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We have injected dish-cultured oncogenic RasV12 cells into adult male flies and analyzed by single cell transcriptomics their destiny within the host after 11 days. We identified in the preinjection samples and in the 11-day postinjection samples in all 16 clusters of cells, of which 5 disappeared during the experiment in the host. The other cell clusters expanded and expressed genes involved in the regulation of cell cycle, metabolism, and development. In addition, three clusters expressed genes related to inflammation and defense. Predominant among these were genes coding for phagocytosis and/or characteristic for plasmatocytes (the fly equivalent of macrophages). A pilot experiment indicated that the injection into flies of oncogenic cells, in which two of most strongly expressed genes had been previously silenced by RNA interference, into flies resulted in a dramatic reduction of their proliferation in the host flies as compared to controls. As we have shown earlier, the proliferation of the injected oncogenic cells in the adult flies is a hallmark of the disease and induces a wave of transcriptions in the experimental flies. We hypothesize that this results from a bitter dialogue between the injected cells and the host, while the experiments presented here should contribute to deciphering this dialogue.

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.respol.2005.07.007 Byline: Ryuzo Furukawa (a), Akira Goto (b)(c) Keywords: Corporate scientist; Core scientist; Co-author; Citation; Innovation Abstract: We conducted empirical analysis of the role of corporate scientists in Japanese pharmaceutical companies using data on published papers and patent applications. We found that scientists with the highest publication performance scores did not apply for a considerably greater number of patents than other researchers in their companies. Instead, we found that these 'core scientists' had a positive effect on the number of patent applications filed by their co-authors. Our findings suggest that core scientists play an important role as central conduits for the in-flow of knowledge from outside their companies, thereby stimulating innovation. Author Affiliation: (a) Graduate School of Environmental Studies, Tohoku University, 6-6-11 Aoba, Aramaki, Aoba-ku, Sendai 980-8579, Japan (b) Research Center for Advanced Science and Technology, University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8904, Japan (c) Research Institute of Economy, Trade and Industry, Japan Article History: Received 7 September 2004; Revised 11 March 2005; Accepted 23 July 2005

Findings of a national survey on research and development in manufacturing sectors of United States and Japan are presented.

Onfarm storage facilities can be useful in satisfying the variable diurnal irrigation demand during a day and, at the same time, can prevent waste of water. In this study, an irrigation pond IP model is developed for the determination of required minimum storage capacity at a desired reliability level with a given intake operating rule to meet the fluctuating water demands. This IP model is applied to determine the storage capacities of several lateral canals of the MaeTaeng Irrigation Project in Chiang Mai Province, Thailand. Simulations are carried out to analyze effects of different cropping patterns and hourly irrigation demand patterns at various reliability levels on the storage capacities of irrigation ponds. The determined storage capacities of irrigation ponds for various lateral canals having different cropping patterns varies from about 1,870m3to 2,525m3for a 100 ha command area at 90% reliability. It is observed that introduction of onfarm irrigation ponds can reduce both water waste and deficit by about 20–30%.

Blow forming accompanied with superplasticity makes possible the forming of complex parts, which cannot be formed by cold press forming. The conventional superplastic AA5083 alloy ‘ALNOVI-1’ developed by the Furukawa-Sky Aluminum Corp. shows high superplasticity because of its fine grain and is widely used for blow forming. However, for mass production of components, an Al-Mg alloy with finer-sized grains is needed. In this research, the newly developed high Mn version of the Al-Mg alloy ‘ALNOVI-U’ is used, and this material possesses grains finer than those of the conventional AA5083 alloy. The effects of finer grain size on the blow formability at high strain rates over 10-2/s and the properties of the resulting moldings were studied.

Background: It is difficult to determine the kinematics of the subtalar joint because of its anatomical and functional complexity. The purpose of the study was to clarify the 3D kinematics of the subtalar joint in vivo. Materials and Methods: Subjects were four healthy female volunteers. Magnetic resonance imaging (MRI) sequences were acquired in seven positions during dorsi-plantarflexion (DPF) and in 10 positions during inversion-eversion (IE) at intervals of 10 degrees. MRI data of the talus and calcaneus in the neutral position were superimposed on images of the other positions using voxel-based registration, and relative motions and axes of rotation were visualized and quantitatively calculated. Results: The calcaneus always rotated from dorsolateral to medioplantar during DPF and IE, and the motion plane was very similar to that of the entire foot in IE. The axes of rotation of the calcaneus relative to the talus during DPF and IE had a very close spatial relationship, running obliquely from antero-dorsomedial to postero-planto-lateral and penetrating the talar neck. The rotation angle around each of these calcaneal axes tended to be greater in IE (20 degrees ± 2 degrees) than in DPF (16 degrees ± 3 degrees). In DPF, motion of the calcaneus relative to the talus occurred predominantly around maximum dorsiflexion and plantarflexion, with little movement observed at intermediate positions. During IE, the calcaneus exhibited uninterrupted motion related to foot movement. Conclusion: The subtalar joint is essentially a uniaxial joint with a motion plane almost identical to that of IE of the entire foot. Clinical Relevance: Knowledge of normal subtalar kinematics may be helpful when evaluating pathologic conditions.

