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Background & Aim Flow cytometry is a useful technique for analyzing phenotypic characteristics of cells and is used for measuring purity of target cells in heterogeneous cell populations for clinical diagnosis, as well as for controlling quality of therapeutic cell products and monitoring them in blood samples. One obstacle for analyzing the cell products by flow cytometry is, however, that it requires multiple cell labeling using specific antibodies/reagents under GMP compliance, and the process is time-consuming and costly. Here we introduce label-free ghost cytometry (LFGC), a novel machine learning-driven label-free flow cytometry that analyzes cell morphology based on cell-specific waveform signals without image production. LFGC can become a simple, fast and cost-effective analysis method of cells. In the present study, we assessed the potential applicability of LFGC to analyze cell characteristics for the quality control of cell products. Methods, Results & Conclusion A supervised machine learning model was developed from the label-free GC waveform signals of cells that were labeled with markers of Annexin V plus PI. The T cells were then classified into live or dead cells by applying the trained model to the label-free GC waveforms without seeing the fluorescent labels. In a similar manner, we assessed the ability of LFGC to identify T cells in human PBMC by labeling them with CD3. Furthermore, we applied LFGC for discriminating in-vitro activated T cells from resting T cells by labeling them with CD25. Our data showed that LFGC can accurately classify the live and dead T cells with area under the receiver operating characteristic curve (AUC) of 0.954. LFGC was also found to discriminate the CD3 positive T cells from PBMC with AUC of 0.967 and also the activated T cells from the resting ones with AUC of 0.987. These results support that LFGC can accurately predict the viability, purity and possibly functions of T cells without any labeling and could be thus beneficial for the quality control of cell products.

Highlights • A novel adenovirus outbreak occurred in a colony of African pygmy hedgehogs (APHs). • An African pygmy hedgehog adenovirus 1 (AhAdV-1) infection was confirmed in 13 APHs. • AhAdV-1 was isolated from one APH. • Five APHs died during the outbreak and AhAdV-1 can cause fatal pneumonia in a case. • AhAdV-1 should be monitored as a potential cause of emerging infections in APHs., An African pygmy hedgehog adenovirus 1 (AhAdV-1) outbreak in a colony of 24 African pygmy hedgehogs (APHs) with a case of fatal pneumonia occurred in Japan. Thirteen out of a colony of 15 APHs with respiratory symptoms were diagnosed with AhAdV-1 infection based on the detection of AhAdV-1 genome in throat/nasal swabs and further one APH was diagnosed on isolation of the virus. Five infected APHs died during the outbreak and AhAdV-1 caused severe pneumonia and death in one case. After the outbreak, persistent AhAdV-1 infection was suggested in one surviving APH. AhAdV-1 is a novel adenovirus and is suspected to be an emerging pathogen.

