The Emi2 Protein of Saccharomyces cerevisiae is a Hexokinase Expressed under Glucose Limitation

Hexokinases catalyze glucose phosphorylation at the first step in glycolysis in eukaryotes. In the budding yeast Saccharomyces cerevisiae , three enzymes for glucose phosphorylation have long been known: Hxk1, Hxk2, and Glk1. In this study, we focus on Emi2, a previously uncharacterized hexokinase-like protein of S. cerevisiae . Our data show that the recombinant Emi2 protein (rEmi2), expressed in Escherichia coli , possesses glucose-phosphorylating activity in the presence of ATP and Mg ²⁺ . It was also found that rEmi2 phosphorylates not only glucose but also fructose, mannose and glucosamine in vitro . In addition, we examined changes in the level of endogenous Emi2 protein in S. cerevisiae in the presence or absence of glucose and a non-fermentable carbon source. We found that the expression of Emi2 protein is tightly suppressed during proliferation in high glucose, while it is strongly upregulated in response to glucose limitation and the presence of a non-fermentable carbon source. Our data suggest that the expression of the endogenous Emi2 protein in S. cerevisiae is regulated under the control of Hxk2 in response to glucose availability in the environment.     

Oxidation of doped europium in BaMgAl10O17 by annealing studied by x‐ray‐absorption fine‐structure measurements

Abstract— We studied the influence of annealing in air on doped europium in BaMgAl₁₀O₁₇ by performing x‐ray absorption fine‐structure measurements. We determined the oxidation of doped divalent europium by annealing in air at over 500°C. The interatomic distance between the europium and the surrounding oxygen atoms was compressed by oxidation. It also appears that the oxidation process of europium is determined by the diffusion of oxygen into BaMgAl₁₀O₁₇.     

An on-chip active decoupling circuit to suppress crosstalk in deep-submicron CMOS mixed-signal SoCs

A decoupling circuit using an operational amplifier is proposed to suppress substrate crosstalk in mixed-signal system-on-chip (SoC) devices. It overcomes the parasitic inductance problem of on-chip capacitor decoupling. The effect of the proposed decoupling circuit is not limited by parasitic fine impedance. A 0.13-/spl mu/m CMOS test chip showed that substrate noise at frequencies from 40 MHz to 1 GHz was incrementally suppressed by sequentially activating three of the proposed circuits in parallel. The power dissipation of each circuit was 3.3 mW at a 1.0-V power supply. The test chip measurement showed that the proposed decoupling reduced crosstalk by 31% at 200 MHz, whereas it was reduced by 4.4% with capacitor decoupling. This 7:1 ratio, or 17 dB, corresponds to the gain of the opamp. Design of the opamp and its feedback loop for active decoupling is simple, making the opamp useful for SoC applications.     

Neoadjuvant intra-arterial chemotherapy based on chemosensitivity tests for locally invasive bladder cancer

We investigated the clinical usefulness of individualization of chemotherapeutic regimen in neoadjuvant intra-arterial chemotherapy for locally invasive bladder cancer. Anticancer drugs were selected according to the results of an in vitro chemosensitivity test (collagen matrix assay or succinic dehydrogenase inhibition test). Nine patients with locally invasive bladder cancer received 1 to 4 courses of neoadjuvant intra-arterial chemotherapy, followed by radical cystectomy. Histopathological responses in the cystectomized specimens were grade 3 in 3 cases, grade 2 in 2, grade 1b in 2 and no response in 2. Pathologically, a complete response and downstaging were observed in 3 and 4 cases, respectively. Seven of the 9 patients were alive no evidence of disease with a mean follow-up period of 38.9 months, whereas 2 patients died of metastasis within 2 years. Six of the 7 patients who showed a complete response or down staging have been free of recurrence. These findings suggest that our chemotherapeutic strategy may improve the prognosis for locally invasive bladder cancer.     

