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Commercial greenhouse growers in both Japan and China are increasingly using reared orange-tailed bumblebees known previously as Bombus hypocrita Pérez as pollinators. Phylogenetic analysis of the DNA (COI) barcodes with Bayesian methods shows that this "species" is a long-standing confusion of two cryptic species. We find that the orange-tailed bumblebees in North China are actually part of the widespread Russian (otherwise white-tailed) B. patagiatus Nylander (as B. patagiatus ganjsuensis Skorikov, n. comb.), whereas the orange-tailed bees in Japan are true B. hypocrita. This situation has been further complicated because two other cryptic species from North China that were previously confused with the Russian B. patagiatus are now recognised as separate: B. lantschouensis Vogt n. stat. and B. minshanensis Bischoff n. stat. As demand for pollination services by greenhouse growers inevitably increases, these bees are more likely to be transported between countries. In order to conserve genetic resources of pollinator species for their option value for future food security, we advocate preventing trade and movement of B. patagiatus from China into Japan and of B. hypocrita from Japan into China. © 2012 Williams et al.

Bumblebees of the subgenus Bombus s. str. dominate (or used to dominate) many north temperate pollinator assemblages and include most of the commercial bumblebee pollinator species. Several species are now in serious decline, so conservationists need to know precisely which ones are involved. The problem is that many Bombus s. str. species are cryptic, so that species identification from morphology may be impossible for some individuals and is frequently misleading according to recent molecular studies. This is the first review of the entire subgenus to: (1) avoid fixed a priori assumptions concerning the limits of the problematic species; and (2) sample multiple sites from across the entire geographic ranges of all of the principal named taxa worldwide; and (3) fit an explicit model for how characters change within an evolutionary framework; and (4) apply explicit and consistent criteria within this evolutionary framework for recognising species. We analyse easily-obtained DNA (COI-barcode) data for 559 sequences from 279 localities in 33 countries using general mixed Yule-coalescent (GMYC) models, assuming only the morphologically distinctive species B. affinis Cresson, B. franklini (Frison), B. ignitus Smith and B. tunicatus Smith, and then recognise other comparable COI-barcode groups as putative species. These species correspond to modified concepts of the taxa B. cryptarum (Fabricius), B. hypocrita Pérez, B. jacobsoni Skorikov, B. lantschouensis Vogt n. stat., B. longipennis Friese, B. lucorum (Linnaeus), B. magnus Vogt, B. minshanensis Bischoff n. stat., B. occidentalis Greene, B. patagiatus Nylander, B. sporadicus Nylander, B. terrestris (Linnaeus) and B. terricola Kirby (a total of 17 species). Seven lectotypes are designated. Our results allow us for the first time to diagnose all of the putative species throughout their global ranges and to map the extent of these geographic ranges.

Scaffold design plays a crucial role in developing graft-based regenerative strategies, especially when intended to be used in a highly ordered nerve tissue. Here we describe a hybrid matrix approach, which combines the structural properties of collagen (type I) with the epitope-presenting ability of peptide amphiphile (PA) nanofibers. Self-assembly of PA and collagen molecules results in a nanofibrous scaffold with homogeneous fiber diameter of 20-30 nm, where the number of laminin epitopes IKVAV and YIGSR can be varied by changing the PA concentrations over a broad range of 0.125-2 mg/ml. Granule cells (GC) and Purkinje cells (PC), two major neuronal subtypes of cerebellar cortex, demonstrate distinct response to this change of epitope concentration. On IKVAV hybrid constructs, GC density increases three-fold compared with the control collagen substrate at a PA concentration of ≥0.25 mg/ml, while PC density reaches a maximum (five-fold vs. control) at 0.25 mg/ml of PA and rapidly decreases at higher PA concentrations. In addition, adjustment of the epitope number allowed us to achieve fine control over PC dendrite and axon growth. Due to the ability to modulate neuron survival and maturation by easy manipulation of epitope density, our design offers a versatile test bed to study the extracellular matrix (ECM) contribution in neuron development and the design of optimal neuronal scaffold biomaterials. © 2011 Elsevier Ltd.

Long-term depression (LTD) was induced in rat cerebellar slices by conjunctive stimulation of parallel fibers (PFs) and climbing fibers (CFs) under perfusion of 20 μM picrotoxin. LTD was estimated by the reduction in the initial rising slope of EPSPs PF-induced in Purkinje cell dendrites. LTD-inducing efficacy was represented by both the average amount of depression and the probability of inducing depression greater than 25%, both measured at 40 min after the onset of conjunctive stimulation. Using 300 regularly recurring pulses given to both CFs and PFs with 0 ms interval, LTD was optimally induced at 1 Hz, and to lesser degrees at other frequencies. When the number of conjunctive stimuli at 1 Hz with zero CF-PF interval was varied from 50 to 500, 300 stimuli induced LTD most robustly. When CF-PF interval was varied while 300 pulses were given at 0.25–4 Hz, LTD was induced even when PF stimuli were delayed after CF stimuli by as much as 2 s, but it was inhibited when PF stimuli preceded CF stimuli by 10–100 ms. LTD was also induced by applying repeated short pulse trains to both CFs and PFs, but repeated application of a PF stimulus train immediately followed by a CF stimulus train as in classical conditioning was effectless. The present results suggest complex processes leading to LTD as a result of conjunctive CF and PF stimulation.

