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Bioconjugation with polyethylene glycol (PEG) is important for protein drug development as it has improved biological stability. In contrast, proteins including PEGylated ones are susceptible to physicochemical stresses. Particularly, protein drugs in solution may form aggregates or subvisible particles if they are exposed to dropping stress during transportation. However, many PEGylation studies have focused on its usefulness, such as the extension of half-life in blood, and changes in the physical properties or biological responses of PEGylated proteins under dropping stress remain unexplored. Here, we prepared four PEGylated ovalbumin (PEG-OVA) molecules conjugated with different lengths (5 or 20 kDa) and numbers (large [L] or small [S]) of PEG, analyzed the formation of subvisible particles under dropping stress, and examined their impact on antibody production and clearance. Under dropping stress, the aggregated particle concentration of 20 kDa PEG-OVA (S) and (L) solutions was approximately 3-fold that of the OVA solution. Moreover, administration of 20 kDa PEG-OVA with dropping stress induced anti-PEG antibody production and clearance of PEG-OVA. As a mechanism, dropping stress could enhance the uptake of 20 kDa PEG-OVA (L) by macrophages. These findings could provide insights into proper transportation conditions to ensure the quality of PEGylated protein drugs.

Public knowledge-based application ("Kouchi-shinsei" in Japanese) is unique to Japan, implemented to eliminate the off-label use of unapproved indications, dosages, and administrations because of drug lag. The guidance for public knowledge-based application was issued in 1999. This study comprehensively investigated the trends of items approved by public knowledge-based application in Japan during the last 2 decades. Prescription drugs approved from January 2000 to December 2019 were surveyed. In Japan, 1,855 drugs were approved within the target survey period. Among them, 219 (11.8%) were approved by public knowledge-based application. Considering the changes in the number of items approved by public knowledge-based application over the years, the number of items approved in 2000 was 7, reaching a maximum of 34 items in 2011, and decreased after that, 8 items were approved in 2019. The regulatory characteristics of drugs approved by public knowledge-based application and those of other drugs were compared. By public knowledge-based application, more anticancer and pediatric drugs were approved (P

Coke undergoes gasification reactions of CO2 and H2O in the blast furnace, and the gasification reactions degrade the coke and affect the operation of the furnace. In this study, to evaluate the factors that govern the strength of coke in the furnace, the fracture behavior of coke under blast furnace conditions was examined both experimentally and numerically. The three-point bending test was performed on coke subjected to the gasification reaction under conditions similar to those inside a furnace. It was found that coke gasified with CO2 is more likely to fracture than that with H2O since the conversions at fracture of the samples gasified with CO2 and H2O were 0.71 and 0.78, respectively. X-ray computed tomography images of coke taken before and after the gasification reactions showed that increase rate of porosity was almost uniform from the central part to the external surface in the case of CO2 gasification reaction, while it increased from the sample center towards the external surface in the case of the H2O gasification reaction. In addition, the finite element analyses using an isotropic damage model were performed. As a result, the damage spread in the analytical object in which the pore structure after the CO2 gasification reaction was reflected, while the damage was localized in the analytical object in which the pore structure after the H2O gasification reaction was reflected. These results suggested that the difference in the spatial distribution of the porosity increase rate within the coke samples owing to the differences in the rate-limiting steps of the gasification reactions greatly affected the strength of the coke samples.

Lepton flavor violating (LFV) process $e^+ e^- \to e^+ \tau^-$ induced by the four-Fermi contact interactions at the International Linear Collider (ILC) is studied. Taking account of the event selection conditions, it is shown that the ILC is sensitive to smaller LFV couplings as compared to the measurement of $\tau\to 3e$ process at the B-factory experiment. The upper bounds on some of the LFV couplings are improved by several factors using polarized $e^-/e^+$ beams at $\sqrt{s}=250~\mathrm{GeV}$ and by an order of magnitude at $\sqrt{s}=1~\mathrm{TeV}$., Comment: 14 pages, 6 figures. Version to appear in Phys.Lett.B

To prevent stress corrosion cracking (SCC) initiation at the inner surface of the butt-welding region of a small-diameter pipe, a residual stress improvement method has been developed. In this method, the butt-welding region is heated up to 325 degrees centigrade in air. After heating, a water quenching process is applied to the inner surface of the butt-welding region. During a water quenching process, a large thermal stress is generated by the temperature difference between inner and outer surfaces. When a thermal stress exceeds the yield stress of piping material, a tensile yield is generated at the inner surface of the butt-welding region. Because a tensile yield changes into a compressive residual stress, a compressive residual stress can be introduced at the inner surface of the butt-welding region after application of this method. In this study, residual stresses at the inner surface of the butt-welding region were measured. The measured residual stresses proved this method effectively reduces residual stresses and prevents SCC initiation.