Background Venous thromboembolism (VTE) is frequently observed in patients with solid tumor. Previous report suggested that the prevalence of VTE varied among different ethnic groups. The objectives of this prospective observational study are to estimate the prevalence of VTE in Japanese patients with advanced solid tumor based on intensive screening, using CT scans, lower-extremity ultrasonography, and D-dimer testing. Methods Eight-hundred sixty Japanese patients were enrolled into this study. Adult (age ≥20 years) patients with metastatic or locally advanced solid tumor without anticoagulant therapy, who are planning to receive chemotherapy during 4 weeks, PS 0-2 were eligible in this study. Evaluations of VTE were performed at enrollment, 12 and 24 weeks. Primary endpoint was the prevalence of VTE for 24 weeks, and secondary endpoints included the incidence of VTE (at enrollment, 12 weeks, 24 weeks), VTE for 24 weeks by primary cancer site, symptomatic VTE for 24 weeks, pulmonary thromboembolism (PE), treatment of VTE. Results The median age was 68 years (range 28-96 years), 34% female, 46/45/8% performance status 0/1/2. Primary cancer site included lung (71%), gastrointestinal (GI) (15%), hepatobiliary and pancreatic (HP) (5%), and gynecological (3%). The prevalence of VTE for 24 weeks was 22.6% (95%CI: 19.8-25.5%). The incidences of VTE were 11.3% (97/858) at enrollment, 16.8% (134/799) at 12 weeks, 14.1% (101/716) at 24 weeks. Among 761 patients without VTE at enrollment, 97 (12.7%) showed VTE after the start of chemotherapy. The prevalence of VTE for 24 weeks was 24.1% in patients with lung, 17.7% with GI, 25.6% with HP, and 32.1% in gynecological cancer. Symptomatic VTE for 24 weeks was observed in 4.0%, and PE was in 1.0%. Treatment of anticoagulant for VTE was started in 15.0% (129/858) after enrollment in this study. Conclusions This prospective observational study showed that prevalence of VTE was relatively high in Japanese patients with advanced solid tumor under intensive screening, compared with previous reports. Although most of patients with VTE were asymptomatic, prophylactic anticoagulant therapies may be considered in this study population. Clinical trial identification UMIN000015243. Legal entity responsible for the study The authors. Funding Has not received any funding. Disclosure All authors have declared no conflicts of interest.

The National Museum of Western Art is the only work of Le Corbusier in Japan and 50 years have already passed since its construction. In order to maintain the museum’s value as a cultural asset, there is an urgent need to draft a thorough retrofit plan both to maintain the building’s function as an art museum and to restore Le Corbusier’s original design concept.This study, which carried out an indoor environmental investigation for the purpose of developing such a retrofit plan in the main building, confirms the current airflow characteristics on the basis of a survey of factors such as the flow rates of the air conditioning system, internal pressures, and the age of air distribution. Because the exhaust flow rate was greater than the supply of fresh air into the building, there was concern about air infiltration. It was found that the age of air was highest in the exhibition space, comparatively lower in the entrance hall, and lowest in the restaurant. The adjacent wall air velocities varied gre...

This study examined the relationship between rapid weight gain during infancy and/or early childhood and anthropometric measurements [body mass index (BMI), percent body fat (%BF), waist circumference (WC) and waist-to-height ratio (WHtR)] in preadolescence by sex.Subjects were fourth-grade school children (aged 9 to 10 years) from elementary schools in Ina-town, Japan, in 2010. Measurements of height, weight, %BF and WC were conducted for each subject. We obtained data on height and weight of subjects at birth, age 1.5 years and age 3 years from the Maternal and Child Health handbook. Rapid weight gain was defined as a change in weight-for-age standard deviation score greater than 0.67 from birth to age 1.5 years (infancy) or from age 1.5 to 3 years (early childhood).All anthropometric variables (BMI, %BF, WC and WHtR) at age 9 to 10 years were significantly higher in the rapid weight gain during both infancy and early childhood period group than in the no rapid weight gain group, regardless of sex. When compared with the no rapid weight gain group, rapid weight gain during early childhood period had significantly higher BMI and WC in boys and BMI, %BF and WC in girls. Compared with the no rapid weight gain group, the rapid weight gain during infancy group had a significantly higher WC in boys and significantly higher BMI and WC in girls.Rapid weight gain during both infancy and early childhood was related to higher anthropometric measurements, including WHtR, among Japanese preadolescents, regardless of sex. This study suggests that rapid weight gain during infancy and early childhood may be a risk factor for general/abdominal obesity later in life.

