Molecular cloning and characterization of rat lin-10

In Caenorhabditis elegans, the vulval induction is mediated by tyrosine kinase receptor/Ras signal transduction pathway composed of the lin-3, let-23, and let-60 products. In addition to these gene products, the lin-2, lin-7, and lin-10 products are also implicated in this pathway. Lin-2 encodes a MAGUK and lin-7 encodes a small protein with one PDZ domain. The lin-10 product has no homology to known proteins. Here, we have cloned a rat homologue of the lin-10 product and characterized it. Rat lin-10 is ubiquitously expressed in various rat tissues and distributed in both the cytosol and membrane fractions. In brain, however, rat lin-10 is distributed only in the membrane fraction and enriched in the synaptic plasma membrane and postsynaptic density fractions. These results suggest that rat lin-10 is involved at least in synaptic functions in brain.     

Development of next generation PWR in Japan

The demand for energy in Japan is expected to increase steadily into the future, and it seems that the importance of nuclear power generation will be heightened more when the situation of our country which is not rich in energy resources is taken into account.

Furthermore, when we consider the present situation that the light water reactors have become common, recent outlook for the supply and demand for uranium resources, trends in the development of the fast breeder reactor technology, etc., the light water reactors are expected to remain dominant in the nuclear power generation of our country until at least the second half of the 21st century.

Based on such a background five PWR utilities in Japan (Hokkaido, Kansai, Shikoku, Kyushu, and the Japan Atomic Power), and Mitsubishi Heavy Industries, Ltd. have jointly started researching the Next Generation PWR which is expected to be the leading nuclear power plant taking place of APWR.     

Protein modelling using a chimera reference protein derived from exons

Bovine pancreatic /S-trypsin (PDB ID-code: 1TPO) which is registeredin the Brookhaven Protein Data Bank (PDB) consists of four exons.The results of homology searches for each exon in the PDB showedthat homologous proteins were tonin (PDB ID-code: 1TON), ratmast cell protease (PDB ID-code: 3RP2_A), kaffikrein A (PDBID-code: 2PKA_B) and kallikrein A (2PKA_B) respectively. Thus,for the three-dimensional structure prediction of 1TPO, a chimeraprotein was constructed from the three proteins mentioned aboveand the 3-D structure prediction was performed using this chimerareference protein. The modelled structure of 1TPO was energeticallyoptimized by molecular mechanics and molecular dynamics simulationand was compared with its X-ray crystal structure registeredin the PDB. The root mean square deviations (r.m.s.d.) of mainchain atoms and the neighbouring active site (5 sphere fromHis57, AsplO2 and Serl95) between the modelled structure andthe X-ray structure were 1.66 and 0.94 respectively. Porcinepancreatic elastase (PDB ID-code: 3EST) which is registeredin the PDB was used as the reference protein and the modelledstructure from 3EST was also compared with the X-ray data. Ther.m.s.d. of main chain atoms and that of the active site were2.14 and 1.18 respectively. These results dearly support thepropriety of this method using the chimera reference protein.     

Dynamics and organization of MAP kinase signal pathways

In the budding yeast, Saccharomyces cerevisiae, four separate but structurally related mitogen-activated protein kinase (MAPK) activation pathways are known. The best understood of these regulates mating. Pheromone binding to receptor informs cells of the proximity of a mating partner and induces differentiation to a mating competent state. The MAPK activation cascade mediating this signal is made up of Ste11 (a MEK kinase [MEKK]), Ste7 (a MAPK/ERK kinase [MEK]), and the redundant MAPK-related Fus3 and Kss1 enzymes. Another MAPK activation pathway is important for cell integrity and regulates cell wall construction. This cascade consists of Bck1 (a MEKK), the redundant Mkk1 and Mkk2 enzymes (MEKs), and Mpk1 (a MAPK). We exploited these two pathways to learn about the coordination and signal transmission fidelity of MAPK activation cascades. Two lines of evidence suggest that the activities of the mating and cell integrity pathways are coordinated during mating differentiation. First, cells deficient in Mpk1 are susceptible to lysis when they make a mating projection in response to pheromone. Second, Mpk1 activation during pheromone induction coincides with projection formation. The mechanism underlying this coordination is still unknown to us. Our working model is that projection formation generates a mobile second messenger for activation of the cell integrity pathway. Analysis of a STE7 mutation gave us some unanticipated but important insights into parameters important for fidelity of signal transmission. The Ste7 variant has a serine to proline substitution at position 368. Ste7-P368 has higher basal activity than the wild-type enzyme but still requires Ste11 for its function. Additionally, the proline substitution enables the variant to transmit the signal from mammalian Raf expressed in yeast. This novel activity suggests that Ste7-P368 is inherently more permissive than Ste7 in its interactions with MEKKs. Yet, Ste7-P368 cross function in the cell integrity pathway occurs only when it is highly overproduced or when Ste5 is missing. This behavior suggests that Ste5, which has been proposed to be a tether for the kinases in the mating pathway, contributes to Ste7 specificity and fidelity of signal transmission.     

