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We investigate the low-temperature magnetic properties of the molecule-based chiral spin chain [Cu(pym)(H_{2}O)_{4}]SiF_{6}·H_{2}O (pym=pyrimidine). Electron-spin resonance, magnetometry and heat capacity measurements reveal the presence of staggered g tensors, a rich low-temperature excitation spectrum, a staggered susceptibility, and a spin gap that opens on the application of a magnetic field. These phenomena are reminiscent of those previously observed in nonchiral staggered chains, which are explicable within the sine-Gordon quantum-field theory. In the present case, however, although the sine-Gordon model accounts well for the form of the temperature dependence of the heat capacity, the size of the gap and its measured linear field dependence do not fit with the sine-Gordon theory as it stands. We propose that the differences arise due to additional terms in the Hamiltonian resulting from the chiral structure of [Cu(pym)(H_{2}O)_{4}]SiF_{6}·H_{2}O, particularly a uniform Dzyaloshinskii-Moriya coupling and a fourfold periodic staggered field.

Cyclin-dependent kinase (CDK)-activating phosphatases, CDC25A and CDC25B, are labile proteins, and their levels vary in a cell cycle-dependent manner. Immediate-early response IER5 protein negatively regulates the cellular CDC25B levels, and stress-induced IER5 expression potentiates G2/M arrest. IER5 binds to protein phosphatase PP2A and regulates the PP2A substrate specificity. We show that IER5 binds to CDC25B and assists PP2A to convert CDC25B to hypophosphorylated forms. Hypophosphorylation at Ser323 results in the dissociation of CDC25B from 14-3-3 phospho-binding proteins. In IER5 expressing cells, CDC25B dissociated from 14-3-3 is unstable but slightly activated, because 14-3-3 inhibits CDC25B polyubiquitination and CDC25B binding to CDK1. The 14-3-3 binding to CDC25A also impedes CDC25A degradation and CDC25A-CDK2 interaction. We propose that 14-3-3 is an important regulator of CDC25A and CDC25B and that PP2A/IER5 controls the stability and activity of CDC25B through regulating the interaction of CDC25B and 14-3-3., (Copyright © 2018 Elsevier Inc. All rights reserved.)

The specific heat of the Kondo insulator YbB_{12} has been measured up to 60 T. The Sommerfeld coefficient γ significantly increases at around 50 T, where the insulator metal transition occurs with a steep increase of the magnetization. γ reaches 67  mJ/(mol K^{2}) at high fields, which directly indicates that the quasiparticles gain a heavy thermodynamic effective mass and transform into a Kondo metal under magnetic fields. The field-induced Kondo metal has a rather high Kondo temperature around 200 K. The strong Kondo coupling proves that the energy gap collapse does not correspond to the breakdown of the Kondo bound state. The steep increase of the magnetization at the transition manifests the sharp density of states at the Fermi energy formed via the Kondo resonance.

We report on magnetization, sound-velocity, and magnetocaloric-effect measurements of the Ising-like spin-1/2 antiferromagnetic chain system BaCo_{2}V_{2}O_{8} as a function of temperature down to 1.3 K and an applied transverse magnetic field up to 60 T. While across the Néel temperature of T_{N}∼5  K anomalies in magnetization and sound velocity confirm the antiferromagnetic ordering transition, at the lowest temperature the field-dependent measurements reveal a sharp softening of sound velocity v(B) and a clear minimum of temperature T(B) at B_{⊥}^{c,3D}=21.4  T, indicating the suppression of the antiferromagnetic order. At higher fields, the T(B) curve shows a broad minimum at B_{⊥}^{c}=40  T, accompanied by a broad minimum in the sound velocity and a saturationlike magnetization. These features signal a quantum phase transition, which is further characterized by the divergent behavior of the Grüneisen parameter Γ_{B}∝(B-B_{⊥}^{c})^{-1}. By contrast, around the critical field, the Grüneisen parameter converges as temperature decreases, pointing to a quantum critical point of the one-dimensional transverse-field Ising model.

Growth arrest and DNA damage-inducible 45 (GADD45) family genes encode related proteins, including GADD45α, GADD45β, and GADD45γ. In HeLa cells, expression of GADD45 members is differentially regulated under a variety of environmental conditions, but thermal and genotoxic stresses induce the expression of all genes. The heat shock response of GADD45β is mediated by the heat shock transcription factor 1 (HSF1), and GADD45β is necessary for heat stress survival. Heat and genotoxic stress-induced activation of c-Jun N-terminal kinase (JNK) is suppressed by the expression of GADD45 proteins. GADD45 proteins bind the JNK kinase mitogen-activated protein kinase kinase 7 (MKK7) and inhibit its activity, even under normal physiological conditions. Our findings indicate that GADD45 essentially suppresses the MKK7-JNK pathway and suggest that differentially expressed GADD45 family members fine-tune stress-inducible JNK activity., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Although high-throughput sequencing can elucidate the genetic basis of hereditary cardiomyopathy, direct interventions targeting pathological mutations have not been established. Furthermore, it remains uncertain whether homology-directed repair (HDR) is effective in non-dividing cardiomyocytes. Here, we demonstrate that HDR-mediated genome editing using CRISPR/Cas9 is effective in non-dividing cardiomyocytes. Transduction of adeno-associated virus (AAV) containing sgRNA and repair template into cardiomyocytes constitutively expressing Cas9 efficiently introduced a fluorescent protein to the C-terminus of Myl2. Imaging-based sequential evaluation of endogenously tagged protein revealed that HDR occurs in cardiomyocytes, independently of DNA synthesis. We sought to repair a pathological mutation in Tnnt2 in cardiomyocytes of cardiomyopathy model mice. An sgRNA that avoided the mutated exon minimized deleterious effects on Tnnt2 expression, and AAV-mediated HDR achieved precise genome correction at a frequency of ~12.5%. Thus, targeted genome replacement via HDR is effective in non-dividing cardiomyocytes, and represents a potential therapeutic tool for targeting intractable cardiomyopathy.

