Staining pattern of the brush border and detection of cytoplasmic granules in the uriniferous tubules of female DBA/2Cr mouse kidney: comparison among various fixations and stains.

The proximal straight tubules of the female mouse kidney exhibit heavy periodic acid Schiff (PAS) staining in their brush borders and numerous cytoplasmic granules. In the present study, the female DBA/2Cr mouse kidney was examined, using various fixatives (formalin, PFA, PLP, Zamboni's, Bouin, or Carnoy solution) and various staining methods (HE, PAS, alcian blue, periodic acid methenamine-silver (PAM), toluidine blue, azan, or Congo red). Under azan and PAM, the staining pattern of the brush border was similar to that of PAS, and few effects of the fixative were observed. Cytoplasmic granules were clearly detected with PAM as well as PAS. However, these granules were not detected with Carnoy solution. Furthermore, distribution of granules differed between PAS and PAM.     

Decreased transport of orally administered protein into the blood circulation of developing juveniles of Japanese eel Anguilla japonica

ABSTRACT: To clarify the quantitative changes in the transport of orally intubated protein into the blood circulation as macromolecules in development, immunoglobulin Y (IgY) extracted from chicken eggs was administered orally to juvenile Japanese eel, Anguilla japonica . For the first experiment, which was performed before the commencement of artificial feeding, the oral delivery of 2.0 μg/0.1 g bodyweight of IgY resulted in a rapid increase in plasma IgY to a maximum of 2.30 μg/mL. However, the transport of IgY into the blood decreased significantly in the experiments that followed, which were performed after 12, 25 and 42 days. During this period, bodyweight increased approximately by a factor of eight, and rapid growth of the stomach was observed histologically. Possible contributions for the development of the alimentary canal to the diminishment of intestinal protein assimilation are discussed.     

Norsalsolinol uptake into secretory vesicles via vesicular monoamine transporter and its secretion by membrane depolarization or purinoceptor stimulation in PC12 cells.

The intracellular dynamics of norsalsolinol, a neurotoxin candidate causing parkinsonism-like symptoms, in PC12 cells was studied. We found that dopamine and norsalsolinol are co-localized to secretory granule layer by sucrose density gradient in norsalsolinol-treated PC12 cells. The norsalsolinol was actively taken up into isolated secretory vesicle fraction from PC12 cells with a Km value of 41.5+/-6.8 microM. The uptake of 10 microM of norsalsolinol was sensitive to reserpine (1 microM), an inhibitor of vesicular dopamine transporter, and dopamine, an endogenous substrate, but insensitive to GBR-12909, an inhibitor of dopamine transporter on plasma membrane. In norsalsolinol-treated PC12 cells, exposure to high K+ or ATP resulted in simultaneous release of norsalsolinol and dopamine. Time course of a release of dopamine and that of norsalsolinol evoked by 50 mM KCl or 100 microM ATP corresponded to each other. These releases were dependent on the concentrations of secretagogues. These data suggest that norsalsolinol is taken up with dopamine into secretory vesicle via vesicular catecholamine transporter.     

Application of acoustic Doppler current profiler combined with a scientific echo sounder for krill Euphausia pacifica density estimation

ABSTRACT:   An application of the acoustic Doppler current profiler (ADCP, 153.6 kHz) in combination with a scientific echo sounder (EK60, 38 and 200 kHz) was investigated to estimate the density of krill Euphausia pacifica . The acoustic backscattering strength from sound scattering layers was compared with biomass estimates from midwater trawls. Euphausia pacifica was targeted among mixed species populations in the sound scattering layer in the offshore Funka Bay area of Hokkaido, Japan. The frequency characteristics of acoustic backscattering by krill were calculated using a distorted wave Born approximation scattering model at three frequencies. Krill aggregations identified from the EK60 data were extracted as the mean volume backscattering strength difference between two frequencies. They were then used to identify similar aggregations in the ADCP data by matching observation times and depths for the two methods, which were applied simultaneously. Results from the comparison of the mean volume backscattering strength and the density calculated from the ADCP and EK60 showed that ADCP can be used to measure density and spatial–temporal distribution of krill aggregations. Current speed and direction at the study site were found to be 16.1 cm/s and 187.0°, respectively, and krill speed and direction (including the current component) were found to be 19.8 cm/s and 172.2°, respectively. Based on the ADCP data, the net speed and direction of the krill aggregations were found to be 5.9 cm/s and 128.0°, respectively.     

