3-(4-Hydroxyphenyl)propionic acid is involved in the biosynthesis of myricanol in Myrica rubra

There is little evidence concerning the biosynthetic pathways for cyclic diarylheptanoids. We previously demonstrated that the cyclic diarylheptanoids myricanol and myricanone were biologically synthesized from two molecules of 4-coumaric acid by the feeding of 4-[8,9-¹³C₂]coumaric acid to young shoots of Myrica rubra. In the present study, using a ¹³C-labeled compound, we revealed that two molecules of 3-(4-hydroxyphenyl)propionic acid could also be a biosynthetic precursor of myricanol in M. rubra. These results indicated that both 4-coumaric acid and its dihydro-derivative were incorporated into myricanol. Competitive feeding experiments with 4-[8,9-¹³C₂]coumaric acid and 3-(4-hydroxyphenyl)-[1-¹³C]propionic acid were performed in M. rubra to determine the preferential incorporation of these two precursors. ¹³C-NMR studies indicated that 3-(4-hydroxyphenyl)-[1-¹³C]propionic acid was preferentially incorporated into myricanol. The data provided evidence for a biosynthetic sequence originating from 4-coumaric acid and leading to myricanol, through 3-(4-hydroxyphenyl)propionic acid, in M. rubra.     

A radio transmission pH measurement system for continuous evaluation of fluid pH in the rumen of cows

We developed a novel wireless radio transmission pH measurement system to continuously monitor ruminal bottom pH in cows, and compared these measurements to pH values determined by a spot-sample method. The wireless system consists of a pH sensor, data measurement receiver, relay unit, and personal computer with special software. The bullet-shaped sensor can be easily administered orally via a catheter into the rumen, without surgery. The glass electrode, using a temperature compensation system, can detect the rumen fluid pH with high accuracy. The ruminal bottom pH in healthy rumen-fistulated cows was measured as 6.52 ± 0.18 by the wireless system and as 6.62 ± 0.20 by the spot-sample method; with a correlation between pH measurements using these different methods (n = 8, 24 samples, r = 0.952, P < 0.01). When measured serially in a cow fed a diet evoking rumen acidosis, the ruminal bottom pH decreased markedly following the morning feeding and then increased gradually by the next morning feeding. This wireless system is a ready-to-use tool for estimating circadian changes in ruminal bottom pH.     

3-Methylthiopropionic acid ethyl ester, isolated from Katsura-uri (Japanese pickling melon, Cucumis melo var. conomon), enhanced differentiation in human colon cancer cells

The fully ripened fruit of Katsura-uri Japanese pickling melon ( Cucumis melo var. conomon) has rarely been used for food because the midripened fruit is utilized for making pickles, but the fully ripened fruit is no longer valuable for pickles due to the fruit body being too soft. We have considered the utilization of the fully ripened Katsura-uri fruit that may be used for nonpickling products, particularly if the fully ripened fruit demonstrated health benefits such as anticarcinogenic properties. The phytochemical extract from the fully ripened fruit of Katsura-uri Japanese pickling melon was purified via a bioassay-guided fractionation scheme, which was based on the induction of differentiation in a RCM-1 human colon cancer cell line. On the criteria of two differentiation markers (duct formation and alkaline phosphatase activity), the most potent fraction contained a compound identified as 3-methylthiopropionic acid ethyl ester, based on GC retention time, EI-MS, (1)H NMR, and (13)C NMR spectra. Previously, the role of 3-methylthiopropionic acid ethyl ester was considered as an odor producing compound in many fruits, but this study indicates potential medical benefits of this compound.     

Molecular epidemiology of VapA-positive Rhodococcus equi in thoroughbred horses in Kagoshima,Japan

The prevalence of virulent R. equi having 15- to 17-kDa antigens (VapA) in fecal isolates from 13 thoroughbred foals and their dams on 5 farms in Kagoshima, Japan, and the plasmid profiles of VapA-positive isolates by restriction fragment digestion patterns were investigated to compare the genotypic variation among virulence plasmids of R. equi isolates from Japan. In total, 218 (24.6%) of 886 isolates from the feces of the 13 foals and 13 (12.5%) of 104 isolates from the feces of their dams demonstrated VapA-positive R. equi. Plasmid DNA preparations of 231 virulent isolates from foals and dams were analyzed by restriction enzyme digestion with endonucleases EcoRI, EcoT22I and HindIII and were divided into 3 types: 172 isolates contained a 90-kb type I plasmid, 57 contained a 90-kb type III plasmid and 2 contained a 90-kb type IV plasmid. This study demonstrates a geographic character in the distribution of virulence plasmids found in VapA-positive isolates from thoroughbred foals in Kagoshima.     