Turmeric ( Curcuma longa ) is a widely used spice that has various biological effects, and aqueous extracts of turmeric exhibit potent antioxidant activity and anti-inflammatory activity. Bisacurone, a component of turmeric extract, is known to have similar effects. Oxidative stress and inflammatory cytokines play an important role in ethanol-induced liver injury. This study was performed to evaluate the influence of a hot water extract of C. longa (WEC) or bisacurone on acute ethanol-induced liver injury. C57BL/6 mice were orally administered WEC (20 mg/kg body weight; BW) or bisacurone (60 µg/kg BW) at 30 min before a single dose of ethanol was given by oral administration (3·0 g/kg BW). Plasma levels of aspartate aminotransferase and alanine aminotransferase were markedly increased in ethanol-treated mice, while the increase of these enzymes was significantly suppressed by prior administration of WEC. The increase of alanine aminotransferase was also significantly suppressed by pretreatment with bisacurone. Compared with control mice, animals given WEC had higher hepatic tissue levels of superoxide dismutase and glutathione, as well as lower hepatic tissue levels of thiobarbituric acid-reactive substances, TNF-α protein and IL-6 mRNA. These results suggest that oral administration of WEC may have a protective effect against ethanol-induced liver injury by suppressing hepatic oxidation and inflammation, at least partly through the effects of bisacurone.

The crystal structures of NiX2(pyz)2 (X = Cl (1), Br (2), I (3), and NCS (4)) were determined by synchrotron X-ray powder diffraction. All four compounds consist of two-dimensional (2D) square arrays self-assembled from octahedral NiN4X2 units that are bridged by pyz ligands. The 2D layered motifs displayed by 1-4 are relevant to bifluoride-bridged [Ni(HF2)(pyz)2]EF6 (E = P, Sb), which also possess the same 2D layers. In contrast, terminal X ligands occupy axial positions in 1-4 and cause a staggered packing of adjacent layers. Long-range antiferromagnetic (AFM) order occurs below 1.5 (Cl), 1.9 (Br and NCS), and 2.5 K (I) as determined by heat capacity and muon-spin relaxation. The single-ion anisotropy and g factor of 2, 3, and 4 were measured by electron-spin resonance with no evidence for zero-field splitting (ZFS) being observed. The magnetism of 1-4 spans the spectrum from quasi-two-dimensional (2D) to three-dimensional (3D) antiferromagnetism. Nearly identical results and thermodynamic features were obtained for 2 and 4 as shown by pulsed-field magnetization, magnetic susceptibility, as well as their Néel temperatures. Magnetization curves for 2 and 4 calculated by quantum Monte Carlo simulation also show excellent agreement with the pulsed-field data. Compound 3 is characterized as a 3D AFM with the interlayer interaction (J⊥) being slightly stronger than the intralayer interaction along Ni-pyz-Ni segments (J(pyz)) within the two-dimensional [Ni(pyz)2](2+) square planes. Regardless of X, J(pyz) is similar for the four compounds and is roughly 1 K.

We describe the construction of a simple, compact, and cost-effective feedback system that produces flat-top field profiles in pulsed magnetic fields. This system is designed for use in conjunction with a typical capacitor-bank driven pulsed magnet and was tested using a 60-T pulsed magnet. With the developed feedback controller, we have demonstrated flat-top magnetic fields as high as 60.64 T with an excellent field stability of ±0.005 T. The result indicates that the flat-top pulsed magnetic field produced features high field stability and an accessible field strength. These features make this system useful for improving the resolution of data with signal averaging.

Conventional, thermally driven continuous phase transitions are described by universal critical behavior that is independent of the specific microscopic details of a material. However, many current studies focus on materials that exhibit quantum-driven continuous phase transitions (quantum critical points, or QCPs) at absolute zero temperature. The classification of such QCPs and the question of whether they show universal behavior remain open issues. Here we report measurements of heat capacity and de Haas-van Alphen (dHvA) oscillations at low temperatures across a field-induced antiferromagnetic QCP (Bc0 ≈ 50 T) in the heavy-fermion metal CeRhIn5. A sharp, magnetic-field-induced change in Fermi surface is detected both in the dHvA effect and Hall resistivity at B0* ≈ 30 T, well inside the antiferromagnetic phase. Comparisons with band-structure calculations and properties of isostructural CeCoIn5 suggest that the Fermi-surface change at B0* is associated with a localized-to-itinerant transition of the Ce-4f electrons in CeRhIn5. Taken in conjunction with pressure experiments, our results demonstrate that at least two distinct classes of QCP are observable in CeRhIn5, a significant step toward the derivation of a universal phase diagram for QCPs.

Objectives: We treated patients experiencing drug-eluting stent (DES) restenosis with plain old balloon angioplasty (POBA), implantation of the same type of DES [homogeneous drug-eluting stent (HOMO-DES)], or implantation of a different type of DES [heterogeneous drug-eluting stent (HETERO-DES)], and compared the efficacy and safety of these procedures for the prevention of repeated in-stent restenosis (ISR)., Background: In patients with de-novo coronary lesions, DES implantation is associated with a markedly reduced restenosis rate as compared with that associated with a bare metal stent and POBA. However, the optimal management strategy for patients with DES ISR remains unknown., Patients and Methods: We identified 191 consecutive DES ISR lesions from 183 patients who required clinically driven revascularization and divided them into three groups according to the treatment: 38 lesions were treated with POBA, 38 with HOMO-DES, and 115 with HETERO-DES., Results: The incidence of target lesion revascularization (TLR) was 42.1% (16/38), 15.8% (6/38), and 16.5% (19/115) in the POBA, HOMO-DES, and HETERO-DES groups (POBA vs. HOMO, HETERO-DES; P=0.002, respectively). Multivariate analysis indicated that diabetes [odds ratio (OR), 3.4], hemodialysis (OR, 7.74), nonfocal ISR patterns (OR, 3.35), previous myocardial infarction (OR, 3.26), and POBA (OR, 8.84) were independent predictors of TLR., Conclusion: A strategy involving repeated DES implantation was superior to POBA for preventing recurrent restenosis. Treatment with a different type or generation of DES does not appear to reduce the incidence of TLR. Moreover, we identified certain useful factors for facilitating appropriate and early triage in the patients with repeated DES ISR.

