Confirmation of nitrogen fixation in two tropical grasses by 15N2 incorporation

Intact soil cores containing plants of Paspalum notatum or Digitaria decumbens were selected with the acetylene reduction method, and then exposed to ¹⁵N₁₅ to confirm nitrogen fixation in tropical grass-bacteria associations. In a preliminary experiment with P. notatum¹⁵N₂ incorporation was slow but progressive during 24 h in roots but translocation to rhizomes and leaves ceased after 17h. With improved assay chambers, enrichments of 0.151 and 0.563 ¹⁵N atom % excess were obtained in roots of D. decumbens cv transvala and P. notatum systems respectively, after 3 days. Enrichments in rhizomes were similar to those of roots; however in the leaves only 8% of root enrichment was observed. The addition of sucrose to the soil doubled N₂-fixation in roots in both grass species studied, but did not result in increased incorporation into the leaves of P. notatum.     

Ruminant forestomach and abomasal mucormycosis under rumen acidosis

Spores of Absidia corymbifera were inoculated orally into sheep with ruminal acidosis produced by feeding barley. Lesions, which developed in forestomachs of all four inoculated cases, included desquamation of superficial layers of the mucosae and focal necrosis from lamina propria to muscular layers. Granulomatous lesions were in the submucosa of three sheep. Lesions in the abomasum (two sheep) included focal necrosis, diffuse hemorrhages, and infiltration of neutrophils. All lesions were accompanied by mycotic proliferation. These results show that A. corymbifera can invade forestomach mucosae through degenerate epithelium resulting from ruminal acidosis.     

Heterogeneous expression of glycoconjugates in the primary olfactory centre of the Japanese sword‐tailed newt (Cynops ensicauda)

Histochemical organization of the Caudata olfactory system remains largely unknown, despite this amphibian order showing phylogenetic diversity in the development of the vomeronasal organ and its primary centre, the accessory olfactory bulb. Here, we investigated the glycoconjugate distribution in the olfactory bulb of a semi‐aquatic salamander, the Japanese sword‐tailed newt (Cynops ensicauda), by histochemical analysis of the lectins that were present. Eleven lectins showed a specific binding to the olfactory and vomeronasal nerves as well as to the olfactory glomeruli. Among them, succinylated wheat germ agglutinin (s‐WGA), soya bean agglutinin (SBA), Bandeiraea simplicifolia lectin‐I (BSL‐I) and peanut agglutinin showed significantly different bindings to glomeruli between the main and accessory olfactory bulbs. We also found that s‐WGA, SBA, BSL‐I and Pisum sativum agglutinin preferentially bound to a rostral cluster of glomeruli in the main olfactory bulb. This finding suggests the presence of a functional subset of primary projections to the main olfactory system. Our results therefore demonstrated a region‐specific glycoconjugate expression in the olfactory bulb of C. ensicauda, which would be related to a functional segregation of the olfactory system.     

Antimicrobial activity of heartwood components of sugi (Cryptomeria japonica) against several fungi and bacteria

Methanol extract of sawdust of sugi (Cryptomeria japonica) heartwood was fractionated with toluene and n-hexane to give solvent-soluble and solvent-insoluble fractions. The n-hexane-soluble fraction showed the most inhibition activity among the fractions against phytopathogenic microorganisms, namely Fusarium oxysporum, Phytophthora capsici, Pythium splendens, and Ralstonia solanacearum. Sandaracopimarinol and ferruginol, isolated from the n-hexane-soluble fraction, showed moderate antifungal activity against the three fungi and strong antibacterial activity against R. solanacearum. The content of sandaracopimarinol (7.07 g/kg based on the dried sawdust) in the heartwood was about twice that of ferruginol. Sandaracopimarinol and ferruginol strongly inhibited the growth of Gram-positive bacteria but did not show inhibitory action against Gram-negative bacteria except for R. solanacearum. The antibacterial effect of sandaracopimarinol was first found in the present study and was stronger than that of ferruginol.     

