Characterization of vitamin B12 compounds from commercially available fish roe products

Although fish roes (or the egg-laden ovary) are seafood products with high nutritional value and are considered abundant in vitamins including vitamin B₁₂, nevertheless, the detailed properties of vitamin B₁₂ have not been fully elucidated in fish roe products. Vitamin B₁₂ content was determined using C18-reversed-phase high-performance liquid chromatography after purification of samples from immunoaffinity columns. Salmon egg-laden ovary products (sujiko), salmon roe products (ikura), dried mullet roe products (bottarga), and white sturgeon roe products (caviar) were found to contain substantial levels of vitamin B₁₂ (more than approximately 15 µg/100 g wet weight). Interestingly, high levels of vitamin B₁₂ per 100 g wet weight have been measured in pollack roe products (tarako) and flying fish roe products (tobiko). Liquid chromatography–electrospray-ionization/tandem mass spectrometry analysis revealed that vitamin B₁₂ was the predominant corrinoid in the evaluated fish roe products examined, while no inactive corrinoid compounds were identified. These results suggest that commercially available fish roe products could be an important source of vitamin B₁₂ for use as a dietary product for humans.

     

Discrete sandwich compounds of monolayer palladium sheets

Despite the abundance of "sandwich" complexes, in which two cyclic aromatic hydrocarbon ligands flank a metal center, this motif has not been extended to sheets of multiple metal atoms. We prepared and isolated two such compounds. In the first, three palladium centers form a planar triangular array, capped by chlorides, between two cycloheptatrienyl ligands. In the second, a pentapalladium sheet adopts an edge-sharing triangle-trapezoid skeleton between two naphthacene rings. The compounds were characterized by x-ray crystallography and nuclear magnetic resonance spectroscopy. The nature of bonding in the clusters was analyzed by quantum calculations.     

Azimuthal variations of sap flux density within Japanese cypress xylem trunks and their effects on tree transpiration estimates

Sap flow techniques are practical tools for estimating tree transpiration. Though many previous studies using sap flow techniques did not consider azimuthal variations of sap flux density (F _d) on xylem trunk to estimate tree transpiration, a few studies reported that ignoring the azimuthal variations in F _d could cause large errors in tree transpiration estimates for some tree species. Therefore, examining azimuthal variations in F _d for major plantation tree species is critical for estimating tree transpiration. Using the thermal dissipation method, we examined azimuthal variations in F _d in six trees of Japanese cypress Chamaecyparis obtusa (Sieb. et Zucc.) Endl., which is one of the most common plantation tree species in Japan. We recorded considerable variations among F _d at four different azimuthal directions. The F _d value for one aspect was more than 100% larger than those for the other aspects. We calculated differences between tree transpiration estimates based on F _d for one to three azimuthal directions and those based on F _d for four aspects. The differences relative to tree transpiration estimates based on F _d for four aspects were typically 30, 20, and 10% in accordance with the F _d for one, two, and three measurement aspects, respectively. This finding indicates that ignoring azimuthal variations could cause large errors in tree transpiration estimates for Japanese cypress.     

Effects of Cd on the fluxes of K+ and H+ in excised roots

As a result of Cd treatment, K concentrations decreased in Cd sensItive maize and kidney bean calli (Obata et al. 1994) and in intact roots of kidney bean plants (Obata et al. unpublished). Potassium may be extruded from the roots or the absorption of K may be depressed by the Cd treatment in these Cd sensitive plants. Obata et al. (1996) observed that Cd inhibited both the efflux of H⁺ and influx of K⁺ following K⁺ addition in intact roots of bean. Thus Cd may affect the activity of proteins essential to ion movement., i.e. ioncarriers, channels and ATPase embedded in the membranes and/or may affect the permeability of the lipids of the membrane.     

Preliminary experiments on mechanical stretch-induced activation of skeletal muscle satellite cells in vivo

We have shown in vitro that mechanical stretch triggers activation of quiescent satellite cells of skeletal muscle to enter the cell cycle through an intracellular cascade of events including nitric oxide (NO) synthesis that results in the release of hepatocyte growth factor (HGF) from its extracellular association and its subsequent presentation to signaling receptors. In order to explore the activation mechanism in vivo, stretch experiments were conducted in the living animal using our suspension model developed. This system used the weight of the hind portion of rats to stretch the inside muscles of the left hind limb suspended for a period of 0.5–2.0 h. At the end of the stretch period, the rats received an intraperitoneal injection of bromodeoxyuridine followed by immunocytochemistry for its incorporation as an index of satellite cell activation in vivo. Depending on the period of stretch, bromodeoxyuridine labeling was increased significantly over the contralateral unstretched leg or control muscle from untreated rats. A stretched muscle extract prepared from the 2 h stretched tissue by incubating it in PBS, showed the active form of HGF as revealed by immunoblotting and it could stimulate the activation of unstretched satellite cells. Also, administering NO synthase inhibitor L‐NAME prior to muscle stretch abolished the stretch activation of satellite cells. Therefore, the results from these experiments demonstrate that stretching muscle triggers NO synthesis and HGF release, which could activate satellite cells in vivo.     