Alkaline phosphatases (APs) are a family of dimeric metalloenzymes that has been utilized in many areas due to its ability to hydrolyze a variety of phosphomonoesters. While mammalian APs have higher specific activity than prokaryotic APs, they are generally less thermostable. To cultivate the possibility to confer mammalian APs with higher thermostability as well as high activity, we focused on human AP isozymes. Among the four isozymes of human APs, placental AP (PLAP) retains the highest thermostability, while intestinal AP (IAP) has the highest specific activity. Since the two APs display high homology, a series of chimeric enzymes were made in a secreted form to analyze their properties. Surprisingly, chimeric APs with IAP residues at the N-terminal and PLAP residues at the C-terminal regions showed higher specific activity than PLAP, while keeping thermostability as high as PLAP. Especially, one showed similar specific activity to IAP, while showing slower inactivation than PLAP after incubation at 75°C. Interestingly, the mutant also showed higher resistance to uncompetitive inhibitors Phe and Leu than their parent enzymes, possibly due to increased hydrophilicity of the active site entrance residues. The obtained chimera will be useful as a novel reporter in various assays including gene hybridization.

The motion of electrons in a solid has a profound effect on its topological properties and may result in a nonzero Berry's phase, a geometric quantum phase encoded in the system's electronic wave function. Despite its ubiquity, there are few experimental observations of Berry's phase of bulk states. Here, we report detection of a nontrivial π Berry's phase in the bulk Rashba semiconductor BiTeI via analysis of the Shubnikov-de Haas (SdH) effect. The extremely large Rashba splitting in this material enables the separation of SdH oscillations, stemming from the spin-split inner and outer Fermi surfaces. For both Fermi surfaces, we observe a systematic π-phase shift in SdH oscillations, consistent with the theoretically predicted nontrivial π Berry's phase in Rashba systems.

Purpose: The purpose of this study was to identify predictors of contrast-induced acute kidney injury (CI-AKI) and the effect of CI-AKI on cardiovascular outcomes after hospital discharge in patients with ST-segment elevation myocardial infarction (STEMI) treated with primary percutaneous coronary intervention (PCI)., Methods and Materials: We retrospectively reviewed 194 STEMI consecutive patients who underwent primary PCI to evaluate the predictors for CI-AKI and 187 survivors to examine all-cause mortality and cardiovascular events. Outcomes were compared between patients with CI-AKI and those without CI-AKI, which was defined as an increase >50% or >0.5mg/dl in serum creatinine concentration within 48hours after primary PCI., Results: CI-AKI occurred in 23 patients (11.9%). Multivariate analysis identified pre-procedural renal insufficiency as a predictor of CI-AKI, and this predictor was independent from hemodynamic instability and excessive contrast volume. Receiver-operator characteristics analysis demonstrated that patients with an estimated glomerular filtration rate (eGFR) of ≤43.6ml/min per 1.73m(2) had the potential for CI-AKI. Patients who developed CI-AKI had higher mortality and cardiovascular events than did those without CI-AKI (27.8% vs. 4.7%; log-rank P=.0003, 27.8% vs. 11.2%; log-rank P=.0181, respectively). Cox proportional hazards model analysis identified CI-AKI as the independent predictor of mortality and cardiovascular events [hazard ratio [HR]=5.36; P=.0076, HR=3.10; P=.0250, respectively]., Conclusions: The risk of CI-AKI is increased in patients with pre-procedural renal insufficiency, and eGFR is clinically useful in the emergent setting for CI-AKI risk stratification before primary PCI., (© 2013 Elsevier Inc. All rights reserved.)

Magneto-caloric effects (MCEs) measurement system in adiabatic condition is proposed to investigate the thermodynamic properties in pulsed magnetic fields up to 55 T. With taking the advantage of the fast field-sweep rate in pulsed field, adiabatic measurements of MCEs were carried out at various temperatures. To obtain the prompt response of the thermometer in the pulsed field, a thin film thermometer is grown directly on the sample surfaces. The validity of the present setup was demonstrated in the wide temperature range through the measurements on Gd at about room temperature and on Gd3Ga5O12 at low temperatures. The both results show reasonable agreement with the data reported earlier. By comparing the MCE data with the specific heat data, we could estimate the entropy as functions of magnetic field and temperature. The results demonstrate the possibility that our approach can trace the change in transition temperature caused by the external field.

The magnetic ground state of Sr2CrO4 with a distorted honeycomb lattice was investigated by means of measurements of the magnetic susceptibility, high-field magnetization process, and electron spin resonance (ESR). Antiferromagnetic ordering was observed clearly below TN = 3.2 K, while the magnetic ground state had been thought to be a spin-singlet state. A two-sublattice model with biaxial anisotropy was applicable for the observed ESR modes. Plateau-like behavior and a sharp kink were detected in the magnetization curve.

BiCu(2)PO(6) is a frustrated two-leg spin-ladder compound with a spin gap that can be closed with a magnetic field of approximately 20 T. This quantum phase transition and its related phase diagram as a function of magnetic field and temperature (H, T) are investigated up to 60 T by means of specific heat, magnetocaloric effect, magnetization, and magnetostriction measurements. In contrast to other gapped quantum magnets, BiCu(2)PO(6) undergoes a series of unexpected first- and second-order phase transitions when an external magnetic field is applied along the crystallographic c axis. The application of a magnetic field along the b axis induces two second-order phase transitions. We propose that the anisotropy and complex phase diagram result from the interplay between strong geometrical frustration and spin-orbit interaction necessary for the description of this fascinating magnetic system.

