Analysis of fumonisin B1 and B2 in beer and raw materials of beer by liquid chromatography/tandem mass spectrometry

A simple method for analysis of fumonisin B1 (FB1) and fumonisin B2 (FB2) in beer and raw materials of beer using liquid chromatography-electrospray ionization tandem mass spectrometry (LC/MS/MS) was developed. Samples were prepared and purified by using solid-phase extraction columns (SAX). The LC separation was performed with an octadecylated silica column at a flow-rate of 0.2 mL/min, using a mobile phase consisting of water and acetonitrile. MS/MS was used in multiple reaction monitoring, employing electrospray ionization (ESI-MRM). Recoveries of spiked FB1 and FB2 from beer were 64% and 52%, respectively, at the level of 5 microg/kg. Recoveries of spiked FB1 and FB2 from malt, cornstarch, and corn grit, at the level of 50 microg/kg, were 87.9% and 86.4%, 89.5% and 87.9%, 86.6% and 81.1%, respectively. This method may have applications in quality assurance.     

Production of vitamin B12 in genetically engineered Propionibacterium freudenreichii

Since the chemical synthesis of vitamin B12 requires more than 70 steps, the production of vitamin B12 has been achieved by microorganism fermentation with additional brief chemical modifications. In an effort to increase the productivity of vitamin B12, we tried to express 10 genes belonging to the hem, cob and cbi gene families involved in the synthesis of vitamin B12 in Propionibacterium freudenreichii, which is a known producer of vitamin B12. In a recombinant P. freudenreichii clone that harbored the expression vector containing a cobA, cbiLF, or cbiEGH, we obtained an increase in vitamin B12 production of 1.7-, 1.9-, and 1.5-fold higher, respectively, than that in the microorganism without any cloned genes in the expression vector pPK705. The cobU and cobS genes caused a slight increase in the production of vitamin B12. Furthermore, we achieved multigene expression in P. freudenreichii. In a recombinant P. freudenreichii clone that harbored an exogenous gene, hemA, from Rhodobacter sphaeroides and endogenous hemB and cobA genes, we successfully achieved the production of about 1.7 mg/l vitamin B12, 2.2-fold higher than that produced by P. freudenreichii harboring pPK705.     

Role of interfacial potential in coagulation of cuprammonium cellulose solution

The electric potential, copper ion flux, and ammonia flux across the interface of cuprammonium cellulose solution (CCS) and various 1.0 equiv/Lelectrolyte solutions (ES) at 25°C were measured. The interfacial potentials were strongly negative (–10 to –35 mV) with H₂SO_4, HCI, and (NH₄)₂SO₄ as ES, weakly positive (6 to 8 mV) with NaCl, KCl, LiCl, CsCL, and RbCl as ES, and strongly positive (19 to 34 mV) with KOH and NaOH as ES, generally showing values similar to the diffusion potentials for electrolyte solutions comprising ions of the same absolute charge. The ammonia flux (about 1 X 10^-4 mol/cm₂/s) was relatively unaffected by the interfacial potential, but the copper ion flux was clearly dependent on it. These results, together with the observed rates of CCS coagulation, indicate that the mechanism of the coagulation was largely determined by the interfacial potential, with strongly negative potential gradients accelerating the Cu²⁺ flux into the ES and CCS coagulation proceeding rapidly by Cu²⁺ removal, strongly positive potential gradients accelerating the Na⁺ flux into the CCS and coagulation proceeding rapidly via the formation of cellulose-Na⁺ complex, and the absence of a strong potential gradient capable of accelerating the ion flux resulting in slow coagulation by ammonia removal. It may therefore be possible to control the interfacial potential and the ion flux by the ES composition, and thus to influence the structure of regenerated cellulosic fibers and membranes. © 1996 John Wiley & Sons, Inc.     

Interactive visualization of three-dimensional magnetic fields

In order to observe the physical values (magnetic flux density and eddy current) in 3D magnetic field analysis, an interactive and highly manipulative visualization system depicting stereo images is intalled in a graphics workstation with high functional graphic processors. The system has the following characteristics:

• 1 An interactive and highly manipulative menu window with many functions: it enables visualization of complex phenomena in a 3D field through observation of various combinations of physical values from various viewpoints.
• 2 Simultaneous display of both magnetic flux density and eddy current by using the appropriate use of two colour display methods for stream lines and distribution maps of density: this function facilitates observation of the mutual relations between two physical values in a 3D field.
• 3 Animations and stereo displays: since they give extremely distinct images, observers can easily understand even highly complex 3D phenomena.

In this paper, the calculation method of density vectors, their display methods, and interactive functions are described. Some examples are also illustrated.     