The maturation of dendritic cells (DCs) in situ by danger signals plays a central role in linking innate and adaptive immunity. We previously demonstrated that the activation of invariant natural killer T (iNKT) cells by administration of α-galactosylceramide (α-GalCer)–loaded tumor cells can act as a cellular adjuvant through the DC maturation. In the current study, we used allogeneic fibroblasts loaded with α-GalCer and transfected with antigen-encoding mRNA, thus combining the adjuvant effects of iNKT-cell activation with delivery of antigen to DCs in vivo. We found that these cells produce antigen protein and activate NK and iNKT cells. When injected into major histocompatibility complex (MHC)–mismatched mice, they elicited antigen-specific T-cell responses and provided tumor protection, suggesting that these immune responses depend on host DCs. In addition, antigen-expressing fibroblasts loaded with α-GalCer lead to a more potent T-cell response than those expressing NK cell ligands. Thus, glycolipid-loaded, mRNA-transfected allogeneic fibroblasts act as cellular vectors to provide iNKT-cell activation, leading to DC maturation and T-cell immunity. By harnessing the innate immune system and generating an adaptive immune response to a variety of antigens, this unique tool could prove clinically beneficial in the development of immunotherapies against malignant and infectious diseases.

We examined the fragmentation and ionization of molecules by low-temperature electrons generated by electron cyclotron resonance (ECR) plasma. We examined several types of metallocene compounds comprising a metal and 1,3-cyclopentadienes as ligands. We performed analyses using an ECR ion source (ECRIS) mass spectrometer. Consequently, we succeeded in ionizing fragments of an organometallic compound by adjusting the input power of the microwave introducing a super high-frequency plasma. Moreover, we succeeded in dynamically generating a significant quantity of fragment ions by continuously varying the input power. Information on the structure of a molecule may be acquired from this operation. Moreover, a molecule that could not be easily ionized thus far may now be ionizable when soft ionization is performed with this technique.

The ionization efficiency of an electron cyclotron resonance ion source (ECRIS) is generally high and all elements can be fundamentally ionized by the high-temperature plasma. We focused our attention on the high potentiality of ECRIS as an ion source for mass spectrometers and attempted to customize a mass spectrometer equipped with an ECRIS. Precise measurements were performed by using an ECRIS that was specialized and customized for elemental analysis. By using the charge-state distribution and the isotope ratio, the problem of overlap, such as that observed in the spectra of isobars, could be solved without any significant improvement in the mass resolution. When the isotope anomaly (or serious mass discrimination effect) was not observed in ECR plasma, the system was found to be very effective for isotope analysis. In this paper, based on the spectrum (ion current as a function of an analyzing magnet current) results of low charged state distributions (2+, 3+, 4+, …) of noble gases, we discuss the feasibility of an elemental analysis system employing an ECRIS, particularly for isotopic analysis. The high-performance isotopic analysis obtained from an ECRIS mass spectrometer in this study suggests that it can be widely applied to several fields of scientific study that require elemental or isotopic analyses with high sensitivity.

Summary: In this paper we examine the role of what we call core scientists in innovation in Japanese electronics companies. Core scientists are those who have the top total scores as measured by the number of their publications and citations received. We find that even though they may not apply for a large number of patents themselves, the scientific knowledge of the core scientists may have a positive effect in stimulating patent applications by their collaborators.

Summary: The paper aims to clarify the extent to which the results of scientific-oriented research conducted by corporations are reflected in their application-oriented research. Focusing on large Japanese manufacturers of electrical machinery, the paper analyses firm-level data on presentations of scientific papers that represent the results of scientific-oriented research activities, citations of scientific papers in patents, and inventions. The electrical machinery industry, a prototypical science-based industry, has been placing a growing emphasis on scientific-oriented research during the 1990's as is evident from trends in R&D expenses, scientific papers, and inventions. Regression analysis results suggest a complementary relationship between citations of basic scientific knowledge as presented in scientific papers on the one hand and acts of invention on the other hand, in the sense that a rise in citations corresponds to a rise in inventions. Moreover, the results suggest that invention efficiency (number of patent claims per unit of R&D expenditure) has been increasing during the 1990's. Furthermore, the results suggest that, given the exogenous influences on the patent system in Japan, it is necessary to include the number of patent claims when attempting to measure corporate technology development activity through the volume of patent applications. However, there was no finding of a clear relationship between the number of scientific papers and inventions. Implications of these results for corporate R&D strategy are examined.

Two ecologically distinct forms, fresh- and brackish-water types, of ninespine stickleback coexist in several freshwater systems on the coast of eastern Hokkaido. Recent genetic analyses of 13 allozyme loci revealed genetic separation between the two types even though their spawning grounds were in close proximity. On the other hand, there is only a small difference in mitochondrial DNA (mtDNA) sequence between the two types suggesting that they diverged quite recently or that mtDNA introgression occurred between them. To test for postzygotic reproductive isolating mechanisms and hybrid mediated gene flow, we examined the viability and reproductive performance of reciprocal F1 hybrids. The hybrids grew to the adult size normally and both sexes expressed secondary sexual characters in the reciprocal crosses. The female hybrids were reciprocally fertile, while the male hybrids were reciprocally sterile. Histological and flow-cytometric analyses of the hybrid testis revealed that the sterility pattern was classified as 'gametic sterility,' with gonads of normal size but abnormal spermatogenesis. To our knowledge, the present finding is a novel example of one sex hybrid sterility in the stickleback family (Gasterosteidae). [ABSTRACT FROM AUTHOR]

