[High-speed liquid chromatographic analysis of drugs. IV. Speration analysis of phyllodulcin and hydrangenol in sweet hydrangea (author's transl)].
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The inhibitory actions of thunberginol A on anti-allergic (type I-IV) activity were examined. Oral administration of thunberginol A 2 hr before the challenge significantly inhibited the PCA reaction (type I) in rats at a dose of more than 300 mg/kg and the ear PCA reaction in mice at a dose of more than 50 mg/kg. Thunberginol A at a dose of more than 300 mg/kg also significantly inhibited the allergic bronchoconstriction in rats. Thunberginol A concentration-dependently (10(7)-10(-4), 10(-5)-10(-4) M) inhibited the allergic contractions of rat trachea sensitized with IgE and those of the guinea pig lung preparation sensitized with IgG. It also inhibited the allergic histamine release from sensitized peritoneal exudate cells in a concentration-dependent manner (10(-5)-10(-4) M). Thunberginol A had anti-serotonic activity on the contraction of smooth muscle, and it increased the ear vascular permeability in mice. Thunberginol A significantly inhibited the primary response of contact dermatitis (type IV) in mice at 100 mg/kg from the day after immunization to the day before challenge, and it also inhibited the delayed type foot pad swelling in mice at a dose of more than 300 mg/kg at 0 and 8 hr after the challenge. These findings suggest that orally administered thunberginol A is effective against type I and type IV allergy.
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The submicroscopic anatomy of the eye in female Eupulvinaria is described. This scale insect has no compound eyes but simple pericerebral eyes composed of one pair of ocelli. The ocelli, located dorsolaterally to the brain, are of rhabdomeric type. They are situated in the subepidermal region underneath the basement membrane of the epidermis. Each ocellus consists of a cup made of a layer of numerous pigmented cells enclosing a few photoreceptive cells. The cuticular lens is facing the opening of the pigment-cup, while the optic nerve is emerging from the bottom of the cup. Pigmented cells show characteristic pigment granules. They are linked to each other by interdigitations or by desmosomes junctions. Both dendrites and cell body of the photoreceptor cell lie inside the eyecup, while the axon lies outside. The dendritic processes consist of large shaft-shaped rhabdomeres with microvilli extending perpendicular to the direction of the shaft. These microvilli are enveloped by digitiform cytoplasmic extensions of pigmented cells. These latter strands are free of pigment granules but contain microtubules orientated to the centre of the cup.
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The photosynthetic light acclimation of fully expanded leaves of tree seedlings in response to gap formation was studied with respect to anatomical and photosynthetic characteristics in a natural cool-temperate deciduous forest. Eight woody species of different functional groups were used; two species each from mid-successional canopy species (Kalopanax pictus and Magnolia obovata), from late-successional canopy species (Quercus crispula and Acer mono), from sub-canopy species (Acer japonicum and Fraxinus lanuginosa) and from vine species (Schizophragma hydrangeoides and Hydrangea petiolaris). The light-saturated rate of photosynthesis (Pmax) increased significantly after gap formation in six species other than vine species. Shade leaves of K. pictus, M. obovata and Q. crispula had vacant spaces along cell walls in mesophyll cells, where chloroplasts were absent. The vacant space was filled after the gap formation by increased chloroplast volume, which in turn increased Pmax. In two Acer species, an increase in the area of mesophyll cells facing the intercellular space enabled the leaves to increase Pmax after maturation. The two vine species did not significantly change their anatomical traits. Although the response and the mechanism of acclimation to light improvement varied from species to species, the increase in the area of chloroplast surface facing the intercellular space per unit leaf area accounted for most of the increase in Pmax, demonstrating the importance of leaf anatomy in increasing Pmax.
Sequence data have been determined for 5 members of subgroup 2 of the genus Ilarvirus. These data support the known serological relationships among accepted members of this group and indicate that the ilarvirus Hydrangea mosaic virus (HdMV) is an isolate of Elm mottle virus (EMoV). The close relationships between members of this subgroup, exhibited through the coat proteins coded on RNA 3, extend to the other genomic molecules. Primers designed from the sequences of RNA 1 and RNA 2 of EMoV amplified fragments from all other subgroup 2 viruses but not from other ilarviruses. Although closely related, members of this subgroup occur naturally in distinctly different host species. The possible origins of the viruses are discussed in relation to similarities among the genomic molecules, in particular RNA 3.
