We evaluated the effects of elevated carbon dioxide concentration ([CO2]) and two nutrient regimes on stem growth rate, annual ring structure and temporal variations in photosynthetic characteristics of seedlings of Japanese larch (Larix kaempferi (Lamb.) Carr.). Seedlings were grown in phytotron chambers in an ambient (360 ppm) or an elevated (720 ppm) [CO2] in two nutrient regimes for one growing season. Elevated [CO2] reduced stem height and increased stem basal diameter compared with ambient [CO2]. The effect of elevated [CO2] on growth tended to be greater at high-nutrient supply than at low-nutrient supply. Elevated [CO2] had no significant effect on ring width or the number of tracheids per radial file. There was no obvious difference in cell wall thickness or the relative area of the cell wall between seedlings grown in ambient or elevated [CO2]. Although growth in elevated [CO2] resulted in a slight increase in cell diameter, the increase had a relatively minor effect on the relative area of the cell wall. Net assimilation rate increased in response to elevated [CO2]; however, the increase in whole-crown photosynthetic rate (Total Agrowth) in seedlings in the elevated [CO2] treatment was minimal because of the smaller specific needle area and acclimation of the photosynthetic characteristics of the needles to the growth [CO2]. In conclusion, we observed no obvious enhancement in the capacity for carbon fixation in Japanese larch seedlings grown in the presence of elevated [CO2] that might be attributable to changes in stem growth. However, elevated [CO2] caused changes in the temporal pattern of stem growth and in some anatomical features of the tracheids.

ABSTRACT. Intrathecal (IT) immunization involves injecting antigens directly into the intraventricular or subarachnoid spaces, or brain, to induce antigen-specific antibodies (Ab) in the cerebrospinal fluid (CSF). In the present study, rabbits were immunized IT with inactivated rabies virus to investigate the origins of CSF Ab. The time course of Ab induction and tumor necrosis factor-alpha expression suggested the possibility that the CSF Ab originated in the serum. In addition, Ab-producing cells infiltrated around the blood vessels of the brain, suggesting local production of Ab within the central nervous system (CNS). Furthermore, subcutaneous (SC) immunization prior to IT immunization induced a rapid and magnified Ab response in the CSF compared with IT immunization alone. These results were confirmed by the fact that mice immunized SC prior to IT were more resistant to intracerebral challenge with rabies virus than mice immunized via the IT route alone. Taken together, these results suggest that combined SC and IT immunization is a more effective vaccination protocol for prophylaxis and treatment of rabies.

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To investigate the efficacy of intracerebral (IC) immunization in preventing viral spread in the brain, we immunized mice with inactivated rabies virus via the subcutaneous (SC) or IC route, followed by administration of a lethal dose of rabies virus (challenge virus standard strain), directly into the brains of immunized mice. Progressive paralytic neurological signs were observed in control and 75% of SC immunized mice, whereas only 20% of IC immunized mice exhibited symptoms. Neutralizing antibody titers in blood plasma were significantly elevated in SC and IC immunized mice, with the highest levels seen in IC immunized mice. Analysis of whole brain lysates revealed a strong induction of immunoglobulin in the brains of IC immunized mice that had virus neutralizing activity. Histopathological examination of brain tissue revealed mild encephalitis and disseminated viral antigen in control and SC immunized mice, but rare in IC immunized mice. These results suggest that IC immunization induces a preventive humoral immune response against intracerebrally inoculated rabies virus. Induction of neutralizing antibody in cerebrospinal fluid represents a putative therapeutic measure for the treatment of rabid animals and humans.

We describe here a method using HPLC for the simultaneous determination of albiflorin, paeoniflorin, glycyrrhizin and six flavanone glycosides (liquiritin, liquiritin apioside, naringin, neohesperidin, hesperidin and narirutin) in the Kampo medicines, Shigyaku-san and Haino-san. All nine components were separated in less than 40 min by linear gradient elution using a mobile phase containing aqueous phosphoric acid and acetonitrile. The dissolution of these components from powders of Shigyaku-san in aqueous solution at pH 1.80, 4.08 and 6.89 was examined. All of the components except glycyrrhizin were dissolved entirely within 5 min regardless of pH. Dissolution of glycyrrhizin was dependent on the pH of the aqueous solution, and increased with increasing pH.