Enhancement of nucleoside phosphorylation activity in an acid phosphatase

Escherichia blattae non-specific acid phosphatase (EB-NSAP)possesses a pyrophosphate-nucleoside phosphotransferase activity,which is C-5'-position selective. Current mutational and structuraldata were used to generate a mutant EB-NSAP for a potentialindustrial application as an effective and economical proteincatalyst in synthesizing nucleotides from nucleosides. First,Gly74 and Ile153 were replaced by Asp and Thr, respectively,since the corresponding replacements in the homologous enzymefrom Morganella morganii reduced the K_m value for inosine andthus increased the productivity of 5'-IMP. We determined thecrystal structure of G74D/I153T, which has a reduced K_m valuefor inosine, as expected. The tertiary structure of G74D/I153Twas virtually identical to that of the wild-type. In addition,neither of the introduced side chains of Asp74 and Thr153 isdirectly involved in the interaction with inosine in a hypotheticalbinding mode of inosine to EB-NSAP, although both residues aresituated near a potential inosine-binding site. These findingssuggested that a slight structural change caused by an aminoacid replacement around the potential inosine-binding site couldsignificantly reduce the K_m value. Prompted by this hypothesis,we designed several mutations and introduced them to G74D/I153T,to decrease the K_m value further. This strategy produced a S72F/G74D/I153Tmutant with a 5.4-fold lower K_m value and a 2.7-fold higherV_max value as compared to the wild-type EB-NSAP.     

Effect of the (Ba + Sr)/Ti ratio on the microwave‐tunable properties of Ba0.6Sr0.4TiO3 ceramics

The impact of the (Ba + Sr)/Ti (A/B) ratio on the microwave‐tunable characteristics of diffuse phase transition (DPT) ferroelectric Ba_0.6Sr_0.4TiO₃ (0.6‐BST) ceramics was investigated. The reduction in the lattice constant with increasing nonstoichiometry was attributed to introduced partial Schottky defects, i.e., and . The magnitude of the dielectric constant, ε′, at room temperature in the absence of an applied electric field was governed by the shift in the dielectric maximum temperature, T_m, because T_m was close to room temperature for the 0.6‐BST. The dielectric loss, tanδ, diminished as the ε′ decreased for 0.98≤A/B≤1.05, while the tanδ was much higher for A/B=0.95 having the greatest A‐site vacancy loading. The negatively charged and were mainly compensated by oxygen vacancies and likely partly compensated by holes, h^?, which contributed to the electrical conduction. The tunability, T, at 100 MHz was almost constant at 20%–25% for A/B≥1.00 despite the reduction of the ε′, whereas T decreased for A/B<1.00 to ca. 10% for A/B=0.95 having the greatest A‐site vacancy loading. The results implied that the for larger A/B values was more efficient in generating nucleation sites in the polar nanoregions (PNRs) than the for smaller A/B values, thereby providing greater dipole polarization. Consequently, the figure of merit, FOM, reached its maximum of 250 at A/B=0.9875, which was ca. 155% higher than that of the stoichiometric BST.     

Optical properties of polystyrene and polyarylate block copolymer and their control by morphology generation

Relationship between phase morphology and optical properties of polystyrene and polyarylate (PS‐PAr) block copolymers synthesized from telechelic polystyrene has been investigated. In the PS‐PAr block copolymers, the PAr domains with higher melt viscosity were dispersed in the PS phase matrix with lower melt viscosity over the wide range of their composition from PS/PAr = 25/75 to 75/25 (wt ratio). The PAr domain size was dependent on the reactive ratio of PAr determined analogously by the mole fraction of the fed telechelic polystyrene. By controlling the mole fraction of the telechelic polystyrene more than 0.016 in synthesizing the PS‐PAr block copolymer, the size of PAr domains could be reduced to the microscopic scale (smaller than 100 nm). Then, the PS‐PAr block copolymers exhibited almost the same transparency as PAr in spite of the large difference in the refractive index between the PS and the PAr phase. Birefringence free condition for the PS‐PAr block copolymers was determined by not only the PS/PAr composition but also the balance in the degree of molecular orientation of these chains. The latter factor suggests that PS and PAr chains undergo inhomogeneous stress and relaxation history during the injection process. By controlling Mn (number average molecular weight) and weight fraction of the fed OH‐PS‐OH around 20 000 and 55 wt %, respectively, in the synthesis of the PS‐PAr block copolymer, the PS‐PAr block copolymer exhibited almost zero birefringence without any sacrifice of transparency. Because in the PS‐PAr block copolymer low birefringence and high transparency can coexist by controlling the adequate feeding condition in the synthesis process, the PS‐PAr block copolymer would be a promising material for optical applications, such as a substrate of optical disks or optical lenses. © 2000 John Wiley & Sons, Inc. J Appl Polym Sci 78: 953–961, 2000     

