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.     

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.     

The metabolism of R-20458 [(E)-6,7-epoxy-1-(4-ethylphenoxy)-3,7-dimethyl-2-octene] in rat hepatocyte suspensions

The metabolism of R-20458 [(E)-6,7-epoxy-1-(4-ethylphenoxy)-3,7-dimethyl-2-octene] by rat hepatocytes has been analyzed and compared with that of juvenile hormone I [methyl-(E,E)-cis-10,11-epoxy-7-ethyl-3,11-dimethyl-2,6-tridecadienoate] under identical conditions. The metabolism of R-20458 is characterized by the predominance of NADPH-dependent cytochrome P-450 and epoxide hydrolase reactions; whereas, JH I is metabolized mainly by carboxylesterase, epoxide hydrolase, and glutathione S-transferases. The metabolites of R-20458 have been shown to correspond to (E)-6,7-epoxy-1-(4-hydroxyethylphenoxy)-3,7-dimethyl-2-octene; (E)-6,7-epoxy-1-(4-acetylphenoxy)-3,7-dimethyl-2-octene; (E)-6,7-dihydroxy-1-(4-ethylphenoxy)-3,7-dimethyl-2-octene; and, (E)-6,7-dihydroxy-1-(4-acetylphenoxy)-3,7-dimethyl-2-octene. The production of the α-hydroxyethyl, p-acetylphenoxy, and acetylphenoxy-6,7-diol metabolites is markedly inhibited by SKF 525-A. No dramatic effects are produced by diethylmaleate and 1,2-epoxy-3,3,3-trichloropropane.     

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.     

Toxigenic Fusarium species and Mycotoxins Associated with Head Blight in Small-Grain Cereals in Europe

The Fusarium species predominantly found associated with Fusarium head blight (FHB) in wheat and other small-grain cereals all over Europe are F. graminearum, F. avenaceum and F. culmorum. Among the less frequently encountered species are several others which are less pathogenic or opportunistic, but also toxigenic. These include F. poae, F. cerealis F. equiseti F. sporotrichioides F. tricinctum and, to a lesser extent, F. acuminatum F. subglutinans F. solani F. oxysporum F. verticillioides F. semitectum and F. proliferatum. The species profile of FHB is due to several factors, primarily climatic conditions, particularly rain and the temperature at flowering stage, but also agronomic factors, such as soil cultivation, nitrogen fertilization, fungicides, crop rotation, and host genotype. The most frequently encountered Fusarium mycotoxins in FHB in Europe has proved to be deoxynivalenol and zearalenone produced by F. graminearum and F. culmorum with the former more common in southern (warmer) and the latter in northern (colder) European areas. Nivalenol was usually found associated with deoxynivalenol and its derivatives (mono-acetyldeoxynivalenols), together with fusarenone-X, formed by F. graminearum F. cerealis F. culmorum and, in northern areas, by F. poae. Moreover, from central to northern European countries, moniliformin has been consistently reported, as a consequence of the widespread distribution of F. avenaceum whereas the occurrence of T-2 toxin derivatives, such as T-2 toxin and HT-2 toxin, and diacetoxyscirpenol have been recorded in conjunction with sporadic epidemics of F. sporotrichioides and F. poae. Finally, beauvericin and various enniatins have recently been found in Finnish wheat colonized by F.avenaceum and F. poae.