Facile synthesis of rubiadin by microwave heating

Rubiadin (4) (1,3-dihydroxy-2-methylanthraquinone), which is one of the natural dyes from the roots of Rubiaceae, was synthesized from phthalic anhydride (1) and 2,6-dihydroxytoluene (3) by two reaction steps, that is, a Friedel-Crafts reaction with AlCl₃ and a cyclization with H₂SO₄/HBO₂. Microwave heating resulted in remarkable acceleration of the reaction rate in the first reaction, and had an effect in reducing the reaction time and increasing the yield of rubiadin (4) in the second reaction.     

Morphological features and dietary functional components in fruit bodies of two strains of <Emphasis Type="Italic">Pholiota adiposa</Emphasis> grown on artificial beds

 The morphological features and dietary functional components of two strains (FPF-13 and Oninome-B) of Pholiota adiposa (numerisugitake mushroom) grown on artificial bed blocks were examined. The components examined were chitin, mannitol, trehalose, guanosine 5′-monophosphate, ergosterol, and β-glucan. There was a significant difference in the external shape of the two strains. However, the differences in the contents of functional components between the two strains, as well as between the pilei and stipes of the strains, were small. In both strains the trehalose content was much higher than the mannitol content, in contrast to those of Lentinula edodes. From a commercial point of view, Oninome-B has a clear advantage over other strains of P. adiposa owing to its less removable scale. Received: May 15, 2002 / Accepted: June 10, 2002 Acknowledgment The authors thank Dr. F. Eguchi (Takasaki University of Health and Welfare) for his technical advice on the β-glucan analysis. Part of this report was presented at the 51st Annual Meeting of the Japan Wood Research Society, Tokyo, April 2001 Correspondence to:K. Shimizu     

Influences of genetic and environmental factors on the concentration of the allergen Cry j 1 in sugi (Cryptomeria japonica) pollen

Pollen from sugi (Japanese cedar, Cryptomeria japonica D. Don), a forest tree species that is widely grown in Japan, causes serious allergic disease. The major allergens from sugi pollen, Cry j 1 and Cry j 2, have been isolated and characterized. It has been reported that Cry j 1 concentration in pollen varies considerably among trees. If Cry j 1 concentration is genetically controlled, the planting of trees with low Cry j 1 concentrations would reduce pollinosis. We investigated genetic and environmental effects on Cry j 1 concentration in eight clones growing at four sites. Concentrations of Cry j 1 in pollen were measured with a monoclonal antibody-based Enzyme Linked Immunosorbent Assay (ELISA). The Cry j 1 concentrations differed significantly among clones and sites, but the site x clone interaction was not significant, suggesting that the Cry j 1 concentration is controlled primarily by genetic factors. We examined correlations between Cry j 1 concentration and temperature and precipitation from July through February. Temperature was not significantly related to Cry j 1 concentration, whereas cumulative precipitation during the 8 months and mean daily precipitation in September showed significant negative correlations with Cry j 1 concentration.     

Screening of Indonesian plants for tyrosinase inhibitory activity

In our efforts to find new tyrosinase inhibitory materials, we investigated 44 Indonesian plants belonging to 24 families for tyrosinase inhibitory activity. The extracts of 5 Artocarpus woods showed potent tyrosinase inhibitory activity (over 80% at 100 μg/ml) similar to a positive control, kojic acid. In Artocarpus woods, the extracts of the sapwoods showed stronger inhibitory activity than those of the heartwoods. Chlorophorin was isolated as one of the active compounds in the sapwood of Artocarpus heterophyllus. The content of chlorophorin in sapwood was higher than that in heartwood. Part of this paper was presented at the 53rd (Fukuoka, March 2003) and 54th (Hokkaido, August 2004) Annual Meetings of the Japan Wood Research Society, and the 5th International Wood Science Symposium (Kyoto, September 2004)     

