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In aquaculture and ornamental industries, copepods are recognized as preferred live feeds for marine fish larvae over commonly used organisms such as Artemia and rotifers. Marine fish larvae fed with copepods show better survival and growth. Despite obvious advantages of copepods as live feed, their use is still limited owing to low productivity and cost-efficiency when mass cultured. Copepods can be divided into free-spawner and sac-spawner (egg-carrying) according to their spawning style. In cultivation of egg-carrying copepod, a simple nauplii collection/harvesting method with low labor has not been established, because females carry the egg-sac until hatching. Manual collection of nauplii using a siphon hose and mesh-filters is a common method in copepod cultivation, but automation is necessary to reduce labor costs if copepods are to be viably cultured at a commercial scale. Recently, we devised a zooplankton bioreactor for rapid separation of free-spawner copepod eggs from adults in a tank. The automation collects eggs on a mesh filter in a water flow system which can also function as water exchange device. In the present study, we apply this bioreactor to egg-carrying copepod, and report the results from cultivation at laboratory-scale for 45 days. Species of Oithona are good candidates for live feed because their body size and biochemical composition are suitable for many marine fish larvae which have small gapes. Oithona oculata is widely distributed and a typical dominant species in coastal waters, and was chosen as target species of egg-carrying copepod in present study. 150 adult individuals of O. oculata were placed into a culture chamber which were immersed in a reproduction tank containing 3 L seawater. The culture chamber to retain the copepods has a 100-μm nylon mesh placed 5 mm above the bottom, while allowing passage of nauplii. The copepods were fed daily with a sufficient amount of Thalassiosira weissflogii and Isochrysis galbana and incubated at 28°C for 45 days. Produced nauplii in the reproduction tank were collected daily using water flow of 1.5 L min-1. The species abundance in each development, ovigerous rate and collected number of nauplii were measured daily. In the semi-continuous culture, the collected nauplii were cultured in maturation vessels and returned to the production tank after their maturation, and 45 days cultivation (four generations) was achieved. Total abundance was gradually increased from day 4 to day 20 and reached at 500 inds. L-1. Ovigerous rate varied from 0 to 88% during the incubation period. The maximum number of collected nauplii was 380 nauplii L-1 at day 5. The results obtained in present study suggest that the bioreactor can also be applied to egg-carrying copepod by optimization of the mesh size and the water flow velocity.

Herbal tea can maintain our health and handle ill health before symptoms appear, and its usage is easier than taking traditional medicines. Depending on individual characteristics, suitable tea should be different for each individual. To provide effective herbal tea, it is desirable to analyze daily personal behaviors. Combinations of herbs can vary for individuals considering the kinds of herbs and amounts that are found to be effective. In traditional medicine, this would typically be very difficult without specialized expertise. We aimed to develop electrical household equipment for preparing herbal teas that can be used even without such expertise. In this study, we present our system, along with case studies of effects of the tea proposed for each.

A mutant of cutinase-like enzyme from Saccharomonospora viridis AHK190, Cut190_S226P/R228S, designated as Cut190*, possesses high-thermal stability and has high polyethylene terephthalate (PET)-degrading activity. The functional characterizations of PET-degrading enzymes are generally conducted using accessible substrates such as poly(butylene succinate-co-adipate) (PBSA) and p-nitrophenyl butyrate (pNPB), even though their structures are different from that of PET. Bis(2-hydroxyethyl) terephthalate (BHET) is a component of PET, and the structure is similar to that of PET compared to PBSA and pNPB. Therefore, the analysis using BHET as the substrate is important to evaluate effective PET degradation. In this study, we analyzed the enzymatic activity of Cut190* using BHET under various conditions including Ca2+ concentration and pressure. Although terephthalate was supposed to be the final product, the intermediate product, mono(2-hydroxyethyl) terephthalate (MHET), was the only product generated, possibly due to the low binding affinity of MHET. The Cut190* activity towards BHET was observed even in the absence of Ca2+ and increased with increasing Ca2+ concentration, which is similar to its activity towards pNPB, but different from its activity towards PBSA. The difference can be attributed to the size of substrates. We also analyzed the activities of Cut190* and another high-activity mutant, Cut190*Q138A/D250C-E296C, under high pressures up to 400 MPa. BHET was non-enzymatically hydrolyzed under high pressure at 37 ℃. Enzyme activities were maintained under high pressures, and degradation of BHET in the presence of enzyme was higher than that in the absence of enzyme. Furthermore, the structural analysis of Cut190* under high pressure using Fourier transform infrared spectroscopy showed that most of the enzyme molecules were populated in the native structure below 400 MPa. These results indicate that BHET can contribute to the effective functional analysis of PET-degrading enzyme, and the combination of enzyme and pressure can lead to eco-friendly PET degradation.