Background. The purpose of this study was to examine whether EP4 as prostaglandin (PG) E2 receptor is involved in lipopolysaccharide (LPS)-induced cervical ripening. Methods. New Zealand White non-pregnant rabbits were randomly allocated to 4 equal groups and treated with vaginal suppositories containing LPS with various concentrations of EP4-selective antagonist once daily for 3 days, and then sacrificed for analysis. Analysis was performed in order to examine the inhibitory effect of EP4 antagonist on LPS-induced cervical ripening. The expression of EP4 in a cervix treated with LPS was examined immunohistochemically. The percent of cervical extensibility in the segment of cervix as the tensile strength was determined with 5.8 g of constant perpendicular stretching. The ripening area of the cervix with edematous changes in H&E sections was calculated by a microscopic system with a computer-assisted digital analyser. Type-1 collagenase activity was determined in cervical tissue using FITC-labelled type-1 collagen. Results. Immunohistochemical study shows the positive staining of EP4 at the interstitial cells in the cervix treated with LPS. The extensibility, cervical edematous area and type-1 collagenase activity are significantly reduced in the cervix treated with LPS in the presence of EP4 antagonist compared with that treated with LPS alone. Conclusions. These data implicates the EP4 as the PGE2 receptor involved in LPS-induced cervical ripening.

Endothelin-1 (ET-1) exists in endothelial cells as well as a variety of other cell types. The presence of ET-1 and its receptors in neurons suggests its possible role as a neurotransmitter and/or neuromodulator. Studies utilizing exogenous ET-1 have suggested that ET-1 affects pain transmission. This study was designed to examine the possible role(s) of neuronal ET-1 in pain processing. We produced neuron-specific ET-1 knockout mice using the Cre/loxP system with a synapsin I promoter and examined the effects produced by the lack of neuronal ET-1 on pain behavior using common pain models and a model of stress-induced analgesia. In acute nociceptive pain models, paw withdrawal thresholds to radiant heat and mechanical stimuli applied with von Frey hairs were significantly lower in the knockout mice compared with control. This indicated that the absence of neuronal ET-1 leads to greater sensitivity to acute nociceptive stimuli. After inflammation was produced by intraplantar injection of carrageenan, there was a significantly greater degree of thermal hyperalgesia and mechanical allodynia in the knockout mice even after the difference in baseline values was compensated. Furthermore, in a neuropathic pain model produced by spinal nerve ligation, there was also a greater degree of mechanical allodynia in the knockout mice. Finally, in a swim-stress model, the magnitude of stress-induced analgesia was less in the knockout mice, indicating the involvement of neuronal ET-1 in stress-induced analgesia. The results suggest that there is a basal release of ET-1 from neurons that affect baseline pain thresholds as well as an additional release during persistent pain states that acts to suppress the pain. The involvement of neuronal ET-1 in stress-induced analgesia further suggests its role in endogenous pain inhibitory systems. To confirm that ET-1 is released in persistent pain states and to determine which part of the CNS is involved, we measured the concentrations of ET-1 before and after inducing peripheral inflammation in different parts of the CNS involved in endogenous pain inhibitory systems in normal mice. We found that ET-1 was increased in the hypothalamus while no significant increase was observed in the midbrain, medulla and spinal cord. The results of the present study suggest that neuronal ET-1 is involved in endogenous pain inhibitory control likely via pathways through the hypothalamus.

