in a 66-year-old male who presented with a large solid tumor within the parenchyma of the middle pole of the left kidney with metastases to the left adrenal gland and right ischium. A fine-needle biopsy was performed and showed a small round cell tumor. Results of immunohistochemical staining suggested this tumor belonged to ESFT. Preoperative VDC-IE (combined vincristine, doxorubicin and cyclophosphamide followed by another combination of ifosfamide and etoposide) chemotherapy and left radical nephrectomy and adrenalectomy were performed. The histopathological findings of the resected tumor were similar to those in the biopsy specimen, but the results of AE1/AE3 were different. For the diagnosis, fluorescence in situ hybridization was performed. Split signals of the EWSR1 gene were detected, and transmission electron microscopy showed neuroendocrine granules and microtubules. The final diagnosis of this tumor was PNET of the kidney.

Mutations in the Autism susceptibility candidate 2 gene (AUTS2), whose protein is believed to act in neuronal cell nuclei, have been associated with multiple psychiatric illnesses, including autism spectrum disorders, intellectual disability, and schizophrenia. Here we show that cytoplasmic AUTS2 is involved in the regulation of the cytoskeleton and neural development. Immunohistochemistry and fractionation studies show that AUTS2 localizes not only in nuclei, but also in the cytoplasm, including in the growth cones in the developing brain. AUTS2 activates Rac1 to induce lamellipodia but downregulates Cdc42 to suppress filopodia. Our loss-of-function and rescue experiments show that a cytoplasmic AUTS2-Rac1 pathway is involved in cortical neuronal migration and neuritogenesis in the developing brain. These findings suggest that cytoplasmic AUTS2 acts as a regulator of Rho family GTPases to contribute to brain development and give insight into the pathology of human psychiatric disorders with AUTS2 mutations.

Generation of new concepts in product design process serves as driving force of innovation. While many methods have been proposed for supporting design concept generation, it has rarely been supported in a systematic way, because its process is done in designer's implicit thought, and therefore, it is difficult to explicitly formalize the process. This paper proposes a model of designers' thinking process in concept generation called ECF cycle. It consists of three steps, i.e., (1) Externalization: the step for representing design concepts with drawings and language, (2) Classification: the step for classifying concepts with function trees, and (3) Finding voids: the step for finding potentials of new concepts called void, which is a defect of the classification. Then, this research builds a new method of concept generation incorporating the theory of solving inventive problems (TRIZ). The method is based on a quick reference table of TRIZ principles, which helps designers to create a new concept corresponding to a void found by ECF cycle. A case study of concept generation of new running shoes is demonstrated in order to verify the ability of the proposed method.

BackgroundUrinary neutrophil gelatinase‐associated lipocalin (U‐NGAL) is an early predictor of acute kidney injury and adverse events in various diseases; however, in acute decompensated heart failure patients, its significance remains poorly understood. This study aimed to investigate the prognostic value of U‐NGAL on the first day of admission for the occurrence of acute kidney injury and long‐term outcomes in acute decompensated heart failure patients. Methods and ResultsWe studied 260 acute decompensated heart failure patients admitted to our department between 2011 and 2014 by measuring U‐NGAL in 24‐hour urine samples collected on the first day of admission. Primary end points were all‐cause death, cardiovascular death, and heart failure admission. Patients were divided into 2 groups according to their median U‐NGAL levels (32.5 μg/gCr). The high‐U‐NGAL group had a significantly higher occurrence of acute kidney injury during hospitalization than the low‐U‐NGAL group (P=0.0012). Kaplan‐Meier analysis revealed that the high‐U‐NGAL group exhibited a worse prognosis than the low‐U‐NGAL group in all‐cause death (hazard ratio 2.07; 95%CI 1.38‐3.12, P=0.0004), cardiovascular death (hazard ratio 2.29; 95%CI 1.28‐4.24, P=0.0052), and heart failure admission (hazard ratio 1.77; 95%CI 1.13‐2.77, P=0.0119). The addition of U‐NGAL to the estimated glomerular filtration rate significantly improved the predictive accuracy of all‐cause mortality (P=0.0083). ConclusionsIn acute decompensated heart failure patients, an elevated U‐NGAL level on the first day of admission was related to the development of clinical acute kidney injury and independently associated with poor prognosis.