Condensation reactions of phenolic resins VII: catalytic effect of sodium bicarbonate for the condensation of hydroxymethylphenols

This article describes the catalytic effect of NaHCO₃ on condensation reactions of monomeric hydroxymethylphenols (HMPs) to elucidate the cure-acceleration mechanism. By comparison of the kinetics of self-condensations of HMPs, NaHCO₃ was proved to increase the reactivity of para-hydroxymethyl groups. The changes of ¹³C nuclear magnetic resonance (NMR) chemical shifts on each HMP system with the additive indicated that the addition of NaHCO₃ enhanced some molecular interactions between HMPs and NaHCO₃, facilitating a resonance effect that might play a similar role in dissociation of the phenolic hydroxyl groups of HMPs. In addition, computational modeling by molecular orbital calculations elucidated that hydrogen carbonate anion (HCO ₃ ⁻ ) forms an interaction between either the para-hydroxymethyl group and the phenolic hydroxyl group or between the two para-hydroxymethyl groups of HMPs by hydrogen bonds. From the experimental results, the authors proposed the mechanism of the catalytic action of NaHCO₃: it appears to be due to the delocalization of an electron initiated by the interaction of the para-hydroxymethyl groups and the phenolic hydroxyl of HMPs with HCO ₃ ⁻ through hydrogen bonds, which results in facilitating the formation of active species.     

Studies on hypersaline-tolerant white-rot fungi IV: effects of Mn 2+ and NH 4 + on manganese peroxidase production and Poly R-478 decolorization by the marine isolate Phlebia sp. MG-60 under saline conditions

Manganese peroxidase (MnP) production by Phlebia sp. MG-60 is strongly regulated by Mn²⁺ and NH₄⁺ at various sea salt incubation conditions. Extra-added Mn²⁺ and NH₄⁺ obviously inhibited MnP production, but sea salts relieved the inhibition partially or completely. Three media were prepared: Kirk medium with addition of 0%–5% sea salts (KS medium), a high level of Mn²⁺ (300mg/l) in KS medium (HM-KS medium), and a high level of NH₄⁺ (430mg/l) in KS medium (HN-KS medium). Without addition of sea salts, the dye Poly R-478 was significantly decolorized by low MnP activity (about 200U/l) and a low level of laccase activity (about 100U/l) in KS and HM-KS media. In the cultures in which laccase activity was almost completely inhibited by 3% and 5% sea salts, MnP activity higher than 400U/l was necessary for Poly R-478 decolorization in all of the three media. We first report the linear correlation of MnP activity and decolorization of Poly R-478 under saline conditions and the effect of laccase on this relation.     

Iron-binding compounds produced by white-rot fungusPhanerochaete sordida YK-624

Iron-binding compounds were isolated from a culture ofPhanerochaete sordida YK-624 and were found to bind to Fe(III) preferentially compared with Fe(II). Two iron-binding compounds were purified to near-homogeneity with gel permeation chromatography. Hydrolysis of the iron-binding compounds with 6N hydrochloric acid gave ninhydrin-negative products. The molecular weight of these compounds was 3–5 kDa. These compounds may play an important role in the reduction of extracellular manganese dioxide to Mn(II) by intracellular ferrireductases for lignin degradation by manganese peroxidase.     

Individual-based measurement and analysis of root system development: case studies for <Emphasis Type="Italic">Larix gmelinii</Emphasis> trees growing on the permafrost region in Siberia