1.4 G, 1.7 G, and 2.0 G of +Gz and 60 minutes centrifugation was adopted to 20 healthy male subjects using 1.8 m radius centrifuge equipped to Nihon University School of Medicine. G was applied from lower G, considering G training effect for the subjects. Effects on performance decline and side effects of such a short-arm centrifugation were especially observed in the experiments, because this size of centrifuge could be used in space station in future for a strong countermeasure of cardiovascular deconditioning, demineralization from bone, etc. G training effect was observed same as higher and rapid G acceleration in fighter pilot. Subjects suffered from many types of discomfort; such as sensation of heaviness of diaphragm, cold sweat, nausea, irritable feeling, arrhythmia, tachycardia, rapid decrease of blood pressure, which sometimes caused interruption of G load. As 2.0 G and 60 minutes centrifugation seemed very tough load to the subjects, there should be necessary some G suit or other countermeasure, if we apply a higher G and/or longer G duration. Performance decline due to the load commonly continued for 1 hour or so. Side effects were observed in relation to neuro-vestibular, cardio-vascular, and autonomic nervous system.

Tachyplesin I, a polycationic antimicrobial peptide isolated from hemocytes of horseshoe crabs, kills bacteria by disrupting the membrane potential of the cytoplasmic membrane. The present study shows that, among 36 oral streptococcal strains, 12 of 21 Streptococcus sanguis, 3 Streptococcus mutans, 9 Streptococcus salivarius and 3 Streptococcus milleri strains were susceptible to tachyplesin I, whereas 9 S. sanguis strains were resistant. Interestingly, these resistant strains include the clinical isolates from both Kawasaki disease and Behçet patients. According to the time-kill study, tachyplesin I inhibited irreversibly the growth of S. sanguis, S. mutans and S. salivarius strains within 20 min and an S. milleri strain within 80 min. Although it has been suggested that Escherichia coli cultured in rich media were more susceptible to tachyplesin I, the present results show that only 3 S. milleri strains were more sensitized to tachyplesin I in a glucose-supplemented medium, and other tested strains were not. Similarly, only 4 strains were more resistant to tachyplesin I in saline than these were in a rich medium.

A 10 bit CMOS A/D converter with 3 V power supply has been developed for being integrated into system VLSI's. In this A/D converter, redundant binary encoders named "twin encoders" enhance tolerance to substrate noise, together with employing differential amplifiers in comparators. The bias circuit using a replica of the amplifier is developed for biasing differential comparators with 3 V power supply. Subranging architecture along with a multilevel tree decoding structure improves dynamic performance of the ADC at 3 V power supply. The A/D converter is fabricated in double-polysilicon, double-metal, 0.8 /spl mu/m CMOS technology. The experimental results show that the ADC operates at 20 MS/s and the twin encoders suppress the influence of substrate noise effectively. This ADC has a single power supply of 3 V, and dissipates 135 mW at 20 MS/s operation. >

Background The interactions between PDZ (PSD-95, Dlg, ZO-1) domains and PDZ-binding motifs play central roles in signal transductions within cells. Proteins with PDZ domains bind to PDZ-binding motifs almost exclusively when the motifs are located at the carboxyl (C-) terminal ends of their binding partners. However, it remains little explored whether PDZ-binding motifs show any preferential location at the C-terminal ends of proteins, at genome-level. Results Here, we examined the distribution of the type-I (x-x-S/T-x-I/L/V) or type-II (x-x-V-x-I/V) PDZ-binding motifs in proteins encoded in the genomes of five different species (human, mouse, zebrafish, fruit fly and nematode). We first established that these PDZ-binding motifs are indeed preferentially present at their C-terminal ends. Moreover, we found specific amino acid (AA) bias for the 'x' positions in the motifs at the C-terminal ends. In general, hydrophilic AAs were favored. Our genomics-based findings confirm and largely extend the results of previous interaction-based studies, allowing us to propose refined consensus sequences for all of the examined PDZ-binding motifs. An ontological analysis revealed that the refined motifs are functionally relevant since a large fraction of the proteins bearing the motif appear to be involved in signal transduction. Furthermore, co-precipitation experiments confirmed two new protein interactions predicted by our genomics-based approach. Finally, we show that influenza virus pathogenicity can be correlated with PDZ-binding motif, with high-virulence viral proteins bearing a refined PDZ-binding motif. Conclusions Our refined definition of PDZ-binding motifs should provide important clues for identifying functional PDZ-binding motifs and proteins involved in signal transduction.

The microcircuit in the cerebellum is characterized by the relative simplicity, precision, and geometric beauty of its arrangement. Its structure is identical throughout the cerebellar cortex except for some regional differences. The cerebellar cortex has three layers (molecular layer, Purkinje cell layer, and granular layer) and can be divided into more than 100 subareas by horizontal grooves and longitudinal bands. Each subarea can be further subdivided into a number of microzones (there could be 10,000 microzones in the human cerebellum). A microzone, in combination with a small portion of the interior olive, and in some regions, also with that of the parvocellular red nucleus, consists of a microcomplex, a functional unit of the cerebellum. Structural and functional data of neuronal elements and their connections in the cerebellar microcircuit have been. Theoretical models have been proposed to explain how the cerebellar microcircuit operates and generates unique functions.