Sensory experience regulates the development of various brain structures, including the cortex, hippocampus, and olfactory bulb (OB). Little is known about how sensory experience regulates the dendritic spine development of OB interneurons, such as granule cells (GCs), although it is well studied in mitral/tufted cells. Here, we identify a transcription factor, Npas4, which is expressed in OB GCs immediately after sensory input and is required for dendritic spine formation. Npas4 overexpression in OB GCs increases dendritic spine density, even under sensory deprivation, and rescues reduction of dendrite spine density in the Npas4 knockout OB. Furthermore, loss of Npas4 upregulates expression of the E3-ubiquitin ligase Mdm2, which ubiquitinates a microtubule-associated protein Dcx. This leads to reduction in the dendritic spine density of OB GCs. Together, these findings suggest that Npas4 regulates Mdm2 expression to ubiquitinate and degrade Dcx during dendritic spine development in newborn OB GCs after sensory experience.

Recent studies have shown that epigenetic alterations lead to oncogenic activation, thus indicating that these are therapeutic targets. Herein, we analyzed the efficacy and therapeutic potential of our developed histone lysine demethylase 1 (LSD1) inhibitor, NCL1, in castration-resistant prostate cancer (CRPC). The CRPC cell lines 22Rv1, PC3, and PCai1CS were treated with NCL1, and LSD1 expression and cell viability were assessed. The epigenetic effects and mechanisms of NCL1 were also evaluated. CRPC cells showed strong LSD1 expression, and cell viability was decreased by NCL1 in a dose-dependent manner. Chromatin immunoprecipitation analysis indicated that NCL1 induced histone H3 lysine 9 dimethylation accumulation at promoters of P21. As shown by Western blot and flow cytometry analyses, NCL1 also dose-dependently induced caspase-dependent apoptosis. The stimulation of autophagy was observed in NCL1-treated 22Rv1 cells by transmission electron microscopy and LysoTracker analysis. Furthermore, WST-8 assay revealed that the anti-tumor effect of NCL1 was reinforced when autophagy was inhibited by chloroquine in 22Rv1 cells. Combination index analysis revealed that a concurrent use of these drugs had a synergistic effect. In ex vivo analysis, castrated nude mice were injected subcutaneously with PCai1 cells and intraperitoneally with NCL1. Tumor volume was found to be reduced with no adverse effects in NCL1-treated mice compared with controls. Finally, immunohistochemical analysis using consecutive human specimens in pre- and post-androgen deprivation therapy demonstrated that LSD1 expression levels in CRPC, including neuroendocrine differentiation cases, were very high, and identical to levels observed in previously examined prostate biopsy specimens. NCL1 effectively suppressed prostate cancer growth in vitro and ex vivo without adverse events via the regulation of apoptosis and autophagy, suggesting that NCL1 is a potential therapeutic agent for CRPC.

A method is proposed to expand the Z-directional measurement range of a fiber-based dual-detector chromatic confocal probe with a mode-locked femtosecond laser source. In the dual-detector chromatic confocal probe, the Z-directional displacement of a measurement target is derived from the peak wavelength in the normalized intensity ratio from the two light intensities obtained by the two identical fiber detectors. In this paper, a new method utilizing the main-lobe and side-lobes of axial responses acquired from both the normalized intensity ratio Ia and the invert normalized intensity ratio In, which is the inverse of Ia, is proposed to obtain the seamless relationship between the peak wavelength and the Z-directional displacement of a measurement target. Theoretical calculations and experimental investigation are carried out to demonstrate the feasibility of the proposed measurement range expansion method.