[Ni(HF(2))(3-Clpy)(4)]BF(4) (py = pyridine) is a simple one-dimensional (1D) coordination polymer composed of compressed NiN(4)F(2) octahedra that form chains with bridging HF(2)(-) ligands. In spite of significant distortion of the HF(2)(-) bridge, a quasi-1D antiferromagnetic (AFM) behavior was observed with J(FHF) = 4.86 K.

[Ni(HF(2))(pyz)(2)]X {pyz = pyrazine; X = PF(6)(-) (1), SbF(6)(-) (2)} were structurally characterized by synchrotron X-ray powder diffraction and found to possess axially compressed NiN(4)F(2) octahedra. At 298 K, 1 is monoclinic (C2/c) with unit cell parameters, a = 9.9481(3), b = 9.9421(3), c = 12.5953(4) Å, and β = 81.610(3)° while 2 is tetragonal (P4/nmm) with a = b = 9.9359(3) and c = 6.4471(2) Å and is isomorphic with the Cu-analogue. Infinite one-dimensional (1D) Ni-FHF-Ni chains propagate along the c-axis which are linked via μ-pyz bridges in the ab-plane to afford three-dimensional polymeric frameworks with PF(6)(-) and SbF(6)(-) counterions occupying the interior sites. A major difference between 1 and 2 is that the Ni-F-H bonds are bent (∼157°) in 1 but are linear in 2. Ligand field calculations (LFT) based on an angular overlap model (AOM), with comparison to the electronic absorption spectra, indicate greater π-donation of the HF(2)(-) ligand in 1 owing to the bent Ni-F-H bonds. Magnetic susceptibility data for 1 and 2 exhibit broad maxima at 7.4 and 15 K, respectively, and λ-like peaks in dχT/dT at 6.2 and 12.2 K that are ascribed to transitions to long-range antiferromagnetic order (T(N)). Muon-spin relaxation and specific heat studies confirm these T(N)'s. A comparative analysis of χ vs T to various 1D Heisenberg/Ising models suggests moderate antiferromagnetic interactions, with the primary interaction strength determined to be 3.05/3.42 K (1) and 5.65/6.37 K (2). However, high critical fields of 19 and 37.4 T obtained from low temperature pulsed-field magnetization data indicate that a single exchange constant (J(1D)) alone is insufficient to explain the data and that residual terms in the spin Hamiltonian, which could include interchain magnetic couplings (J(⊥)), as mediated by Ni-pyz-Ni, and single-ion anisotropy (D), must be considered. While it is difficult to draw absolute conclusions regarding the magnitude (and sign) of J(⊥) and D based solely on powder data, further support offered by related Ni(II)-pyz compounds and our LFT and density-functional theory (DFT) results lead us to a consistent quasi-1D magnetic description for 1 and 2., (© 2011 American Chemical Society)

Several quantum paramagnets exhibit magnetic-field-induced quantum phase transitions to an antiferromagnetic state that exists for H c1 ≤ H ≤ H c2. For some of these compounds, there is a significant asymmetry between the low- and high-field transitions. We present specific heat and thermal conductivity measurements in NiCl2-4SC(NH2)2, together with calculations which show that the asymmetry is caused by a strong mass renormalization due to quantum fluctuations for H ≤ H c1 that are absent for H ≥ H c2. We argue that the enigmatic lack of asymmetry in thermal conductivity is due to a concomitant renormalization of the impurity scattering.

A new calorimeter for measurements of the heat capacity and magnetocaloric effect of small samples in pulsed magnetic fields is discussed for the exploration of thermal and thermodynamic properties at temperatures down to 2 K. We tested the method up to μ(0)H=50 T, but it could be extended to higher fields. For these measurements we used carefully calibrated bare-chip Cernox(®) and RuO(2) thermometers, and we present a comparison of their performances. The monotonic temperature and magnetic field dependences of the magnetoresistance of RuO(2) allow thermometry with a precision as good as ±4 mK at T=2 K. To test the performance of our calorimeter, heat capacity and magnetocaloric effect for the spin-dimer compound Sr(3)Cr(2)O(8) and the triangular lattice antiferromagnet RbFe(MoO(4))(2) are presented.

Single crystals of the spin dimer system Sr(3)Cr(2)O(8) have been grown for the first time. Magnetization, heat capacity, and magnetocaloric effect data up to 65 T reveal magnetic order between applied fields of H(c1) approximately 30.4 T and H(c2) approximately 62 T. This field-induced order persists up to T(c)(max) approximately 8 K at H approximately 44 T, the highest observed in any quantum magnet where H(c2) is experimentally accessible. We fit the temperature-field phase diagram boundary close to H(c1) using the expression T(c) = A(H-H(c1))(nu). The exponent nu = 0.65(2), obtained at temperatures much smaller than T(c)(max), is that of the 3D Bose-Einstein condensate (BEC) universality class. This finding strongly suggests that Sr(3)Cr(2)O(8) is a new realization of a triplon BEC where the universal regimes corresponding to both H(c1) and H(c2) are accessible at (4)He temperatures.

We report the Hall resistivity, ρ(xy), of polycrystalline SmFeAsO(1-x)F(x) for four different fluorine concentrations from the onset of superconductivity through the collapse of the structural phase transition. For the two more highly doped samples, ρ(xy) is linear in magnetic field up to 50 T with only weak temperature dependence, reminiscent of a simple Fermi liquid. For the lightly doped samples with x<0.15, we find a low temperature regime characterized as ρ(xy)(H) being both nonlinear in magnetic field and strongly temperature-dependent even though the Hall angle is small. The onset temperature for this nonlinear regime is in the vicinity of the structural phase (SPT)/magnetic ordering (MO) transitions. The temperature dependence of the Hall resistivity is consistent with a thermal activation of carriers across an energy gap. The evolution of the energy gap with doping is reported.

