Method for analysis of tannic acid and its metabolites in biological samples: application to tannic acid metabolism in the rat

A new analytical method for measuring tannic acid (TA) using tannase was developed and applied to the investigation of TA metabolism in the rat following oral administration at a dose of 1.0 g/kg. The proposed method for TA determination was based on the enzymatic hydrolysis of TA to gallic acid (GA) and subsequent determination by HPLC. TA metabolites were determined by HPLC. 4-O-Methylgallic acid (4-OMGA), pyrogallol (PY), and resorcinol (RE) were detected in serum. TA was excreted into urine as GA (0.01%), 4-OMGA (0.10%), PY (0.24%), and RE (2.06%) and into feces as TA (62.74%), GA (0.19%), PY (0.02%), and RE (0.76%) within 54 h after oral administration. It was suggested that >60% of TA remained unchanged but that some was hydrolyzed to GA by tannase in the intestine and further metabolized to 4-OMGA, PY, and RE.     

The effects of long-term conservation tillage, crop residues and P fertilizer on soil conditions and responses of summer and winter crops on an Andosol in Japan

A field experiment was conducted from 1983 to 1992 in Tsukuba, Japan to investigate the effects of tillage on soil conditions and crop growth in a light-colored Andosol. Three tillage methods (NT: no-tillage, RT: no-tillage for summer cropping and moldboard plowing for winter cropping, and CT: conventional rotary tillage to a depth of 15 cm) were employed in combination with crop residue application (+R, −R) and fused magnesium phosphate (FMP) fertilization (+P, −P). Under the combination of NT and +R, diurnal variation of soil temperature at a depth of 5 cm was smaller during the summer cropping season and soil temperature in the daytime was lower during the winter cropping season than under CT. Soil inorganic N concentration at a depth of 0–30 cm was +R > −R and NT > RT > CT. The early growth of summer crops was accelerated under NT in comparison with CT, and yields were higher under NT and RT in comparison with CT. On the other hand, winter crop yields were significantly reduced under NT, while they were still higher under RT in comparison with CT. Yields were higher with +R and +P application, respectively, and these effects were more pronounced in winter cropping. The positive effect of FMP fertilization was greater in combination with NT, and that of residue treatment was greater in combination with RT and NT than with CT. In conclusion, the best tillage practice for Andosols on the Kanto Plain is RT, i.e. a combination of NT for summer cropping and CT for winter cropping. The application of NT for winter cropping is not recommended, although the application of phosphate and crop residues could reduce the risk of yield reduction, because of improved soil nutrient status and moderation of diurnal soil temperature.     

Effects of testosterone on rat placental development

We investigated the morphological effects of testosterone on placental development in a rat model of polycystic ovarian syndrome (PCOS). Testosterone propionate (TP), which was subcutaneously administered to pregnant rats with 5 mg/animal from gestation day (GD) 14 to GD 18, induced a maternal weight reduction without mortality or clinical signs from GD 19 onwards. A decrease in fetal and placental weight, an increase in intrauterine growth retardation (IUGR) rates, and histological changes in the placenta were observed on GD 21 but not on GD15 or 17. Histopathologically, on GD 21, the trophoblast septa thickened, and the maternal sinusoids were narrowed in the labyrinth zone, resulting in a small placenta. Additionally, the placental weight, thickness, and histological morphology in the labyrinth zone on GD 21 in the TP-treated group were nearly identical to those on GD 17 in the control and TP-treated groups. Therefore, it was assumed that the testosterone-induced small placenta was induced in association with the developmental inhibition of the fetal part of the placentas from GD 17 onwards.     

DNA polymorphism of srRNA gene among Eimeria tenella strains isolated in Japan

DNA polymorphism in twelve starains of Eimeria tenella isolated from various places in Japan was examined using 1.l kb small subunits ribosomal RNA amplified by PCR. Genetic variation was evaluated by random amplification of polymorphic DNA (RAPD) analysis. DNA fingerprint patterns were grouped into two, indicating that at least two DNA polymorphisms exist in Japanese E. tenella strains.     

Erythrocyte-replaced mouse model for Haemoparasite studies: comparison of NOD/shi-scid and C.B-17/Jcl-scid mouse upon acceptability of human erythrocytes