Polymer/ceramic pressure-sensitive paint (PC-PSP), which incorporates a high percentage of particles in the binder layer, is proposed in order to improve the characteristics of PSP. The procedure for embedding particles into the binder layer was modified. In the conventional procedure, dye is adsorbed onto a polymer/ceramic coating film (denoted herein as a dye-adsorbed (D-adsorbed) PSP). In the new procedure, the mixture of a dye and particles is adsorbed onto a polymer coating film (denoted herein as the particle/dye-adsorbed (PD-adsorbed) PSP). The effect of particle mass content on PSP characteristics was investigated. In addition, the effect of solvent on PSP characteristics and film structure were evaluated for the PD-adsorbed PSP. As a result, the difference in the PSP characteristics between the two types of PSP was clarified. Although surface roughness and time response increase with increased mass content of particles for both D- and PD-adsorbed PSPs, the critical pigment volume concentration (CPVC) for the PD-adsorbed PSP is smaller than that of the D-adsorbed PSP (88 wt% and 93 wt%, respectively). The PD-adsorbed PSP has a higher frequency response comparing with the D-adsorbed PSP while maintaining the same surface roughness. Observation by scanning electron microscope showed that the CPVC of the PC-PSP is governed primarily by surface structure. The coating film structure can be roughly classified into two states depending on the particle mass content. One is a state in which the coating film consisted of two layers: a lower particle-rich layer and an upper polymer-rich layer. This type of structure was observed in the PD-adsorbed PSP as well as in the D-adsorbed PSP. In the other state, polymer and particles are homogeneously distributed in the film, and pores are formed. This difference in the coating structure results in a change in the time response.

Schizophrenia is a severe and common psychiatric disease with a lifetime prevalence of 0.5% to 1% globally. Because of limitations of the experimental approach in humans, valid animal models are essential in the effort to identify novel therapeutics for schizophrenia. In most animal models of schizophrenia, second generation antipsychotic drugs are reported to be effective in ameliorating behavioral abnormalities, while clinical evidence indicates that some of the patients are resistant to the antipsychotic drug therapy. Accordingly, animal models of antipsychotic drug-resistant schizophrenia are needed for screening of novel agents that may be more effective than the existing antipsychotic drugs. Furthermore, utilization of appropriate behavioral tasks with reference to human testing is essential to facilitate the development of novel pharmacotherapeutic approaches for the treatment in schizophrenia. Experimental data suggest that there are different types of potential candidate molecules as novel antipsychotic drugs with some therapeutic effects on negative symptoms and cognitive deficits in schizophrenia. It is proposed that to develop novel antipsychotic drugs the efficacy of potential candidate molecules should be evaluated using animal models for treatment-resistant schizophrenia with appropriate behavioral tasks in reference to human testing.

Transforming growth factor-β (TGF-β) is central during the pathogenesis of pulmonary fibrosis, in which the plasminogen activator inhibitor-1 (PAI-1) also has an established role. TGF-β is also known to be the strongest inducer of PAI-1. To investigate the link between PAI-1 and TGF-β in fibrotic processes, we evaluated the effect of SK-216, a PAI-1-specific inhibitor, in TGF-β-dependent epithelial-mesenchymal transition (EMT) and fibroblast to myofibroblast differentiation. In human alveolar epithelial A549 cells, treatment with TGF-β induced EMT, whereas co-treatment with SK-216 attenuated the occurrence of EMT. The inhibition of TGF-β-induced EMT by SK-216 was also confirmed in the experiment using murine epithelial LA-4 cells. Blocking EMT by SK-216 inhibited TGF-β-induced endogenous production of PAI-1 and TGF-β in A549 cells as well. These effects of SK-216 were not likely mediated by suppressing either Smad or ERK pathways. Using human lung fibroblast MRC-5 cells, we demonstrated that SK-216 inhibited TGF-β-dependent differentiation of fibroblasts to myofibroblasts. We also observed this inhibition by SK-216 in human primary lung fibroblasts. Following these in vitro results, we tested oral administration of SK-216 into mice injected intratracheally with bleomycin. We found that SK-216 reduced the degree of bleomycin-induced pulmonary fibrosis in mice. Although the precise mechanisms underlying the link between TGF-β and PAI-1 regarding fibrotic process were not determined, PAI-1 seems to act as a potent downstream effector on the pro-fibrotic property of TGF-β. In addition, inhibition of PAI-1 activity by a PAI-1 inhibitor exerts an antifibrotic effect even in vivo. These data suggest that targeting PAI-1 as a downstream effector of TGF-β could be a promising therapeutic strategy for pulmonary fibrosis.

