Ryosuke Miyauchi

High-resolution scanning electron microscopic evaluation of cell-membrane porosity by ultrasound

The effect of low-intensity ultrasound on HL-60 cells (human promyelocytic leukemia cells) in the presence of the photo sensitizing drug merocyanine 540 (MC 540) was evaluated morphologically, using a high-resolution scanning electron microscope and a transmission electron microscope. Exposure of HL-60 cells to ultrasound without MC 540 resulted in a decrease of finger-like processes in the cells. The cells showed many undulating ruffles on the surface. Distinct pits or holes in the membrane were not observed in these cells. The surface of HL-60 cells treated only with MC 540 was relatively smooth compared with that of control cells. HL-60 cells exposed to ultrasound in the presence of MC 540 showed apparent surface deformation. Numerous crater-like depressions of heterogeneous dimensions were observed in many cells. In addition, various-sized pores were noted in the cell membranes of more damaged cells. These results indicate that cell degeneration was induced by a rapid change in cell membrane porosity during sonication in the presence of MC540.

Staining of pancreatic centroacinar cells, liver bile canaliculi and testicular Leydig cells with a monoclonal antibody against adrenocortical cells

The immunoreactivity of a monoclonal antibody against cell suspensions from guinea pig adrenal glands was examined at light- and electron-microscopic levels. In addition to the cell surface membrane of adrenocortical cells, the antibody labeled specific sites in the pancreas, liver and testis, but did not label any of the other tissues examined. In the pancreas, microvilli-like processes and the cell surface membrane of centroacinar cells were immunoreactive to the antibody. The microvilli of interlobular duct cells and pancreatic duct cells were also immunoreactive. In the liver, bile canalicular microvilli of hepatocytes were exclusively labeled. Membrane structures of cell organelles, mainly mitochondria, in testicular Leydig cells were also labeled. Immunoblot analysis showed that the monoclonal antibody bound to two common bands at molecular weights of approximately 62 kDa and 110 kDa in the pancreas, liver, testis, and adrenal gland. The two bands reacted with the digoxigenin-conjugated lectin, Sambucus nigra agglutinin (SNA), which recognizes sialic acid linked α(2–6) to galactose. Reaction patterns of SNA in the pancreas, liver and testis were similar to those of the monoclonal antibody; pancreatic centroacinar cells and interlobular duct cells, hepatocyte bile canaliculi and testicular Leydig cells were densely stained with SNA. Thus, the monoclonal antibody recognizes two common membrane glycoproteins containing sialic acids in the pancreas, liver, testis and adrenal cortex.

A study of the myofibrous organization of the biceps brachii muscles from the crab-eating macaque

TheM. biceps brachii ofMacaca fascicularis was examined, noting the total number of muscle fibers, the number of muscle fibers per square millimeter, the cross sectional area of Venter musculi and the thickness of individual fibers in the cross sectional area of the right brachial biceps specimens of 10 adult crab-eating macaques, five males and five females.The results of this investigation are compared with a previous study of a similar type made on the brachial biceps of the adult human. Comparative analysis shows that the mean ventral cross sectional area of the macaque biceps muscle is 1/7 to 1/3 that of the cross sectional area in the human muscle. Macaque brachial biceps muscle shows approximately one half the total number of muscle fibers of the human specimens, though the number of fibers per square millimeter is one to two times greater than in human specimens. The macaque muscle fibers were 1/2 to 5/6 as thick as those in the human specimens.The relations between the ventral cross sectional area and thickness of individual muscle fibers is discussed. Comparisons are made between the macaque and human specimens. It is suggested that such factors as age, sex, and the nutritional history of the specimen donors may have influenced the myofibrous organization in both human and macaque specimens. It is suggested also, that differences in myofibrous organization may be related to more continuous or sustained muscular activity in the macaque and more forceful muscular contraction in humans.