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We propose a highly selective, sensitive, accurate, and high-throughput bioanalysis method for bevacizumab utilizing an anti-idiotype DNA aptamer. With this method, bevacizumab in a plasma sample was reacted in a 96-well plate immobilized with the aptamer and further reacted with a protein A–HRP conjugate. The resulting HRP activity was colorimetrically detected using a microplate reader. The calibration curve of bevacizumab ranged from 0.05 to 5.0 μg/mL, and showed a good correlation coefficient (r2= 1.000). The limit of detection was 2.09 ng/mL. We also demonstrated both the possibility of highly sensitive detection using luminol chemiluminescence and the repeated use of affinity plates. The proposed method is applicable for planning optimal therapeutic programs and for an evaluation of the biological equivalencies in the development of biosimilars.

The linear ubiquitin chain assembly complex (LUBAC) is required for optimal gene activation and prevention of cell death upon activation of immune receptors, including TNFR11. Deficiency in the LUBAC components SHARPIN or HOIP in mice results in severe inflammation in adulthood or embryonic lethality, respectively, owing to deregulation of TNFR1-mediated cell death2–8. In humans, deficiency in the third LUBAC component HOIL-1 causes autoimmunity and inflammatory disease, similar to HOIP deficiency, whereas HOIL-1 deficiency in mice was reported to cause no overt phenotype9–11. Here we show, by creating HOIL-1-deficient mice, that HOIL-1 is as essential for LUBAC function as HOIP, albeit for different reasons: whereas HOIP is the catalytically active component of LUBAC, HOIL-1 is required for LUBAC assembly, stability and optimal retention in the TNFR1 signalling complex, thereby preventing aberrant cell death. Both HOIL-1 and HOIP prevent embryonic lethality at mid-gestation by interfering with aberrant TNFR1-mediated endothelial cell death, which only partially depends on RIPK1 kinase activity. Co-deletion of caspase-8 with RIPK3 or MLKL prevents cell death in Hoil-1−/−(also known as Rbck1−/−) embryos, yet only the combined loss of caspase-8 with MLKL results in viable HOIL-1-deficient mice. Notably, triple-knockout Ripk3−/−Casp8−/−Hoil-1−/−embryos die at late gestation owing to haematopoietic defects that are rescued by co-deletion of RIPK1 but not MLKL. Collectively, these results demonstrate that both HOIP and HOIL-1 are essential LUBAC components and are required for embryogenesis by preventing aberrant cell death. Furthermore, they reveal that when LUBAC and caspase-8 are absent, RIPK3 prevents RIPK1 from inducing embryonic lethality by causing defects in fetal haematopoiesis.

Salmon myofibrillar protein (Mf) was investigated as a source of edible anti-inflammatory products. Peptides produced by stepwise digestion of Mf (without carbohydrate) with pepsin and trypsin had little effect on the secretion of inflammation-related compounds from lipopolysaccharide-stimulated RAW 264.7 macrophage cells. However, peptides prepared from Mf conjugated with alginate oligosaccharide (AO; 19 μg/mg protein) (dMSA) through the Maillard reaction in the presence of sorbitol significantly reduced the secretion of the pro-inflammatory mediators nitric oxide, tumor necrosis factor (TNF)-α and interleukin (IL)-6, as well as mRNA expression of TNF-α, IL-6, inducible nitric oxide synthase and cyclooxygenase-2. Additionally, dMSAinhibited acute inflammation in a carrageenan-induced model of paw edema in mice, but had no effect on natural killer cell cytotoxic activity or macrophage phagocytosis. These results suggest that fish Mf conjugated with AO may be a potential food material with anti-inflammatory function.

Background:Human dedifferentiated fat cells (HDFATs) may be a new cell type suitable for regenerative therapies. The aim of this study is to assess the potential of HDFATs for vascular regeneration of periodontal tissue. To do this, HDFATs and human gingival endothelial cells (HGECs) were cocultivated, and vascular regeneration was examined in vitro. Methods:HDFATs were isolated from subcutaneous adipose tissue, and HGECs were isolated from gingival cells using anti‐cluster of differentiation 31 antibody‐coated magnetic beads. HDFATs were cocultured with HGECs in microvascular endothelial cell growth medium‐2 (EGM‐2MV) for 7 days. Expression of endothelial cell (EC) markers, the formation of capillary‐like tubes, and the expression of pericyte markers were determined. Results:HDFATs, cultured in EGM‐2MV or cocultured with HGECs, expressed EC markers. HDFATs in both conditions initiated tube formation within 5 hours of seeding and formed extensive capillary‐like structures within 12 hours. These structures disintegrated within 24 hours when cells were cultured in EGM‐2MV alone, whereas cocultured HDFATs maintained tubes for >24 hours. Cocultured HDFATs significantly increased expression of pericyte markers, a cell type associated with microvasculature. Conclusion:HDFATs possess the ability to express EC markers, and coculture with HGECs promotes differentiation into pericytes involved in the maturation and stabilization of the microvasculature.