Mesenchymal stem cells (MSCs) have been used for cell therapy in various experimental disease models. However, the regenerative potential of MSCs from different tissue sources and the influence of the tissue niche have not been investigated. In this study, we compared the regenerative potential of dental pulp, bone marrow and adipose tissue-derived CD31(-) side population (SP) cells isolated from an individual porcine source. Pulp CD31(-) SP cells expressed the highest levels of angiogenic/neurotrophic factors and had the highest migration activity. Conditioned medium from pulp CD31(-) SP cells produced potent anti-apoptotic activity and neurite outgrowth, compared to those from bone marrow and adipose CD31(-) SP cells. Transplantation of pulp CD31(-) SP cells in a mouse hindlimb ischemia model produced higher blood flow and capillary density than transplantation of bone marrow and adipose CD31(-) SP cells. Motor function recovery and infarct size reduction were greater with pulp CD31(-) SP cells. Pulp CD31(-) SP cells induced maximal angiogenesis, neurogenesis and pulp regeneration in ectopic transplantation models compared to other tissue sources. These results demonstrate that pulp stem cells have higher angiogenic, neurogenic and regenerative potential and may therefore be superior to bone marrow and adipose stem cells for cell therapy.

Introduction Pulp stem/progenitor cells have been successfully transplanted after pulpectomy in dogs and have led to pulp regeneration. The regenerated pulp tissue was investigated by the qualitative and quantitative protein expression patterns in comparison with those of normal pulp. There is an unmet need for a quality standard for regenerated pulp tissue. Methods Three distinct human CD105 + stem/progenitor cells from dental pulp, bone marrow, and amnion were compared by 2-dimensional electrophoresis and the differential proteomic expression profiles. The protein identified with high confidence in pulp CD105 + cells was examined by immunohistochemistry and real-time reverse-transcription polymerase chain reaction in regenerated pulp tissue compared with normal pulp. Its migration effect was further examined in pulp CD105 + cells using small interfering RNA techniques to knock down the protein. Results Nine protein spots were detected solely in pulp CD105 + cells; one of these was identified as vimentin. The expression of vimentin messenger RNA was highest in pulp tissue among a variety of human tissues and higher in pulp CD105 + cells compared with other CD105 + cells and unfractionated total pulp cells. Pulp cells and endothelial cells were positively stained with vimentin in regenerated pulp tissue similarly as those in normal pulp tissue. The expression of vimentin in regenerated pulp was similar to normal pulp. RNA interference knock down of vimentin expression in pulp CD105 + cells significantly reduced the migration activity. Conclusions The highest expression of vimentin in pulp tissue among other tissues and its migration effect in pulp stem cells suggest that it is a quality standard for pulp regeneration and pulp cell function.

PukaPuCam is an application service that utilizes a camera attached to balloons, to capture users' photo continuously from a third-person view. Then, users can glance through their photos by using PukaPuCam Viewer. PukaPuCam records the interaction between users and their surrounding objects or even with the people they meet. As balloon experiences air resistance, it can change its inclination according to the user's speed and thus, capture pictures from different direction or angles. This gives rise to interesting and unusual records to be added to the user's collection. As compare to other similar devices, PukaPuCam uses a common design people are familiarize with — a balloon; making it an interesting application to be used at tourist spots. As balloons are cute, we aim to give users a more enjoyable, delightful experience.