Characteristics of cytotoxic cells induced by Toxoplasma lysate antigen in mouse spleen.

Spleen cells from Toxoplasma lysate antigen (TLA)-sensitized BALB/c mice showed the strong cytotoxic activity against both natural killer (NK)-sensitive cells (YAC-1 and RL male-1) and NK-insensitive cells (P-815), when incubated with TLA or recombinant human IL-2 (rhIL-2). The increment of TLA concentration in culture medium increased the cytotoxic activity. Treatment of effector cells; spleen cells from TLA-sensitized mice incubated with TLA, with anti-asialo GM1 or anti-Thy-1 plus complement inhibited the cytotoxic activity of effector cells, whereas treatment with anti-mouse Lyt-2.2 serum plus complement had no effect on the cytotoxic activity. Treatment of spleen cells from TLA-sensitized mice with anti-asialo GM1 and/or anti-Thy-1 plus complement inhibited cytotoxic activities of effector cells. These results suggested that spleen cells sensitized with TLA both in vivo and in vitro were asialo GM1 positive and Thy-1 positive, and the majority of cytotoxic cells induced by TLA were similar to lymphokine-activated killer (LAK) cells induced by IL-2.     

Purification and properties of pyrethroid carboxyesterase in rat liver microsome

Pyrethroid carboxyesterase which hydrolyzes the esters of chrysanthemumic acid was purified from rat liver microsome by cholic acid solubilization, ammonium sulfate fractionation, heat treatment, and DEAE-Sephadex A-50 column chromatography. The 45-fold purified enzyme (38% yield) is likely to consist of single protein, as evidenced by polyacrylamide gel disc electrophoresis and Sephadex G-100 column chromatography, and had a molecular weight of approximately 74,000 and a K_m of 0.21 mM. It is susceptible to inhibition by organophosphates and carbamate insecticides and insensitive to pCMB, mercuric ion, and cupric ion. It is capable of hydrolyzing trans isomers of synthetic pyrethroids much more rapidly (five to ten times) than the cis counterparts. The purified pyrethroid carboxyesterase is apparently identical in nature with malathion carboxyesterase and with p-nitrophenyl acetate carboxyesterase.     

Competitive enzyme-linked immunosorbent assay for the detection of the antibodies specific to akabane virus.

A competitive enzyme-linked immunosorbent assay (C-ELISA) using neutralizing monoclonal antibodies (MAbs) against Akabane virus (AKAV) was developed to detect antibodies to AKAV in cattle sera. The performance of the test using 7 different competitor MAbs was evaluated in sequential serum samples and sera from cattle infected with various bovine arboviruses. The dynamics of the antibody response expressed by percentage of inhibition (PI) in C-ELISA coincided with those of neutralizing antibody titers in sequential serum samples from 2 cattle experimentally infected with AKAV. The value of PI in C-ELISA for convalescent sera from cattle infected with arboviruses correlated with the neutralizing antibody titer to AKAV but was unaffected by the antibodies to other arboviruses. In the validation experiment of C-ELISA using 286 bovine sera previously examined for the AKAV antibody by serum neutralization (SN) test, the relative specificity of C-ELISA was more than 98%, whereas the relative sensitivities of individual MAbs ranged from 49% to 82.2%. Overall agreement between C-ELISA and the SN test varied from 72% to 90% depending on the MAb. These results suggest that the C-ELISA is acceptable as a rapid and specific method for detecting antibodies to AKAV and is a potential alternative to the SN test.