CD133 (prominin-1) is a transmembrane glycoprotein expressed on the surface of normal and cancer stem cells (tumor-initiating cells), progenitor cells, rod photoreceptor cells and a variety of epithelial cells. Although CD133 is widely used as a marker of various somatic and putative cancer stem cells, its contribution to the fundamental properties of cancer cells, such as tumorigenesis and differentiation, remains to be elucidated. In the present report, we found that CD133 was expressed in several neuroblastoma (NB) cell lines/tumor samples. Intriguingly, CD133 repressed NB cell differentiation, for example neurite extension and the expression of differentiation marker proteins, and was decreased by several differentiation stimuli, but accelerated cell proliferation, anchorage-independent colony formation and in vivo tumor formation of NB cells. NB cell line and primary tumor-sphere experiments indicated that the molecular mechanism of CD133-related differentiation suppression in NB was in part dependent on neurotrophic receptor RET tyrosine kinase regulation. RET transcription was suppressed by CD133 in NB cells and glial cell line-derived neurotrophic factor treatment failed to induce RET in CD133-expressing cells; RET overexpression rescued CD133-related inhibition of neurite elongation. Of note, CD133-related NB cell differentiation and RET repression were mainly dependent on p38MAPK and PI3K/Akt pathways. Furthermore, CD133 has a function in growth and RET expression in NB cell line- and primary tumor cell-derived tumor spheres. To the best of our knowledge, this is the first report of the function of CD133 in cancer cells and our findings may be applied to improve differentiation induction therapy for NB patients.

飽和するとコラプス沈下によって強度を失う脆弱層が多く存在する斜面で，崩壊現象を議論するために必要な降雨・土壌水分・斜面移動量の観測を行った。観測期間中に崩壊は生じなかったが，豪雨時にわずかな斜面移動が観測され，小さな亀裂も生じた。斜面移動は上部，中部，下部の順で開始し，上部は引張ひずみ，中部はほぼ平行移動，下部は圧縮ひずみとなるすべり変位が生じた。この移動時に斜面内で飽和帯が不均一に生じた。試験地には，複数の露岩帯が同一走向で分布し，既発生の15個の崩壊位置を区分している。この露岩帯が中間流の流下を遮り飽和帯の不均一な分布を生じさせていた。調査・観測結果を組み込んだ臨界すべり面解析結果は観測結果とよく一致し，すべり面は脆弱層の底面深度になり，すべり面頭部は露岩帯下方の実際に亀裂が生じた位置に計算された。以上より，本試験地では脆弱層の存在がすべり面深度を決定しており，露岩帯の存在が斜面上下方向の崩壊発生位置を規制していることがわかった。

Severely rolled sheets of Al-4.5%Mg-0.7%Mn based alloys have been prepared to achieve grain refinement leading to low temperature superplasticity. Rolling is one of straining technique which has a high possibility of industrial application. The alloys used were T1: base one adding 0.08%Zr and 0.05Ti for suppression of grain growth, F1: 0.80%Fe addition and F2: 1.92%Fe addition to increase nucleation sites of recrystallization. For comparison, M1: Al-4.2%Mg was also prepared. All of samples were 99.8% cold rolled to thin sheet 0.1mm thick. The maximum elongation of 150%, which was markedly large in consideration of specimen thickness of 0.1mm, was obtained at 533K and 1.4×10-3s-1 for T1 sample while the elongations for F1 and F2 showed less than 100%. On the other hand, M1 exhibited larger elongation of 190% at 533K and 1.4×10-3s-1. Further, difference in superplastic deformation of these alloys was investigated by using SEM/ EBSP analysis of intragranular misorientation, which reflects strain or stored energy generated during deformation with dislocation glide, to discuss the deformation mechanism.

The infiltration of rainfall reduces shear strength on the slip surface by increasing pore water pressure, resulting in increased landslide movement. Although pore water pressure fluctuation is an important factor influencing landslide activity, it is difficult to observe its mechanism at various depths of the slip surface because of the complex hydrogeological structure. This study aims to examine the pore water pressure fluctuation in shallow and deep strata of the slip surface. The rainfall, pressure head and drainage rate were observed at the Tairasawa landslide before and after the drainage well work. The pressure heads were measured in the vertical observation borehole with a screened section at the depth of slip surface. The horizontal drain borings of drainage well were drilled for confining groundwater in a screened section at the same depth of slip surface. After drilling a horizontal drain boring, the pore water pressure on the slip surface was reduced and the decreasing volume in the deep stratum was larger than that in the shallow stratum. The effects of horizontal drain boring could be easily evaluated by comparing the decrease in pore water pressure before and after the works. Furthermore, the decrease in pore water pressure on the slip surface shows two significant characteristics. First, the variation in pore water pressure corresponds to drilling depth and seepage water in a horizontal drain boring. Second, the decreasing effects of pore water pressure can be clearly seen when the drilling direction of a horizontal drain boring is near the vertical observation borehole.