In this study, we show the crucial roles of lipid signaling in long-term depression (LTD), that is, synaptic plasticity prevailing in cerebellar Purkinje cells. In mouse brain slices, we found that cPLA 2 α knockout blocked LTD induction, which was rescued by replenishing arachidonic acid (AA) or prostaglandin (PG) D 2 or E 2 . Moreover, cyclooxygenase (COX)–2 inhibitors block LTD, which is rescued by supplementing PGD 2 /E 2 . The blockade or rescue occurs when these reagents are applied within a time window of 5–15 min following the onset of LTD-inducing stimulation. Furthermore, PGD 2 /E 2 facilitates the chemical induction of LTD by a PKC activator but is unable to rescue the LTD blocked by a PKC inhibitor. We conclude that PGD 2 /E 2 mediates LTD jointly with PKC, and suggest possible pathways for their interaction. Finally, we demonstrate in awake mice that cPLA 2 α deficiency or COX-2 inhibition attenuates short-term adaptation of optokinetic eye movements, supporting the view that LTD underlies motor learning.

The growth of the Japanese steel industry has been supported by three factors: operation, equipment, and management technology. Computer-integrated manufacturing (CIM) is the core of management technology. CIM's purpose is to improve production efficiency through computer integration of data covering the staff (design and production control) and management department as well as the production line process department (order entry, manufacturing, and shipment). In this paper, an overview of the Japanese steel industry's system development process is given and the steps taken in order to establish the CIM system are described. The organization, operation, and improvement of operational capability, along with marketing and production-related systems, are integrated. Furthermore, an integrated system model is given as a CIM conceptual model. In closing, reference is also made to the challenges facing CIM in the Japanese steel industry as well as future directions to be taken.

In general, cancer treatment is evaluated based on the parameters of tumor reduction rate, remission period, and survival rate. Recently, the quality of life (QOL) in cancer patients has become increasingly important in medical treatment. The patients with head and neck cancer usualy suffer from symptoms of oral dysfunction such as dysgeusia, dysmasesis, dysphagia and allolalia after cancer treatment. In addition cosmetic disturbances are caused by surgical treatment. In advanced cases, severe pain is involved. We devised a QOL questionnaire for head and neck cancer patients. In this study, we analysed changes in QOL scores and oral function scores caused by chemotherapy and surgery. Furthermore, the relationships between pain and QOL, oral function and QOL, ingestive function and QOL, appetite and QOL, and speech and QOL were analysed through the questionnaire.

Synaptic potentials were recorded intracellularly from Purkinje cells in guinea pig cerebellar slices. EPSPs evoked by stimulation of parallel fibers were effectively blocked by perfusion of a slice with the synthetic analog of Joro spider toxin, 1-naphthylacetyl-spermine (NAS) at 250 microM. However, it did not influence those responses evoked by stimulation of climbing fibers. This action of NAS is in contrast to other commonly used glutamate antagonists, CNQX or APV: CNQX (5 microM) blocked both parallel fiber- and climbing fiber-induced responses, while APV (up to 1 mM) did not influence either except for a weak reduction observed in climbing fiber responses. NAS thus provides a useful tool for pharmacologically distinguishing parallel fiber and climbing fiber synapses.

Seven patients were reconstructed their occlusion with osseointegrated implant (Branemark System) in Department of Oral Surgery, Nagoya University Hospital. Three cases ofpatients have been analysed for their mandibular bone quality with Digital Image Processing (DIP) method preoperatively. Results obtained were as follows:1. Seven patients who had an extreme difficulty in wearing conventional removal dentureswere treated in this study. The subjects consisted of 3 males and 4 females, and their agesranged from 46 to 66 years.The lesion was an edentulous mandible in 3 cases, an edentulous maxilla in one case andpartially edentulous in 3 cases.2. Twenty one fixtures in maxilla and 23 fixtures in mandible, totally 44 fixtures wereinstalled in 7 cases.One fixture in mandible and one in maxilla failed to osseointegrate, giving the fixture survivalrate of 95.2% in mandible and 95.7% in maxilla.3. Three edentulous mandibular patients received the bone quality analysis (DIP method) using occlusal film preoperatively. Results of bone density were 2.98, 1.98, 3.10Almm in eachcase.Case 2 who had the lowest bone density failed one fixture to osseointegrate.4. DIP method was very useful as a bone quality measuring method. Because it could beeasily carried out to analyse the jaw bone quality for dental implant without any variation offilm conditions.

Three mandibular edentulous patients treated with osseointegrated implants were reported. They were treated in the department of Oral Surgery, Nagoya University Hospital and followed for three years. In this paper the relationship between bone quality and degree of bone resorption around the fixtures was analysed.Results were summarized as follows:1. The cases were one male and two female and aged 66, 64 and 50 years old.2. The bone qualities of each case were analysed by DIP method preoperatively. The bone densities were 2.98, 1.98 and 3.10.3. Six fixtures were installed in each case and one fixture of case 2 failed to osseointegrate. Final survival rate of the fixtures was 95%.4. The degree of bone resorption in the low DIP case was greater than that in the high DIP case.5. The degree of bone resorption were almost equal in any site of mandible.6. The degree of bone resorption in high Plaque Index case was greater than that in the low Plaque Index case.