Volatile organic compounds from wood and their influences on museum artifact materials II: Inference of causal substances of deterioration based on intercomparison of laser Raman spectra of deteriorated products

Volatile organic compounds (VOCs) from wood and their effects on artifacts are of intense interest to museum staff who must strictly manage the preservation environment. In this study, the causal substances of deterioration were inferred by comparison with laser Raman (LR) spectra of deterioration products. The artifact samples were deteriorated using VOCs from woods (WV test), and using specific substances that occur in wood VOCs (SV test). The deteriorated samples were analyzed by LR spectroscopy and the results of the WV and SV tests were compared. Hinokitiol and acetic acid were found to be two of the main causal substances of deterioration in western red cedar. Deterioration of iron and copper by spruce did not appear to be caused by acetic acid, as is generally assumed, but by some other unknown compound(s). Sensitivity to wood-based VOC components depended on the type of artifact. In western red cedar, mitsudasou (litharge) was very sensitive to acetic acid, while other artifacts were sensitive to hinokitiol. The LR method used in this study is very useful for the inference of causal substances of deterioration based on the detection of small amounts of deteriorated products such as those generated by deterioration due to wood VOCs. Part of this article was presented at the 25th Annual Meeting of the Japan Society for the Conservation of Cultural Property, Kyoto, June 2003     

alpha-Glucosidase inhibitory profile of catechins and theaflavins

To clarify the postprandial glucose suppression effect of flavonoids, the inhibitory effects of catechins and theaflavins against alpha-glucosidase (AGH) were examined in this study. It was initially demonstrated that theaflavins and catechins preferentially inhibited maltase rather than sucrase in an immobilized AGH inhibitory assay system. For the maltase inhibitory effects of theaflavins, the effects were observed in descending order of potency of theaflavin (TF)-3-O-gallate (Gal) > TF-3,3'-di-O-Gal > TF-3'-O-Gal > TF. This suggests that the AGH inhibition induced by theaflavins is closely associated with the presence of a free hydroxyl group at the 3'-position of TF as well as the esterification of TF with a mono-Gal group. In addition, the R-configuration at the 3'-position of TF-3-O-Gal showed a higher inhibitory activity than the S-configuration. As a result of a single oral administration of maltose (2 g/kg) in rats, a significant reduction in blood glucose level was observed at a dose of 10 mg/kg of TF-3-O-Gal, demonstrating for the first time that TF-3-O-Gal can suppress glucose production from maltose through inhibition of AGH in the gut.     

A cooperative action of endothelin-1 with prostaglandin F(2alpha) on luteal function in the cow

Prostaglandin F(2alpha) (PGF(2alpha)) is the primary luteolysin in the cow, and luteal endothelin-1 (ET-1) interacts with PGF(2alpha) during the process of luteolysis. In contrast, a developing corpus luteum (CL) is refractory to exogenous administration of PGF(2alpha). Thus, the present study was aimed to investigate the functional relationship between ET-1 and PGF(2alpha) in the mid-CL (PGF(2alpha)-sensitive) and early-CL (PGF(2alpha)-refractory). In the mid-CL model, cows (n = 6/treatment) were assigned to receive one of five types of treatments on day 10 of the estrous cycle: (1) an injection of saline; control, (2) a 500 microg of PGF(2alpha) analogue (sufficient dose to induce luteolytis); full-PG, (3) an intraluteal injection of 0.25 mg ET-1; ET-1, (4) a 125 micro g of PGF(2alpha) (insufficient dose to induce luteolytis); 1/4PG or (5) an intraluteal injection of 0.25 mg ET-1 after administration of a insufficient dose of PGF(2alpha) analogue; 1/4PG/ET. In the early-CL model, cows were assigned to receive one of two types of treatments on day 5 of the estrous cycle: (1) a sufficient dose of PGF(2alpha) analogue; PG (n = 5) or (2) an intraluteal injection ET-1 after a sufficient dose of PGF(2alpha); PG/ET (n = 7). In the mid-CL model, 1/4PG/ET resulted in a rapid reduction of progesterone (P) concentrations similar to that in full-PG from the next day. However, the levels of P in 1/4PG/ET (1.5-2.5 ng/ml) kept significantly higher than that in full-PG (< 0.5 ng/ml). ET-1 or 1/4PG did not decrease plasma P concentrations (4-6 ng/ml). The plasma ET-1 levels increased with the full-PG administration. In the early-CL model, both treatments had no effect on plasma P increase and ET-1 levels. The overall results indicate that the intraluteal ET-1 injection after administration of insufficient dose of PGF(2alpha) induces the depression of P secretion in vivo during the mid luteal phase in the cow, supporting the concept that ET-1 is one of a local mediator of functional luteolysis in the cow. The result further indicates that the early-CL is not only PG-refractory but also ET-1-refractory.