表層崩壊発生場の地盤特性を解明するために0.46haの試験地を設け,簡易貫入試験を169地点実施した後に深さ75cmをすべり面とする表層崩壊が発生した。すべり面は,Nc

Nc値が1.0未満の層を風化花崗岩斜面の表層崩壊すべり面深さに見出し,これを脆弱層と名付けた。表層崩壊現象を理解する鍵になるであろうこの層について,さまざまな土質試験を行った。脆弱層の飽和土の内部摩擦角は11~14°だった。この低角度はせん断試験に先立つ飽和に際してコラプス沈下と呼ばれる構造破壊が起こった結果だと考えられた。吸水過程にある脆弱層の土は飽和に達した時点で急激に沈下した。脆弱層の構造は,空隙率が上下の土層と比較して格段に高いだけではなく,0.85~2.0mmの粗砂の容積率が高く,2.0~4.75mmの空隙の容積率がそれよりさらに高い特性をもっていた。そのため脆弱層の骨格を作る粗砂は容易に空隙中へ転がり込める。飽和化に伴ってそのような粒子の転がり込みが生じることを顕微鏡下で観察した。この現象は,結果的に総体積を減少させコラプス沈下となる。斜面に潜在する脆弱層で起こるコラプス沈下は表層崩壊現象に重要な役割を果たしている。

Objective: To study the effect of a new fermented milk product containing GABA (FMG) on the blood pressure (BP) of patients with mild hypertension. Design: A randomized, placebo-controlled, single-blind trial. Setting: The study was carried out at the outpatient clinic of the Cardiovascular Disease Center, Tokyo Metropolitan Police Hospital, Japan. Subjects: The study population comprised 39 mildly hypertensive patients (16 women and 23 men) aged 28–81 y (mean, 54.2 y). Interventions: The study consisted of a 12-week period of daily intake of FMG or placebo (weeks 1–12) followed by 2 weeks of no intake (weeks 13 and 14). We measured the peripheral BP and heart rate of seated patients at weeks 0, 2, 4, 8, 12 and 14. Routine blood study and urinalysis were performed before and after the intake. Results: There was a significant decrease of BP within 2 or 4 weeks, and it remained decreased throughout the 12-week intake period. For the FMG recipients, the mean decrease after 12 weeks was 17.4±4.3 mmHg in the systolic BP (SBP) and 7.2±5.7 mmHg in the diastolic BP (DBP). Both of these values differed statistically from baseline levels (P

We have investigated i) the formation of gravitationally bounded pairs of gas-giant planets (which we call "binary planets") from capturing each other through planet-planet dynamical tide during their close encounters and ii) the following long-term orbital evolution due to planet-planet and planet-star {\it quasi-static} tides. For the initial evolution in phase i), we carried out N-body simulations of the systems consisting of three jupiter-mass planets taking into account the dynamical tide. The formation rate of the binary planets is as much as 10% of the systems that undergo orbital crossing and this fraction is almost independent of the initial stellarcentric semi-major axes of the planets, while ejection and merging rates sensitively depend on the semi-major axes. As a result of circularization by the planet-planet dynamical tide, typical binary separations are a few times the sum of the physical radii of the planets. After the orbital circularization, the evolution of the binary system is governed by long-term quasi-static tide. We analytically calculated the quasi-static tidal evolution in later phase ii). The binary planets first enter the spin-orbit synchronous state by the planet-planet tide. The planet-star tide removes angular momentum of the binary motion, eventually resulting in a collision between the planets. However, we found that the binary planets survive the tidal decay for main-sequence life time of solar-type stars (~10Gyrs), if the binary planets are beyond ~0.3 AU from the central stars. These results suggest that the binary planets can be detected by transit observations at >0.3AU., 16 pages, 7 figures. Accepted for publication in the Astrophysical Journal