Urinary bladder carcinogenicity of uracil and N-butyl-N-(4-hydroxybutyl)nitrosamine (BBN) in combination was investigated in male F344 rats. Two experiments were performed: in the first animals were treated with 3.0, 1.0 and 0.3% uracil in the diet and/or 0.005% BBN in the drinking water for 36 weeks followed by a 4 week period without chemicals, the total observation time being 40 weeks; in the second rats received 3.0% uracil in the diet or 0.05% BBN in the drinking water for 4 weeks. In the first experiment, a markedly high incidence of carcinomas was observed in the urinary bladder with both 3.0% uracil alone (11/15, 73%) and 3.0% uracil plus 0.005% BBN (18/20, 90%) treatments. The multiplicity, i.e. numbers of carcinomas induced per 10 cm of basement membrane, was significantly higher with the latter treatment (P less than 0.05), as were proliferative lesions in the renal pelvis. Stone formation was observed in all 3.0% uracil treatment animals. In the second experiment, prostaglandin E2 levels in bladder tissue of uracil-treated animals proved to be significantly higher than in BBN-treated or untreated animals. Thus, the results clearly indicated that calculi associated with 3.0% uracil ingestion can themselves cause carcinoma development in the urinary bladder, while also acting as a potent co-carcinogen agent for BBN carcinogenesis.

Various treatments for hepatic metastasis of gastric cancer have been attempted, but problems remain with respect to long-term effectiveness and recurrence. Case reports have indicated the tumor regression effect of polysaccharide K(PSK)combined with chemotherapy, and meta-analysis has shown that PSK combined with chemotherapy improves the prognosis compared to chemotherapy alone. However, marked improvement of disease following PSK administration is rarely reported. We report a case of hepatic metastasis of gastric cancer in which low-dose UFT and PSK therapy resulted in regression of metastatic hepatic lesions and improvement of tumor markers. A 78-year-old man had synchronous hepatic metastasis of gastric cancer. Gastrectomy and microwave coagulation therapy using Microtase were conducted, followed by postoperative adjuvant chemotherapy with UFT 300 mg/day. Recurrences of metastatic hepatic lesion and new hepatic lesion were observed 6 months after surgery. Addition of PSK to UFT chemotherapy was selected as the treatment for recurrences, resulting in disappearance of the hepatic lesions and normalization of tumor markers. The patient is alive without recurrence at this writing, 38 months after surgery.

The intricate neuronal circuitry of the cerebellum is thought to encode internal models that reproduce the dynamic properties of body parts. These models are essential for controlling the movement of these body parts: they allow the brain to precisely control the movement without the need for sensory feedback. It is thought that the cerebellum might also encode internal models that reproduce the essential properties of mental representations in the cerebral cortex. This hypothesis suggests a possible mechanism by which intuition and implicit thought might function and explains some of the symptoms that are exhibited by psychiatric patients. This article examines the conceptual bases and experimental evidence for this hypothesis.

Desensitization of quisqualate (QA)-specific glutamate receptors of Purkinje cells was examined by 'wedge' recording from cerebellar slices. The desensitization was induced by conditioning with bath application of 100 microM QA for 4 min, and was represented by reduced responsiveness to testing QA applications. Bath application of 10 microM alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) for 1 min at a time had no effect on inducing desensitization, but AMPA responsiveness was reduced by the 4-min QA conditioning to the same extent as QA responsiveness. These results indicate that QA-induced desensitization in Purkinje cells takes place specifically in the ionotropic subtype of QA receptors which AMPA stimulates selectively, and further suggest that another metabotropic subtype of QA receptors which AMPA does not stimulate plays a role in inducing the desensitization. By using AMPA applications as neutral testings, the desensitization was followed for 2 h without sign of recovery. For intervals longer than 2 h, AMPA responsiveness was compared with responsiveness to a 1-min application of 3 mM aspartate (AS) as a control, since the latter was not affected by QA conditioning. The AMPA/AS responsiveness ratio remained reduced throughout 10 h after 4-min QA conditioning without sign of recovery.

Recently, mice and embryonic stem (ES) cells with allelic polymorphisms have been used extensively in the field of genetics and developmental biology. In this study, we examined whether intersubspecific hybrid mice and ES cells with these genotypes can be efficiently produced by intracytoplasmic sperm injection (ICSI). Frozen-thawed spermatozoa from wild-derived strains, JF1 (Mus musculus molossinus), MSM (M. m. molossinus), HMI (M. m. castaneus), and SWN (M. m. spp.), were directly injected into mature oocytes from laboratory mice ((C57BL/6 � DBA2)F1; M. m. domesticus). The in vitro and in vivo developmental capacity of F1 embryos was not significantly different among the groups (P >0.05), and term offspring were efficiently obtained in all groups (27%-34% of transferred embryos). However, the mean body and placental weights of the offspring differed significantly with genotype (P