Analysis of T-cell receptor beta of the T-cell clones reactive to the human PDC-E2 163-176 peptide in the context of HLA-DR53 in patients with primary biliary cirrhosis

T-cell-mediated autoimmune mechanisms are considered to be involved in the pathogenesis of primary biliary cirrhosis (PBC). In the previous study, we identified the immunodominant T-cell epitope on the E2 component of pyruvate dehydrogenase complex (PDC-E2) in patients with PBC who have HLA-DRB4*0101. In this report, we revealed that the frequency of the T cells reactive to the human PDC-E2 163-176 peptide is significantly increased in the peripheral blood of patients with PBC as compared with healthy subjects. We also confirmed that these T cells were all restricted with HLA-DRB4*01 (DR53) by using HLA-DR-transfected L cells. These results together with the evidence that the immunodominant B-cell epitope overlaps with the human T-cell epitope of the PDC-E2 antigen indicate that the T cells reactive to this epitope are closely associated with the pathogenesis of PBC at least in patients who have HLA-DR53. Therefore, we analyzed the T-cell receptor (TCR) Vbeta sequence of the five different T-cell clones and the three T-cell clones derived from three patients with PBC and healthy subjects, respectively, which are reactive to the human PDC-E2 163-176 peptide in the context of HLA-DR53. The Vbeta- and the Jbeta-gene usages were diverse among the T-cell clones (Vbeta11-Jbeta1.4, Vbeta8-Jbeta1.2, Vbeta12-Jbeta2.1, Vbeta10-Jbeta1.5, and Vbeta20-Jbeta2.1) in patients with PBC. By contrast, in the third complementarity determining region (CDR3), G was frequently found and GXG or GXS motif was identified in all T-cell clones. Moreover, RGXG motif was found in three clones generated from two patients. In healthy subjects, the Vbeta- and the Jbeta-gene usages were also diverse, and GXG and RGXG motif were found. These results indicate that the T cells may recognize the ligand (the human PDC-E2 163-176 peptide/HLA-DR53 complex) using the limited motif in the CDR3 region and that the design of CDR3-specific immunotherapy would be possible using these motifs.     

Effect of sulfasalazine on B cell hyperactivity in patients with rheumatoid arthritis

OBJECTIVE: We investigated the in vitro immunomodulatory effects of sulfasalazine on B cells in rheumatoid arthritis (RA). METHODS: Reversed hemolytic plaque assay and 3H-thymidine incorporation were measured. RESULTS: B cells from patients with RA showed hyperactivity to stimulation by Staphylococcus aureus Cowan I. Sulfasalazine significantly inhibited this B cell hyperactivity in a dose dependent manner. The kinetic study and a decrease in 3H-thymidine incorporation on Day 3 indicate that sulfasalazine inhibited the early phase (0-48 h) of B cell proliferation in these patients. Sulfapyridine also inhibited B cell hyperactivity in these patients, but 5-aminosalicylic acid and N-acetylsulfapyridin had no significant effect. CONCLUSION: Sulfasalazine exhibited a direct immunosuppressive effect on B cell hyperactivity in patients with RA, which may be responsible for its therapeutic effectiveness in this disorder.     

Structural study of degraded ZnMgSSe blue light emitters

Using electroluminescence (EL) topography and transmission electron microscopy (TEM), we investigated the nonluminescent regions which form while current is being injected into ZnMgSSe/ZnSSe/ZnCdSe-based blue light emitters. Small dark spots were observed just after turn-on and spread out forming rough nonluminescent triangles in the <100> directions in the EL image of the active region. TEM studies showed that the small dark spots are pre-existing stacking faults originating at the substrate/epitaxial layer interface. The nonluminescent triangles were found to be a dense region of dislocation dipoles and dislocation loops. Each dipole was aligned along two <110> directions in the {111} planes. The Burgers vectors were of the type a/2<011> inclined at 45° to the (001) junction plane.     

Interferon therapy after ablation of Kent bundle for a patient with chronic hepatitis type B complicated with WPW syndrome

Cardiotoxicity of interferon-alpha or gamma, such as fatal arrhythmia and myocardial infarction, has been reported. Therefore cardiotoxicity of interferon should be seriously considered before administration for patients with a pre-existing heart disease. We treated a patient with chronic active hepatitis type B, coexisted with Wolff-Parkinson-White syndrome, who has had frequent attacks of paroxysmal atrial fibrillation. To prevent the occurrence of fatal arrhythmia with an interferon therapy in this patient, we performed radiofrequency catheter ablation of the Kent bundle. After the successful ablation, we could safely administered recombinant interferon alpha-2b for chronic hepatitis type B.