We have previously shown that the LG4 (laminin G-like) domain of the laminin α4 chain is responsible for the significantly higher affinity of the α4 chain to heparin than found for other α chains [Yamaguchi, Yamashita, Mori, Okazaki, Nomizu, Beck and Kitagawa (2000) J. Biol. Chem. 275, 29458–29465]; four basic residues were identified to be essential for this activity [Yamashita, Beck and Kitagawa (2004) J. Mol. Biol. 335, 1145–1149]. By creating GST (glutathione S-transferase)-fused LG1, LG2, LG4 and LG5 proteins, we found that only LG4 is active for the adhesion of human HT1080 cells, human umbilical vein endothelial cells and Drosophila haemocytes Kc167 with a half-saturating concentration of 20 μg/ml. Adhesion was counteracted by treatment of the cells with heparin, heparan sulphate and heparitinase I. Upon mutating the four basic residues essential for heparin binding within LG4, the adhesion activity was abolished. Pull-down experiments using glutathione beads/GST-fusion proteins indicate a direct interaction of LG4 with syndecan-4, which might be the major receptor for cell adhesion. Neither the release of glypican-1 by treating human cells with phosphatidylinositol-specific phospholipase C nor targeted knockdown of dally or dally-like protein impaired the cell-adhesion activity. As the LG4–LG5 domain of the α4 chain is cleaved in vivo from the main body of laminin-8 (α4β1γ1), we suggest that the heparan sulphate proteoglycan-binding activity of LG4 is significant in modulating the signalling of Wnt, Decapentaplegic and fibroblast growth factors.

In northeast Thailand, the rapid expansion of rainfed paddy fields has decreased the stability of rice production. This paper describes a model that computes rice production on the basis of the hydrologic conditions of rainfed paddy fields on hill slopes. The model well expressed the hydrologic processes, rice yield, and production at the study site. We simulated rice production as uphill paddy plots are abandoned, increasing catchment area, under various rainfall conditions. The simulation showed that rice yield and stability increase as uphill plots are abandoned, although the total rice production decreases. Thus, the effect of catchment size on rice production in each plot was quantified. The model proved to be useful for analyzing rainfed rice production under various land and water conditions.

Abstract:  Both growth and migratory history of the Pacific Ocean forms of the threespine stickleback Gasterosteus aculeatus collected in Otsuchi Bay, northeastern Japan, were examined using otolith microstructure and analysis of strontium (Sr) and calcium (Ca) concentrations with wavelength dispersive X-ray spectrometry by an electron microprobe. Age of the juveniles (21.6–25.9 mm in total length) examined ranged from 101 to 128 days (115 ± 8.5 days; mean ± SD), hatching being estimated as having occurred between March and April 2001. The Sr : Ca ratios in the otoliths changed with both ontogenic development and salinity of the habitat. The otolith Sr : Ca ratios increased gradually from 4.1 × 10−3 around the core to 7.5 × 10−3 around the edge of the otolith. The fluctuation pattern of otolith Sr : Ca ratios was different from those observed in both freshwater resident and anadromous forms in previous studies. These results suggested that the fish sampled spend their lives in the estuarine and sea environment without freshwater life after hatching.

Abstract:  Migratory histories of three types of Cottus pollux, the small-egg type (SE type), middle-egg type (ME type), and large-egg type (LE type), were studied by examining strontium (Sr) and calcium (Ca) in their otoliths with wavelength dispersive X-ray spectrometry on an electron microprobe. The Sr : Ca ratios in the otoliths changed both with ontogenetic development and with salinity of the habitat. Otolith Sr : Ca ratios of LE-type samples and the ME-type samples from the Honmyo River, Kyushu Island, showed consistently low ratios, averaging 1.8 × 10−3 and 2.4 × 10−3 from the core to the edge, respectively. In contrast, otolith Sr : Ca ratios of SE-type samples and the other four ME-type samples from Hokkaido and Honshu Islands fluctuated strongly along the life history transects in accordance with migration patterns from freshwater to the sea and vice versa. The otolith Sr : Ca ratios of SE-type samples showed low ratios from the core to a point around 15 μm, averaging 1.5 × 10−3, and subsequently increased sharply with a high Sr : Ca ratio phase to a point around 400 μm, averaging 5.5 × 10−3, and followed again a low ratio phase to the edge with averages of 3.1 × 10−3. Similar fluctuation patterns in otolith Sr : Ca ratios were found for the four ME-type samples. These findings clearly demonstrated that otolith Sr : Ca ratios reflected the sculpin's life histories, as being fluvial for the LE type and the Honmyo River ME type and amphidromous for the SE type and the other four populations of ME type.