Various magnetic levitation systems using oxide superconductors are developed as strong pinning forces are obtained in melt-processed bulk. Lots of studies have especially been made on superconducting magnetic bearings, and the analysis of flux creep phenomena is important for developing them. It is important to determine the dynamic behavior of internal current distribution in the superconductor. Moreover, other applications including magnetic shields, superconducting motors, etc. also need similar kind of analysis to resolve the irreversible magnetization or hysteresis characteristics. This paper propose the application of inverse problem solution to examine the dynamic analysis of the internal current of bulk high- T c superconductor. The method infers and determines the internal superconducting current from the exterior flux density distribution. The paper reported the applicability of the proposed analysis method.

We examined a chloride (Cl-)-dependent K+ transport (K(+)-Cl- cotransport) and regulatory volume decrease in dog red blood cells with high K+, low Na+, and high glutathione (GSH) content (HK/HG) due to the presence of an Na(+)-K+ pump. The HK/HG cells were separated according to their density, and the age-marker enzyme activities, such as glucose-6-phosphate dehydrogenase and cholinesterase, were determined. Unexpectedly, we found that young cells were heavier (more dense) and smaller in size compared with the old cells, which were lighter (less dense) and larger. The K(+)-Cl- cotransport was nearly 10-fold higher in the most dense cells, representing a 12% fraction of the total population compared with the lightest cohort. Although K(+)-Cl- cotransport in both the dense and the light cells was activated by N-ethylmaleimide, swelling and depletion of cellular divalent cations and the activation of the transport in the dense cells was greater. Both the dense and light cells regulated their volume when they were isosmotically swollen. Therefore, the lower activity of K(+)-Cl- cotransport might not explain the relative large volume in old HK/HG cells. The concentration of GSH and glutamate was higher in the light cells. Thus the higher the GSH and glutamate concentration, the greater the cell volume and the lower the K(+)-Cl- cotransport.

Models of uranium-uptake at the early stage and at a constant rate (linear-uptake) were reaxamined for ESR dating of fossil bones and teeth. An analytical equation for total dose of natural radiation ( D T ) was obtained for uranium uptake in the form of a saturation function, C U ( t ) = C US (1−e − λt ), where C US is the saturation level of the uranuim and λ is the uranium uptake rate, presumably associated with the decomposition rate of organic materials and the diffusion of uranium. The present uranium content C U (T) = C U ∗ at the unknown age T gives C US -C U ∗ /1− e −glT ) and so λ = −(1/T) ln (1−C U ∗ /C US ) . The D T calculated by taking 238 U-series secular disequilibrium into into account is reduced to that of an early uptake for λT ⪢1 and to that of a linear uptake for λT ⪡1. The activity ratios of 234 U/ 238 U and 230 Th/ 238 U for the saturation-uptake model are also given. The model of linear uptake is applicable for C U ∗ US . The curves of D T as a function of T were calculated for C US as a parameter.

This paper summarizes the theory and practice of geosynthetic reinforcement of fills over extremely soft ground in Japan. Emphasis is placed on artificially reclaimed soft ground composed of marine soils which are characterized by high water contents and low shear strengths. The use of geosynthetics facilitates the movement of construction equipment and the placement of fill. The bearing capacity of the ground is improved by using high strength geosynthetics. Classical bearing capacity theory, cable theory combined with modulus of subgrade reaction theory, and plate theory are found to be useful in designing embankments on soft ground. The empirical parameters used in these theories are available for most projects. The unique geotechnical conditions of the soft ground typically inhibit the construction procedures specified in the design. The experience gained from past geosynthetic reinforced fill projects, one of which is highlighted in the paper, facilitates the improvement of fill placement and spreading techniques on soft ground.

