COMPARISON OF METHODS FOR THE DETERMINATION OF CELL VIABILITY IN STORED BAKER'S YEAST

The classical plate method for discriminating between viable and nonviable yeast cells in stored baker's yeast was compared with dead cell staining techniques using methylene blue and three fluorochrome stains. The increase of dead yeast cells during storage of baker's yeast for up to 16 days at 5°C, 20°C and 35°C was determined. During prolonged storage, especially at 35°C, the death rate increased rapidly and the yeast cake began to soften because of autolysis. In these conditions the choice of method for determining the proportion of dead cells proved to be of great importance. Useful results for yeast stored for some time at 35°C could be obtained only by the fluorochrome technique using primuline, acridine orange or acriflavine as fluorochromes.     

Ionic liquid enables simple and rapid sample preparation of human culturing cells for scanning electron microscope analysis

Ionic liquid is a kind of salt that stays in a molten state even at room temperature. It does not vaporize at all in vacuum and facilitates electrical conductivity to the sample surfaces for observations with a scanning electron microscope (SEM). In this study, we used an ionic liquid in SEM for the first time to observe fixed human culture cells. The condition for the cell culture using wrapping sheets and SEM settings were varied to elucidate the optimized protocol. Compared to samples prepared by the conventional way, the ionic liquid‐treatment of samples gave SEM images of the cellular ultra structures in more detail, enabling observation of microvilli that made bridges between separated cells. In addition, the ionic liquid treatment is less time consuming as well as less laborious compared with the conventional way that includes dehydration, drying, and conductivity treatments. Totally, we concluded the ionic liquid is a useful reagent for SEM sample preparation. Microsc. Res. Tech., 2011. © 2010 Wiley‐Liss, Inc.     

Interactive segmentation of the cerebral lobes with fuzzy inference in 3T MR images.

Measurement of volume and surface area of the frontal, parietal, temporal and occipital lobes from magnetic resonance (MR) images shows promise as a method for use in diagnosis of dementia. This article presents a novel computer-aided system for automatically segmenting the cerebral lobes from 3T human brain MR images. Until now, the anatomical definition of cerebral lobes on the cerebral cortex is somewhat vague for use in automatic delineation of boundary lines, and there is no definition of cerebral lobes in the interior of the cerebrum. Therefore, we have developed a new method for defining cerebral lobes on the cerebral cortex and in the interior of the cerebrum. The proposed method determines the boundaries between the lobes by deforming initial surfaces. The initial surfaces are automatically determined based on user-given landmarks. They are smoothed and deformed so that the deforming boundaries run along the hourglass portion of the three-dimensional shape of the cerebrum with fuzzy rule-based active contour and surface models. The cerebrum is divided into the cerebral lobes according to the boundaries determined using this method. The reproducibility of our system with a given subject was assessed by examining the variability of volume and surface area in three healthy subjects, with measurements performed by three beginners and one expert user. The experimental results show that our system segments the cerebral lobes with high reproducibility.     

Selective and in-situ determination of carbonate and oxide particles in aqueous solution using laser-induced breakdown spectroscopy (LIBS) for wearable information equipment

Laser-induced breakdown spectroscopy (LIBS) has been applied for quantitative analysis of Al₂O₃, CaCO₃ and MgO particles suspended in water. In the single elemental system, the plasma emission intensities of Al, Ca and Mg were linearly increased with concentration of elements in the range of 1.0×10^?5–1.0×10^?3  M, 1.0×10^?4–2.0×10^?3 M and 8.0×10^?5 –4.0×10^?3 M, respectively. We also investigated the concentration dependence of breakdown spectra for suspended mixtures of Al₂O₃, CaCO₃ and MgO particles. The emission lines, such as Al I, Ca I, Ca II and Mg I, were appeared in the LIBS spectrum simultaneously, and each emission peak could be deconvoluted. The plasma emission intensities of Al, Ca and Mg in the multielemental system were also linearly increased with their concentrations in the range of 1.0×10^?5–1.0×10^?3 M, 1.0×10^?4–2.0×10^?3 M and 4.0×10^?4–2.0×10^?3 M, respectively. LIBS was found to be available for quantitative and qualitative measurement of the concentrations of Al₂O₃, CaCO₃ and MgO particles suspended in water. The present results suggest that LIBS is a potentially useful tool for in-situ analysis on particles composition and concentrations for environmental monitoring by the wearable information equipments.     