PLANGENT: an approach to making mobile agents intelligent

Network environments give computer users the option of employing distributed information and services to complete a task. However, gathering information and using services distributed in networks requires knowing exactly what kinds of information and services are required for a task, where they are, and how they can be obtained or utilized. Tracking down the answers to these questions can be difficult, time consuming tasks for users. Mobile agent technology is expected to release them from having to do so. Instead, “intelligent” mobile agents will comprehend the user's requirements, search network nodes autonomously for appropriate information and services, and return with the answers. But several problems must be solved before we can expect agents to perform such actions effectively. We focus on the question of intelligence as a prerequisite for agent functions. What sort of intelligence is expected of agents? We have adopted a model based on the ability to make flexible plans. Specifically, we think mobile agents must be able to: understand user requirements; plan actions that will satisfy the requirements act according to the plan; modify the plan according to actual conditions when they differ from those initially expected; and execute the modified plan. We have implemented these functions in the Plangent system and validated their effectiveness in several example applications. We describe how we combined these planning functions with mobile agent facilities, and show how the agents behave intelligently in an example application of personal travel assistance     

Serum Amyloid A3 Promoter-Driven Luciferase Activity Enables Visualization of Diabetic Kidney Disease

The early detection of diabetic nephropathy (DN) in mice is necessary for the development of drugs and functional foods. The purpose of this study was to identify genes that are significantly upregulated in the early stage of DN progression and develop a novel model to non-invasively monitor disease progression within living animals using in vivo imaging technology. Streptozotocin (STZ) treatment has been widely used as a DN model; however, it also exhibits direct cytotoxicity to the kidneys. As it is important to distinguish between DN-related and STZ-induced nephropathy, in this study, we compared renal responses induced by the diabetic milieu with two types of STZ models: multiple low-dose STZ injections with a high-fat diet and two moderate-dose STZ injections to induce DN. We found 221 genes whose expression was significantly altered during DN development in both models and identified serum amyloid A3 (Saa3) as a candidate gene. Next, we applied the Saa3 promoter-driven luciferase reporter (Saa3-promoter luc mice) to these two STZ models and performed in vivo bioluminescent imaging to monitor the progression of renal pathology. In this study, to further exclude the possibility that the in vivo bioluminescence signal is related to renal cytotoxicity by STZ treatment, we injected insulin into Saa3-promoter luc mice and showed that insulin treatment could downregulate renal inflammatory responses with a decreased signal intensity of in vivo bioluminescence imaging. These results strongly suggest that Saa3 promoter activity is a potent non-invasive indicator that can be used to monitor DN progression and explore therapeutic agents and functional foods.     

Lower Tropospheric Aerosol Measurements by MAX-DOAS During Severe Asian Dust Period

A recently developed aerosol retrieval algorithm based on O₄ slant column densities (SCDs) measured at a visible wavelength (476 nm) was utilized to derive aerosol information (e.g., aerosol optical depth (AOD) and vertical distributions of aerosol extinction coefficients (AECs)) in the lower troposphere during a severe Asian dust period. The MAX-DOAS measurements were carried out at Gwangju, Korea for nearly three months from February through May 2008. Comparison with AOD and surface PM₁₀, measured by collocated sunphotometer and beta gauge sampler, were made to validate the retrieved AODs and AECs in the atmospheric layer surface to 1 km height above ground. On the Asian dust days, temporal variations of the AODs retrieved from MAX-DOAS measurements show similar patterns, but with reduced magnitudes, to those measured by sunphotometer whereas similar AOD magnitude and temporal variation was observed between MAX-DOAS and sunphotometer measurements during the non-episodic days. Smaller correlation was observed between the surface PM₁₀ and AECs at 0.5 km during the Asian dust period compared to the correlation obtained for the non episodic days. This study demonstrates the ability of MAX-DOAS as a remote sensing technique for surface aerosol measurements under conditions of homogeneously distributed pollution in the planetary boundary layer. However, for the measurement of significantly enhanced aerosol loads with heterogeneous vertical distribution (e.g., Asian dust), measured AODs and AECs are underestimated at altitudes above 1 km due to decreased sensitivity of MAX-DOAS measurements at high altitudes.     

Calcitonin gene-related peptide (CGRP)-containing nerve fibers in bone tissue and their involvement in bone remodeling

Bone remodeling is a process of bone renewal accomplished by osteoclastic bone resorption and osteoblastic bone formation. These two activities are regulated by systemic hormones and by local cytokines and growth factors. Moreover, the nervous system and certain neuropeptides seem to be involved in regulation of bone remodeling. In this paper, we focus on the distribution of CGRP-containing nerve fibers and their dynamics, and discuss the role of these fibers as a possible mechanism for nervous system involvement in regulation of bone remodeling. CGRP-immunoreactive nerve fibers are widely distributed in bone tissue, such as periosteum and bone marrow, and show apparent regional distribution with different densities. They are often associated with blood vessels and show a beaded appearance. The wide distribution of CGRP-immunoreactive nerve fibers in bone tissue and the changes in distribution during bone development and regeneration suggest the involvement of these fibers in bone remodeling. The effect of CGRP on bone remodeling could partly be through its action on blood vessels, thereby regulating local blood flow. Moreover, in vitro biochemical data and the localization of CGRP-immunoreactive nerve fibers in the vicinity of bone cells suggest that they are directly involved in local regulation of bone remodeling by elevating the concentration of CGRP in the microenvironment around bone cells, especially during bone growth or repair.     

The measurement of hydrogen activities in molten copper using an oxide protonic conductor

We have developed a hydrogen sensor for in situ measurements of hydrogen activities in molten copper. The sensor consists of a concentration cell utilizing a proton conductor, CaZr_0.9In_0.1O_3-δ, as the solid electrolyte. The electromotive force (emf) of the cell was generated by both hydrogen and oxygen activity gradients across the cell in a high-temperature region simulating the fire refining processes of copper. However, accurate hydrogen activity in molten copper could be evaluated from the emf if oxygen activity in molten copper was determined simultaneously by another concentration cell and if the hydrogen and the oxygen activities at the reference electrode were known. The performance of the sensor was studied under various conditions. The observed good response and reliability of the sensor show that it should be a powerful tool for improvement of the fire refining process of the molten copper. Theoretical treatment of the calculation of the emf of the concentration cells using a mixed ionic conductor, i.e., protonic and oxide ionic conductor, as solid electrolytes is also discussed briefly.