Hypophosphatasia (HPP) is an inherited disease caused by genetic mutations in the gene encoding tissue-nonspecific alkaline phosphatase (TNALP). This results in defects in bone and tooth mineralization. We recently demonstrated that TNALP-deficient (Akp2−/−) mice, which mimic the phenotype of the severe infantile form of HPP, can be treated by intravenous injection of a recombinant adeno-associated virus (rAAV) expressing bone-targeted TNALP with deca-aspartates at the C-terminus (TNALP-D10) driven by the tissue-nonspecific CAG promoter. To develop a safer and more clinically applicable transduction strategy for HPP gene therapy, we constructed a self-complementary type 8 AAV (scAAV8) vector that expresses TNALP-D10 via the muscle creatine kinase (MCK) promoter (scAAV8-MCK-TNALP-D10) and examined the efficacy of muscle-directed gene therapy. When scAAV8-MCK-TNALP-D10 was injected into the bilateral quadriceps of neonatal Akp2−/− mice, the treated mice grew well and survived for more than 3 months, with a healthy appearance and normal locomotion. Improved bone architecture, but limited elongation of the long bone, was demonstrated on X-ray images. Micro-CT analysis showed hypomineralization and abnormal architecture of the trabecular bone in the epiphysis. These results suggest that rAAV-mediated, muscle-specific expression of TNALP-D10 represents a safe and practical option to treat the severe infantile form of HPP.

Immunohistochemistry (IHC) for detecting key signal molecules involved in programmed cell death (PCD) in archival human pathology specimens is fairly well established. Detection of cleaved caspase-3 in lymphocytes in rheumatoid arthritis (RA) and gastric surface foveolar glandular epithelia but not in synoviocytes in RA, gastric fundic glandular epithelia, or nasal NK/T-cell lymphoma (NKTCL) cells suggests anti-apoptotic mechanisms in cell differentiation and in oncogenesis such as the induction of survivin. Enzymatically pretreated and ultra-super sensitive detection of beclin-1 in synoviocytes in RA and gastric fundic glandular epithelia suggests enhanced autophagy. The deposition of beclin-1 in fibrinoid necrosis in RA and expression of beclin-1 in detached gastric fundic glandular cells suggest that enhanced autophagy undergoes autophagic cell death (ACD). NKTCL exhibited enhanced autophagy through LC3 labeling and showed densely LC3 labeled cell-debris in regions of peculiar necrosis without deposition of beclin-1, indicating massive ACD in NKTCL and the alternative pathway enhancing autophagy following autophagic vesicle nucleation. Autophagy progression was monitored by labeling aggregated mitochondria and cathepsin D. The cell-debris in massive ACD in NKTCL were positive for 8-hydroxydeoxyguanosine, suggesting DNA oxidation occurred in ACD. Immunohistochemical autophagy and PCD analysis in archival human pathology specimens may offer new insights into autophagy in humans.

Polyriboinosinic-polyribocytidilic acid (polyI:C) is a synthetic analog that elicits viral-like immune responses in mammals. We have recently found that polyI:C treatment in neonatal mice induced abnormalities of emotional, cognitive, and sensorimotor gating and dysfunction of glutamatergic neurotransmission in adulthood. In this study, we investigated the effect of the NMDA-receptor co-agonist d-serine on polyI:C-induced behavioral abnormalities in mice. Neonatal ICR mice were repeatedly injected with polyI:C for 5 days from postnatal day 2 to 6. At 10 weeks, sensorimotor gating function was analyzed in the prepulse inhibition (PPI) test. Emotional function was analyzed in open field and social interaction tests. Cognitive function was analyzed by novel object recognition tests. d-Serine dose-dependently improved polyI:C-induced impairment of emotional and cognitive behaviors whereas it had no effect on PPI deficit in adults. The ameliorating effects of d-serine were antagonized by pretreatment with an NMDA-receptor antagonist, MK-801. Although the mRNA level of d-amino acid oxidase (DAAO) was increased in the prefrontal cortex and hippocampus of neonatal polyI:C-treated mice in adulthood, no changes were observed in d-serine content and DAAO enzymatic activity. These results suggest that d-serine ameliorates emotional and cognitive impairments of the polyI:C-treated mice through potentiating NMDA receptor activity. Keywords:: cognition, emotion, d-serine, polyI:C, d-amino acid oxidase