Previous reports have shown that exposure to hypergravity decreases rat body mass during the initial phase, with this decrease and level of gravity showing a dose-response relationship. The present study examined whether rate of body mass gain after the initial phase of exposure is attenuated by hypergravity in a dose-dependent manner and sought to identify any threshold.Male 10-wk-old rats (n = 64) were used, with 16 rats serving as 1.0-G controls, and 48 rats exposed to hypergravity for 14 d in 4 groups (1.5, 2.0, 2.5, and 3.5 G; n = 12 each). Body mass gain was evaluated according to slope of change in body mass from day 7 of exposure to hypergravity, as both absolute and relative values.Slopes of body mass gain did not differ between the 1.0- and 1.5-G groups (6.09 and 5.75 g x d(-1), respectively), but were significantly less for the 2.0-, 2.5-, and 3.5-G groups (4.91, 3.03 and 1.99 g x d(-1), respectively) than for the 1.0- and 1.5-G groups. Body mass gain as a relative value did not differ between the 1.0-, 1.5-, and 2.0-G groups (1.5 +/- 0.2, 1.6 +/- 0.6 and 1.4 +/- 0.3 g x d(-1) x 100 g(-1) body mass, respectively), but was significantly less for the 2.5- and 3.5-G groups (1.1 +/- 0.6 and 0.8 +/- 0.3 g x d(-1) x 100 g(-1) body mass, respectively) than for the 1.0-, 1.5-, and 2.0-G groups. Absolute values and rate of body mass gain were reduced with increases in gravity.Exposure to hypergravity attenuates body mass gain in a dose-dependent manner, with a threshold possibly existing between 1.5- and 2.5-G for 10-wk-old male rats.

To establish reproductive biological techniques in mammals, it is important to understand the growth environment of the embryo. Oviduct epithelial cells are in close proximity to the embryo during pre-implantation development. We, therefore, established an immortalized oviduct epithelial cell line from the cynomolgus monkey, evaluated the usefulness of these cells as feeder cells for embryo culture, and investigated the gene expression of several growth factors and cytokines in the cells. The immortalized cells were positive for the anti-cytokeratin antibody, as determined by immunocytochemistry, indicating that they are epithelial. They also expressed oviductin, which is specific to oviduct epithelial cells, glyceraldehyde-3-phosphate dehydrogenase (control), leukemia inhibitory factor, vascular endothelial growth factor, epidermal growth factor, insulin-like growth factor 1, transforming growth factor beta-2, and interleukin 4. Mouse embryo development was improved when the immortalized cells were used as feeder cells. This cell line is also useful for studying the factors secreted by oviduct epithelial cells.

To examine the effects of buffalo rat liver (BRL) cells on the preimplantation development of mouse embryos in vitro, we first cultured two-cell mouse embryos alone in serum-free Dulbecco modified Eagle medium. As expected, the embryos did not develop to subsequent stages. However, when cocultured with BRL cells, the embryos developed to the blastocyst stage efficiently. Direct contact of embryos with BRL cells was not necessary for development: the medium conditioned by BRL cells contained soluble factors that supported the preimplantation development of mouse embryos. Embryos cultured with BRL-conditioned medium that was replaced at various intervals had a further increased rate of development to the blastocyst stage. This finding indicated that the activities of the factors were maintained only briefly. Seven proteins between 35 and 44 kDa that were detected in the medium were highly beneficial to the development of the embryos. Follistatin-related protein and pigment epithelium-derived factor are believed to be the factors supporting embryo development. The other five proteins also may improve the environment for the development of mouse embryos cultured in vitro.

Publisher Summary The cerebellum–pons–medulla block preparation provides material suitable for investigating the neuronal circuitry of the cerebellum and its early postnatal development. This chapter describes the development of a unique block preparation of the cerebellum isolated from neonatal rats; the isolated block contains the pons and the medulla oblongata. Cerebellar long-term depression (LTD) is a major type of synaptic plasticity expressed in the unique structure of the cerebellar cortex and is considered to represent a major memory process underlying the error-driven learning in the cerebellum. The chapter shows that in the P5 block preparation, Purkinje cells are already functionally mature in terms of synaptic transmission and synaptic plasticity; however, they undergo rapid changes in their morphology and molecular mechanisms, for example, while LTD induction requires protein phosphatase 2A in Purkinje cells cultured for 22–35 days, protein phosphatase 1 plays a major role in Purkinje cells cultured for 12–15 days. Hence, LTD required for cerebellar function appears to be supported by molecular mechanisms, which dynamically change with time during postnatal development. The chapter also presents the advantages of block preparation; the block preparations provide a convenient material for the pharmacological investigation of cerebellar circuits, and the functional development of cerebellar neuronal circuits during the first postnatal week can be followed.

The cerebellum has a fine compartment structure, which represents various discrete functions. Evolutionarily old, medial parts of the cerebellum are involved in the adaptive control of various brainstem and spinal cord functions, and long-term depression (LTD) plays a key role in this adaptive control. To extend these views to evolutionarily newer, lateral parts of the cerebellum involved in cerebral cortical functions such as voluntary movement, perception and language, the internal model hypothesis is instrumental. This hypothesis explains not only the characteristic cerebellar symptom, dysmetria, but also a number of otherwise unexplainable phenomena displayed in movements and mental actions.