The erythrocyte-exchanged chimera mouse model has become to be a significant tool for studying animal and human (hu) protozoan haemoparasites, though the usefulness of this model varies depending primarily on the acceptability of xenogeneic red blood cells (RBC). To find a superior recipient in comparison with C.B-17/Jcl mouse with severe combined immuno-deficiency (scid) mutation, we examined in this report the non-obese diabetes (NOD)/shi-scid mouse, a recently available strain of SCID. When 2.5 x 10(8) of fluorescent dye-labeled hu-RBCs were transfused, C.B-17scid mouse eliminated them logarithmically by a simple linear regression, while NOD-scid mouse eradicated hu-RBCs by a unique two-step fashion, i.e., a potent but only briefly functioning RBC eradication followed by a weak steadily functioning step. The means of regression line constance +/- their standard deviations (SD) of 205 C.B-17scid and of 213 NOD-scid mice for their short- and long-lasting steps were -0.73 +/- 0.63, -0.53 +/- 0.25 and -0.16 +/- 0.10, respectively. Hu-RBC half-lives determined from these means of C.B-17scid mice and of NOD-scid mice for the short- and long-living steps were 3.6, 4.9 and 16.3 hr, respectively. Higher hu-RBC acceptability of NOD-scid mouse, especially at their long-lasting step, was also demonstrated under at an activated state of mouse innate immunity. Treatment with 1.0 mg heat-killed Candida cells caused an acceleration of hu-RBC elimination in both mouse strains but the magnitudes for the short- and long-living steps of NOD-scid mice evaluated by "stimulation index" were only 1/2.6 and 1/7.6 of C.B-17scid mice, respectively.     

Comparison of macrophage scavenger receptor-A knockout mice with wild type ones in the immune response against repeated infestation with Haemaphysalis longicornis

Using macrophage scavenger receptor-A knockout (SRKO) mice, we examined the role of macrophage class A scavenger receptors (MRS-A) on the immune response and acquisition of host resistance against repeated infestation with Haemaphysalis longicornis. Except for one batch of nymphs that infested one of the SRKO (SR-/-) mice and showed no appreciable reduction in body weight, all the other groups of nymphs manifested significant decrease in body weight. Both SR-/- and wild type (SR+/+) mice showed a sustained increase in anti-tick antibody titers, but SR+/+ mice showed significantly higher titers. The IFN-gamma assayed in SR-/- mouse immune sera was substantially less compared with that in SR+/+ mice. Immune sera from SR-/- and SR+/+ mice recognized the 51 and 44 kDa, and 44 kDa proteins, respectively, of the salivary gland antigen. The difference in the level of anti-tick resistance manifested by both groups of mice may be influenced by less efficient trapping and processing of tick antigens by macrophages in mice lacking for the macrophage scavenger receptors, and consequently affected the cascade of Th1 and Th2 responses. We have thus obtained valuable data that strongly infer the role of MSR-A in enhancing host defense against repeated infestation with H. longicornis.     

Biotin-labeled proteins of Eimeria tenella sporozoite

To identify surface proteins of eimerian sporozoite, Eimeria tenella sporozoites were labeled with biotin-LC-hydrazide, and analyzed by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and Western blotting. More than 200 sporozoite proteins were recognized with silver staining after 2D-PAGE. Among them, 7 biotin-labeled proteins were detected as surface molecules after Western blotting and reaction with horseradish peroxidase conjugated streptavidin. Their molecular masses ranged from 23 to 117 kilo daltons and their isoelectric points from 4.3 to 6.3. It appears that the biotin-labeling technique can be used to analyze of surface proteins of eimerian parasites.     

Effects of in vivo administration of anti-IL-10 or anti-IFN-gamma monoclonal antibody on the host defense mechanism against Plasmodium yoelii yoelii infection

Our previous reports indicated that C57BL/6 mice infected with a lethal variant of Plasmodium yoelii 17X (P. yoelii 17XL) produced high levels of interleukin 10 (IL-10) and interferon-gamma (IFN-gamma) while mice infected with the nonlethal variant of the parasite did not produce detectable levels of IL-10. In the present study, the involvement of IL-10 and IFN-gamma in exacerbation or regulation of blood-stage malaria was investigated by using the lethal variant of P. yoelii 17XL and monoclonal antibodies (mAb) against the cytokines. C57BL/6 mice were injected intraperitoneally with a neutralizing anti-IL-10 mAb or anti-IFN-gamma mAb after inoculation with P. yoelii 17XL. Treatment of mice with anti-IL-10 mAb resulted in substantial prolongation of survival and 60% of treated mice survived while 100% of control mice died by day 11. On the contrary, treatment of mice with anti-IFN-gamma mAb exacerbated infection and all mice died after an earlier period than those treated with normal rat Ig. No differences in parasitemias were found between treated and untreated mice. To elucidate the involvement of nitric oxide in the host protection or exacerbation, mice were treated with aminoguanidine, an inhibitor of nitric oxide synthetase, after inoculation of P. yoelii 17XL. Neither mortality nor parasitemia was influenced by the treatment. These results indicate that an IFN-gamma response is associated with protective immunity in mice infected with P. yoelii 17XL, while an IL-10 response is associated with disease exacerbation during the infection.     

Appearance of race Pfs:5 of spinach downy mildew fungus, Peronospora farinosa f. sp. spinaciae, in Japan

The races for the causal agent of spinach downy mildew Peronospora farinosa f. sp. spinaciae were identified by inoculation of race-differential cultivars. One isolate was identified as Pfs:5s and the others belonged to a new race. This is the first report of race Pfs:5 and another new race in Japan.