ZrO2–phenolic resin and SiO2–ZrO2–phenolic resin hybrid fibres have been prepared by sol–gel processing using zirconium tetrakis(2,4-pentanedionate) (ZTP), tetraethoxysilane (TEOS), and novolac-type phenolic resins as raw materials. ZTP and the phenolic resins dissolved in a mixture of ethanol and 2,4-pentanedione have undergone a sol–gel reaction to give ZrO2–phenolic resin hybrid fibres when H2O and H2SO4 have been added to the solutions at a H2O/Zr molar ratio of 0.45 and H2SO4/Zr molar ratio of 0.66. On the basis of the molar ratios and the H2O/Si and H2SO4/Si molar ratios of solutions, which were demonstrated to be optimal for preparing SiO2–phenolic resin hybrid fibres from TEOS and the phenolic resins, amounts of H2O and H2SO4 to be added to TEOS-ZTP-phenolic resin–H2O–H2SO4–ethanol–2,4–pentanedione solutions with numerous Si-to-Zr atomic ratios have been estimated. The solutions containing the estimated amounts of H2O and H2SO4 have been found to give SiO2–ZrO2–phenolic resin hybrid fibres with Si-to-Zr ratios which correspond to those of the solutions, meaning that the hybrid fibres with a variety of Si-to-Zr atomic ratios can be prepared by this estimation method. The hybrid fibres have been convertible into ZrC or Si–Zr–C fibres upon firing at 1500°C for 4 h in an Ar flow.

Synthesis conditions for producing SiO 2 -TiO 2 -phenolic resin hybrid fibers with a wide range of Si-to-Ti atomic ratios from tetraethoxysilane, titanium tetrakis(2,4-pentanedionate) and novolak-type phenolic resins by sol-gel processing have been investigated. It was found that 2,4-pentanedione was an effective solvent for the sol-gel reaction in order to produce the hybrid fibers. For favorable spinn-ability of the hybrid fibers from the solutions it was necessary to adjust critically the amounts of H 2 O and of acid, which were added to promote hydrolysis and polycondensation of the Si and Ti compounds in the solutions, according to their Si-to-Ti ratios. The amounts found to be optimal for spinning the hybrid fibers were in proportion to the Si-to-Ti ratio of the solutions. Using these combinations, SiO 2 -TiO 2 -phenolic resin hybrid fibers with various Si-to-Ti ratios could then be produced, which could then be converted into Si-Ti-C fibers by carbothermal reduction at 1500 °C in Ar.

By the use of molecular dynamics simulation (MD), varieties of properties including structural, transport, optical and thermodynamic properties can be calculated. Since this feature is very attractive to the analyses of refining slags, its applicability was studied. CaO-SiO2 melts with CaO contents ranging from 40 to 60 at% were simulated at 1873 K. The calculated pair distribution functions and general tendency of silicon ion fractions with various number of non-bridging oxygen ions agreed well with experiments reported elsewhere. The calculated diffusion coefficient of every ion as a function of CaO content was also in good agreement with the measured values. These demonstrate the promising applicability of MD to slag analyses.The melt of 45at%CaO-45at%SiO2-10at%CaF2 was also simulated by MD to study the effects of fluorine addition to silicate melt. The hypothesis of the existence of di-fluorine ions in some thermodynamic modeling of slags was not verified due to the lack of appropriateness of the interatomic potentials used in this study.

Some cases of Stress Corrosion Cracking (SCC) failures in the proximity of welded zone of core internals and pipes in Primary Loop Recirculation, which are made of austenitic stainless steel, were reported in existing boiling water reactors, from the late 1970s to 1980s. As a countermeasure against SCC, low-carbon stainless steel was developed in order to reduce susceptibility to SCC, and used as standard material at that time. However, SCC failures were still observed in the core internals and pipes made of low-carbon stainless steel in recent years. It is well understood that residual tensile stress due to welding largely affects occurrence and growth of SCC in low-carbon stainless steel. Based on these observations, stress reduction methods for the pipe welded zone have already been developed such as Induction Heating Stress Improvement (IHSI) and Heat Sink Welding (HSW). However, these stress reduction methods are applied to only large-bore, thick-wall pipes, because it is difficult to apply these established countermeasures to the small-bore, thin-wall pipes which diameter is 50A and below. Thus stress reduction method for small-bore pipe has not been established. In this study, residual tensile stress reduction method that is applicable to the welded zone of small-bore pipe has been developed. The stress reduction method uses rapid quenching of inner surface of the pipe by cooling water after heating outer surface of the pipe by a heating device. Just after starting to cool the inside of the pipe, the temperature of the inner surface is low and tensile stress is generated. On the other hand, since temperature of outer surface is high, large temperature difference between inside and outside surface of the pipe develop and the high through-wall thermal stress is generated. When the temperature difference between the inner and outer surfaces of the pipe is large, thermal stress exceeds the yield stress on the pipe inner surface, and plastic deformation occurs on the inner surface of the pipe. The residual stress of the inner surface becomes compressive after the heat treatment due to the residual layer of plastic strain. In this study, the effectiveness of this method is shown by comparing the residual stress on the inner surface of the pipe before and after the application of this method by mock-up tests.Copyright © 2010 by ASME

