Analysis of a 36·2 kb DNA sequence including the right telomere of chromosome VI from Saccharomyces cerevisiae

The nucleotide sequence of a 36·2-kb distal region containing the right telomere of chromosome VI was determined. Both strands of DNA cloned into cosmid clone 9965 and plasmid clone pEL174P2 were sequenced with an average redundancy of 7·9 per base pair, by both dye primer and dye terminator cycle sequencing methods. The G + C content of the sequence was found to be 37·9%. Eighteen open reading frames (ORFs) longer than 100 amino acids were detected. Four of these ORFs (9965orfR017, 9965orfF016, 9965orfR009 and 9965orfF003) were found to encode previously identified genes (YMR31, PRE4, NIN1 and HXK1, respectively). Six ORFs (9965orfR013, 9965orfF018, 9965orfF006, 9965orfR014, 9965orfF013 and 9965orfR020) were found to be homologous to hypothetical 121·4-kDa protein in the BCK 5′ region, Bacillus subtilis DnaJ protein, hypothetical Trp-Asp repeats containing protein in DBP3-MRPL27, putative mitochondrial carrier YBR291C protein, Salmonella typhimurium nicotinate-nucleotide pyrophosphorylase, and Escherichia coli cystathionine β-lyase, respectively. The putative proteins encoded by 9965orfF018, 9965orfR014 and 9965orfR020 were found to be, respectively, a new member of the family of DnaJ-like proteins, the mitochondrial carrier protein and cystathionine lyase. The nucleotide sequence reported here has been deposited in the DDBJ/GenBank/EMBL data library under Accession Number D44597.     

Characteristics of the thiobarbituric acid reactivity of human urine as a possible consequence of lipid peroxidation

A 532 nm red pigment formed in the thiobarbituric acid (TBA) assay of human urine was characterized after separation of the pigment by high-performance liquid chromatography. The yield of the red pigment was some-what higher at pH 2 than at pH 5; its development was not inhibited by ethylenediaminetetraacetic acid. The characteristics of the pigment were similar to those of the pigment derived from standard malonaldehyde. The amount of the pigment formed was roughly equal to the content of malonaldehyde derivatives estimated as 1-(2,4-dinitrophenyl)pyrazole. Pigment formation was significantly enhanced byt-butyl hydroperoxide (t-BuOOH) and ferric ions, which may be due to pigment formed from aldehydes other than malonaldehyde; the presence of these aldehydes was confirmed by the formation of the corresponding 2,4-dinitrophenylhydrazones. The amount of pigment produced from 24-h urine samples of 12 healthy subjects was estimated to be 26–95 nmol/kg, and 65–182 nmol/kg in the presence oft-BuOOH. These values are lower than those for urine of rabbit or rat. The TBA reactivity in the absence and presence oft-BuOOH of human urine was not related to age or sex. The TBA reactivity of human urine collected in the afternoon and in the evening was higher than that of urine collected in the morning.     

Increase of Trap Catches by a Combination of Male Sex Pheromones and Floral Attractant in Longhorn Beetle,Anaglyptus subfasciatus

A combination of the synthetic sex pheromone [a 125:1 blend of (R)-3-hydroxy-2-hexanone and (R)-3-hydroxy-2-octanone] with methyl phenylacetate captured significantly moreAnaglyptus subfasciatus Pic females than either the sex pheromone or methyl phenylacetate did alone.     