From specific heat measurements in high quality H2@C60 samples performed over a broad temperature range, we obtain the smallest yet observed splitting of rotational energy sublevels of encapsulated single H2 molecules, 0.1-0.2 meV, in the nearly spherical potential well provided by highly isotropic C60 cages. Additionally, we find evidence of the quantized oscillation state of isolated H2 in the C60 cage. The minuscule splitting indicates that H2@C60 provides unprecedented opportunities to study free-molecule quantum dynamic properties.

Ba&#95;{3}Mn&#95;{2}O&#95;{8} is a spin-dimer compound based on pairs of S = 1, 3d;{2}, Mn;{5+} ions arranged on a triangular lattice. Antiferromagnetic intradimer exchange leads to a singlet ground state in zero field, with excited triplet and quintuplet states at higher energy. High field thermodynamic measurements are used to establish the phase diagram, revealing a substantial asymmetry of the quintuplet condensate. This striking effect, all but absent for the triplet condensate, is due to a fundamental asymmetry in quantum fluctuations of the paramagnetic phases near the various critical fields.

We present resistivity measurements of the complete superconducting upper critical field (H{c2}) phase diagram as a function of angle (theta) and temperature (T) for cobalt-doped SrFe2As2 epitaxial films to 0.5 K and 50 T. Although H{c2}(theta) at 10 K is indistinguishable from that derived from a single-band anisotropy model, the apparent anisotropy H{c2}{ perpendicularc}/H{c2};{ parallelc} linearly decreases to 1 at low T, with H{c2}(0)=47 T. The data are well described by a two-band model with small, opposing anisotropies for the bands. This unusual relationship is confirmed by the observation of a local maximum for H{c2};{ parallelc} at low T.

An athymic mouse-derived immature T-cell clone, N-9F, was not maintained by interleukin-2 alone but required another soluble factor, contained in concanavalin A-stimulated rat splenocyte culture supernatant, namely T cell growth factor (TCGF), for its proliferation. An N-9F-proliferation factor (NPF) was isolated in a pure form from TCGF. N-9F cells and immature thymocytes proliferated in the presence of N-9F at 10(-12)-10(-9)M in a dose-dependent manner, but adult thymocytes were not stimulated by NPF. NPF increased DNA synthesis of N-9F. NPF increased CD4 and CD8 double negative, single positive and double positive thymocytes in fetal thymus organ culture. A hamster anti-NPF antiserum possessing the capacity to neutralize N-9F proliferation activity of NPF neutralized the increasing effect of NPF on immature thymocytes. All effects of NPF was inhibited by mAb QR6.6 to recognize a 100 kDa surface molecule of N-9F. The amino-terminal 20 amino acid sequence of NPF was identified and identical to that of rat saposin A. The apparent molecular weight of NPF, 16000, was comparable to that of saposin A. A Hitrap-mouse recombinant His-tag-saposin A antibody column bound NPF, pulled down the NPF activity in TCGF, and the antibody recognized a 16kDa molecule in western-blotting of TCGF. Thus, NPF in TCGF was a saposin A-like protein possessing the capacity for growth and differentiation of immature thymocytes. The physiological significance of NPF in the growth and differentiation of immature thymocytes was discussed in view of the characteristic distributions of NPF and the molecule recognized by its mAb QR6.6 in fetal thymi.

Fetal liver (FL) hematopoiesis is thought to be important for expanding the cell number during ontogeny. In order to investigate the cellular interaction molecules among FL stromal and hematopoietic cells, we established a monoclonal antibody, Ndk-10, that reacts with FL stromal cells but not with dish non-adherent cells. When Ndk-10 was added to an FL stromal and hematopoietic cell-coculture, it inhibited the survival of c-kit+ cells. The inhibitory activity of Ndk-10 was also observed in the fetal liver organ culture. The Ndk-10 recognized a 150 kD molecule in the adherent cells of FL and kidney, and the N-terminal amino acid sequence was identical to that of mouse aminopeptidase N/CD13. The peptidase activity of CD13 was inhibited by Ndk-10, and addition of its specific inhibitor resulted in the same inhibitory activity as Ndk-10. We propose that aminopeptidase N/CD13 is a critical molecule that regulates the survival of c-kit+ cells in the FL microenvironment.

A novel monoclonal antibody, SM/C-2.6, specific for mouse muscle satellite cells was established. SM/C-2.6 detects mononucleated cells beneath the basal lamina of skeletal muscle, and the cells co-express M-cadherin. Single fiber analyses revealed that M-cadherin+ mononucleated cells attaching to muscle fibers are stained with SM/C-2.6. SM/C-2.6+ cells, which were freshly purified by FACS from mouse skeletal muscle, became MyoD+ in vitro in proliferating medium, and the cells differentiated into desmin+ and nuclear-MyoD+ myofibers in vitro when placed under differentiation conditions. When the sorted cells were injected into mdx mouse muscles, donor cells differentiated into muscle fibers. Flow cytometric analyses of SM/C-2.6+ cells showed that the quiescent satellite cells were c-kit-, Sca-1-, CD34+, and CD45-. More, SM/C-2.6+ cells were barely included in the side population but in the main population of cells in Hoechst dye efflux assay. These results suggest that SM/C-2.6 identifies and enriches quiescent satellite cells from adult mouse muscle, and that the antibody will be useful as a powerful tool for the characterization of cellular and molecular mechanisms of satellite cell activation and proliferation.

