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Biomedical subjects

T Nagae

Publications and source records attributed to T Nagae.

69 records · Page 4Linked to original sources

Acrosome reaction in human spermatozoa.

Human spermatozoa were allowed to undergo the acrosome reaction in vitro by incubation in media with or without reagents known to accelerate the onset of the acrosome reaction. The first observable change before the acrosome reaction was a partial decondensation of the acrosomal matrix. This was followed by invaginations of the outer acrosomal membrane alone or of both the plasma and outer acrosomal membranes, which resulted in formation of many vesicles within the acrosomal cap. Subsequently, the plasma and outer acrosomal membranes fused, but the fusion was seldom seen in the acrosomal cap region. On the other hand, fusion of the two membranes was observed consistently at the anterior end of the equatorial segment of the acrosome. Soon after the membranes over the acrosomal cap disappeared, many vesicles, which were originally within the cap, were seen on or in the vicinity of the inner acrosomal membrane. These vesicles dispersed eventually, leaving the inner acrosomal membrane completely exposed. Thus the acrosome reaction in human spermatozoa seems to proceed in a way somewhat different from that in spermatozoa of most other species, although the end result of the reaction is the same.

Acrosome↗

[The effects of calmodulin and it's antagonist, W-7, on the acrosome reaction and fertilization of human spermatozoa].

The acrosome reaction of mammalian spermatozoa has been shown to be dependent upon an influx of Ca2+ following capacitation. Recently it was shown that calmodulin which activates various enzymatic activities in a calcium-dependent manner is contained in the acrosomal portion of sperm obtained from diverse species including human. Therefore calmodulin appears to be the primary target for Ca2+-dependent regulatory process involved in the acrosome reaction. This report examines the effects of calmodulin and it's antagonist, W-7, on the acrosome reaction in human spermatozoa and the fertilization with zona free hamster eggs. The results are as follows: 1) In the experiment on fertilization in mBWW medium with 30nM of calmodulin, there was no significant difference between the calmodulin and the control. (2) In fertilization in mBWW medium with 2.5 to 25 microM of W-7, there were no significant changes in the fertilization rate, but there was a significant decrease in the fertilization rate when the concentration of W-7 was elevated up to 50 microM. (3) When fertilization in mBWW was performed using spermatozoa pretreated with W-7, the fertilization rates were more markedly and promptly elevated with insemination times than the control. (4) When the eggs and spermatozoa were pretreated with W-7 before insemination and then placed in mBWW medium, the fertilization rate was markedly decreased. (5) Triple stain method and transmission electron microscopy showed that a large proportion of spermatozoa had undergone the acrosome reaction in mBWW medium containing W-7 with normal manner. From the results given above, it appears that calmodulin plays an important role in the acrosome reaction and fertilization in human spermatozoa.

Acrosome↗

Relation between cardiotoxic effect of adriamycin and superoxide anion radical.

The investigation was undertaken to study a possible mechanism for adriamycin cardiotoxicity. The activities of superoxide dismutase and catalase in the heart of mice were increased significantly by the intraperitoneal administration of 15 mg/kg of adriamycin. In contrast, these enzymes in the liver and kidney were unaffected by this dose of adriamycin. In vitro studies revealed that adriamycin inhibited the NADH-cytochrome c oxidoreductase activity of mitochondria in the guinea pigs heart. Moreover adriamycin stimulated the formation of superoxide anion radical in mitochondria isolated from guinea pigs. Particularly, the formation of superoxide anion radical in the heart mitochondria was 5 times higher than that in the liver mitochondria particle. On the other hand, the contents of superoxide dismutase in the heart were significantly lower than that in the liver. These results suggest that the cardiotoxic effect of adriamycin is caused by the following mechanism: adriamycin directly stimulates the formation of superoxide anion radical, particularly in the heart mitochondria. In spite of the induction of defence enzymes such as superoxide dismutase and catalase, their abilities seem to be swamped by enhanced active oxygen radicals.