Creative communication for chance discovery in shopping

This study considered an information presentation method to help customers make a concept-articulation type of purchase. When customers follow the concept-articulation type of thinking, they only have vague requirements and try to make a gradual clarification of what they really want through the interaction with salesclerk(s). Skillful salesclerks can communicate with their customers appropriately to discover an opportunity for bringing the customers to their concept-articulation, which often leads to a successful sale. Based on the observations from our analysis of these human behavior in actual purchase activities, we constructed a system called S-Conart (Concept Articulator for Shoppers) to support the concept-articulation type of purchase. The user study conducted using S-Conart shows that using a different method of presenting information can cause a change in the user’s mental world, which is also observed in a real-life shopping situation, although in a different form. The result of this user study suggests the possibility of chance discovery by customers themselves, which is expected to be useful for building support systems for concept-articulation type of shoppers. Hiroko Shoji: She is an associate professor in the Department of Information and Communication Sciences at Kawamura Gakuen Women’s University, and also a doctoral student in the Department of Advanced Interdisciplinary Studies at the University of Tokyo. She received her B.S. in mechanical engineering in 1989 and her M.Eng. in mechanical engineering and engineering synthesis in 1991 from the University of Tokyo. Her current research interests include creativity in real life and chance discovery. She was awarded a Research Promotion Award from Japanese Society for Artificial Intelligence (JSAI) in 2000. Koichi Hori, D.Eng.: He received his B.Eng, M.Eng, and Dr.Eng. degrees in electronic engineering from the University of Tokyo, in 1979, 1981, and 1984, respectively. In 1984, he joined National Institute of Japanese Literature, where he developed AI systems for literature studies. Since 1988, he has been with the University of Tokyo. He is currently a professor with Department of Advanced Interdisciplinary Studies, the University of Tokyo. From September 1989 to January 1990, he also held a visiting position at University of Compiegne, France. His current research interests include AI technology for supporting human creative activities, cognitive engineering and Intelligent CAD systems. Professor Hori is a member of IEEE, ACM, IEICE, IPSJ, JSAI, JSSST, and JCSS.     

Microfabricated on-chip-type electrochemical flow immunoassay system for the detection of histamine released in whole blood samples

This paper describes an on-chip-type electrochemical flow immunoassay system with a multichanneled matrix column. The multichanneled matrix column was functionally coated with cation-exchange resin and used for separation of proteins. Antihistamine immunoglobulin G (IgG) antibody conjugated with ferrocenemonocarboxylic acid (Fc) was also prepared and used as a novel analytical reagent. Antibody-antigen complexes were separated from free Fc-conjugated IgG antibody (Fc-IgG) on the basis of differences in isoelectric point (pI) using the multichanneled matrix column coated with cation-exchange resin. The assay yields a good relationship between current and histamine concentration in the range of 200-2000 ng/mL. This simple technique enables the assay of histamine released in whole blood within 2 min. Furthermore, a good correlation was found between the response of the electrochemical immunoassay described in this paper and the conventional RIA (radioimmunoassay). This on-chip-type electrochemical flow immunoassay requires only minute quantities of whole blood samples and generates highly reproducible results.     

Dynamic structures of motor proteins myosin and kinesin, and switch protein troponin as detected by SDSL-ESR

We have studied biological nano-machines, motor and switch proteins operating as supramolecular complexes by electron spin resonance (ESR) and found key features of their molecular movements. In all the systems, the specific movements of elements or domains were detected and quite dynamic at nanometer scale. We have observed two broad but distinct orientations, separated by a 25 degrees axial rotation, of a spin label attached specifically to the light chain (LC) domain of myosin motor in the muscle fibers. The distribution became only narrower upon muscle activation. ESR spectrum from the spin label of the neck-linker of dimeric kinesin motor consisted of immobilized and mobilized components and did not exhibit nucleotide-dependent mobility change. The distance between two labels of kinesin dimer was also measured by spin dipole-dipole interaction, showing a broad distribution and a nucleotide-dependent change on the nanometer scale (>1.5 nm). These results suggest that two LC domains of myosin and two neck linkers of kinesin play a similar role for sliding movement using two conformations. The spin label of the skeletal (Tn)-I regulatory domain (TnIreg) showed a large mobility change by Ca2+ ion suggesting a Ca-induced switch movement of TnIreg. Spin dipole-dipole interaction showed that in reconstituted muscle fibers both skeletal and cardiac TnC undergo Ca2+-induced structural change that is thought to be essential for TnIreg movement. We also succeeded in fixing the newly-synthesized bifunctional spin label rigidly on the TnC molecule in solution, indicating that we can determine the precise coordinate of the spin principal axis of troponin on the oriented filament.     

Mechanochemistry and permeability of polyelectrolyte complex membranes composed of poly(vinyl alcohol) derivatives

Two polyanions and two polycations were prepared by the esterification of poly(vinyl alcohol). Properties of their neutral polyelectrolyte complex membranes were compared with those of polyelectrolyte membranes. The charged state on the membranes was related closely to the mechanochemical reaction, the salt rejection, and the solution permeability. Both the salt rejection and the water flux of neutral polyelectrolyte complex memberanes were lower than those of the corresponding acidic or basic complex membranes.     