Abstract:  The migratory history of two highly divergent forms (the Japan Sea and Pacific Ocean forms) of the threespine stickleback Gasterosteus aculeatus collected from Japanese brackish water (seawater) and freshwater was studied by examining strontium (Sr) and calcium (Ca) concentrations in their otoliths using wavelength dispersive X-ray spectrometry on an electron microprobe. The Sr : Ca ratios in the otoliths changed with salinity of the habitat. The otolith Sr : Ca ratios of the freshwater resident-type samples of the Pacific Ocean form showed consistently low Sr : Ca ratios, averaging 0.85–0.96 × 10−3 from the core to the edge. In contrast, the otolith Sr : Ca ratios of the anadromous type of both the Japan Sea and Pacific Ocean forms fluctuated strongly along the life history transects in accordance with their migration patterns from seawater to freshwater. The higher ratios in the anadromous type, averaging 5.4 × 10−3, in the otolith region from the core to 200 μm, corresponded to the seagoing period, suggesting that otolith Sr : Ca ratios are affected by ambient water salinity. These findings clearly indicate that otolith Sr : Ca ratios reflect individual life histories, and that these two highly divergent forms of stickleback have a flexible migration strategy.

Abstract:  Specimens of the Cottus pollux species' group collected from the upper part of the Honmyo River, Nagasaki Prefecture, Japan, were subjected to morphological and allozyme analyses to place them into one of the recognized valid taxa, viz. small-egg type (SE type), middle-egg type (ME type), or large-egg type (LE type). They were identified as ME type on the basis of specific morphological characteristics, such as laterally depressed cross-sectional shape at posterior half of the body and deep caudal peduncle, and by having a diagnostic allozyme allele (MEP * 54). This ME-type population is the first recorded from Kyushu Island, Japan. An ecological survey of the population revealed that females spawned larger eggs (2.8–3.2 mm in diameter) than those of other amphidromous populations, from which well-developed yolk-sac larvae of about 8.0 mm TL were hatched out. In addition, ME-type specimens collected on 25 and 26 May 2001 included 15 sex-unknown juveniles ranging from 18.8 to 30.2 mm SL, suggesting that they represented larvae hatched out on a nearby spawning ground, with no experience of downstream migration into Isahaya Bay. This observation strongly suggests that the ME-type population in the Honmyo River has a fluvial lifestyle, being different from other amphidromous populations. The former population may have arisen from an amphidromous ancestor through changes in egg size and early ontogenetic development.

Abstract:  The fluvial sculpin, Cottus nozawae, is a coldwater-adapted fish distributed in Hokkaido Island and the northeastern part of Honshu Island (Tohoku District), Japan. Mitochondrial DNA (mtDNA) control region sequencing was used to investigate the geographic distribution of genetic variation and phylogeography of C. nozawae. Most populations possessed unique haplotypes, few being shared across river systems. Phylogenetic analysis of the sequences of the mtDNA control region and adjacent regions of C. nozawae revealed three distinct phylogenetic groups that differed by 3.05% to 3.11%, corresponding to distinct geographic regions, Hokkaido Island, northern Tohoku District, and Yamagata Prefecture (southwestern Tohoku District), respectively. The divergence times of three groups were estimated to be about 1.5 million years ago by applying a general rate for mtDNA, suggesting that the divergence among them might have occurred in the early Pleistocene. Divergence among the haplotypes within the group from the northern Tohoku District was also high (1.84%), no haplotypes being shared by local populations in different river systems in this region. Local populations from a single river system in this region comprise a distinct lineage that differed from other river systems. Such genetically divergent population structures among the different regions and river systems are considered to have resulted mainly from long-term isolation and restricted gene flow among river systems, probably promoted by the fluvial benthic life history and low dispersal ability of this species.

Abstract: Habitat selection by fluvial lamprey larvae, Lethenteron reissneri (Petromyzontidae), was studied in a natural stream and an experimental aquarium to clarify microhabitat requirements for future conservation of natural populations. A gross collection survey of lamprey larvae in the Monbetsu River, southeastern Hokkaido, Japan, revealed a remarkable bias toward distribution in sandy-mud beds. An analysis using Jacobs' electivity index showed that the larvae selectively utilized spaces having regard to shallow water, weak current, deep sandy-mud, and fine substrate particles. A comparison of microhabitat use between small- (≤5 cm) and large-sized larvae (>5 cm) indicated that the latter utilized the space with greater ranges in both water depth and substrate particle size than the former. Both the field survey and laboratory experiments on larval selectivity of physical habitat variables clearly demonstrated that substrate particle size was the most important variable for small-sized larvae whereas both water depth and substrate depth were more important for large larvae. These findings should be applicable in directing attempts at fluvial habitat restoration for conservation of this endangered lamprey species.

Drosophila laminin αβγ trimer assembly in Kc167 cells was perturbed by chain-specific RNA interference (RNAi). The intracellular pool of α and γ chains remained unchanged under β-chain RNAi by lipofection of double-stranded RNA encoding a β-chain partial sequence. This was also the case for the intracellular pool of α and β chains under γ-chain-specific RNAi. Nonetheless, the intracellular pool of β and γ chains increased markedly under α-chain-specific RNAi. Non-reducing SDS/PAGE revealed that some of the increased β and γ chains migrated as disulphide-linked βγ dimers but that the rest migrated as monomers. Since the monomeric β and γ bands detected under α-chain RNAi were denser than the β band under γ-chain RNAi and the γ band under β-chain RNAi, respectively, β and γ also appeared to accumulate by forming βγ dimers without the disulphide linkage. We suggest that interconversion of these βγ dimers is crucial for the replaceable and selective assembly of the α chain for αβγ trimer formation.