Weld residual stress of a small bore pipe was evaluated using three-dimensional thermal elasto-plastic analysis, which was based on the finite element method (FEM), taking into consideration the effect of a moving heat source during the butt-weld process. A moving heat source during the weld process complicates analysis process due to the necessity of considering the set welding conditions at each time step and this increases computation time. Therefore, there are remarkably few studies on the effect of a moving heat source during the weld process on the analytical results of weld residual stress for small bore pipes. We used this analysis to find the effect of the welding start/end on weld residual stress. Weld residual stress is not a circumferentially uniform state. Increased tensile and compressive residual stress occurred near the end position of the final welding pass. In addition, an accelerated stress corrosion cracking (SCC) test was carried out using a boiling 42% magnesium chloride (MgCl2 ) solution. In particular, butt-weld joints of low-carbon austenitic stainless steel (Type 316L) pipe, which is often used in boiling water reactors (BWRs), were examined. Comparison between three-dimensional thermal elasto-plastic analysis and accelerated SCC testing showed the tensile stress zone indicated by analysis agreed well with SCC occurrence indicated by examination.Copyright © 2009 by ASME

Stress Corrosion Cracking (SCC) failures have occurred in the vicinity of austenitic stainless steel pipe welds used in boiling water reactors, since the late 70s. One of the initial countermeasures against SCC has been to use low-carbon stainless steel. However, in older plants, SCC failures in low-carbon stainless steel pipe were still observed in recent years. It is well understood that residual tensile stress due to welding largely affects occurrence and growth of SCC in low-carbon stainless steel. Because of this, it is important to reduce the residual tensile stress in the welded zone in addition to utilizing less susceptible material. However, a countermeasure to reduce residual tensile stress in small-bore pipe has not been established. In this study, a new stress reduction method is developed in order to reduce residual tensile stress in the welded zone, even for small-bore pipe. This method is applied to the butt-welded zone while the pipe is filled with water. First, the pipe is frozen at two points centering around the welded zone by cooling the outer surface of the pipe to transform the water to plugs of ice. The volume inside the pipe between the ice plugs becomes watertight and pressurized by the expansion of the plugs up-on freezing. The pipe expands near the zone of the weld groove, because the wall thickness of the butt-welded zone is thinner locally because of the weld counter bore. In this process, tensile strain occurs near the welded zone due to the pipe expansion beyond its plastic range, and then hoop tensile plastic strain of inner surface is larger than that of outer surface. Once the pressure load is removed, the difference in these tensile strains induces the hoop residual tensile stress on the inner surface to reverse to a compressive stress. On the other hand, the local bending deformation that occurs in the welded zone results in an axial residual compressive stress on inner surface. Both experimental studies and finite element analysis confirmed that the hoop and axial residual tensile stress at the inner surface of the welded zone are reversed to compressive stress by the pipe expansion.

Using cultured neonatal rat atrial cardiocytes, we have studied the effect of synthetic porcine endothelin (pET), a novel potent vasoconstrictor isolated from endothelial cells, on the release of immunoreactive (IR) rat atrial natriuretic peptide (rANP). pET stimulated IR-rANP secretion in a dose-dependent manner (10(-10)-10(-7) M) with an approximate half-maximally stimulatory dose of 2 x 10(-10) M. The pET-induced IR-rANP secretion was attenuated by Ca2+-channel blocker nicardipine, but no further stimulation was induced when combined with a Ca2+-channel agonist BAY-K 8644. pET in combination with tetradecanoyl-phorbol-acetate resulted in a synergistic effect on IR-rANP secretion. These data suggest that ET may play as an endogenous secretagogue for rANP by modulating Ca2+ influx through the voltage-dependent Ca2+-channels in atrial cardiocytes.