Linear ubiquitination is crucial for innate and adaptive immunity. The linear ubiquitin chain assembly complex (LUBAC), consisting of HOIL-1, HOIP, and SHARPIN, is the only known ubiquitin ligase that generates linear ubiquitin linkages. HOIP is the catalytically active LUBAC component. Here, we show that both constitutive and Tie2-Cre-driven HOIP deletion lead to aberrant endothelial cell death, resulting in defective vascularization and embryonic lethality at midgestation. Ablation of tumor necrosis factor receptor 1 (TNFR1) prevents cell death, vascularization defects, and death at midgestation. HOIP-deficient cells are more sensitive to death induction by both tumor necrosis factor (TNF) and lymphotoxin-α (LT-α), and aberrant complex-II formation is responsible for sensitization to TNFR1-mediated cell death in the absence of HOIP. Finally, we show that HOIP’s catalytic activity is necessary for preventing TNF-induced cell death. Hence, LUBAC and its linear-ubiquitin-forming activity are required for maintaining vascular integrity during embryogenesis by preventing TNFR1-mediated endothelial cell death.

In 2007 development and assessment of a precise point positioning (PPP) software for land vehicular navigation have been reported. This paper presents one phase of the continued study at NDA for further extension of the software to helicopter navigation. For 3D users, the height-dependent tropospheric delay is a critical factor, and so the sophisticated correction models and parameter estimation strategies have to be investigated to mitigate the delay effect. Since the precise positioning results become available only after the filter convergence and the filter generally converges rather slowly, speeding up the convergence has to be taken into special attention. In this paper we propose some new ideas to cope with this problem, and report the test using GPS helicopter flight data in post-processing. With proposed techniques including the variable Q and pseudo-observation concept, we show that point positioning accuracy at about decimeter level both horizontally and vertically can be achieved with improved convergence within about half an hour. We also show that the use of the high-rate (30-second) satellite clocks rather than the 5-minute clocks further improves positioning accuracy at sub-decimeter level in each 3D coordinate.

Cellulose derivative resin (CDR) suspensions containing resin particles of cellulose nitrate (CDR(CN)), cellulose acetate (CDR(CA)), or cellulose triacetate (CDR(CTA)) were prepared as the sorbent for resin suspension injection (RSI) electrothermal atomic absorption spectrometry (ETAAS), in which fine resin particles holding a desired trace element were directly injected into the graphite tube as the suspension. To compare the sorption ability, the distribution ratios of the Mo(VI)-phenylfluorone complex were measured between the aqueous phase and the sorbents, including the CDRs mentioned above and the finely divided anion exchange resin (AR). The results showed that the sorption ability decreases in the following order: CDR(CN) ≥ AR > CDR(CA) ≥ CDR(CTA). It was concluded that CDR(CN) was able to extract more than 98% of Mo(VI), and was suitable for RSI-ETAAS as well as AR. CDR(CN) was used for the determination of Mo in NIES certified reference materials, No. 10 Rice flour-unpolished; the results showed fairly good agreements between the analytical values and the certified values.

The changes in the oxidation state of vanadium in artificial and natural seawater samples were studied by electrothermal atomic absorption spectrometry (ETAAS) with a direct injection of a resin suspension. V(IV) and V(V) were extracted as the complex with Chromazurol B and with N-cinnamoyl-N-2,3-xylylhydroxylamine, respectively, using a suspension of an anion-exchange resin and determined by ETAAS independently. The detection limits of both methods were 0.02 ng ml-1for 40 ml of a sample solution. The recovery tests for an artificial seawater sample spiked with V(IV) and/or V(V) were carried out carefully. The results showed that the recoveries of V(IV) or V(V) were 99.2 - 109% and the standard deviations were 1 - 6%. The total V was also determined after V(V) was reduced by ascorbic acid. In artificial seawater at pH 7.8, V(V) was stable but V(IV) was oxidized rapidly. In acidified artificial seawater (pH 2.0), V(IV) was oxidized slowly but only a small tendency of such reduction of V(V) was observed. In a natural seawater sample, V(IV) was not detected. The acidification of the natural seawater sample resulted in the reduction of V(V).