We identified a novel Drosophila protein of ≈ 400kDa, hemolectin (d-Hml), secreted from haemocyte-derived Kc167 cells. Its 11.7kbp cDNA contains an open reading frame of 3843 amino acid residues, with conserved domains in von Willebrand factor (VWF), coagulation factor V/VIII and complement factors. The d-hml gene is located on the third chromosome (position 70C1-5) and consists of 26 exons. The major part of d-Hml consists of well-known motifs with the organization: CP1-EG1-CP2-EG2-CP3-VD1-VD2-VD′-VD3-VC1-VD′′-VD′′′-FC1-FC2-VC2-LA1-VD4-VD5-VC3-VB1-VB2-VC4-VC5-CK1 (CP, complement-control protein domain; EG, epidermal-growth-factor-like domain; VB, VC, VD, VWF type B-, C- and D-like domains; VD′, VD′′, VD′′′, truncated C-terminal VDs; FC, coagulation factor V/VIII type C domain; LA, low-density-lipoprotein-receptor class A domain; CK, cysteine knot domain). The organization of VD1-VD2-VD′-VD3, essential for VWF to be processed by furin, to bind to coagulation factor VIII and to form interchain disulphide linkages, is conserved. The 400kDa form of d-Hml was sensitive to acidic cleavage near the boundary between VD2 and VD′, where the cleavage site of pro-VWF is located. Agarose-gel electrophoresis of metabolically radiolabelled d-Hml suggested that it is secreted from Kc167 cells mainly as dimers. Resembling VWF, 7.9% (305 residues) of cysteine residues on the d-Hml sequence had well-conserved positions in each motif. Coinciding with the development of phagocytic haemocytes, d-hml transcript was detected in late embryos and larvae. Its low-level expression in adult flies was induced by injury at any position on the body.

Evolutionary processes in East Asian ninespine sticklebacks ( Pungitius spp.), including both extremes of armor morphology in the genus, were demonstrated with mitochondrial DNA control region (CR) phylogeny. Entire CR sequences (830-930 bp long) were determined for three species: the most heavily armored ( P. sinensis ), the most reduced ( P. tymensis ), and an intermediate ( P. pungitius ). The former two species are endemic to East Asia, the latter being circumpolar. Three major lineages (A, B, and C) were revealed, whereas both the phylogenetic trees and the insertion sequence dynamics supported the polyphyly of P. sinensis. Haplotypes of the mainland populations of P. sinensis possessed lineage B, being the sister group of P. tymensis lineage A. Island populations of P. sinensis, however, possessed lineage C, along with all P. pungitius haplotypes. A molecular clock hypothesis was clearly rejected for the CR sequences, significantly slower evolutionary rates being observed in the P. tymensis lineage. The split of mainland P. sinensis and P. tymensis was considered to have preceded that of the lineage C colonization in East Asia. The contrasting morphology is probably attributable to adaptation of P. tymensis to island freshwater environments and an ecological interaction between P. tymensis and lineage C emigrants.

Abstract: The freshwater type of ninespine sticklebacks, genus Pungitius, is widely distributed in northern Japan and reproductively isolated from other genetically divergent types endemic to small regions in Japan. This type expresses dimorphism in its lateral plate morphology: complete and partial row morphs. The two morphs show a parapatric distribution in Japan. To clarify the process involving the distribution of these two morphs, we examined their phylogeography based on restriction fragment length polymorphism of an entire mitochondrial DNA (mtDNA). The survey was carried out with seven restriction enzymes on the populations of the freshwater type collected from 41 localities across the distribution range in Japan, and 6 further Pungitius populations from the Okhotsk Sea coast of Russia were appended. An unweighted pair-group method with arithmetic averages (UPGMA) tree among 54 mtDNA haplotypes resolved eight clustering groups that differed in sequence divergence by approximately 1.3%–2.1%. Two of the eight groups were found only in Russia. mtDNA phylogenies constructed by neighbor-joining and Wagner parsimony methods suggested that the haplotypes of each plate morph were polyphyletic. The geographic distribution pattern of these groups suggests that they should be classified into two broad categories, one with extensive distribution and the other with localized distribution of the constituent haplotypes within a group. The former groups were found mainly in the populations with the completely plated morph and the latter groups with the partially plated morph. It is supposed that twice dispersals of dimorphic or complete plated ancestors and genetic differentiation during the interglacial played an important role in the formation of the present distribution of the two morphs in Japan.

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We have cloned cDNA encoding Drosophila mitochondrial (mt) transcription factor A (d-TFAM). RNA interference (RNAi) of d-TFAM by lipofection of haemocyte-derived Kc167 cells with double-stranded RNA reduced d-TFAM to less than 5% of the normal level. Reflecting the ability of TFAM to stabilize mtDNA, RNAi of d-TFAM reduced mtDNA to 40%. Nonetheless, transcription of the ND2 and ND5 genes and their mRNAs remained unchanged for 8 days of the duration of RNAi. We thus show that d-TFAM is not essential for the transcription of Drosophila mtDNA.

The distribution pattern of the freshwater sculpin,Cottus nozawae, was investigated in the Gakko River system, Tohoku District, northern Japan, that region representing the southern limit of the species distribution.C: nozawaewas not found in the lower sections of the river, being instead restricted mainly to the upstream segments of some tributaries. The relationships between habitat variables and density ofC. nozawaeanalysed at reach scale, revealed that fish density increased with cover area, decreasing stream width and lower maximum water, temperature. Additionally, maximum water temperature was the best predictor of fish density according to stepwise multiple regression analysis. These data indicated that tributary populations ofC. nozawaewere isolated from thermally unsuitable environments. Accordingly,C. nozawaewas distributed patchily in headwater streams of this river system, in contrast to those on Hokkaido Island, where the species is distributed continuously throughout middle to upper stream courses.

