Antioxidative compounds from leaves of Tahongai (<Emphasis Type="Italic">Klienhovia hospita</Emphasis>)

In our effort to find antioxidant agent, we focused on Tahongai (Kleinhovia hospita) which have been used traditionally in Indonesia as medicinal herbal to cure liver disease. Based on the biologically guided fractionation using DPPH radical scavenging assay, eleutherol and kaempferol 3-O-β-d-glucoside was isolated from the leaves of K. hospita. Kaempferol 3-O-β-d-glucoside (1) and eleutherol (2) scavenged the radical with IC₅₀ of 71.4 and 491.8 μM, respectively. In addition, both of the compounds did not exhibit cytotoxicity on HepG2 cells.     

No evidence of horizontal infection in horses kept in close contact with dogs experimentally infected with canine influenza A virus (H3N8)

Background

Since equine influenza A virus (H3N8) was transmitted to dogs in the United States in 2004, the causative virus, which is called canine influenza A virus (CIV), has become widespread in dogs. To date, it has remained unclear whether or not CIV-infected dogs could transmit CIV to horses. To address this, we tested whether or not close contact between horses and dogs experimentally infected with CIV would result in its interspecies transmission.

Methods

Three pairs of animals consisting of a dog inoculated with CIV (10^8.3 egg infectious dose₅₀/dog) and a healthy horse were kept together in individual stalls for 15 consecutive days. During the study, all the dogs and horses were clinically observed. Virus titres in nasal swab extracts and serological responses were also evaluated. In addition, all the animals were subjected to a gross pathological examination after euthanasia.

Results

All three dogs inoculated with CIV exhibited clinical signs including, pyrexia, cough, nasal discharge, virus shedding and seroconversion. Gross pathology revealed lung consolidations in all the dogs, and Streptococcus equi subsp. zooepidemicus was isolated from the lesions. Meanwhile, none of the paired horses showed any clinical signs, virus shedding or seroconversion. Moreover, gross pathology revealed no lesions in the respiratory tracts including the lungs of the horses.

Conclusions

These findings may indicate that a single dog infected with CIV is not sufficient to constitute a source of CIV infection in horses.     

Isolation and genetic analysis of Japanese encephalitis virus from a diseased horse in Japan

Japanese encephalitis (JE) developed in an unvaccinated half-bred horse kept in Tottori Prefecture, Japan. The animal showed ataxia with pyrexia and low appetite, and ultimately died. A viral strain was isolated from the cerebrum of the horse and was identified as JE virus (JEV) by RT-PCR using JEV specific primers. The isolated JEV was classified into genotype I by nucleotide sequence analysis of the viral envelope gene. We believe that this is the first report of the genotype I strain being isolated from a horse.     

Efficacy of oseltamivir phosphate to horses inoculated with equine influenza A virus

We investigated the efficacy of the oral administration of oseltamivir phosphate (OP) in horses experimentally infected with equine influenza A virus (H3N8). Nine horses were divided into three horses each of control, treatment and prophylaxis groups. An administration protocol for the treatment group (2 mg/kg of body weight, twice a day for five days) was started immediately after the onset of pyrexia (above 38.9 degrees C). An administration protocol for the prophylaxis group (2 mg/kg of body weight, once a day for five days) was started on a day before viral inoculation. In the treatment group, periods of virus excretion (mean days +/- standard deviation, 2.3 +/- 0.6) and pyrexia (2.0 +/- 0.0) were apparently shorter than those of the control group (6.0 +/- 0.0 and 8.0 +/- 1.0, respectively). In the prophylaxis group, although virus excretion and pyrexia were not prevented, the periods of virus excretion (5.0 +/- 0.0) and pyrexia (4.7 +/- 1.5) were shorter than those of the control group. Moreover, in the treatment and prophylaxis groups, bacterial counts of Streptococcus equi subsp. zooepidemicus known as the common pathogen of secondary bacterial pneumonia in bronchoalveolar lavage fluids collected seven days after inoculation were significantly fewer than that of the control group. The results indicated that the oral administration of OP to horses affected with equine influenza would contribute to reduce the magnitude of virus excretion, pyrexia and consequent secondary bacterial pneumonia.     

Cutaneous plasmacytoma in three golden hamsters (Mesocrietus auratus)

Spontaneously occurring cutaneous tumours in three golden hamsters were characterized using histological, immunohistochemical and ultrastructural methods. Histologically, the tumours were composed of sheets of round to oval plasmacytoid cells with eccentrically placed nuclei. Tissue sections were weakly positive for anti-B lymphocyte antigen (BLA) staining. Ultrastructurally, large amounts of rough endoplasmic reticulum in the cytoplasm were observed. BLA positivity and characteristics of ultrastructure showed the plasma cell origin.     