Urinary N-acetyl-β-D-glucosaminidase (NAG) was determined in infants during the first year of life to clarify its evolutional change and the reliability of this enzyme as a marker of renal tubular damage in the newborn period. During the first 2 weeks of life, there was no difference in the urinary levels between preterm (gestational age of 33–36 weeks) and full-term infants. The urinary NAG index (NAG is expressed as the ratio of the enzyme activity to milligrams of creatinine) during the first month of life was significantly elevated when compared with that of any other succeeding period and gradually decreased during the first year of life. The index in newborn infants on tobramycin was significantly elevated and maintained a higher value during the 5 days after withdrawal of the medication. Infants with asphyxia had a higher urinary NAG index during the first week of life. These data suggest that the urinary NAG index is a useful indicator of renal tubular damage during the newborn period.

The effect of 12-O-tetradecanoylphorbol-13-acetate (TPA) on serotonin-induced inositol phosphate (IP) accumulation and intracellular free Ca2+ concentrations ([Ca2+]i) was investigated in cultured rat vascular smooth muscle cells. Pretreatment with TPA had no effect on basal levels of both IP production and [Ca2+]i, whereas it significantly attenuated serotonin-induced increases in both IP production and [Ca2+]i. These data suggest that protein kinase C is involved in the negative feedback control of serotonin-induced rises in both IP production and [Ca2+]i.

Summary Specific binding sites for the endothelium-derived vasoconstrictor endothelin (ET) and its effect on cytosolic free Ca2+ concentrations ([Ca2+]i) were studied in a primary culture of cardiocytes from neonatal rats. Binding studies using 125I-labeled-porcine ET as a radioligand revealed the presence of a single class of high-affinity binding sites for ET in cardiocytes with an apparent Kd of 6−9×10−10 M and a Bmax of 50,000–80,000 sites/cell. Neither various vasoconstrictors nor Ca2+-channel blockers affected the binding. Pretreatment with ET substantially reduced the total number of ET receptors without changing their affinity. ET dose- dependently increased [Ca2+]i in fura-2-loaded cardiocytes. These data indicate that cardiocytes have specific ET receptors that are controlled by a down-regulation mechanism, and that ET induces a receptor-mediated increase in [Ca2+]i in cardiocytes.

Using 125I-labeled-Tyr0-rat(r)-calcitonin gene-related peptide (CGRP), a potent vasodilatory neuropeptide, we have identified and characterized specific binding sites for CGRP in cultured rat vascular smooth muscle cells (VSMC) and bovine endothelial cells (EC). rCGRP and human (h) CGRP equipotently inhibited 125I-rCGRP binding to both cells, but human calcitonin (hCT) was less potent and other unrelated polypeptides were ineffective. Both rCGRP and hCGRP, but not hCT, equally stimulated intracellular cAMP generation in both cells distinct from beta-adrenergic receptor-mediated mechanism, although they had no effect on cGMP generation in either cell or synthesis of prostacyclin in EC. Autoradiograph of affinity-labeled cell membranes revealed that 125I-rCGRP interacts with a single binding component of almost identical molecular size (approximately 60-kDa) in both cells under reducing and nonreducing conditions. The present study demonstrates for the first time the presence of CGRP receptors in cultured VSMC and EC, functionally coupled to adenylate cyclase system distinct from beta-adrenergic receptors. It is suggested that CGRP-induced vasorelaxation may be mediated partly by cAMP-dependent and/or endothelium-dependent mechanism.