Kinking deformation by buckling of a PVD (Prefabricated Vertical Drain) due to consolidation settlement of the clay layer around it is discussed in this paper. A new testing device is manufactured for the purpose, and the deformation properties of two-kinds of PVD during consolidation of the surrounding clay are compared. It is concluded that a fibredrain made only of natural fibres can sustain the vertical permeability during a settlement strain up to 24%, while a PVD made of plastic loses its permeability before 19% strain. The field performance of both materials is also referred to.

Tactile sensitivity to shifts of a point stimulus was determined at the forehead for spatially filled (continuous) and spatially separated (discrete) stimulation. The minimal limen was between 12 and 30 mm/sec. for continuous stimulation and at a 200-msec. interstimulus interval for discrete stimulation. The continuous thresholds were always lower than the discrete thresholds at the examined time rates. Furthermore, the continuous thresholds were slightly higher for oblique directions than for both vertical and horizontal directions, while the discrete thresholds were quite low for both vertical and horizontal directions.

Two experiments examined the temporal effect on tactile letter recognition of sequentially tracing a stimulus in the order of making the necessary strokes. The effect of time interval between letter-strokes was examined in Exp. 1, with the three measures, recognition accuracy, latency, and readability. Accuracy of recognition did not differ by change in time intervals, but latency showed a downward trend, and readability showed an upward trend with an increase in the time interval. The effect of duration of exposure (number of pulses which activates each vibratory stimulator defining a letter) was examined in Exp. II with two measures, accuracy of recognition and latency. Both accuracy of recognition and latency reached optimum with 8 pulses per stimulator. The time interval between letter-strokes contributed strongly at the shorter exposure. The minimum was about 80 msec, for the interval between letter-strokes, and 4 pulses per stimulator considering the practical demands of fast exposure and easy identification.

This study reports on social isolation of the frail elderly using a representative sample of 441 persons living in downtown Tokyo. The average SII (Social Isolation Index) scores of the aged who were living with their families were 7.92 and S.D. 4.15 and those of persons who were living alone and elderly couples 9.14, 4.26, respectively. The relationships between length of residence and SII score was statistically significant at the 5% level. Analysis by stepwise regression disclosed that variables which genuinely influenced the scores of SII were: 1) whether older persons were physically and/or mentally impaired, 2) whether they had children who were living separately, 3) length of residence, 4) number of family members, and 5) amount of household income. These results indicated that socioeconomic factors had substantial impact on social isolation of the impaired elderly in Japan.

The effect of stimulus orientation of letters presented either visually or vibrotactually was examined to obtain basic information on sensory substitution using the tactile sense. The reaction time (RT) to identify the letters F and R presented in normal or mirror-image form at four orientations each was measured. In addition, conditions of 0° and 270° of head rotation from vertical and arm rotation from the midline axis were employed. Data from 5 trained subjects showed that vibrotactile RTs were always longer than visual RTs. Stimulus rotation away from normal orientation increased visual RTs significantly but not vibrotactile RTs. Visual orientation effect then seemed to be determined by the body-coordinate system but not the vibrotactile orientation. Although further studies are warranted, from the results of this experiment, any convenient and constant stimulus orientation could be used with a wearable vibrotactile display system to exploit passive touch.

An interferometric technique for the determination of the radial distribution of the refractive index in light-focusing plastic rods (LFR) has been developed using the shearing method of Interphako interference microscopy. We have derived mathematical expressions for the shape of interference fringes across the rod assuming the index distribution of LFR is described by n(r) = n_0(I − ar^2 + br^4 + cr^6 + dr^8) in which a, b, c, and d are distribution constants. The dsitribution constants were determined using a least-squares technique on a computer from the actual shape of fringes. Results for the representative cases are shown.

A computer-controlled vibrotactile display against the abdomen was used to investigate recognition performance of Katakana characters as a function of 3 display modes. The superior performance (about 95% correct) is achieved when the letter is presented sequentially as if it were being written. The other two modes (the entire letter is presented at once, the letter is moved horizontally across the display) allow only 20 to 40% correct recognition.

Catalytic allylation of aryl halides with allyltributyltin in the presence of tetrakis(triphenylphosphine)palladium(O) was investigated. The reaction with aryl bromides proceeded efficiently to yield allylbenzenes.

Using an apparent-motion mode, the psychophysical effect of apparent movement was introduced for tactile letter reading. This mode reduces time for the tracing mode. Five temporal parameters were examined for application to actual communication. 4 subjects recognized words composed of 3 to 7 Japanese letters in about 5 seconds.

Alkylation, arylation, and vinylation of acyl chlorides by means of organotin compounds in the presence of catalytic amounts of tetrakis(triphenylphosphine)palladium(O) were demonstrated. The corresponding ketons were obtained in fairly good yields.