The level of reproductive isolation and reproductive characteristics were examined in sympatric populations of the northern and southern forms ofLethenteron reissneri, in the Ushiwatari Stream, a tributary of the Gakko River, Yamagata Prefecture, Japan, to elucidate the reproductive isolating mechanisms existing between them. Extensive allozyme data showed no gene exchange between the two forms in the study area, evidence of their strict reproductive isolation from each other. The spawning seasons overlapped between the northern and southern forms, the spawning period of the latter being slightly later than that of the former. The size at maturity also overlapped between the two forms, although the southern form indicated significantly larger in average size than the northern one. These results suggested that neither seasonal reproductive isolation nor size assortative mating occurred. On the other hand, the nesting assemblages were observed to comprise only males and females of each respective form, suggesting that a recognition system restricted to each form may operate as a premating isolating mechanism.

Kc 167 is a cell line established from Drosophilaembryonic hemocytes and has been shown to express many extracellular matrix (ECM) and other proteins important during development. We have screened monoclonal antibodies (mAbs) raised against heparin affinity purified proteins from conditioned medium of Kc 167 cells to identify novel proteins with important roles for development. One mAb recognized a protein expressed with temporary and tissue specific patterns during Drosophilaembryogenesis and larval development. This approach is an alternative to screening of Expression Sequence Tag (EST) clones by in situhybridization to initiate reverse genetics. In addition, a number of mAbs recognizing ECM proteins were also identified. These mAbs will be useful for biochemical and cell biological analyses of DrosophilaECM proteins.

This report describes symmetry properties of tetragonal vortex lattice solutions of the Bogoliubov–de Gennes equation in a two-dimensional square lattice in a uniform magnetic field. The invariance group of a tetragonal vortex lattice solution is expressed in a form of G(l)= (e+ tC2x) $$\widetilde C_4^l L$$ (l= 0, 2, ± 1), where tC2xis a space π rotation around the x-axis accompanied with time reversal, $$\widetilde C_4^l$$ is a kind of fourfold rotation group, and Lis the magnetic translational group of the vortex lattice state. We give a new, refined definition of local symmetric order parameters (OPs) (s-wave, d-wave, and p-wave), which have a well-defined nature such that the OP (e.g., s-wave OP) at the translated site by a lattice vector (of the vortex lattice) from a site (m, n) is expressed by the OP (e.g., s-wave) at the site (m, n) times a phase factor. Winding numbers around the origin of s-wave and d-wave OPs are obtained for four types of solutions G(l)(l= 0, 2, ± 1). It is shown that all energy bands of quasiparticles of a vortex lattice state are doubly degenerate.

The relationships between marine growth and survival, and smolt sizes were examined for white-spotted charr (Salvelinus leucomaenis) populations in the Nairo River, Rebun Island, off northern Hokkaido Island, and the Haraki River, southern Hokkaido Island, Japan. Smolts in the Haraki River were much larger in size than those in the Nairo River, despite being similar size among age cohorts and between the sexes in each population. From scale analyses, smolt size-specific growth rates and survival were estimated by utilizing both observed and back-calculated size-frequencies. Inverse correlations between specific growth rates at sea and smolt size were found in both populations. However, the expected size increments at a given size of smolts in the Haraki River were significantly larger than of those in the Nairo River. For both populations, there were no significant effects of smolt ages (2 to 5 or 6 years) on growth rates at sea. Larger smolts had a consistent survival advantage, although the degree of size-dependent survival function seemed to differ between populations. The results suggested that growth and survival of individuals at sea are determined ultimately by size-, rather than age-, dependent factors depending upon local environmental conditions, supporting the hypothesis of a threshold size for smolting within populations.

Reproductive behavior, distribution of spawning grounds and possibility of reproductive homing were studied and examined in the river sculpin. Cottus hangiongensis. The courtship, spawning behavior and parental care of this species, observed under artificial stream conditions using a fiber-scope, were quite similar to those of several congenous species already reported. In a natural river, the spawning grounds were restricted to the lower reaches within the distribution range of the species. Slightly before the spawning period, mature individuals inhabiting more upper reaches than the spawning area migrated downstream to the spawning area. The upstream movement of many individuals, apparently downstream migrants only, was observed just after the spawning period. By tracing the location at recapture, it was detected that a total of 11 marked individuals (10 females and 1 male) migrated downstream before spawning, and after spawning they migrated upstream and returned to the original home sections. This suggests the ability for reproductive homing.

This paper analyzes vertical restraints in relation to market access issues. In Section II we briefly review the recent three major trade conflicts between Japan and the USA in light of vertical restraints. In Section III we review the major policy lessons of the economics literature of vertical restraints, and analyze whether we need a special treatment for vertical restraints abroad for a market access reason. In Section IV we evaluate the policy options towards foreign foreclosure and suggest that nondiscriminatory application of foreign or home competition policy should be the mechanism to address such concern instead of trade policy actions such as “voluntary” import expansions.