Using fura-2-loaded rat vascular smooth muscle cells (VSMCs) in culture, we have attempted to partially purify and characterize yet unidentified factor(s) from normal human plasma that stimulates cytoplasmic free Ca2+ concentration ([Ca2+]i). The plasma extract caused an immediate and transient increase of [Ca2+]i in a dose-dependent manner, of which effect was not prevented by pretreatment with either any of receptor antagonists for -adrenergic agonist, angiotensin II, arginine vasopressin, serotonin, thromboxane A2, or with EGTA and nifedipine. This novel plasma factor(s) was heat-stable and completely inactivated by pronase E, suggesting its protein nature. Furthermore, plasma extracts dose-dependently stimulated the accumulation of [3H] inositol phosphates in rat VSMCs. Sephadex G-50 gel chromatography of plasma extracts resolved one major component (mol wt 13,000) and two minor components with larger (greater than 30,000) and smaller (3,000) mol wt. Present study demonstrates the presence of hitherto unidentified plasma factor(s) with size heterogeneity capable of stimulating both mobilization of [Ca2+]i and breakdown of phosphatidylinositol-4,5-biphosphates in rat VSMCs.

The effect of endothelin (ET), a novel endothelium-derived vasoconstrictive peptide, on DNA synthesis was studied in cultured rat vascular smooth muscle cells (VSMC). ET stimulated incorporation of [3H]thymidine into DNA of the quiescent VSMC in a dose-dependent manner; the approximate half-maximal and maximal stimulation for DNA synthesis was induced with 2 x 10(-10) M and 10(-9) M, respectively. The stimulatory effect by ET on DNA synthesis was completely inhibited by the calcium channel blocker nifedipine. ET combined with epidermal growth factor and transforming growth factor-alpha, but not with platelet-derived growth factor, had synergistic effects. These data indicate that ET is a potent mitogen as well as a constrictor for VSMC, suggesting its potential role in the development of vascular disease.

The effects of H-7 and ML-9, inhibitors of protein kinase C and myosin light-chain kinase, respectively, on DNA synthesis stimulated by platelet-derived growth factor (PDGF) and epidermal growth factor (EGF) were studied in cultured rat vascular smooth muscle cells (VSMC). H-7 and ML-9 significantly inhibited PDGF-stimulated DNA synthesis in lower concentrations, while both compounds were only effective in inhibiting EGF-induced DNA synthesis in higher concentrations. These data suggest that protein kinase C and myosin light-chain kinase activated by PDGF play a more important role in cell proliferation of VSMC than EGF.

The response of cadmium(II) and copper(II) ion-selective electrodes was evaluated in methanol, ethanol, acetonitrile solutions and their mixtures with water. The electrodes showed an almost Nernstian behavior in nonaqueous solutions and their mixtures with water except in acetonitrile.

In order to clarify the characteristics of urinary human epidermal growth factor (hEGF) excretion in the newborn period, we examined hEGF in the 41 newborn infants; 12 healthy preterm infants (group A), 10 healthy full-term infants (group B), 12 full-term infants with neonatal asphyxia (group C), 7 full-term infants treated with tobramycin (group D) during the first week of life. Renal function tests, i.e. creatinine clearance (Ccr), and N-acetyl-β-D-glucosaminidase (NAG) as a parameter of renal tubular damage, were examined in 29 full-term infants. Urinary hEGF excretion showed a linear increase with gestational age. During the first week of life, urinary hEGF excretion in group B increased with age. However, urinary hEGF excretion in group A remained at the constant level through the study period. Ccr in group C was significantly decreased through the study period when compared with that in group B. NAG indices in group C during the first week of life and those in group D on days 5–7 of life were more elevated than those in group B. Urinary hEGF excretion in group C on days 4–7 of life and that in group D on day 7 of life were significantly decreased when compared with that in group B. These results suggest that urinary hEGF excretion is related to the maturation and that the source of urinary hEGF may be renal tubular cells.

Using primary culture of atrial cardiocytes from neonatal rats, we have demonstrated that alpha 1-adrenergic and muscarinic cholinergic agonists have a direct stimulatory effect on secretion of atrial natriuretic factor (ANF) and that ANF secretion is stimulated by phorbol ester, Ca2+ ionophore, high K+-induced depolarization and Ca2+-channel agonists. These data suggest that receptor-mediated mobilization of intracellular Ca2+ and activation of protein kinase C as well as Ca2+ influx via voltage-dependent Ca2+ channels are involved in the secretory mechanism of ANF.

The response of cadmium(II) and copper(II) ion-selective electrodes was evaluated in methanol, ethanol, acetonitrile solutions and their mixtures with water. The electrodes showed an almost Nernstian behavior in nonaqueous solutions and their mixtures with water except in acetonitrile.