The development of γ-enolase immunoreactive cells in five human embryos (Carnegie stages: 13-16 and 21; crown-rump length: 6.3-23.4mm) was investigated by immunohistochemistry, using the basal plate of the neural tube, dorsal root ganglion at the cervical level, trigeminal and vestibulocochlear ganglia, and para-esophageal and para-tracheal ganglia.
In the basal plate of the stage 13 embryo, not only the motor neurons but also the anterior root was immunoreactive at the level of the cervical spinal cord. Some of these reactive neurons were in the rhombencephalon. The motor neurons in the basal plate of mesencephalon and thoracic spinal cord of the stage 14 embryo were immunoreactive, while those of the lumbar spinal cord were immunoreactive in the stage 15 embryo. In the dorsal root ganglion, trigeminal and vestibulocochlear ganglia, and para-esophageal and para-tracheal ganglia, immunoreactive cells were observed first in the stage 15 embryo. In the stage 21 embryo, immunoreactivity was observed around the peripheral respiratory tract.
As the appearance of γ-enolase in neurons is related to functional and morphological maturation, our study will provide descriptive information for the evaluation of neuronal development in early human embryos.

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Localization of amylase and lysozyme within the serous secretory granule of the human parotid and submaxillary glandular acinocytes was studied with immuno-electron microscopy, using the protein A-gold technique. Antigenic sites of amylase were observed to be localized diffusely throughout the secretory granule and those of lysozyme in its central core of the granule. Therefore, it was concluded that amylase disperses within and throughout the granule, and lysozyme tends to concentrate in the central core.

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The purpose of this paper is to describe the immunocytochemical localization of envelope G protein of vesicular stomatitis virus in the cells throughout the infectious cycle, in comparison with pure morphological observation. Vero cells, MDBK cells and Swiss 3T3 cells were examined at various times after infection. They were fixed with PLP fixative and treated with 0.05% Triton X before incubation with mouse monoclonal antibody and rabbit antimouse IgG coupled to rhodamine. The G protein was seen around nuclear membranes at 2.5hr and on Golgi areas at 3hr. At 4-6hr, it was detected not only on Golgi areas but also on cell surfaces. For electron microscopic immunostaining, Vero cells were fixed first in the mixture of glutaraldehyde and paraformaldehyde and then with PLP fixative followed by 0.05% saponin treatment. They were immunostained with the monoclonal antibody and rabbit anti-mouse IgG coupled to peroxidase. The G protein could be identified in the luminar sides of Golgi membranes and smooth vesicles. For pure morphological observation, Vero cells were fixed in glutaraldehyde and paraformaldehyde and routinely embedded in Epon. At 5-6hr, Golgi apparatus became larger than normal in uninfected cells and contained fuzzy materials in the luminar sides of Golgi membranes and smooth vesicles that corresponded to the immunostained reaction products. It is concluded that smooth vesicles are possible candidates for the transportation of G protein, instead of clathrin coated vesicles.

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In order to clarify the mechanism of development of ischemic brain edema, the blood-brain barrier in Wistar rats was observed by the electron microscopic histochemical technique. In experimental models, the bilateral carotid arteries were occluded for 2hr with or without 30min exposure to nitrogen. The brains were perfused with a fixative containing 2% paraformaldehyde and 0.25% glutaraldehyde for 15min.
In controls, Mg-ATPase activity (Ogawa and Mayahara 1969) was localized on the basal surface of capillary endothelium and the astrocytic plasma membrane, and alkaline phosphatase (ALPase) (Mayahara et al. 1967) was demonstrated on both the luminal and basal plasma membranes of capillary endothelium with some variations. The reaction products of Mg-ATPase were coarsely distributed and increased in the ischemic group without nitrogen exposure. However, it markedly decreased in the anoxic-ischemic group which showed marked swelling of astrocytic foot processes. ALPase tended to be well preserved in both experimental groups, but showed focal dispersion in the anoxic-ischemic group.
The decreased activities of Mg-ATPase as well as ALPase may change the membrane permeability, and may induce intracytoplasmic fluid accumulation which causes brain edema in the early stage of ischemia.

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Immunoperoxidase techniques were used to investigate the localization patterns and distribution of hepatitis B virus (HBV)-associated antigens in hepatitis B surface antigen (HBsAg)-positive patients with liver cirrhosis. The membranous type of HBsAg was rarely seen in liver cirrhosis unless there was concomitant evidence of chronic active hepatitis. HBsAg was stained mainly throughout the hepatic cytoplasm or its periphery in liver cirrhosis regardless of the presence of the hepatitis B e antigen (HBeAg) or hepatitis B e antibody (HBeAb) in the serum. In all cases of liver cirrhosis with hepatocellular carcinoma, the inclusion body type of HBsAg was detected in hepatocytes. Simultaneous staining of HBsAg and hepatitis B core antigen (HBcAg) in biopsy tissues of liver cirrhosis revealed that the HBcAg was mainly stained in the nuclei of hepatocytes, and that almost all of the HBcAg-positive hepatocytes contained either cytoplasmic or festoon-like immunoreaction patterns of HBsAg. HBcAg-positive hepatocytes were distributed spottedly, diffusely or in groups in pseudolobules, and clusters of HBcAg-positive hepatocytes were detected in the non-tumor areas adjacent to hepatomas, where liver cell dysplasia was also observed.

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