We examined the severity of experimental colitis induced by dextran sulfate sodium (DSS) using immunologically manipulated mice. C57BL/6 mice showed more severe colitis than BALB/c mice, but mice of both strains recovered fully from the disease after the removal of DSS from their drinking water. The infiltrated cells at the lesions were mainly granulocytes in normal littermates. However, C.B-17 scid, IL-7Ralpha deficient, and TCR-Cbetadelta double-deficient mice showed severe colitis and did not recover from the disease even after the removal of DSS. It was found that the infiltrated cells at the lesions in the lethal strains were monocytes. Although both TCR-Cdelta(-/-) and TCR-Cbeta(-/-) mice showed severe colitis phenotypes, infiltration in the former is monocyte-dominant while that in the latter is granulocyte-dominant. Thus the type of cells that infiltrate at the lesions of DSS-induced experimental colitis may be controlled by functional T cell subsets. Immunohistological and RT-PCR analyses of the inflamed colon revealed that the murine homologue of human GROalpha released by some cells under the control of gammadeltaT cells is a possible candidate determining the severity of DSS-induced experimental colitis.

An athymic mouse-derived immature T-cell clone, N-9F, was not maintained by interleukin-2 alone but required another soluble factor, contained in concanavalin A-stimulated rat splenocyte culture supernatant, namely T cell growth factor (TCGF), for its proliferation. An N-9F-proliferation factor (NPF) was isolated in a pure form from TCGF. N-9F cells and immature thymocytes proliferated in the presence of NPF at 10-11-10-8 g/ml in a dose-dependent manner, but adult thymocytes were not stimulated by NPF. NPF increased DNA synthesis of N-9F. NPF increased CD4 and CD8 double negative thymocytes and CD8 single positive thymocytes in fetal thymus organ culture. A hamster anti-NPF antiserum possessing the capacity to neutralize N-9F proliferation activity of NPF decreased double negative thymocytes. The amino-terminal amino acid sequence of NPF was identified to be Ser-Leu-Pro-Cys-Asp-Ile-Cys-Lys-Thr-Val-Val-Thr-Glu-Ala-Cys-Asn-Leu-Leu-Lys-Asp- and was identical to that of rat saposin A. The apparent molecular weight of NPF, 16000, was comparable to that of saposin A. A rabbit anti-mouse recombinant His-tag (mrH)-saposin A antibody recognized a 16000 MW molecule in TCGF. A Hitrap-saposin A antibody column bound NPF and pulled down the NPF activity in TCGF. Thus, NPF in TCGF was a saposin A-like protein possessing the capacity for growth and differentiation of immature thymocytes.

We have cloned a human counterpart to a guinea pig STE20-like kinase cDNA, designated human SLK (hSLK), from a human lung carcinomatous cell line A549 cDNA library. hSLK cDNA encodes a novel 1204 amino acid serine/threonine kinase for which the kinase domain located at the N-terminus shares considerable homology to that of the STE20-like kinase family. The C-terminal domain of hSLK includes both the coiled-coil structure and four Pro/Glu/Ser/Thr-rich (PEST) sequences, but not the GTPase-binding domain (GBD) that is characteristic of the p21-activated kinase (PAK) family, polyproline consensus binding sites, or the Leu-rich domain seen in the group I germinal center kinases (GCKs). Northern blot analysis indicated that hSLK was ubiquitously expressed. hSLK overexpressed in COS-7 cells phosphorylates itself as well as myelin basic protein used as a substrate. On the other hand, hSLK cannot activate any of the three well-characterized mitogen-activated protein kinase MAPK (ERK, JNK/SAPK and p38) pathways. Moreover, hSLK kinase activity is not upregulated by constitutive active forms of GTPases (RasV12, RacV12 and Cdc42V12). These structural and functional properties indicate that hSLK should be considered to be a new member of group II GCKs.

Background: Immunotherapy for cedar pollinosis makes use of multiple injections of allergens, but its effectiveness remains controversial. Recent studies indicate that immunization with certain protein antigens and immunostimulatory DNA sequence (ISS) oligodeoxynucleotides (ODNs) represent a potential approach to allergen-specific immunotherapy., Objective: We determined whether the coadministration of 2 major protein allergens, Cry j 1 and Cry j 2, of Japanese cedar pollen and ISS-ODN (5'-TGACTCTGAACGTTCGAGATGA-3') improves the immune responses induced by protein allergens in BALB/c mice., Methods: Mice were primed intradermally with allergens or ISS-ODN in saline solution and boosted with allergens in alum, and other mice were primed with allergens in alum and boosted with allergens/ISS-ODN. Allergen-specific IgG2a and IgG1 antibody responses were measured by means of ELISA in sera after ODN injection, and allergen-specific IgE antibody production was measured by the passive cutaneous anaphylaxis reaction. IFN-gamma and IL-4 releases were also measured by ELISA in the supernatants of allergen-stimulated spleen cells., Results: The coadministration of allergens/ISS-ODN increased IgG2a titers and IFN-gamma release in both groups of mice, whereas it decreased IgG1 titers and IL-4 release in comparison with control mice injected with allergens/mutant ODN. The coadministration additionally inhibited IgE antibody production., Conclusion: The data demonstrate that the coadministration of cedar pollen allergens and ISS-ODNs before secondary T(H2) and IgE responses or during ongoing primary T(H2) and IgE responses brings about a T(H1)-shifted immune response and inhibition of IgE antibody production, suggesting that this coadministration strategy may provide a novel type of immunotherapy for cedar pollinosis.

Green fluorescent protein (GFP) transgenic (GFP+) mice express GFP in most tissues except erythrocytes and hair. Immune responses of GFP+ mouse and their application to studies of lymphocyte development were investigated. Flow cytometric analyses revealed that differentiation patterns of lymphocytes from GFP+ mice are equivalent to those from parental C57BL/6 mice. There was no difference in mature T-cell proliferative ability in response to allogeneic stimulator cells or anti-CD3epsilon stimulation between GFP+ and C57BL/6 mice. Furthermore, the anti-OVA antibody response of GFP+ mice was also the same as that of C57BL/6 mice. Taken together, these results show no immunological differences between GFP+ and C57BL/6 mice. Bone marrow transplantation and in vitro thymus reconstitution experiments were performed in an attempt to apply the GFP+ mice to the analysis of lymphocyte development. When bone marrow cells from GFP+ mice were transplanted. T and B lymphocytes containing GFP developed normally in scid recipients. Next we examined intrathymic T-cell development by hanging drop culture methods. GFP+ and CD4+8+ immature T-cells developed normally from bone marrow cells in the reconstituted thymus. The experimental system using hematopoietic cells from GFP+ mice is a powerful tool for visualizing lymphocyte development.