Characterization on the Conformation of Glucoamylase from Rhizopus niveus and Rhizopus delemar

Using the pH-jump (neutral to above 11) for Tyr-OH ionization as a probe, two glucoamylases from Rhizopus niveus and Rhizopus delemar were found to be caused the change in conformation, which is in a kinetically single step. The pH titration at pHs below 12 was carried out on a multidimensional correlation spectrophotometer. Correlations among the spectrophotometric properties: fluorescence, absorption and circular dichroism are almost identical for the two enzymes. These results suggest that conformational characteristics of the niveus enzyme is almost the same as that of the delemar one.     

Glucose Production from Maltohexaose as a Substrate by the Bacterial Alpha-amylase-catalyzed Reaction. Its Time Courses Obtained by a Theoretical and an Experimental Approaches

Based on the subsite structure of an α-amylase from Bacillus subtilis, production of glucose from a substrate maltohexaose in the amylase-catalyzed reaction was theoretically predicted as a function of time. The theoretical time course was found consistent with the experimental results, which were obtained by the determination of a product glucose with a commercial assay-kit. Hence, glucose, product from a substrate maltohexaose, will be a useful probe for the amylase-catalyzed reaction.     

Aging and Clonal Behavior of Hematopoietic Stem Cells

Hematopoietic stem cells (HSCs) are the only cell population that possesses both a self-renewing capacity and multipotency, and can give rise to all lineages of blood cells throughout an organism’s life. However, the self-renewal capacity of HSCs is not infinite, and cumulative evidence suggests that HSCs alter their function and become less active during organismal aging, leading ultimately to the disruption of hematopoietic homeostasis, such as anemia, perturbed immunity and increased propensity to hematological malignancies. Thus, understanding how HSCs alter their function during aging is a matter of critical importance to prevent or overcome these age-related changes in the blood system. Recent advances in clonal analysis have revealed the functional heterogeneity of murine HSC pools that is established upon development and skewed toward the clonal expansion of functionally poised HSCs during aging. In humans, next-generation sequencing has revealed age-related clonal hematopoiesis that originates from HSC subsets with acquired somatic mutations, and has highlighted it as a significant risk factor for hematological malignancies and cardiovascular diseases. In this review, we summarize the current fate-mapping strategies that are used to track and visualize HSC clonal behavior during development or after stress. We then review the age-related changes in HSCs that can be inherited by daughter cells and act as a cellular memory to form functionally distinct clones. Altogether, we link aging of the hematopoietic system to HSC clonal evolution and discuss how HSC clones with myeloid skewing and low regenerative potential can be expanded during aging.     

Re-evaluation of the phase relationship between plutonium and zirconium dioxides

The phase relationship between ZrO₂ and PuO₂ was examined in a low PuO₂ content region, from 3.1 to 11.2 mol% PuO₂, at temperatures between 1273 K and 1473 K, by high temperature X-ray diffractometry. The measurements were carried out in air. At 1273 K, the samples in this composition range consisted of two phases, monoclinic and cubic. Another phase, tetragonal, was observed at 1373 K. The low temperature monoclinic phase disappeared at 1473 K. It was confirmed that the monoclinic phase disappears around 1463 K; the disappearance temperature does not depend on the composition of the sample. It was, thus, inferred that there should be a eutectoid line in the phase diagram. Though the eutectoid point is not clear, the PuO₂ content at the point should be less than 3.1 mol%.     

An Enzymatic Assay Method for Bacterial Alpha-amylase Using a Glucose Determination-kit and Maltohexaose as the Substrate

Using maltohexaose G₆ as a substrate, a convenient analytical method for amylase-catalyzed hydrolytic reaction was developed by application of an enzymatic assay-kit (glucose oxidase-peroxidase) to measurement of glucose, produced on the reaction: G₆→G₅ + G. The initial velocity was confirmed to be proportional to the initial concentration of amylase, examined, ∼ 100 μm. The new method was found in a good correlation with the Nelson and Somogyi method. Rate assay was also examined: The initial velocity is proportional to the enzyme concentration, ∼ 30 μm. Hence, this enzymatic assay method is recognized to be convenient and useful for the measurement of α-amylase.     

Radiotherapy for carcinoma of the esophagus in aged patients

One hundred fifty-four patients with esophageal carcinoma were treated with either irradiation alone or irradiation combined with surgery at the University of Occupational and Environmental Health Hospital between January 1980 and February 1992. The number of patients 75 years old and older was 25. In patients 74 years old and younger, the overall five-year survival rate by Kaplan-Meier method was 24.5%. The survival rate was best in the patients who were treated by a combination of irradiation and surgery. In patients 75 years old and older, the one-year survival rate was 59%, and the three-year rate was 20%. Aged patients had a tendency to be worse in performance status, and there was no correlation between treatment modality and survival time. We conclude that radiotherapy is useful for treating esophageal cancer in aged patients particularly when maintenance of the quality of life is considered.