The rate of complex formation between calix[6]arene-p-hexasulfonate and uranyl ion is studied over a wide range of carbonate ion concentrations. The presence of carbonate ion decreases the complexation rate. The distribution of various uranyl species is calculated from a set of mass balances of participating ions with their stability constants. UO2(CO3)34- has the highest concentration, followed by UO2(OH)3- and UO2(CO3)22-. Other uranyl species are negligible. The complexation rate is proportional to the 0.27-1.0 power of the total concentration of uranyl species other than UO2(CO3)34-. This implies that the rate-determining step of the complexation is the reaction between calix[6]arene-p-hexasulfonate and UO2(OH)3- or UO2(CO3)22-.

An amidoxime fibrous adsorbent is contacted with uranium-enriched seawater (10 ppm); about 10 mg uranium is loaded per 1 g dry fiber. Then the rate and yield of uranium desorption from the fiber are determined with various eluents. Acid solutions are superior to alkali carbonate solutions as eluents. With a 0.1 mol·L-1 HCl solution, desorption is completed in 2 hours regardless of the presence of uranium in the leaching solution up to 15 ppm (≈ 6 × 10-5 mol·L-1). Serial operation of the adsorption-desorption cycle four times does not affect desorption efficiency, but the addition of heavy metal ions to the eluent at a level of 1.8 × 10-3 mol·L-1 significantly decreases desorption efficiency.

We have investigated an interorganelle communication pathway between the nucleus and mitochondria. We loaded a stress specific to mitochondria of human fibroblast cells by antimycin A (AA), an inhibitor of the mitochondrial cytochromebc1complex. AA inhibited cellular respiration in a dose-dependent manner. When the respiratory capacity was reduced to 50–70% of the original one, mRNA levels of cytochromec1as well as cytochromebincreased at 24 h after AA treatment, resulting in maintenance of the cell viability. In contrast, the cells retaining less than 40% of the original capacity showed no increase in either mRNA level and were targeted for death. Intracellular H2O2level monitored by the fluorescence of dichlorofluorescein increased within 3 h in both the cases, although this increase was higher in the cells where the mRNA levels increased. An antioxidantN-acetylcysteine repressed the increases of not only H2O2but also cytochromec1mRNA levels. These results suggest that the cells can respond to a limited impairment of electron transfer by promoting expression of nuclear and mitochondrial genes, probably through an H2O2-dependent signaling pathway.

Polypatric speciation from an amphidromous ancestor has been suggested for a fluvial gobyRhinogobius flumineus; a similar history may explain life-history and genetic variation inCottusspecies. We used starch-gel electrophoresis to examine intra- and interspecific genetic differentiation of two siblingCottusspecies, amphidromousC. amblystomopsisand fluvialC. nozawae, which are presumed to be in an ancestor-descendant phyletic relationship. Genetic differentiation was markedly less withinC. amblystomopsis, than inC. nozawae. One group ofC. nozawaepopulations was closer toC. amblystomopsis(by Nei's genetic distance) than it was to other populations ofC. nozawae. This suggests that the differentiation of presently identifiable groups ofC. nozawaepopulations might have occurred polypatrically from an amphidromous ancestral population at different times. The two groups are presumed to be different species in spite of their quite similar morphological and ecological characteristics: we propose to call this type of speciation ‘Polypatric and parallel speciation’. This model of ‘Polypatric and parallel speciation’ might be generalized to explain evolution of amphidromous or anadromous species to fluvial or lacustrine land-locked species.

Japanese freshwater sculpins consist of 6Cottusand 1Trachidermusspecies, with various types of life-cycles, namely, catadromous, amphidromous, lacustrine land-locked and fluvial land-locked. Among them,C. amblystomopsisandC. nozawaehave been regarded as sibling species. Because of very similar morphological and ecological adult characteristics they were formerly classified as a single species.C. amblystomopsismainly inhabits the lower courses of rivers and produces many small eggs from which pelagic larvae are formed. In contrast,C. nozawaelives in the middle or upper courses of rivers and deposits few, large eggs, from which well-developed benthic young emerge, well on the way to the definitive phenotype as fully formed juveniles. These two species have distinctly different life-cycles: amphidromous forC. amblystomopsisand fluvial forC. nozawae. A comparison of the early ontogeny shows that, amongCottusspecies, the small-egg, amphidromousC. amblystomopsisis altricial, while the large-egg, fluvialC. nozawaeis precocial. Assuming thatC. nozawaehas been derived phylogenetically fromC. amblystomopsisor its ancestral relatives, it is reasonable to consider that the fluvial land-locked life-history style ofC. nozawaehas evolved from the ancestral arnphidromous one through adapting to the upstream habitat by an increase in egg size and consequent truncation of the larval period.

Body size at maturity, individual growth, upstream migration and spatial stability in males of the amphidromous sculpin,Cottus hangiongensis(Cottidae), were investigated in the Daitobetsu River of southern Hokkaido, Japan. The body size of males at first maturity varied gradually along the course of the river, from approximately 70 mm SL in the lowest reaches (0.2 km up from the river mouth) to about 120 mm SL in the uppermost reaches (about 6 km up from the river mouth). The age at first maturity was estimated as 2 to 3 years in the lower reaches and as 4 to 5 years in the upper reaches. Individual growth rate tended to increase upstream, while population density decreased. These results suggest that the male life-history pathway varies along the course of the river and so allows one to consider the relationships to the polygynous mating system of this species in terms of current hypotheses regarding alternative reproductive tactics.