Control of antibody-antigen interaction using anion-induced conformational change in antigen peptide

The binding of a monoclonal antibody to an epitope peptide wascontrolled by the conformational change of the epitope peptideinduced by anions. We synthesized peptides in which the epitopesequence DTYRYI for the monoclonal antibody AU1 is located betweenamphiphilic peptides (KKLL)_n and (LLKK)_n. In the absence ofan appropriate anion, the peptide was in a random coil stateand the epitope was linear. In contrast, in the presence ofan appropriate anion, the peptide exhibited an anti-parallel-helical structure and the epitope was subsequently `bent'.In the presence of 41 µM triphosphate, the associationconstant between the antibody and the peptide was decreasedby one order of magnitude in the case of n = 3 and at leastthree orders of magnitude in the case of n = 4 or 5. Oligo-DNAs,as anions, dissociated the antibody–peptide complex, whereastriphosphate did not. The DNA concentrations required for 50%dissociation of the antibody–peptide complex at pH 7.5were 4x10^–8, 1x10^–7 and 6x10^–6 M for decamer,octamer and hexamer DNA, respectively.     

High-Voltage Screening of Unidirectionally Surface-Ground Titania Ceramics

The effects of high-voltage screening were examined for perpendicularly and parallelly surface-ground titania rectangular bars. A screening field at or below which 30% of titania samples break electrically was applied to each group samples. After high-voltage screening, the surviving samples were subjected to mechanical strength measurement and the resultant strength distribution was compared with the original distribution. After screening, Weibull plots of perpendicularly ground samples bent to become convex curves while plots in the high-strength region remained almost the same, indicating that low-strength samples were selectively eliminated by the high-voltage screening. On the other hand, the screening effect on parallelly surface-ground samples was very small.     

Research on traversability of tracked vehicle on slope with unfixed obstacles: derivation of climbing-over,tipping-over,and sliding-down conditions

When a tracked robot explores a volcanic environment, it faces difficulty in climbing over unfixed obstacles such as loose rocks on the ground. Such unfixed obstacles sometimes cause the sliding-down or tipping-over of the robot. Although such phenomena should be avoided for the success of the mission, they have not been sufficiently studied yet. Therefore, this research aims at understanding the phenomena for a tracked vehicle climbing an unfixed obstacle on a slope, and considers the conditions of climbing-over, tipping-over, and sliding-down. To simplify the problem, a model of a single track and circular cross-section obstacle is used in this research. The climbing-over and tipping-over conditions are derived from the geometric relationship, and the sliding-down condition is derived from statics. Moreover, some experiments using an actual robot are conducted to verify the validity of the conditions. The results show that the derived conditions are reasonable. Furthermore, it is revealed that unfixed obstacles typically tend to slide down more than fixed obstacles because of the number of contact points that can support a robot.     