Chalcone (CHS), stilbene (STS) synthases, and related proteins are key enzymes in the biosynthesis of many secondary plant products. Precursor feeding studies and mechanistic rationalization suggest that stilbenecarboxylates might also be synthesized by plant type III polyketide synthases; however, the enzyme activity leading to retention of the carboxyl moiety in a stilbene backbone has not yet been demonstrated. Hydrangea macrophylla L. (Garden Hortensia) contains stilbenecarboxylates (hydrangeic acid and lunularic acid) that are derived from 4-coumaroyl and dihydro-4-coumaroyl starter residues, respectively. We used homology-based techniques to clone CHS-related sequences, and the enzyme functions were investigated with recombinant proteins. Sequences for two proteins were obtained. One was identified as CHS. The other shared 65-70% identity with CHSs and other family members. The purified recombinant protein had stilbenecarboxylate synthase (STCS) activity with dihydro-4-coumaroyl-CoA, but not with 4-coumaroyl-CoA or other substrates. We propose that the enzyme is involved in the biosynthesis of lunularic acid. It is the first example of a STS-type reaction that does not lose the terminal carboxyl group during the ring folding to the end product. Comparisons with CHS, STS, and a pyrone synthase showed that it is the only enzyme exerting a tight control over decarboxylation reactions. The protein contains unusual residues in positions highly conserved in other CHS-related proteins, and mutagenesis studies suggest that they are important for the structure or/and the catalytic activity. The formation of the natural products in vivo requires a reducing step, and we discuss the possibility that the absence of a reductase in the in vitro reactions may be responsible for the failure to obtain stilbenecarboxylates from substrates like 4-coumaroyl-CoA.
TGF-beta plays important roles in the induction of catagen during the hair cycle. We examined whether TGF-beta2 could activate a caspase in human hair follicles. Using active caspase-9 and -3 specific antibodies, we found that TGF-beta2 activated these caspases in two regions, the lower part of the hair bulb and the outer layer of the outer root sheath. In addition, we searched for a plant extract that can effectively suppress TGF-beta action. We found that an extract of Hydrangea macrophylla reduced synthesis of a TGDbeta-inducible protein. We confirmed that the extract has a potential to promote hair elongation in the organ culture system. Furthermore, it delayed in vivo progression of catagen in a mouse model. Our results suggest that the induction of catagen by TGF-beta is mediated via activation of caspases and that a suppressor of TGF-beta could be effective in preventing male pattern baldness.
We determined the effects of short-term cultivation with various amounts of available nitrate nitrogen (NO3-) on NO3- use by woody shrub species. Nitrate concentration ([NO3-]) and nitrate reductase activity (NRA) were measured in leaves and roots of seedlings of Hydrangea hirta (Thunb.) Siebold, Lindera triloba (Sieb. et Zucc.) Blume and Pieris japonica (Thunb.) D. Don. Root [NO3-] increased with increasing NO3- supply in all species, whereas leaf [NO3-] remained low. There were significant correlations between [NO3-] in roots and leaves in all species, but no correlation was found between root NRA and leaf NRA. The low proportion of leaf NO3- assimilation to total NO3- assimilation in all species can be ascribed to the lack of NO3- transport from roots to leaves. In all species, root NRA increased with increasing NO3- supply until reaching a plateau. Species ranking based on maximum root NRA was H. hirta > L. triloba > P. japonica. Root NRA in P. japonica was low, even though root [NO3-] increased with NO3- supply, indicating that NO3- was not an effective N source for this species. The ranking also suggested that H. hirta depended more on NO3- as an N source than L. triloba. The increase in root NRA with increasing NO3- supply was greater in H. hirta than in L. triloba, possibly indicating that a change in NO3- availability has a stronger influence on NO3- use in H. hirta than in L. triloba.