Animals↗

Superoxide dismutase levels following liver and kidney intoxication.

The behavior of superoxide dismutase (SOD) in inflammatory lesions is interesting, because SOD is a scavenger of superoxide anion radical which is produced to excess in inflammation. In order to clarify the relationship between diseases and SOD levels in serum, we induced renal damage in the rabbit or liver damage in the rabbit and guinea pig, and then monitored the SOD levels and other clinical values. We observed the increase of serum cuprozinc-superoxide dismutase (Cu, Zn-SOD) concentration following kidney intoxication caused by the ligation of renal artery, and the increase of manganese-superoxide dismutase (Mn-SOD) concentration following liver intoxication caused by the injection of carbon tetrachloride. From these results, we confirmed the reports that serum Cu, Zn-SOD and Mn-SOD concentrations were markedly increased in patients with renal diseases and liver diseases, respectively.

Acute Kidney Injury↗

Determination of adriamycin by enzyme immunoassay.

A solid phase immunoassay for the determination of adriamycin has been developed. Specific antibody against adriamycin was raised in rabbits after coupling adriamycin to bovine serum albumin. beta-D-Galactosidase labelled adriamycin was prepared by m-maleimidobenzoyl-N-hydroxysuccinimide ester (MBS) as a coupling reagent. This assay system followed the double antibody solid phase method for the B/F separation of antibody-bound and free adriamycin. By this method, from 0.5 to 100 ng of adriamycin could be detected. Using this assay system, drug levels were determined in serum of mice, following administration of adriamycin in a single dose of 15 mg/kg i.p., the peak appearing at 20 min after administration. The change of serum levels was found to be biphasic, and the half-life of beta-phase was about 5 h.

Animals↗

Effects of oviducal cells on the survival and fertilizing ability of fowl spermatozoa.

Oviducal epithelial cells were cultured and the effects of the cells on the survival and fertilizing ability of fowl spermatozoa were investigated. When spermatozoa were incubated at 41 degrees C with cells from the infundibulum, shell gland or uterovaginal junction, their motility, assessed at room temperature (20-23 degrees C), was maintained for 5 to 6 days. Fertilizing ability was also retained after culture at 41 degrees C for at least 4 days. During incubation at this temperature, the spermatozoa were almost immotile and rested in close proximity to the oviducal cells. At 23 degrees C, the same spermatozoa were highly motile.

Animals↗

Enhanced neointima formation and attachment on the high-porosity inner surface of modified PTFE vascular grafts.

Highly porous PTFE arterial prostheses form endothelium more extensively than the low-porosity grafts in clinical use, but are subject to seroma formation. PTFE vascular grafts were modified to produce a highly porous inner layer (inner layer 60 or 90 microns, outer layer 20 microns). The effect of this modified, composite design on the histology of graft healing was investigated. Twenty-five modified and 25 control grafts, each 4 mm in diameter by 5 cm in length, were implanted into carotid and femoral arteries of dogs. No late seroma formation was observed. After 12 and 18 weeks, the neointima of the grafts was examined by light microscopy and scanning electron microscopy. At 2- and 4-mm distances from the proximal and distal anastomoses, intimal thickness of the control grafts was 238.7 and 96.1 microns, respectively; for the modified grafts it was 236.2 and 202.8 microns at 18 weeks. Thus, the neotinima of modified grafts was thicker than that of control grafts when measured at 4 mm from the anastomoses. Neointimal coverage was less extensive in the control grafts than in modified grafts (26.8 +/- 6.1% vs. 58.8 +/- 13.2%; p < .05). Smooth muscle cells were seen on light-microscopy to penetrate the highly porous inner layer; on scanning electron microscopy, the PTFE fibrils appeared to anchor the neointima of the modified graft. The results suggest that modified PTFE grafts with an inner surface of 60 or 90 microns internodal distance have enhanced formation and anchoring of neointima while remaining impervious to blood.

Animals↗