Extractives of <Emphasis Type="Italic">Quercus crispula</Emphasis> sapwood infected by the pathogenic fungi <Emphasis Type="Italic">Raffaelea quercivora</Emphasis> I: comparison of sapwood extractives from noninfected and infected samples

The extracts of Quercus crispula infected by the ambrosia fungus, Raffaelea quercivora, were investigated. Phenol and tannin analyses indicated that normal sapwood (NS) contained a considerable amount of hydrolysable tannins, while infected colored sapwood (IS) contained less hydrolysable tannins and more phenols than NS. In treating pentagalloyl glucose (PGG), which is a model compound of hydrolysable tannins, with a culture medium of R. quercivora, PGG was rapidly hydrolyzed to produce gallic acid. The resulting gallic acid decreased in concentration over the subsequent cultivation period eventually disappeared. Measuring tannase and laccase activities of the culture medium of R. quercivora, tannase activity increased gradually from the beginning, while laccase activity increased rapidly at 5 days of incubation and disappeared at 8 days. An oxidative product from gallic acid treated with laccase was isolated by preparative high performance liquid chromatography, and was identified as purprogallincarboxylic acid (PGCA) by nuclear magnetic resonance spectroscopy and electron-impact mass spectrometry. PGCA was present in a 70% aqueous acetone extract of IS, and showed slight growth inhibition against R. quercivora. Part of this study was presented at the 57th Annual Meeting of the Japan Wood Research Society, Hiroshima, Japan, 2007     

Isolation of Molecular Markers Linked to the <Emphasis Type="Italic">Cry</Emphasis> Locus Conferring Resistance to <Emphasis Type="Italic">Cucumber mosaic cucumovirus </Emphasis>Infection in Cowpea

We previously demonstrated that cowpea [Vigna unguiculata (L.) Walp.] cultivar Kurodane-Sanjaku contains the Cry gene, which confers resistance against Cucumber mosaic cucumovirus infection. In this paper, randomly amplified polymorphic DNA (RAPD) analysis was carried out to tag the Cry locus. Bulked segregant analysis for RAPD resulted in many polymorphisms in amplified DNA patterns. Candidates were further screened using parental and/or F₂ cowpea DNAs. As a result, we obtained three RAPD markers, D13/E14-350, WA3-850 and OPE3-500, flanking the Cry locus. In addition, we amplified cowpea sequences coding for the putative nucleotide-binding site (NBS). Degenerate primers based on NBS sequences of tobacco N and Arabidopsis RPS2 disease resistance genes were used for polymerase chain reaction, and resultant products were cloned and sequenced. Among eight independent clones, cowpea resistance gene analog (CRGA) 5 showed a distinct polymorphism when used as a probe for restriction fragment length polymorphism analysis against the susceptible cowpea cultivar PI 189375 and a near-isogenic line for the Cry. Linkage analyses of these molecular markers showed that genetic distances of CRGA5, D13/E14-350, WA3-850 and OPE3-500 to the Cry locus were 0.7, 5.2, 11.5 and 24.5 cM, respectively. Received 16 December 1999/ Accepted in revised form 22 March 2000     

The Effect of Light on the Accumulation of Lipids in Seeds

To investigate the effects of light on the accumulation of lipids in seeds, fruits of both peanut and soybean plants were allowed to grow under different types of light conditions (natural, red, blue, green, or far-red light) and in darkness. Irradiation with each type of light from the 20th or 30th day, except for green light from the 30th day, resulted in lower dry weight of peanut seeds. The peanut seeds irradiated with natural, blue or far-red light from the 30th day onward contained a smaller amount of lipids, lower levels of triacylglycerides (TG), and higher levels of total sugar and diacylglycerides (DG), than the seeds grown in darkness. In the case of soybean seeds grown in darkness, the lipid content increased, but the total sugar and protein contents decreased. Exposure of soybean seeds to far-red light did not result in an increase in the lipid content. In vitro, ¹⁴C-glycerol-3-phosphate was converted to DG rather than to TG under irradiation with light compared with dark conditions. In contrast, the effect of light on the incorporation of ¹⁴C-oleoyl-CoA into TG was not appreciable. These findings suggest that the accumulation of lipid in the seeds of leguminous plants is depressed by light, with far-red light affecting most the accumulation of lipids, which may regulate the synthesis of TG via DG from glycerol-3-phosphate.     

Comparison of humoral and cellular immune responses to inactivated swine influenza virus vaccine in weaned pigs

Humoral and cellular immune responses to inactivated swine influenza virus (SIV) vaccine were evaluated and compared. Fifty 3-week-old weaned pigs were randomly divided into the non-vaccinated control group and vaccinated group containing 25 pigs each. Pigs were vaccinated intramuscularly twice with adjuvanted UV-inactivated A/SW/MN/02011/08 (MN/08) H1N2 SIV vaccine at 6 and 9 weeks of age. Whole blood samples for multi-parameter flow cytometry (MP-FCM) and serum samples for hemagglutination inhibition (HI) assay were collected at 23 and 28 days after the second vaccination, respectively. A standard HI assay and MP-FCM were performed against UV-inactivated homologous MN/08 and heterologous pandemic A/CA/04/2009 (CA/09) H1N1 viruses. While the HI assay detected humoral responses only to the MN/08 virus, the MP-FCM detected strong cellular responses against the MN/08 virus and significant heterologous responses to the CA/09 virus, especially in the CD4+CD8+ T cell subset. The cellular heterologous responses to UV-inactivated virus by MP-FCM suggested that the assay was sensitive and potentially detected a wider range of antigens than what was detected by the HI assay. Overall, the adjuvanted UV-inactivated A/SW/MN/02011/08 H1N2 SIV vaccine stimulated both humoral and cellular immune responses including the CD4-CD8+ T cell subset.     