In vitro reduction of manganese dioxide by a ferrireductase system from the white-rot fungus <Emphasis Type="Italic">Phanerochaete sordida</Emphasis> YK-624

Reduction of manganese dioxide is demonstrated for an in vitro ferrireductase system that includes NADPH-dependent ferrireductase and the iron-binding compound (IBC) isolated from the white-rot fungus Phanerochaete sordida YK-624. The Fe(II)–IBC complex was more effective in reducing manganese dioxide to Mn(II) than were complexes of Fe(II) and organic acids of low molecular weight such as nitrilotriacetate, although IBC also reduced manganese dioxide to Mn(II) in the absence of Fe(II). The generated Fe(III)–IBC complex was a better substrate for NADPH-dependent ferrireductase than were other ferric chelates, suggesting that the Fe(III)–IBC complex is reduced to an Fe(II) complex by NADPH-dependent ferrireductase. Moreover, production of the Fe(III)–IBC complex by the reduction of manganese dioxide in a reaction system containing Fe(II) and IBC was observed to be coupled to reduction of the Fe(III)–IBC complex by NADPH-dependent ferrireductase. These results indicate that the ferrireductase system of P. sordida YK-624 plays an important role in the reduction of manganese dioxide, which is necessary for the production and function of manganese peroxidase.     

In vitro screening of angiotensin I-converting enzyme inhibitors from Japanese cedar (Cryptomeria japonica)

Screening and isolation of angiotensin I-converting enzyme (ACE) inhibitors from Japanese cedar (Cryptomeria japonica) based on the in vitro ACE inhibitory assay were attempted. The ethanol extract from outer bark showed the highest inhibitory activity (IC₅₀ is 16g/ml) among 24 extracts prepared from roots, leaves, heartwood, sapwood, inner bark, and outer bark by successive extraction with four solvents. The fractionation of the outer bark ethanol extract followed by the bioassay resulted in the isolation of two strong ACE inhibitors, catechin and dimeric procyanidin B3. The bioassay of three flavan-3-ols including (+)-catechin and six flavones revealed that most of these compounds have high ACE inhibitory activity. The results suggest that the phenolic hydroxyl group at the C7 position and heterocyclic oxygen atom of these compounds are important for expressing the inhibitory activity.     

Structural changes of residual lignin in softwood kraft pulp treated with manganese peroxidase

Structural changes of residual lignin in unbleached softwood kraft pulp (SWKP) during manganese peroxidase (MnP) treatment were investigated to obtain some understanding of the biobleaching action of SWKP with MnP treatment. Alkaline-extracted lignin from darkened SWKP by MnP showed more intense color and contained moreo-quinone than that from control SWKP. However, no difference in the conjugated-carbonyl was observed between the lignins from MnP-treated and control SWKP. The nitrobenzene oxidation analysis revealed that oxidative condensation of non-condensed lignin in SWKP occurs during an early stage of MnP treatment. These observations were supported by the model experiment in which the lignin prepared from control SWKP was subjected to MnP treatments three times, and the changes of color and functional groups in the lignin were determined after each treatment. These results suggested that an increase ino-quinone and the condensation reaction of non-condensed lignin in SWKP are responsible for the characteristic darkening of SWKP during MnP treatment. It was also ascertained that darkened lignin was degraded and brightened by repeated MnP treatments.     

Polyethylene degradation by lignin-degrading fungi and manganese peroxidase

Degradation of high-molecular-weight polyethylene membrane by lignin-degrading fungi, IZU-154, Phanerochaete chrysosporium, and Trametes versicolor, was investigated under various nutritional conditions. IZU-154 showed the most significant polyethylene degradation among the three lignin-degrading fungi under nitrogen- or carbon-limited culture conditions. Furthermore, for T. versicolor and P. chrysosporium, the addition of Mn(II) into nitrogen- or carbon-limited culture medium enhanced polyethylene degradation. These results suggest that polyethylene degradation is related to ligninolytic activity of lignin-degrading fungi. Treatment of polyethylene membrane with partially purified manganese peroxidase (MnP) caused significant degradation in the presence of Tween 80, Mn(II), and Mn(III) chelator. This result demonstrates that MnP is the key enzyme in polyethylene degradation by lignin-degrading fungi.This study was presented in part at the 9th International Symposium on Wood and Pulping Chemistry, Montreal, Canada, June 9–12, 1997