Characteristics of alpha-glucosidase production from recombinant Aspergillus oryzae by membrane-surface liquid culture in comparison with various cultivation methods

alpha-Glucosidase was produced using recombinant Aspergillus oryzae by membrane-surface liquid culture (MSLC), a method previously developed by the authors and the results compared with other methods, including shaking flask culture (SFC), agar-plate culture (APC), culture on urethane sponge supports (USC), and liquid surface culture (LSC) to determine possible reasons for the advantageous features of MSLC. When yeast extract was used as a nitrogen source, the amount of enzyme produced by MSLC was 5 or more times higher than those for SFC and LSC, but similar to that using APC. Enzyme production in USC was slightly lower than in MSLC and APC. Cell growth was similar irrespective of the cultivation method used. When NaNO3, a typical inorganic nitrogen source was used, enzyme production in all the cultures was lower than that using yeast extract. However, even using NaNO3, the amount of the enzyme produced by MSLC was 8 to 20 times higher than those by SFC, APC, USC, and LSC. Although cell growth using NaNO3 was similar to that for yeast extract in MSLC, it was markedly decreased in SFC, APC, and LSC. The reason for the difference in enzyme productivity for various cultivation methods using yeast extract and NaNO3 as a nitrogen source is discussed, on the basis of the experimental findings. The role of the oxygen transfer effect and gene expression levels in enzyme production were also examined.     

2 Gbit/s small form factor fiber-optic transceiver for single modeoptical fiber

Small form factor (SFF) optical transceivers are expected to be commonly used in the near future for high-end (high bit-rate, single mode) applications as well as for low-cost applications. SFF optical transceivers require new packaging techniques for the optical portion, because the distance between input and output optical axes is much smaller than those of conventional transceivers, e.g., the “1by9” type. Two types of optical packaging are introduced, both of which are suitable for transceivers which have an MT-based receptacle for single mode applications. One of the packaging types, a fiber-bulk structure, takes full advantage in cost and reliability of standard, hermetically sealed TO-canned optical devices. A conventional TO-can packaged laser diode (LD) and photodiode (PD) are held with short optical fibers by low-cost, injection-molded polymer retainers. The short fibers are connected to an MT ferrule inside the transceiver. An alternative packaging type, a surface mount structure, is intended for drastic cost reduction in large volume production by using advanced highly integrated assembly. An LD chip with an integrated spot-size converter and a waveguide-structured PD chip are mounted on a Si-substrate. Optical coupling between the optical devices and fibers on V-groove is accomplished by passive alignment, which reduces assembling time. A prototype transceiver with an MPO receptacle has been developed using the former packaging structure. Evaluated results show performance which complies with the 2.125 Gbit/s Fiber Channel standard. SFF transceivers with these new packaging technologies are expected to increase the optical port density and also to reduce the cost of high-end systems with bit rate of 2 Gbit/s or higher per port     

Neuroimmunological Implications of AQP4 in Astrocytes

The brain has high-order functions and is composed of several kinds of cells, such as neurons and glial cells. It is becoming clear that many kinds of neurodegenerative diseases are more-or-less influenced by astrocytes, which are a type of glial cell. Aquaporin-4 (AQP4), a membrane-bound protein that regulates water permeability is a member of the aquaporin family of water channel proteins that is expressed in the endfeet of astrocytes in the central nervous system (CNS). Recently, AQP4 has been shown to function, not only as a water channel protein, but also as an adhesion molecule that is involved in cell migration and neuroexcitation, synaptic plasticity, and learning/memory through mechanisms involved in long-term potentiation or long-term depression. The most extensively examined role of AQP4 is its ability to act as a neuroimmunological inducer. Previously, we showed that AQP4 plays an important role in neuroimmunological functions in injured mouse brain in concert with the proinflammatory inducer osteopontin (OPN). The aim of this review is to summarize the functional implication of AQP4, focusing especially on its neuroimmunological roles. This review is a good opportunity to compile recent knowledge and could contribute to the therapeutic treatment of autoimmune diseases through strategies targeting AQP4. Finally, the author would like to hypothesize on AQP4’s role in interaction between reactive astrocytes and reactive microglial cells, which might occur in neurodegenerative diseases. Furthermore, a therapeutic strategy for AQP4-related neurodegenerative diseases is proposed.