High-throughput mapping of a dynamic signaling network in mammalian cells

Signaling pathways transmit information through protein interaction networks that are dynamically regulated by complex extracellular cues. We developed LUMIER (for luminescence-based mammalian interactome mapping), an automated high-throughput technology, to map protein-protein interaction networks systematically in mammalian cells and applied it to the transforming growth factor-beta (TGFbeta) pathway. Analysis using self-organizing maps and k-means clustering identified links of the TGFbeta pathway to the p21-activated kinase (PAK) network, to the polarity complex, and to Occludin, a structural component of tight junctions. We show that Occludin regulates TGFbeta type I receptor localization for efficient TGFbeta-dependent dissolution of tight junctions during epithelial-to-mesenchymal transitions.     

Permeability of the fish intestinal membrane to bulky chemicals

The ability to predict the environmental behavior of chemicals precisely is important for realizing more rational regulation. In this study, the bioaccumulation of nine chemicals of different molecular weights absorbed via the intestinal tract was evaluated in fish using the everted gut sac method. The amounts of chemicals that passed through the intestinal membrane after a 24-hr exposure were significantly decreased for chemicals with MW≥548 and D_{max min}≥15.8 Å (or D_{max aver}≥17.2 Å). These thresholds are consistent with those previously proposed in terms of MW (>800) and molecular size (D_{max min}>15.6 Å or D_{max aver}>17.1 Å) for the limit of permeable chemicals through the gill membrane. The results show that the same MW and D_max criteria can be used to predict low bioaccumulation through both the gill membrane and the intestinal tract. These findings are helpful in reducing the need to conduct animal tests in environmental safety studies.     

Establishing a ready biodegradability test system using OxiTop® to evaluate chemical fate in a realistic environment

The purpose of this study is to propose the use of OxiTop® for measuring biochemical oxygen demand (BOD) under the Japanese Chemical Substances Control Law in order to properly evaluate chemical fate in a real environment. In our previous study, the biodegradation of test chemicals was accelerated by both adsorbing the chemical to silica gel with chloroform and increasing the medium volume from 300 to 3900 mL in the OECD 301F test using a coulometer. However, the biodegradability of these chemicals could not be evaluated based on BOD due to chloroform residue in the silica gel, or the medium volume could not be increased further due to the oven size of the coulometer. In this study, we established an evaluation system using OxiTop® based on BOD by increasing the medium volume to 9000 mL. Based on triplicate testing, increasing the medium volume accelerated biodegradation and decreased variation in BOD.     

Stand-scale transpiration estimates in a Moso bamboo forest: (I) Applicability of sap flux measurements

The applicability of sap flux (F_d) measurements to bamboo forests has not been studied. This study was undertaken to establish an optimal and effective design for stand-scale transpiration (E) estimates in a Moso bamboo forest. To this aim, we validated F_d measurements in Moso bamboos in a cut bamboo experiment. In addition, we analyzed how sample sizes affect the reliability of E estimates calculated from F_d and conducting culm area (A_{S_b}). In the cut bamboo experiments, we found that F_d measurement using a 10 mm probe was a valid means of determining the water-use behavior of a Moso bamboo, although a specific correction was needed. Furthermore, we calculated E from stand A_{S_b} (A_{S_stand}) and mean stand F_d (J_S). Employing Monte Carlo analysis, we examined potential errors associated with sample size in E, A_{S_stand}, and J_S using an original dataset with A_{S_b} and F_d measured for 40 and 16 individuals, respectively. Consequently, we determined the optimal sample size for both A_{S_stand} and J_S estimates as 11. The optimal sample sizes for J_S were almost the same under different vapor pressure deficit and soil moisture conditions. The optimal sample size for J_S at the study site was less than that of a coniferous plantation in the same region probably owing to small individual-to-individual variations in sap flux in the Moso bamboo forest. Our study concludes that sap flux measurements are an applicable technique for assessing water use in Moso bamboo forests.