The taxonomic position of a halo- and organo-sensitive, oligotrophic soil bacterium, strain S213T, was investigated. Cells were Gram-negative, non-motile, strictly aerobic, yellow-pigmented rods of short to medium length on diluted nutrient broth. When 0·1–0·4 % (w/v) NaCl was added to diluted media composed of peptone and meat extract, growth was inhibited with increasing NaCl concentration and the cells became long aberrant forms. When 6 mM CaCl2was added, the cells grew quite normally and aberrant cells were no longer found at 0·1–0·5 % (w/v) NaCl. Chemotaxonomically, strain S213Tcontains chemical markers that indicate its assignment to theSphingomonadaceae: the presence of ubiquinone Q-10 as the predominant respiratory quinone, C18 : 1and C16 : 0as major fatty acids, C14 : 02-OH as the major 2-hydroxy fatty acid and sphingoglycolipids. 16S rRNA gene sequence analysis indicated that strain S213Tbelongs to the genusSphingomonas, exhibiting high sequence similarity to the 16S rRNA gene sequences ofSphingomonas maliIFO 15500T(98·3 %),Sphingomonas pruniIFO 15498T(98·0 %),Sphingomonas asaccharolyticaIFO 15499T(97·9 %) andSphingomonas echinoidesDSM 1805T(97·8 %). The results of DNA–DNA hybridization experiments and its phenotypic characteristics clearly distinguished the strain from its nearest neighbours and demonstrate that strain S213Trepresents a novelSphingomonasspecies, for which the nameSphingomonas oligophenolicasp. nov. is proposed. The type strain is S213T(=JCM 12082T=CIP 107926T).

Non-small cell lung cancer frequently shows loss of heterozygosity of the chromosome 3p21.3 region and several genes such as RASSF1A, BLU, and SEMA3B have been identified as candidate tumor suppressor genes at this region since their downregulation and hypermethylation at their promoter regions were frequently detected in lung cancer. To determine whether these three genes are simultaneously inactivated during lung cancer development, we studied 138 primary non-small cell lung cancers for the promoter methylation status of these genes and allelic loss of the chromosome 3p21.3 region. We found promoter hypermethylation at 32% in RASSF1A, 30% in BLU, and 47% in SEMA3B. Allelic loss of 3p21.3 was detected in 54 (58%) of 93 informative tumors. Despite the weak association of methylation status among these three genes, there was no correlation between the methylation status of each gene and loss of heterozygosity. We also studied possible genes downstream of RASSF1A in 16 primary non-small cell lung cancers and found that the expressions of SM22 and SPARC were significantly downregulated in RASSF1A-hypermethylated tumors. Our results showed that, while candidate tumor suppressor genes at this locus can be simultaneously inactivated by epigenetic alterations, loss of heterozygosity without any hypermethylation of the three genes can also occur in some cases, suggesting that just one allelic loss might also be sufficient for the inactivation of any of these genes for lung cancer development.

Even though inhibition had long been recognized as a distinct process in the nervous system (see Sherrington1), the discovery of inhibitory synapses had to wait until the middle of the 20th century. In mammalian motoneurons, Eccles and his associates recorded inhibitory postsynaptic potentials (IPSPs) that form a mirror image of excitatory postsynaptic potentials (EPSPs)2.

Activating mutations of RAS gene families have been found in a variety of human malignancies, including lung cancer, suggesting their dominant role in tumorigenesis. However, several studies have shown a frequent loss of the wild-type KRAS allele in the tumors of murine models and an inhibition of oncogenic phenotype in tumor cell lines by transfection of wild-type RAS, indicating that wild-type RAS may have oncosuppressive properties. To determine whether loss of wild-type KRAS is involved in the development of human lung cancer, we investigated the mutations of KRAS, NRAS and BRAF in 154 primary non-small cell lung cancers (NSCLCs) as well as 10 NSCLC cell lines that have been shown to have KRAS mutations. We also determined the loss of heterozygosity status of KRAS alleles in these tumors. We detected point mutations of KRAS in 11 (7%) of 154 NSCLCs, with 10 cases at codon 12 and 1 at codon 61, but no mutations of NRAS or BRAF were found. Using the laser capture microdissection technique, we confirmed that 9 of the 11 tumors and 7 of the 10 NSCLC cell lines retained the wild-type KRAS allele. Among the cell lines with heterozygosity of mutant and wild-type KRAS, all of the cell lines tested for expression were shown to express more mutated KRAS than wild-type mRNA, with higher amounts of KRAS protein also being expressed compared to the cell lines with a loss of wild-type KRAS allele. In addition, among 148 specimens available for immunohistochemical analysis, 113 (76%) showed positive staining of KRAS, indicating that the vast majority of NSCLCs continue to express wild-type KRAS. Our findings indicate that the wild-type KRAS allele is occasionally lost in human lung cancer, and that the oncogenic activation of mutant KRAS is more frequently associated with an overexpression of the mutant allele than with a loss of the wild-type allele in human NSCLC development. © 2003 Wiley-Liss, Inc.