We have recently found that combination of ovariectomy (OVX) and chronic restraint stress causes cognitive dysfunction and reduces hippocampal CA3 neurons in female rats and mice and that estrogen replacement and chronic treatment with Ginkgo biloba extract EGb 761 suppress the OVX/stress-induced behavioral and morphological changes. In this study, we examined the effect of placental extract on the memory impairment and neuromorphological change in OVX/stress-subjected mice. Female Slc:ICR strain mice were randomly divided into four groups: vehicle-treated OVX, porcine placental extract (120 and 2160 mg/kg)-treated OVX, and sham-operated control groups. Two weeks after surgical operation, OVX mice underwent restraint stress for 21 days (6 h/day), and all animals were then subjected to a contextual fear conditioning test followed by morphological examination by Nissl staining. Placental extract was orally administered once daily until the behavioral analysis was carried out. Chronic treatment with both doses of placental extract improved the OVX/stress-induced fear memory impairment and Nissl-positive cell loss of the hippocampal CA3 region, although it did not affect the loss of bone mineral density and increase in body weight after OVX. These results have important implications for the neuroprotective and cognition-enhancing effects of placental extract in postmenopausal women. Keywords:: postmenopausal animal model, chronic stress, ovariectomy, cognition, hippocampus

Background: Chronic hepatitis C virus (HCV) infection causes intra- and extra-hepatic complications. The elimination of HCV has been reported to be beneficial for asthmatic patients with HCV infection. Therefore, we hypothesized that chronic HCV infection might be associated with the severity of asthma. Methods: Asthmatic patients were prospectively enrolled from 13 outpatient settings. Hepatitis B surface (HBs) antigen and HCV-RNA were measured at the time of enrollment and evaluated along with the clinical characteristics of the patients including the age, sex, duration of asthma, atopic status, smoking history, and treatment step according to the Global Initiative for Asthma guideline. Results: Of 1327 asthmatic patients, 1258 patients (94.8%) were treated with inhaled corticosteroids, 18 patients were positive for HBs antigen (1.4%), and 32 patients (2.4%) were positive for HCV-RNA. When compared with HCV-RNA-negative patients, HCV-RNA-positive patients required significantly more drugs for the treatment of asthma. No such relationship was observed in patients with positive HBs antigen. A multivariate logistic regression analysis showed that the male sex, a long duration of asthma, status as a current smoker, and HCV-RNA positivity were independently associated with more severe asthma. Conclusions: These results suggest that chronic HCV infection is an independent factor that predisposes asthmatic patients to more severe asthma. The evaluation of chronic HCV infection may be helpful for the management of severe asthmatic patients without obvious factors associated with severe asthma.

In May 2005 a 68-year-old woman received a VDD pacemaker implantation in the right pectoral region at our hospital for the treatment of complete atrioventricular block. In July 2008, she was diagnosed with dilated cardiomyopathy based on histological testing. In November 2008, she developed syncope due to ventricular tachycardia while at another hospital. She underwent external electrical cardioversion with an anterior-lateral paddle position using a single shock of 100 J. This shock led to severe bradycardia resulting in a transfer to our hospital. The physician who provided the shock could not have been aware that the patient had an implanted pacemaker. The skin above the pulse generator was burned. The electrocardiogram showed no pacing spikes or ventricular escape rhythm. Investigation of the pacemaker 3 hours after cardioversion revealed reprogramming of the device and a marked rise in the lead impedance (>3,000 ohm). Removal of the generator and implantation of a biventricular cardioverter defibrillator were required. The emergency situation, the small size of the generator, the small incision made using the buried suture method, and the patient's obesity all probably contributed to the physician's not noticing the implanted pacemaker. It is important to increase awareness of the severe consequences that may follow if the physician administering external defibrillation does not know about the patient's implanted pacemaker.