Autophagy defends cells against invading group A Streptococcus

We found that the autophagic machinery could effectively eliminate pathogenic group A Streptococcus (GAS) within nonphagocytic cells. After escaping from endosomes into the cytoplasm, GAS became enveloped by autophagosome-like compartments and were killed upon fusion of these compartments with lysosomes. In autophagy-deficient Atg5-/- cells, GAS survived, multiplied, and were released from the cells. Thus, the autophagic machinery can act as an innate defense system against invading pathogens.     

Food habits of great cormorant Phalacrocorax carbo hanedae at Lake Biwa, Japan, with special reference to ayu Plecoglossus altivelis altivelis

ABSTRACT:   The recent increase in numbers of individuals of a piscivorous bird, the great cormorant Phalacrocorax carbo hanedae , around Lake Biwa has affected the commercial catch of ayu Plecoglossus altivelis altivelis . The food habits of the bird were studied based on 66 stomachs in 1998 and 2001–2002, and a seasonal variation in food contents was observed. The proportion of ayu in stomachs increased after April, and became most dominant during July and August (0.62 in proportion of weight, 0.85 in frequency of occurrence). In contrast, ayu was not found in the stomachs after late October. The increase of ayu in individuals and density in the surface layer of the pelagic zone of the lake during spring was considered as a trigger for the change of the food habits of the birds. The feeding impact of great cormorant on ayu in Lake Biwa was estimated to be approximately 46 t during April to June, and 310 t in July and August. During the main period for the commercial ayu catch (November to the following June), the estimated mass consumed by the bird is much smaller in comparison with the commercial catch.     

Identification of metabolites in plasma and urine of Uruguayan propolis-treated rats

Propolis is a resinous substance collected by honeybees from various plant sources. It is extensively used in food and beverages to improve health and prevent diseases such as heart disease, diabetes, and cancer. To investigate the absorption and metabolism of the components in propolis, in the present study, we administered ethanol extracts of Uruguayan propolis (poplar type propolis) orally to rats and analyzed their plasma and urine by high-performance liquid chromatography with photodiode array and mass spectrometric detection. After deconjugation of the components by beta-glucuronidase/sulfatase treatment of the specimen, pinobanksin 5-methyl ether, pinobanksin, kaempferol, chrysin, pinocembrin, and galangin were detected in plasma of rats orally administered propolis. These compounds were detected also in urine after beta-glucuronidase/sulfatase treatment. Furthermore, pinobanksin 5-methyl ether, pinobanksin, chrysin, pinocembrin, and galangin were present in the urine also in free form. These results suggest that flavonoids in propolis are metabolized and circulate in the body after oral administration of propolis.     

Emulsifier content and side effects of oil-based adjuvant vaccine in swine

Side effects caused by the excessive emulsifier in oil-based adjuvant vaccine were examined practically in swine using one oil-in-water type adjuvant vaccine against swine pleuropneumonia. The vaccine was prepared from cell-free-antigen of Actinobacillus pleuropneumoniae, liquid paraffin, and several polyoxyethylenesorbitan and sorbitan oleates. Based on findings about safety in mice and emulsion stability, 2 vaccines containing either 11.25% or 6.25% emulsifier content were injected intramuscularly twice in swine, as the highest and lowest limits, respectively, within the practical range. All pigs showed temporary fever and malaise with anorexia for several days after each injection. The fever of the higher emulsifier content group took significantly longer to recover than the lower. Malaise also showed a similar tendency. On the other hand, antibody response was sufficiently induced with no significant difference between the 2 groups. Lowering the emulsifier content is a very simple but effective solution for mitigation of side effects without the reduction of adjuvanticity. For safe and high-quality oil-based adjuvant vaccines, not only antigen and base-oil, but emulsifier content must be optimized.     

Inhibitory effects of CIDR-based ovulation-synchronization protocols on uterine PGF2alpha secretion at the following luteal phase in early postpartum non-cycling beef cows

We investigated whether CIDR-based ovulation-synchronization protocols inhibit secretion of prostaglandin (PG) F2alpha from the uterus in the following luteal phase in non-cycling beef cows. Ten early (a month) postpartum non-cycling Japanese Black beef cows were treated with (1) Ovsynch (GnRH analogue on Day 0, PGF2alpha analogue on Day 7, and GnRH analogue on Day 9; n=3), (2) Ovsynch+CIDR (Ovsynch protocol plus a CIDR for 7 days from Day 0; n=4), or (3) estradiol benzoate (EB) Ovsynch+CIDR (EB on Day 0 in lieu of the first GnRH treatment followed by the Ovsynch+CIDR protocol; n=3). An oxytocin challenge was administered on Day 24 to examine uterine PGF2alpha secretion. Plasma concentrations of 13,14-dihydro-15-keto- PGF2alpha were lower at 30-120 min after oxytocin administration in the Ovsynch+CIDR group and 75 min after administration in the EB Ovsynch+CIDR group than in the Ovsynch group (P<0.05). Plasma progesterone concentrations were higher from Days 1 to 7 in the Ovsynch+CIDR group and from Days 1 to 5 in the EB Ovsynch+CIDR group than in the Ovsynch group (P<0.05). The progesterone concentrations were higher on Days 27 and 29 in both CIDR-treated groups than in the Ovsynch group (P<0.05). In conclusion, in non-cycling beef cows, CIDR-based ovulation-synchronization protocols inhibit uterine PGF2alpha secretion in the following luteal phase and prevent premature luteolysis as is seen with the Ovsynch protocol.