"Nurturing the Brain" is a new research field aiming at facilitating development and maintenance of healthy brains and keeping their learning capabilities at full display throughout life. It is based on recent remarkable progress in developmental neuroscience and non-invasive technologies for visualizing brain activities in humans, even infants and children. "Nurturing the Brain" research will help us to cure or prevent various types of developmental disorders such as ADHD and autism. It will also help us in choosing an appropriate timing for child care and education on the basis of new knowledge of the critical period of development for various brain functions.

At the opportunity of celebrating the 100th Anniversary of the Japanese Society of Psychiatry and Neurology, I am filled with deep emotion when I recall the time only a few decades ago, when mental disorders were believed to be an illness of the mind without any disorder in the brain. It has now become apparent that a mental disease emerges from the malfunction of the brain's unique biological mechanisms, so that any mental disease can be cured or prevented if the cause of the malfunction is clarified. Analyses of intercellular as well as intracellular signal transduction and the mechanisms underlying gene regulation in brain cells have recently advanced markedly and have brought a marked improvement in the methods and technology for investigating pathogenesis of neurological and psychiatric diseases. These impressive results have been derived in the search for the cause of Parkinson's disease, Alzheimer's disease and spinocerebellar degeneration. Certain minor changes in the molecular structure of sodium channels have been found to cause epilepsy. It has also been revealed that ten particular sites in chromosomes harbor the remote cause of schizophrenia and depression. These sites are probably responsible for production of certain synaptic transmitters and modulators. Another major challenge in the field of psychiatry is to understand the unique symptoms exhibited by patients with psychiatric disorders, which requires research to head in a direction different from molecular and cellular brain science. It is a challenge similar to the very fundamental challenge of determining how our brain, which is composed of numerous brain cells, yields our mind (or, should I say, supports our mind). To understand the brain as a complex system is more difficult than researching its molecular and cellular machinery, and accordingly, the progress of research in this field has been slow. However, it is gradually gaining speed thanks to the improvement in non-invasive methods for measuring activities in conscious human brains as well as that to the establishment of computational models. For example, it is a major mystery why a schizophrenic patient experiences auditory hallucinations, but an interesting hypothesis is derived by considering activities of the thinking brain as a neural system. Under this hypothesis, we gather information from the outside world by using our sensory systems and establish its internal model in our brain. Thus, manipulation of the internal model to run a simulation is an important part of our thinking activities. When an internal model behaves abnormally, abnormal information is generated, which makes the patient mistake it for sound from the outside, thus causing auditory hallucinations. It is highly expected that substantial progress will be made in psychiatry through proposals of new hypothetical models such as this one as well as verification of them through non-invasive measurement of brain activities. In the near future, patients with mental disorders will most likely benefit from the results of this two-directional research in psychiatry. The expectation is high for the strong leadership of the Japanese Society of Psychiatry and Neurology to promote research in Japan. Effective research effort requires efficient collection of human materials and systematic survey of patients, which will inevitably increase the involvement of patients in basic research. Improvement of research environments such as by developing a method for assessing research ethics and upgrading brain banks is urgently needed.

Classic studies of the cerebellum before the middle of the twentieth century established the structural entity of the cerebellum and characterized its function as enabling animals and humans to carry out smooth and accurate movements, even at a high speed and without visual feedback. In the 1960s, neuronal circuit structures of the cerebellum were analyzed in detail, which promoted computational approaches toward the study of neuronal network principles of the cerebellum. In the 1970s and 1980s, vestibulo-ocular reflex adaptation, adaptive locomotion, eye blink conditioning, and learning in hand/arm movement were established as effective experimental paradigms for investigating neural mechanisms of cerebellar functions. In the 1980s, long-term depression (LTD) was discovered and considered as a memory process in the cerebellum; in the 1990s, complex signal transduction processes underlying LTD were revealed. It was also in the 1980s that computational approaches were advanced for modeling control system functions of the cerebellum. Currently, there are two alternative models proposed for VOR adaptation. In this decade, we envisage new developments toward the fusion of knowledge of the cerebellum at molecular and cellular levels and those in systems and computation. Studies of LTD will play a key role in pursuing this direction.

When oviduct epithelial cells are collected by using enzymatic methods that involve protease such as trypsin, other cells unintentionally are collected as well, especially from small laboratory animals such as mice and rats, thus contaminating the collected sample. We therefore developed a simple nonenzymatic method that involves using a glass micropipette to aspirate the oviduct epithelial cells from the oviduct. This aspiration method easily removed oviduct epithelial cells from mice, rats, and cynomolgus monkeys. Culture of the collected oviduct epithelial cells confirmed that the cells could adhere and grow on a petri dish. Although this aspiration technique was developed for use in small animals, our results show that it can be applied to cynomolgus monkeys. The advantages of this technique are its simplicity and its success in collecting oviduct epithelial cells from oviducts.