Objective: We present two cases of common carotid artery occlusion that were treated by vascular reconstruction using the transverse cervical artery., Methods: Two patients with common carotid artery occlusion presented with transient ischemic attacks resulting from decreased cerebral blood flow on the affected side. Both patients underwent vascular reconstruction using the transverse cervical artery. The transverse cervical artery was anastomosed to the ipsilateral external carotid artery at its origin, as a pedicle graft. A superficial temporal artery-middle cerebral artery anastomosis was then performed., Results: The postoperative courses were uneventful. The transverse cervical artery bypass grafts were patent, and cerebral blood flow increased to normal levels., Conclusion: Transverse cervical artery grafting provides a less tedious alternative to saphenous vein interposition grafting for revascularization of the internal carotid artery domain.

26,26,26,27,27,27-Hexafluoro-1,25-dihydroxyvitamin D3 (ST-630) is a newly developed agent to maintain the levels of calcium and phosphorus in blood. Herein, we investigated the effect of this compound on the expression of vitamin-D-responsive genes in vitamin-D-deficient mice. ST-630 was more effective than 1, 25-dihydroxyvitamin D3 [1,25(OH)2D3] with respect to the induction of Cyp24 and calbindin-D9k mRNAs in the kidney and in the small intestine. Moreover, the increase in mRNA levels of vitamin-D-responsive genes induced by ST-630 lasted longer than that induced by 1,25(OH)2D3. These results indicate that ST-630 was more effective in inducing Cyp24 and calbindin-D9k gene expression than 1, 25(OH)2D3 when both compounds were injected into vitamin-D-deficient mice.

Protein kinases play a key role in cell growth and differentiation. We have isolated the cDNA of a novel protein serine/threonine kinase (referred to as STE20-like kinase (SLK) from a guinea pig liver cDNA library with a probe generated by a cloning approach based on the polymerase chain reaction. The encoded polypeptide (1231 amino acids, M(r) 141,079) contains all conserved subdomains characteristic of the protein serine threonine kinase family. A hemagglutinin-tagged SLK expressed artificially in COS7 cells was hyperphosphorylated by anisomycin. By Northern blot analysis, SLK mRNA was detected in all organs examined: brain, lung heart, liver, kidney, spleen, testis, and eosinophils. Sequence comparisons of its catalytic domain related SLK to p21-activated kinase family of protein serine/threonine kinases. Its noncatalytic domain comprises several intriguing structural features, including the acidic region and the nuclear targeting sequence. This noncatalytic domain exhibited no extended similarity with other proteins. Thus, SLK is a protein serine/threonine kinase which contains an unknown regulatory domain(s).

cDNA fragments encoding mouse ferredoxin and ferredoxin reductase were simultaneously introduced into COS7 cells by using an expression vector, pUC-SR alpha plasmid. When using the mitochondrial fraction prepared from the transfected cells, cytochrome-c reductase activity was detected. This activity was highest when 7.5 micrograms of the ferredoxin expression plasmid (pSR alpha F) and 2.5 micrograms of the ferredoxin reductase expression plasmid (pSR alpha FR) were transfected into COS7 cells. In this system, NADPH could be replaced by NADH as a cofactor for the reduction of cytochrome-c although the cytochrome-c reductase was more dependent on NADPH than NADH at a low concentration. When CYP24 expression plasmid was transfected into COS7 cells along with both pSR alpha F and pSR alpha FR, the transfected cells revealed a 3-fold higher 25-hydroxyvitamin D3-24-hydroxylase activity than COS7 cells transfected with CYP24 expression plasmid.

The effect of a cytotoxic substance (bis(2-hydroxyethyl) trisulfide; BS-1), isolated from Bacillus stearothermophilus UK563, on messenger ribonucleic acid (mRNA) level in mouse macrophage-like J774A.1 cells was investigated by the method of differential display. The treatment of J774A.1 cells with BS-1 led us to detect the inducible gene. The sequence analysis revealed that the gene was identical to mouse mitochondrial cytochrome b. In fact, Northern blot analysis showed that cytochrome b mRNA in J774A.1 cells was increased by BS-1.

A case of spinal epidermoid caused by lumbar punctures is reported. The 50-year-old man, who had been diagnosed as having poliomyelitis and who had been treated with lumbar puncture 24 times when he was 1 year old, came to our clinic complaining of gradual worsening of atrophy and weakness in his right leg during the last 10 years. Neuroradiologically, intraspinal tumor with calcification was detected in the lumbar region and partial removal of the tumor was performed. Pathological diagnosis was that it was epidermoid. During frequent lumbar puncture at the age of 1, some small skin grafts had been implanted intrathecally and we concluded that, changing its original nature, it had grown slowly to become epidermoid. We want to emphasize especially here that, when we performe lumbar puncture, we should, as far as possible, use a small needle with stilette so as not to induce future spinal epidermoid.

Tuna muscle-derived angiotensin-converting enzyme inhibitory peptide, PTHIKWGD, inhibited the injury of bovine aorta endothelial cells (BAECs) induced by N-formyl-methionyl-leucyl-phenylalanine-activated polymorphonuclear leukocytes and the adhesion of a leukemia cell line, THP-1 cells, to BAECs stimulated by lipopolysaccharides. It is possible that this peptide directly affects BAECs, protecting them from activated leukocytes.