New multilocus genotypes of <Emphasis Type="Italic">Phytophthora infestans</Emphasis> in Japan

Twelve isolates of Japanese Phytophthora infestans, which differed from the major genotypes US-1, JP-1, JP-2, and JP-3, were analyzed for RG57 fingerprints, mtDNA haplotypes, two allozyme genotypes, and mating types. Genotypes JP-1.1, JP-2.1, JP-2.2, JP-3.1, and JP-4 were newly defined. JP-1.1 and JP-2.1 were isolated discontinuously from potato fields in several years, and JP-1.1 was found in Hokkaido and Kagoshima. These results show that some minor genotypes can overwinter and disperse from their original site.     

Effects of auxin and jasmonates on 1-aminocyclopropane-1-carboxylate (ACC) synthase and ACC oxidase gene expression during ripening of apple fruit

1-Aminocyclopropane-1-carboxylate (ACC) synthase and oxidase activities, their gene expression, and ethylene production in apple fruit [Malus sylvestris (L.) Mill. Var. domestica (Borkh.) Mansf.] treated with a synthetic auxin 2,4-dichlorophenoxy-propionic acid (2,4-DP) and n-propyl dihydrojasmonate (PDJ), a jasmonic acid derivative, has been investigated to clarify the action of auxin and jasmonates on ethylene production. The fruit was harvested at 103 d after full bloom (preclimacteric). The expression of MdACS4 messenger RNA (mRNA) at 48 and 96 h after treatment was higher in fruit treated with 2,4-DP than in the untreated control, but those of MdACS1 and MdACO1 were not affected by treatment. The ethylene production in 2,4-DP-treated fruit increased at 96 h after treatment. In contrast, expression of mRNAs hybridized with MdACS1 and MdACO1 probes in the skin of PDJ-treated fruit were higher than those in the untreated control. In addition, ACC synthase activity and ethylene production also increased after treatment. These results show that the ethylene production rate may differ with the kind of genes which were stimulated by auxin or jasmonates.     

NADPH-dependent ferrireductase produced by white-rot fungusPhanerochaete sordida YK-624

An intracellular, soluble ferrireductase thought to be involved in the reduction of manganese dioxide by white-rot fungusPhanerochaete sordida YK-624 was purified for the first time. Two isoenzymes, NAD(P)H-dependent and NADPH-dependent, respectively, were detected by hydrophobic chromatography. The NADPH-dependent ferrireductase was purified to homogeneity by ammonium sulfate fractionation, hydrophobic interaction, gel permeation, and anion-exchange chromatography. The purified protein, which is monomeric, has a molecular mass of 35 kDa (determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis) and pl 5.1 (determined by isoelectric focusing). The purified protein did not use cellobiose as an electron donor. The purified protein reduced Fe(III)-nitrilotriacetate complex, Mn(III)-malonate complex, methoxy-p-benzoquinone, and cytochrome c; veratraldehyde, 2-hydroxy-1,4-naphthoquinone, phenazine methosulfate, and plumbagin could not be reduced. Particularly, the protein showed the highest reducing rate for Fe(III)-organic acid complexes, such as Fe(III)-nitrilotriacetate, among these electron acceptors.     

Effects of co-overproduction of sedoheptulose-1,7-bisphosphatase and Rubisco on photosynthesis in rice

Overproduction of Rubisco did not proportionately increase the rate of CO₂ assimilation (A) in rice probably because the capacity of regeneration of the substrate of Rubisco, ribulose-1,5-bisphosphate (RuBP), could not afford the increased Rubisco capacity. Since sedoheptulose-1,7-bisphosphatase (SBPase) is thought to be one of the limiting factors of RuBP regeneration capacity, SBPase and Rubisco were co-overproduced in rice in order to improve photosynthesis. Although SBPase and Rubisco contents increased by 82–102% and 20–30%, respectively, A did not increase under the conditions of high irradiance and different [CO₂] partial pressures. Thus, co-overproduction of SBPase and Rubisco did not improve photosynthesis in rice. Overproduction and antisense suppression of SBPase did not greatly affect A at high [CO₂], although it is thought to be determined by RuBP regeneration capacity. These results strongly suggest that SBPase does not determine RuBP regeneration capacity in rice. This is likely to be the reason for the unimproved photosynthesis by co-overproduction of SBPase and Rubisco.     

Isoprenoid-substituted flavonoids from wood of Artocarpus heterophyllus on B16 melanoma cells: Cytotoxicity and structural criteria

As a result of cytotoxicity-guided fractionation, nine flavonoids, artocarpin (1), cudraflavone C (2), 6-prenylapigenin (3), kuwanon C (4), norartocarpin (5), albanin A (6), cudraflavone B (7), brosimone I (8) and artocarpanone (9) were identified from the methanol extract of the wood of Artocarpus heterophyllus, known commonly as Nangka in Indonesia. A structure–activity investigation of the effect of these isolated compounds (1–9) and structurally related compounds on B16 melanoma cells indicated that isoprenoid moiety substitutions in flavonoids enhance their cytotoxicity, and that the position of attachment and the number of isoprenoid-substituent moieties per molecule influence flavonoid cytotoxicity.