We present results of individual-based root system measurement and analysis applied for Larix gmelinii trees growing on the continuous permafrost region of central Siberia. The data of root excavation taken from the three stands were used for the analyses; young (26 years old), mature (105 years old), and uneven-aged over-mature stand (220 years old). In this article, we highlight two topics: (1) factors affecting spatio-temporal pattern of root system development, and (2) interactions between aboveground (i.e., crown) and belowground (i.e., root) competition. For the first topic, the detailed observation of lateral roots was applied to one sample tree of the overmature stand. The tree constructed a superficial (<30 cm in depth) and rather asymmetric root system, and each lateral root expanded mainly into elevated mounds rather than depressed troughs. This indicated that spatial development of an individual root system was largely affected by microtopography (i.e., earth hummocks). For these lateral roots, elongation growth curves were reconstructed using annual-ring data, and annual growth rates and patterns were compared among them. The comparison suggested that temporal root system development is associated with differences in carbon allocation among the lateral roots. For the second topic, we examined relationships between individual crown projection area (CA) and horizontal rooting area (RA) for the sample trees of each stand. RA was almost equal to CA in the young stand, while RA was much larger (three or four times) than CA in the mature and overmature stands. Two measures of stand-level space occupation, crown area index (aboveground: CAI; sum of CAs per unit land area) and rooting area index (belowground: RAI; sum of RAs), were estimated in each stand. The estimates of RAI (1.3–1.8 m² m₋₂) exceeded unity in all stands. In contrast, CAI exceeded unity (1.3 m² m₋₂) only in the young stand, and was much smaller (<0.3 m² m₋₂) in the two older stands. These between-stand differences in RAI–CAI relationships suggest that intertree competition for both aboveground and belowground spaces occurred in the young stand, but only belowground competition still occurred in the two older stands. Based on this finding, we hypothesized that competition below the ground may become predominant as a stand ages in L. gmelinii forests. Methodological limitations of our analysis are also discussed, especially for the analysis using the two indices of space occupation (CAI, RAI).     

The effect of 7.2% hypertonic saline solution on the duration of sodium gradient between the cerebrospinal fluid and the venous circulation in the dog

To determine the duration of water movement from cerebrospinal fluid (CSF) into venous blood by the infusion of 7.2% hypertonic saline solution (HSS), the sodium gradient between venous blood and CSF were examined. Venous sodium concentrations remained higher than that in CSF for duration of 60 min following HSS infusion. By 90 min, the CSF sodium concentration reached the equilibrium with venous sodium concentration. Those data suggests that the duration of time during which water moved from CSF into capillaries in brain by the gradient of sodium concentration was less than 90 min.     

The histopathogenesis of paralytic rabies in six-week-old C57BL/6J mice following inoculation of the CVS-11 strain into the right triceps surae muscle

A fatal encephalomyelitis was developed after intracerebral and hind limb inoculation of in 6-week-old C57BL/6J mice by the inoculation of fixed rabies virus (CVS-11 strain), intracerebrally and into hind. After the intracerebral inoculation, virus antigens were detected in the cerebral cortex and hippocampus at 2 days postinoculation (PI), and later spread centrifugally to thalamus, brain stem, cerebellum, spinal cord and spinal ganglia. At 4 days PI, severe apoptosis and DNA fragmentation were observed in the hippocampus and cerebral cortex. All mice infected intracerebrally were dead without limb paralysis at from 10 to 11 days PI. In contrast, mice infected with virus intramuscularly were persistently observed virus antigens in the myocytes at the site of inoculation from 2 days PI. At 4 days PI, the antigens were demonstrated in the spinal dorsal root ganglia, spinal cord and muscle spindles without their detection in the cerebrum and hippocampus. There were no apoptosis in the spinal cord and dorsal root ganglia, however hind limb paralysis was found in all infected mice. Hind limb paralysis was progressed to quadriparalysis, and mice were dead from 11 to 13 days PI. From 4 days PI, necrosis of neuron was observed in the the spinal and dorsal ganglia with infiltration of lymphocyte. This study suggested that the necrosis of spinal neurons was more important to cause the paralysis of hind limb rather than the severe cerebral infection and apoptosis in C57BL/6J mice infected with CVS-11 strain. The virus primarily replicated in the muscles was ascended the spinal cord via afferent fibers and retrogradely invaded the cerebrum, and with subsequent spread to muscle spindles.