Regeneration of dental pulp after pulpectomy was accomplished recently by autologous transplantation of dental pulp stem/ progenitor cells into the root canal. The identical patterns of qualitative and quantitative protein and mRNA expression in the regenerated pulp and normal pulp demonstrated complete pulp regeneration. The lack of tissue specific markers in the dental pulp is a major challenge in dental regenerative medicine. In order to identify specific markers in dental pulp we undertook a comparison of the gene expression profile of the dental pulp with that of periodontal ligament and gingiva. This systematic investigation identified thyrotropinreleasing hormone (TRH)-degrading enzyme (DE) as a marker of dental pulp. Expression of TRH-DE mRNA in human dental pulp was higher than that in any other tissue except brain as analyzed by real time RT-PCR. Induction of neural cells enhanced the expression of TRH-DE mRNA in dental pulp stem/progenitor cells (CD105+ and CD31- side population (SP) cells) in vitro. Immunohistochemical and in situ hybridization analyses demonstrated that TRH-DE in the neuronal processes in dental pulp. In canine pulp cells, TRH down-regulated TRH-DE mRNA expression, while neuropeptide Y up-regulated it, suggesting that TRH-DE has functional role in the neuropeptide signaling in dental pulp tissue. It is noteworthy that TRH-DE mRNA was expressed in the regenerated pulp 28 days after transplantation of CD31- SP cells into root canals after pulpectomy. These results demonstrate the utility of TRH-DE as a novel dental pulp biomarker during regeneration of pulp.

In some real world applications, the data can be represented naturally in a special kind of graphs in which each vertex consists of a set of (structured) data such as item sets, sequences and so on. One of the typical examples is metabolic pathways in bioinformatics. Metabolic pathway is represented in a graph structured data in which each vertex corresponds to an enzyme described by a set of various kinds of properties such as amino acid sequence, enzyme number and so on. We call this kind of complex graphs multi-structured graphs. In this paper, we propose an algorithm named FMG for mining frequent patterns in multistructured graphs. In FMG, while the external structure will be expanded by the same mechanism of conventional graph miners, the internal structure will be enumerated by the algorithms suitable for its structure. In addition, FMG employs novel pruning techniques to exclude uninteresting patterns. The preliminary experimental results with real datasets show the effectiveness of the proposed algorithm.

The cyclooxygenase–prostanoid pathway regulates myometrial contractility through activation of prostanoid receptors on uterine smooth muscles. However, the possible expression of prostanoid receptors on autonomic nerves cannot be excluded completely. The aim of the present study was to clarify the presence of neural prostanoid receptors on adrenergic nerves in the porcine uterine longitudinal muscle. In [ 3 H]-noradrenaline-loaded longitudinal muscle strips of porcine uterus, electrical field stimulation (EFS) evoked [ 3 H]-noradrenaline release in a stimulation frequency-dependent manner. The EFS-evoked release was completely abolished in Ca 2+ -free (EGTA, 1 mM) incubation medium and by tetrodotoxin or ω-conotoxin GVIA, suggesting that [ 3 H]-noradrenaline was released from neural components. The EFS-evoked [ 3 H]-noradrenaline release was significantly enhanced by treatment with indomethacin. In the presence of indomethacin, PGE 2 and PGF 2α , but not PGD 2 , inhibited the EFS-evoked [ 3 H]-noradrenaline release. Of synthetic prostanoid receptor agonists examined, both U46619 (TP) and sulprostone (EP 1 /EP 3 ) decreased the EFS-evoked [ 3 H]-noradrenaline release in a concentration-dependent manner, while fluprostenol (FP), BW245C (DP) and butaprost (EP 2 ) were almost ineffective. SQ29548 (TP receptor antagonist) blocked the effect of U46619, but SC19220 (EP 1 receptor antagonist) did not change the inhibition by sulprostone or PGE 2 . Double immunofluorescence staining using protein gene product 9.5, tyrosine hydroxylase, EP 3 receptor and TP receptor antibodies suggested the localization of EP 3 or TP receptors on adrenergic nerves in the porcine uterus. These results indicated that neural EP 3 and TP receptors are present on adrenergic nerves of the porcine uterine longitudinal muscle. Endogenous prostanoid produced by cyclooxygenase can regulate noradrenaline release in an inhibitory manner through activation of these neural prostanoid receptors.