A cDNA encoding ferredoxin reductase has been isolated from a mouse kidney cDNA library using human ferredoxin reductase cDNA as a probe. Mouse ferredoxin reductase coded for 494 amino acid residues. The mouse mature enzyme which comprises 460 amino acid residues shared 87.8-89.1% amino acid identities with the bovine and human enzyme. Northern blot analysis showed that ferredoxin reductase mRNA was expressed in the adrenal, testis and ovary and to a lesser extent in the liver and kidney. However, this mRNA in the adrenal cell line, Y-1 cell, was not induced by adenosine 3',5'-cyclic monophosphate (cAMP) in contrast with ferredoxin mRNA.

A cDNA encoding a 25-hydroxyvitamin D-3 24-hydroxylase, Cyp-24, has been isolated from mouse kidney cDNA library by hybridization screening. Mouse Cyp-24, coding for 514 amino acid residues, shared 82.1 and 94.7% amino acid identity with human and rat CYP24s, respectively. Among mouse organs examined, Cyp-24 mRNA could be detected in the kidney. When mice were treated with vitamin D-3, Cyp-24 mRNA was induced in the kidney.

Carboxamide-methylated light chain (G1L) from human serum IgG inhibited the secretion of tumor necrosis factor (TNF-alpha), one of the inflammatory cytokines, from adherent splenocytes and thioglycolate-induced peritoneal macrophages. The inhibition of TNF-alpha secretion by G1L was associated with disappearance of tyrosine phosphorylation on about 40 kDa protein when thioglycolate-induced peritoneal macrophages were stimulated with lipopolysaccharide (LPS). It is possible that this G1L anti-inflammatory activity occurs through the blockage of the phosphorylation of about 40 kDa protein.

T lymphocyte unresponsiveness, induced in mice by a single gastric intubation of 0.2 ml cedar pollen extract (CPE, containing 4 micrograms protein/ml)/mouse daily for 3 to 28 consecutive days, was evaluated by the absence of a proliferative response of popliteal lymph node (PLN) T lymphocytes to CPE in vitro. T lymphocyte unresponsiveness increased with the period of gastric intubation of CPE and reached more than 80% of the control on day 28. The unresponsiveness to CPE was antigen-specific and T lymphocyte-mediated. In vitro CPE-specific T lymphocyte proliferation was significantly suppressed by intestinal intraepithelial lymphocytes (IELs) and hepatic mononuclear cells (MNCs), but not by spleen cells or PLN T lymphocytes from mice fed CPE for 28 days. The effector activity of IELs and MNCs was obviously antigen-specific. These results suggest that lymphocytes in the intestine and liver of mice fed CPE would be involved in the induction and maintenance of CPE-specific T lymphocyte unresponsiveness.

Two full-length cDNAs (F1-1 and F41-1) complementary to mouse kidney mRNA coding for cytochrome P-450 (P450) linked ferredoxin were isolated and completely sequenced. The coding sequences between F1-1 and F41-1 were identical. However, the 3' untranslated regions of F1-1 and F41-1 were 228 and 27 bases long due to the presence of alternative polyadenylation sites, respectively. The deduced amino acid sequence of mouse cytochrome P-450 linked ferredoxin showed 92.5, 75.0, 71.2 and 71.0% identities with those of rat, human, pig and bovine cytochrome P-450 linked ferredoxin, respectively. The cytochrome P-450 linked ferredoxin mRNA was detected in adrenal, kidney and ovary among the organs examined. The treatment of Y-1 cells with dibutyryladenosine 3',5'-cyclic monophosphate or forskolin induced the transcript of cytochrome P-450 linked ferredoxin mRNA.

A new fraction, Fr. 8-A, which consists of a phosphatidylethanolamine with C14:0-C18:0 isofatty acids was obtained from Bacillus stearothermophilus UBT8038. The fraction inhibited major histocompatibility complex class II (Ia) antigen expression and antigen presentation on mouse macrophages. The effect of Fr.8-A on macrophage functions related to antigen presentation was investigated. Fr. 8-A increased arachidonate release, prostaglandin (PG) E2 release and nitrite production from peritoneal macrophages. It increased further the levels of PGE2, nitrite and tumor necrosis factor in the culture supernatant of the macrophages induced by the supernatant from concanavalin A-stimulated spleen cell cultures. Fr. 8-A augmented the activity of peritoneal macrophages to suppress Con A-stimulated T cell proliferation. Addition of either indomethacin or Ng-methyl-L-arginine had no effect on the augmentation of suppressor macrophage activity or the inhibition of antigen presentation by Fr. 8-A, while simultaneous addition of both inhibitors abrogated the effect of the fraction. These results indicate that Fr. 8-A inhibits Ia expression and antigen presentation, and augments suppressor macrophage activity at least partly via the activation of both cyclooxygenase and nitric oxide synthase pathways.

Isofatty acid-containing phosphatidylglycerol (Fr. 7-C), isolated from Bacillus stearothermophilus UBT8038, enhances the induction of concanavalin A (Con A)-activated suppressor T (Ts) cells in a dose dependent manner (0.01-1 microgram/ml). Its further biological properties on mouse mixed lymphocyte reaction (MLR) has been demonstrated. Fr. 7-C (0.01-1 microgram/ml) suppressed the MLR at 4 d in a dose-dependent manner when added at the start of splenocyte cultivation. Moreover, Fr. 7-C was effective in preventing the generation of cytotoxic T lymphocytes after the MLR. On the other hand, this fraction significantly enhanced the induction of Ts cells in the MLR carried out in any of the antigen-specific, antigen-nonspecific and major histocompatibility complex antigen-nonrestricted fashions. Fr. 7-C increased prostaglandin E2 (PGE2) release approximately 2-fold in the culture supernatant of Con A-activated splenocytes, and PGE2 release decreased dose-dependently when cultured with indomethacin. The inhibitory effect by Fr. 7-C on the MLR was abrogated by the addition of indomethacin. The enhancement by Fr. 7-C on Ts cell induction was blocked by indomethacin in a dose dependent manner. These results strongly suggest that Fr. 7-C suppresses the MLR via the enhancement of antigen-nonspecific Ts cell induction mediated at least partly by PGE2.