On‐farm storage facilities can be useful in satisfying the variable diurnal irrigation demand during a day and, at the same time, can prevent waste of water. In this study, an irrigation pond (IP) model is developed for the determination of required minimum storage capacity at a desired reliability level with a given intake operating rule to meet the fluctuating water demands. This IP model is applied to determine the storage capacities of several lateral canals of the Mae‐Taeng Irrigation Project in Chiang Mai Province, Thailand. Simulations are carried out to analyze effects of different cropping patterns and hourly irrigation demand patterns at various reliability levels on the storage capacities of irrigation ponds. The determined storage capacities of irrigation ponds for various lateral canals having different cropping patterns varies from about 1,870 m3 to 2,525 m3 for a 100 ha command area at 90&percent; reliability. It is observed that introduction of on‐farm irrigation ponds can reduce both water waste and deficit by about 20–30&percent;.

• A survey was conducted of 89 households in each of which at least one patient with culture-confirmed pertussis had been detected. The source of infection was found to be an adult in 10 (11.2%) of the 89 households, and the rate of secondary attack was 19(10.3%) of 185. Furthermore, a laboratory study disclosed 17 adults with subclinical pertussis; the subclinical infection rate was 17 (25.0%) of 68. When compared with pertussis in young children, the adult illness was generally less severe and had different clinical features. Adult pertussis showed neither leukocytosis nor lymphocytosis, but it produced anti—pertussis toxin anti-body more quickly and higher levels of anti—filamentous hemagglutinin and agglutinin antibodies, and showed stronger growth inhibition of Bordetella pertussis. Although adult pertussis is usually unrecognized because of its different clinical and laboratory features, it is a significant health threat that requires some measures for disease control.(AJDC. 1992;146:163-166)

A 12-month-old girl with delayed developmental milestones, due to muscle hypotonia and weakness from early infancy, exhibited type 2 fiber hypoplasia. A muscle biopsy specimen disclosed type 1 fiber predominance and type 2B fiber deficiency compatible with congenital myopathy. During the following 4 years, she continued to have mild muscle weakness, but no mental retardation. Her mother had similar symptoms from early infancy with minimal progression. Although type 2 fiber hypoplasia is a non-specific finding in various diseases, it may be a specific finding in a limited number of patients with hereditary congenital non-progressive myopathy.

Abstract: Growth patterns of the 1982 year-class, individual growth patterns, age and size at sexual maturity and longevity in females of the river-sculpin,Cottus hangiongensis (Cottidae), were examined along the course of the Daitobetsu River of southern Hokkaido, Japan. Growth of females slightly varied both along the river course and among individual fishes: slow growth occurs in females from the lower reaches, while more rapid growth occurs in females from upstream areas. Body size and age at the first sexual maturity of females slightly increased towards the upstream, from 52 mm SL and 2 years in the most downstream area to 72 mm SL and 2–3 years in the uppermost site. Longevity was estimated to be 7 years in the downstream areas and 8–9 years in the upstream sites. These results suggest that female life history varies along the course of the river and thus allow us to consider the following alternative reproductive tactics: when females stay in the lower reaches, they attain sexual maturity at a smaller body size and younger age, and have a small clutch size, but when females migrate into the upper reaches, their maturity is delayed until they reach a larger body size and older age, and have a greater clutch size.

Abstract: An electrophoretic study on the biochemical genetics of two sibling river-sculpinsCottus amblystomopsis andC. nozawae was undertaken with the primary objective of clarifying the reproductive isolation between the sympatric populations in three rivers around Cape Erimo of Hokkaido, where their distributions overlap widely along the river courses. At the 3 lociAcp,Ldh and6Pgd, out of 20 examined loci, evident displacement of alleles were observed between the two species. In addition, no genetical evidences for hybridization between the two species were detected in the three rivers examined. These results strongly suggest that the two species are reproductively isolated from each other even when they are distributed sympatrically and their distributions overlap widely along the course of a river.

Abstract: A brackish water type ofPungitius pungitius (Gasterosteidae) is newly reported from Japan. Genetic differences, based on 13 loci of 6 enzymes and muscle protein, were examined between fresh and brackish water types in the Biwase River in the eastern part of Hokkaido. The alleles onGpd, Pgi andPgm were displaced between them, even though their distributions partially overlapped in the river. The brackish water type was also statistically different from the fresh water type on the basis of three morphological characters, such as number of dorsal spines, gill-rakers and vertebrae, although ranges overlapped. Segregation of spawning sites and differing nuptial colors of the male were found between fresh and brackish water types. These facts suggest that the brackish water type is reproductively isolated from the fresh water type and that both types should be regarded as independent species.

Abstract: The frequency distribution of the fork length of 0+ aged masu salmon,Oncorhynchus masou, changed from unimodal to bimodal distribution in autumn of the years from 1982 to 1984 in the Mogusa River of southern Hokkaido, Japan. The bimodal distribution consists of two (upper and lower) modal groups. These two groups resulted from a difference in growth rate of 0+ aged individuals in autumn. Fish belonging to the upper modal group are assumed to be potential 1+ smolts. Whether 0+ aged parr transform into smolt or remain as parr in the following spring may be related to the growth rate of fish in the first autumn.