The 64-kDa membrane-bound glycoprotein of the cellular slime mold Polysphondylium pallidum (referred to as gp64), seems to be implicated in cell-cell adhesion. Previously we have isolated a full-length gp64 cDNA, determined its nucleotide sequence, and found that all cysteine residues in the protein are involved in the formation of disulfide bonds. The disulfide arrangement of the 36 cysteines in gp64 was established by analysis of proteolytically cleaved protein and sequence analysis of cystine-containing fragments. Since gp64 has 36 Cys residues, 18 disulfide bonds must exist and the positions of 15 of them were determined. The 15 disulfide bonds in gp64 constitute five characteristic, so-called Sushi domains. In a Sushi domain, the first Cys in a sequence is connected to the third one and the second Cys to the fourth one. This is the first report describing the presence of Sushi domains in a cellular slime mold protein. From these data, gp64 appears to be distinct from all other previously described cell-adhesion proteins.

γ-Aminobutyric acid (GABA)-like immunoreactive neuronal profiles and their synaptic relations in the area postrema of the rat were studied by ultrastructural immunocytochemistry using an antibody directed against GABA. GABA-like immunoreactive (GABA-LI) perikarya and dendrites usually received synaptic inputs from nonimmunoreactive axon terminals. These synaptic contact zones were mostly asymmetrical. Conversely, GABA-LI axon terminals were found to make synapses with nonimmunoreactive perikarya and dendrites, usually of the symmetrical type. Furthermore, GABA-LI axon terminals in synaptic contact with GABA-LI perikarya and dendrites were sometimes found. Although some GABA-LI axon terminals were in close vicinity of other nonimmunoreactive axon terminals, no clear synaptic contacts could be found. On the other hand, GABA-LI profiles could be found close to the capillaries, although they were always separated by a thin layer of glial processes. The above results suggest that the GABAergic neuron receives information from both the circulating blood and other neurons and then performs its depressor function through other neurons.

Recent advances in neuroblastoma (NB) research addressed that epigenetic alterations such as hypermethylation of promoter sequences, with consequent silencing of tumor-suppressor genes, can have significant roles in the tumorigenesis of NB. However, the exact role of epigenetic alterations, except for DNA hypermethylation, remains to be elucidated in NB research. In this paper, we clarified the direct binding of MYCN to Bmi1 promoter and upregulation of Bmi1 transcription by MYCN. Mutation introduction into an MYCN binding site in the Bmi1 promoter suggests that MYCN has more important roles in the transcription of Bmi1 than E2F-related Bmi1 regulation. A correlation between MYCN and polycomb protein Bmi1 expression was observed in primary NB tumors. Expression of Bmi1 resulted in the acceleration of proliferation and colony formation in NB cells. Bmi1-related inhibition of NB cell differentiation was confirmed by neurite extension assay and analysis of differentiation marker molecules. Intriguingly, the above-mentioned Bmi1-related regulation of the NB cell phenotype seems not to be mediated only by p14ARF/p16INK4a in NB cells. Expression profiling analysis using a tumor-specific cDNA microarray addressed the Bmi1-dependent repression of KIF1Bbeta and TSLC1, which have important roles in predicting the prognosis of NB. Chromatin immunoprecipitation assay showed that KIF1Bbeta and TSLC1 are direct targets of Bmi1 in NB cells. These findings suggest that MYCN induces Bmi1 expression, resulting in the repression of tumor suppressors through Polycomb group gene-mediated epigenetic chromosome modification. NB cell proliferation and differentiation seem to be partially dependent on the MYCN/Bmi1/tumor-suppressor pathways.