Rat G-substrate cDNA was isolated from a cerebellar library and characterized. The deduced amino acid sequence of rat G-substrate contained two putative phosphorylation sites for PKG at Thr72 and Thr123; the amino acid sequences (KPRRKDT(p)PA) around these sites are conserved in human, mouse and rabbit. G-substrate phosphorylated by PKG inhibited the catalytic subunits of both protein phosphatase-1 (IC 50 14.1 nM) and -2A (IC 50 5.9 nM). Mutation of Thr123 (site 2) to Ala significantly reduced the inhibition of both PP-1 and PP-2A, while mutation of Thr72 (site 1) to Ala had little effect on inhibitory activity. In situ hybridization analysis revealed that G-substrate mRNA was localized exclusively in cerebellar Purkinje cells. Immunoperoxidase staining showed that in Purkinje cells, G-substrate was present in somata, dendrites and axons. In rat cerebellar slices, activation of PKG with a nitric oxide (NO) donor, NOR3, or 8-Br-cGMP, increased phosphorylation of G-substrate, as demonstrated with a phosphorylation-specific antibody. These results characterize further the inhibition of PP-1 and PP-2A by phospho-G-substrate, and demonstrate its physiological phosphorylation in rat Purkinje cells.

In consumer video-system markets, A/D converters (ADCs) are desired to be integrated with digital signal-processing circuitry to reduce system cost and size. Operation with low supply voltage (V/sub DD/) is also desired in personal equipment. In addition, circuit techniques for low-V/sub DD/ operation are essential for ADC integration into 0.5 /spl mu/m LSIs. The first low-V/sub DD/ 10b video ADC to be published uses CMOS inverters as amplifiers in "chopper" type comparators. However, experimental results show that this type of circuit has poor noise immunity when integrated into mixed-signal LSIs. A 10b 20 Ms/s CMOS ADC with 3 V power supply and high noise immunity for integration into mixed-signal LSIs is described by the authors. >

Synaptic plasticity is an important cellular mechanism for the formation of memory in neuronal circuits of the brain. Research during the past two decades has revealed surprisingly complex signal-transduction processes that underlie various forms of synaptic plasticity. More than 30 molecules are involved in the induction of long-term depression (LTD) — a unique form of synaptic plasticity in the cerebellum. Here, I review recent data on these molecules, defining their roles as mediators or modulators, coincidence detectors or components of a self-regenerating circuit, and show how they are organized to form an efficient molecular machinery for LTD induction.

'Nurturing the brain' is an emerging research field integrating brain science, child care and education, which also involves child neurology. It has emerged from the recent remarkable progress in brain science and strong social demands for improvements in child care and education. This article reviews the current status of three major research themes in this field. First, developmental disorders represented by attention deficit/hyperactivity disorder, autism and Asperger syndrome often introduce difficulties in child care and education, which are to be addressed by appropriate assessment and treatment of affected children based on new knowledge of the pathogenesis of these disorders. Second, recent progress in research on the critical/sensitive periods of development of brain structures and functions promises useful advice for teachers and parents regarding optimal timing and ways of teaching various subjects. Third, the development of the brain throughout infancy, childhood and adolescence is paralleled by the growth and maturation of the mind. Neuronal mechanisms underlying the theory of mind, mirror neurons, internal model, cognitive control, and cognitive emotion regulation are important themes that bridge our understandings of the brain and the mind.

Evidence is presented indicating that the induction of long-term depression (LTD) in Purkinje cells (PCs) requires a rapidly turned over protein(s) during a critical time period within 15 min after the onset of LTD-inducing stimulation and that synthesis of this protein is maintained by mRNAs supplied via transcription. LTD was induced in granule cell axon (GA)-to-PC synapses by stimulation of these synapses at 1 Hz for 5 min in conjunction with the climbing fibers (CFs) forming synapses on the same PCs and represented by a persistent reduction in the GA-induced excitatory postsynaptic potentials (EPSPs). Not only a prolonged but also a brief (5 min) pulse application of translational inhibitors (anisomycin, puromycin, or cycloheximide) effectively blocked the LTD induction. Pulses applied during the period from 30 min before to 10 min after the onset of conjunctive stimulation blocked the LTD induction, but those applied 15 min after were ineffective. The three translational inhibitors blocked the LTD induction similarly, suggesting that the effect is due to their common action of inhibiting protein synthesis. Infusion of a mRNA cap analogue (7-methyl GTP) into PCs also blocked LTD induction, ensuring that the postsynaptic protein synthesis within PCs is required for LTD induction. Transcriptional inhibitors, actinomycin D and 5,6-dichloro-l-β-d-ribofuranosyl-benzimidazole, also blocked the LTD induction, but this effect was apparent when 5-min pulses of the transcriptional inhibitors preceded the conjunctive stimulation by 30 min or more. This time lag of 30 min is presumed to be required for depletion of the protein(s) required for LTD induction. The presently observed effects of translational and transcriptional inhibitors on the LTD induction are of temporal characteristics corresponding to their depressant effects on the type-1 metabotropic glutamate-receptor (mGluR1)-mediated slow EPSPs in PCs as we have reported recently. An antagonist of mGluR1s [(RS)-1-aminoindan-1,5-dicarboxylic acid], however, did not block LTD induction when it was applied during the 10-min period following conjunctive stimulation, where translational inhibitors effectively blocked LTD induction. This discrepancy in time course suggests that the rapidly turned over protein(s) required for LTD induction is involved in a process occurring downstream of the activation of mGluR1s.