Enhanced expression of a new class of liver-enriched b-Zip transcription factors, hepatocarcinogenesis-related transcription factor, in hepatocellular carcinomas of rats and humans

Urinary incontinence (UI) is an unpleasant problem for many adults. This study determined the importance of demographic, health and incontinence variables for the generic and incontinence-specific quality of life (QoL) of older adults (age > or = 60 years). Telephone surveys of adults reporting at least weekly episodes of UI (n = 435) were conducted as part of a randomized, controlled trial. Logistic regression analyses showed that the predictors of generic and incontinence-specific QoL differed. Life satisfaction, a generic outcome, was predicted by education, the number of days in bed due to health problems, the number of days not feeling well and the amount of urine lost. Generic health was related to education, the number of days sick in the previous 30 days and the number of days health issues restricted activities. The incontinence-specific QoL outcomes were predicted by age, mobility difficulties, the amount of urine lost, the frequency of UI, and the number of daytime and night-time voids. The specific QoL measures provide a different profile of those most affected in this sample than that obtained by the generic measures. The most affected are younger persons with severe urine loss. Older persons may have other conditions impinging on QoL and may have adapted behaviourally and psychologically to urine loss.     

Postnatally induced inactivation of gp130 in mice results in neurological, cardiac, hematopoietic, immunological, hepatic, and pulmonary defects

The pleiotrophic but overlapping functions of the cytokine family that includes interleukin (IL)-6, IL-11, leukemia inhibitory factor, oncostatin M, ciliary neurotrophic factor, and cardiotrophin 1 are mediated by the cytokine receptor subunit gp130 as the common signal transducer. Although mice lacking individual members of this family display only mild phenotypes, animals lacking gp130 are not viable. To assess the collective role of this cytokine family, we inducibly inactivated gp130 via Cre-loxP-mediated recombination in vivo. Such conditional mutant mice exhibited neurological, cardiac, hematopoietic, immunological, hepatic, and pulmonary defects, demonstrating the widespread importance of gp130-dependent cytokines.     

Changes of immunological markers after autologous peripheral blood stem cell transplantation

PURPOSE: Recently high-dose chemotherapy with peripheral blood stem cell transplantation (PBSCT) has become an important treatment for hematological and solid tumors. METHODS: Immunological parameters were examined before and after PBSCT in 9 patients with lung cancer and 13 patients with malignant lymphoma. Findings were compared with those for bone marrow transplantation (BMT). Peripheral blood cells were analyzed for phenotype and the levels of cytokines and soluble factors were measured. RESULTS: After PBSCT, activated T cells (CD3+HLA-DR+ cells, CD8+HLA-DR+ cells) and suppressor/cytotoxic T cells (CD8+CD11b- cells) were significantly higher in the patients with lung cancer than in those with malignant lymphoma. Serum levels of interleukin-4 and soluble interleukin-2 receptor were also significantly higher in the patients with lung cancer than in those with lymphoma. On the other hand, the serum levels of interferon gamma, tumor necrosis factor alpha, interleukin-6, soluble human leukocyte antigen class 1, and soluble thrombomodulin were significantly increased after bone marrow transplantation. The transfused peripheral stem cells of lung cancer and lymphoma patients had a similar number of granulocyte/macrophage-colony-forming units, but lung cancer patients had significantly more CD34-positive cells. CONCLUSION: By reinfusing large numbers of autologous immune cells, PBSCT may accelerate immune reconstitution, with T cells being likely to have a marked therapeutic potential. The changes after PBSCT were greater in patients with lung cancer than in lymphoma patients. These blood cells are potent mediators of anticancer activity and could play an important role in the elimination of autologous malignant cells.     

Identification and characterization of a novel SH3-domain binding protein, Sab, which preferentially associates with Bruton''s tyrosine kinase (BtK)

We describe the isolation and sequence of a gene encoding 4-hydroxyphenylpyruvate dioxygenase (HPPD) (EC 1.13.11.27)) from the wheat leaf-spot fungal pathogen Mycosphaerella graminicola (Septoria tritici), that directs the synthesis of 2,5-dihydroxyphenylacetate (homogentisic acid, HGA). The sequence of the deduced peptide showed homology to HPPDs from other organisms; the greatest identity was to a T-cell reactive protein, also identified as HPPD, from the human fungal pathogen Coccidioides immitis. As observed for HPPD from other sources, expression of the M. graminicola HPPD gene in Escherichia coli cells could be detected by the gradual development of a brown pigment in cultures as a result of the spontaneous oxidation and polymerisation of HGA. Pigment development in these cultures was prevented by the HPPD inhibitor sulcotrione.