Characteristics of neutralization of acids by newly isolated fungal cells

Soil microorganisms play an important role in maintaining soil pH at levels suitable for other soil organisms. To clarify the biological neutralization mechanism in soil, we isolated soil microorganisms showing a high ability to neutralize acids and studied their characteristics. From our taxonomic study, three isolated strains were identified as filamentous fungi, namely Mucor sp., Aspergillus fumigatus, and Aureobasidium pullulans. These strains could secrete basic materials, such as ammonia, for neutralization, grow in the medium at pH 4.0 and increase the pH of the medium to approximately 8.0. These microbial cells could neutralize not only nitric acid but also sulfuric and hydrochloric acids. The strains could also grow by utilizing nitric acid as a sole nitrogen source. In the soil containing these organisms, the pH was maintained in the neutral range by the buffering action of basic materials that they secrete. These results suggest that these fungal cells are useful for protecting the soil from acidification by acid rain.     

Gel permeation chromatography using controlled-porosity glass. II. Polyacrylamide aqueous solutions

In a study of polyacrylamide in solutions we have required the rapid characterization on chromatographies. The application to the aqueous solution of the GPC using controlled-porosity glass has been examined from both the viewpoint of the effect of salt addition and the GPC mechanism. An adequate addition to neutral salt, 0.005M KCI to the eluent, gave rise to the elution behaviors being in accord with the hydrodynamic volume concept of the GPC separation.     

Cloning of a fatty acid synthase component FAS1 gene from Saccharomyces kluyveri and its functional complementation of S. cerevisiae fas1 mutant

A gene encoding a fatty acid synthase component, FAS1, has been cloned from a genomic library of the polyunsaturated fatty acid (PUFA)-producing yeast Saccharomyces kluyveri. This gene (named Sk-FAS1) was found to contain an open reading frame of 6150 bp, coding for 2049 amino acids. The deduced Sk-FAS1 protein showed significant (75-59%) homology with FAS proteins from the other yeasts, including S. cerevisiae, Candida albicans and Yarrowia lipolytica. The substrate-binding sites of the acetyl transferase and malonyl/palmitoyl transferase domains, and the FMN- and NADPH-binding sites of the enoyl reductase domain, were all highly conserved. Expression of the Sk-FAS1 gene in S. cerevisiae complemented genetic disruption of the S. cerevisiae FAS1 gene (Sc-FAS1), suggesting the formation of a heterogeneous complex of Sk-FAS1 (beta) and Sc-FAS2 (alpha), which is able to function to synthesize fatty acids. Compared with the isogenic wild-type of S. cerevisiae, as well as S. kluyveri, the S. cerevisiae fas1 mutant carrying the Sk-FAS1 gene showed an increase in the relative amount of 16-carbon fatty acids and a decrease in 18-carbon fatty acids.     

Personalising speech-to-speech translation: Unsupervised cross-lingual speaker adaptation for HMM-based speech synthesis

In this paper we present results of unsupervised cross-lingual speaker adaptation applied to text-to-speech synthesis. The application of our research is the personalisation of speech-to-speech translation in which we employ a HMM statistical framework for both speech recognition and synthesis. This framework provides a logical mechanism to adapt synthesised speech output to the voice of the user by way of speech recognition. In this work we present results of several different unsupervised and cross-lingual adaptation approaches as well as an end-to-end speaker adaptive speech-to-speech translation system. Our experiments show that we can successfully apply speaker adaptation in both unsupervised and cross-lingual scenarios and our proposed algorithms seem to generalise well for several language pairs. We also discuss important future directions including the need for better evaluation metrics.     

Genomics of industrial Aspergilli and comparison with toxigenic relatives

Aspergillus oryzae has been used in Japanese fermentation industries for more than a thousand years. The species produces large amounts of various hydrolytic enzymes and has been successfully applied to modern biotechnology. The size of the A. oryzae genome (37.5 Mb) is very close to that of A. flavus and A. niger, and 20-30% larger than that of either A. nidulans or A. fumigatus. A. oryzae and A. flavus have exactly the same number of aspartic proteinase genes, of which each orthologous pair shares highly conserved amino acid sequences. Synteny analysis with A. fumigatus and A. nidulans showed that the A. oryzae genome has a mosaic structure consisting of syntenic and non-syntenic blocks. In the microorganisms to be compared, the density of the genes having homologs was obviously higher on the syntenic than on the non-syntenic blocks. Expression analysis by the DNA microarray supported the significantly lower expression of genes on the non-syntenic than on the syntenic blocks.