Hiroko Kawano

Studies on Ergothioneine. IX. Ergothioneine Stimulates Mouse Spermatozoa and Fertilization in vitro

The effects of ergothioneine on spermatozoa and ova were investigated in vitro and in vivo. Spermatozoa were treated with ergothioneine in vitro, and injected into the uterine cavity of female mice immediately after the induction of superovulation. The ova were recovered 24 hr later and assessed for fertilization. Preincubation of spermatozoa with ergothioneine resulted in a significant increase in the fertilization rate. When ova were inseminated in the same manner in vitro with spermatozoa treated with 0.1 or 1.0 mM of ergothioneine, the penetration rate was significantly increased. These results suggest that ergothioneine is effective in inducing both capacitation and the acrosome reaction of mouse spermatozoa. Ergothioneine at concentrations of 0.1 and 1.0 mM in the preincubation medium was also effective in inducing the acrosome reaction of guinea pig spermatozoa. However, it had no significant effect on the development of 2‐cell ova in vitro.

Involvement of inflammatory leukocytes in hepatic induction of T-kininogen in rat.

Hepatic synthesis as well as plasma levels of T-kininogen, a protein precursor of T-kinin (Ile-Ser-bradykinin), increase in rats following the induction of acute inflammation. Studies have been undertaken to elucidate an involvement of inflammatory leukocytes in the acute-phase response of T-kininogen. Peritoneal exudate cells (PEC) were prepared from rats six days after the intraperitoneal injection of Freunds complete adjuvant. By transfer of these leukocytes into the peritoneal cavity of normal rats, plasma kininogen levels of these recipients increased about threefold after one day. Secretion of kininogen from rat hepatocytes in primary culture was enhanced about twofold by coculturing with PEC. A similar effect was also obtained by adding culture supernatant of these leukocytes into hepatocytes, and the increased levels of kininogen in culture medium of hepatocytes was due to the increased levels of T-kininogen. From these results, it was concluded that leukocytes in the inflammatory site, probably macrophages, release some substance(s) which stimulate(s) the hepatic synthesis of T-kininogen.

Study on 14C-BHA absorption and distribution in rat

BHA was administered to rats at doses of 5 or 500 mg/kg for seven days. 14C-BHA absorption was investigated on the eighth day and compared with animals receiving a single dose of 5 or 500 mg/kg BHA. Absorption of 14C-BHA was proportionately greater at the high dose rats. 14C distribution was 3–6 times higher in the forestomach than in the pyloric region of the stomach. The urinary excretion of BHA conjugates was not dependent on pretreatment with BHA or on the dose.