Hydrangea febrifuga (syn. Dichroa febrifuga) is a traditional medicinal plant distributed in China and Southeast Asia, and febrifugine, one of its principal bioactive constituents, has served as an important lead compound for antimalarial drug development. Viral infections may adversely affect the quality of medicinal plants; however, no emaravirus has previously been reported from H. febrifuga. Here, high-throughput sequencing was performed on H. febrifuga leaves exhibiting mosaic symptoms collected in Yunnan Province, China. Combined with RT-PCR, Sanger sequencing, and 5'/3' rapid amplification of cDNA ends (RACE), five full-length genomic RNA segments of a putative novel emaravirus, tentatively designated Dichroa emaravirus (DEV), were identified and characterized. The five negative-sense single-stranded RNA (-ssRNA) segments have a combined length of 12,971 nt and encode an RNA-dependent RNA polymerase (RdRp), glycoprotein precursor (GP), nucleocapsid protein (NP), movement protein (MP), and an uncharacterized accessory protein, P5. The maximum amino acid sequence identities of DEV P1-P4 with recognized emaraviruses were 73.90%, 51.82%, 65.60%, and 81.30%, respectively, whereas P5 showed a maximum identity of 49.16% with its closest homolog. Thus, three of the four core proteins had maximum identities below 80%, consistent with the current ICTV species demarcation criterion for the genus Emaravirus. Maximum-likelihood phylogenetic analyses based on the four core proteins further supported the placement of DEV within the genus Emaravirus (family Fimoviridae). These results support DEV as a putative novel emaravirus and represent the first report of an emaravirus associated with H. febrifuga.
We investigated spatio-temporal genetic variation in allele frequency and estimated gene flow among sympatric populations of Tetranychus kanzawai on different host plants by the use of microsatellite markers. In the analysis of spatial genetic variation, no isolation by distance was detected among the populations. Gene flow between populations on Hydrangea macrophylla and those on other host plants was relatively restricted, whereas the populations on Akebia quinata and Clerodendrum trichotomum were almost panmictic. Our study on temporal genetic variation showed (1) that population differentiation was slightly reduced during the period from April to May owing to frequent gene flow among populations; and (2) that population differentiation was greatly enhanced from May to October because of bottleneck effects. Genetic differentiation among T. kanzawai populations was caused by the effect of host plants rather than by the effect of geographic distance among populations, suggesting possibility of sympatric host race formation in this species.
The effects of crude drugs on fungi have been used for a thousand years in China and Japan. These drugs include: Saussureae radix, Magnoliae cortex, Cinnamomi cortex, Hydrangeae dulcis folium, and Artemisiae capillarius flos. The activity of Coptidis rhizoma and Phellodendri cortex was stronger than other crude drugs against the three fungi. Berberine hydrochloride, which is a component of the two crude drugs, was investigated. Minimal inhibitory concentration values of berberine hydrochloride were 1, 0.125, and 0.5 mg/ml against Candida albicans, C. tropicalis, and C. glabrata, respectively. In C. glabrata, compared with C. albicans and C. tropicalis, berberine hydrochloride greatly inhibited the growth of fungi.
The inhibitory effects of 151 natural products, representing most of the frequently occurring types, on the cytotoxicity to MM2 tumor cells of polymorphonuclear leucocytes (PMN) induced by TAK, a polysaccharide immunomodulator, were examined. Forty-two compounds inhibited the TAK-induced activation of PMN. Among them, some naturally occurring quinones and various alkaloids (nicotine, Cinchona alkaloids, isoquinoline alkaloids such as cepharanthine, and indole alkaloids such as ajmaline) exhibited potent inhibitory effects. Using the inhibition assay for monitoring, the extracts of Hydrangea Dulcis folium, Scopoliae rhizoma, Cinchona cortex, Magnoliae cortex, Stephania tuber, and Rauwolfia radix were analysed to characterize the active constituents.
Phytotoxicity of aluminum ion (Al3+) is a serious problem limiting crop production on acid soils. Organic acids with Al-chelating ability play an important role in the detoxification of Al both externally and internally. Al is detoxified externally by the secretion of organic acids such as citric, oxalic, and/or malic acids from the roots. The secretion of organic acids is highly specific to Al and the site of secretion is localized to the root apex. The kind of organic acids secreted as well as secretion pattern differ among plant species. There are two patterns of Al-induced secretion of organic acids: In pattern I, there is no discernible delay between the addition of Al and the onset of the release of organic acids. Activation of the anion channel seems to be involved in this pattern; In pattern II, there is a marked lag phase between the addition of Al and the onset of organic acid release. The action of genes related to the metabolism and secretion of organic acids seems to be involved in this pattern. Internal detoxification of Al in Al-accumulating plants is achieved by the formation of Al-organic acid complex. For instance, the complex of Al-citrate (1:1) in hydrangea and Al-oxalate (1:3) in buckwheat has been identified.