The effects of 2,3-butanedione 2-monoxime (BDM) on mechanical responses of glycerinated fibers and the ATPase activity of heavy meromyosin (HMM) and myofibrils have been studied using rabbit skeletal muscle. The mechanical responses and the ATPase activity were measured in similar conditions (ionic strength 0.06-0.2 M, 0.4-4 mM MgATP, 0-20 mM BDM, 2-20 degrees C and pH 7.0). BDM reversibly reduced the isometric tension, shortening speed, and instantaneous stiffness of the fibers. BDM also inhibited myofibrillar and HMM ATPase activities. The inhibitory effect on the relative ATPase activity of HMM was not influenced by the addition of actin or troponin-tropomyosin-actin. High temperature and low ionic strength weakened BDM's suppression of contraction of the fibers and the ATPase activity of contracting myofibrils, but not of the HMM, acto-HMM and relaxed myofibrillar ATPase activity. The size of the initial phosphate burst at 20 degrees C was independent of the concentration of BDM. These results suggest that the suppression of contraction of muscle fibers is due mainly to direct action of BDM on the myosin molecules.

The cytochrome P-450(17 alpha)-hydroxylase, 17----20 lyase (P-450(17 alpha)) is the key enzyme responsible for the biosynthesis of androgens in steroidogenic organs. Its cellular localization has been examined with an immunohistochemical technique. In immature rat ovary, P-450(17 alpha) was first detected in sparse interstitial cells on postnatal Day 8. The number of immunoreactive interstitial cells increased thereafter and the intensity of P-450(17 alpha) staining in these cells was highest at 3 weeks of age. The intensity of staining then started to decline and was very faint at Day 35. From 6 weeks on, the distribution of immunoreactive P-450(17 alpha) was of the adult type: it was detected exclusively in the thecal cells of the large antral, preovulatory, follicles. P-450(17 alpha) was not detectable during pregnancy except on the day of parturition, when thecal cells were transiently immunoreactive. The staining had vanished 24 h after delivery. Human chorionic gonadotrophin (hCG), injected into immature females on Days 24 to 26, induced P-450(17 alpha) prematurely in thecal cells. When injected on Days 12 to 14 of pregnancy, hCG also induced P-450(17 alpha) in the thecal cells surrounding the largest follicles, whereas the interstitial and luteal cells were not immunostained. The antiprogestin RU486, injected on Day 16 of pregnancy, reinstated P-450(17 alpha) (and P-450scc) immunoreactivity in the thecal cells. Oestradiol selectively suppressed P-450(17 alpha) expression in the thecal cells of RU486-treated females. In immature guinea-pig ovary, P-450(17 alpha) was immunostained in thecal cells, not in interstitial cells, although the interstitial cells expressed the delta 5-3 beta-hydroxysteroid dehydrogenase. P-450(17 alpha) was also immunolocalized in the Leydig cells of rat and guinea-pig testes, and in the guinea-pig adrenal cortex (zonae fasciculata and reticularis), but not in the rat adrenal cortex. P-450(17 alpha) was not detectable in the brain of either rat or guinea-pig.

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Structural changes of guinea pig taenia coli and rat anococcygeus muscle during contraction were studied by X-ray diffraction. The diffraction pattern of the taenia coli showed the 14.4-nm myosin reflection, the 5.9-nm actin layer-line and a diffuse equatorial peak at 1/11.4 nm-1. On application of carbachol, the muscle contracted and the intensity of the 14.4-nm reflection showed a concentration-dependent decrease: the maximum decrease was 24% at 2 x 10(-5) M. Such an intensity decrease was not observed in K-contracture (154 mM). The intensity of the 5.9-nm actin layer-line did not change appreciably on activation. The equatorial peak became broader during contraction. The 14.4-nm myosin reflection of the anococcygeus muscle was weak. Its intensity increased by 106% during contraction induced by 2 x 10(-5) M phenylephrine and by 75% during K-contracture. These results suggest that the number of myosin filaments may increase during contraction of rat anococcygeus muscle but not guinea pig taenia coli.

We studied the effects of BDM (2,3-butanedione monoxime) on the tetanic contraction of frog skeletal muscles using an X-ray diffraction technique. BDM significantly increased the resting equatorial intensity ratio (I1,0/I1,1). In sartorius muscle, 3 mM BDM suppressed tetanic tension by 40-70% whereas the equatorial intensity ratio, which is 2.6 at rest, decreased to 0.75 during tetanus, close to the value in normal contraction (about 0.50). BDM (3 mM) reduced the intensity increase of the 5.1-nm layer-line to 41%, that of the 5.9-nm layer-line to 24%, and the intensity decrease of the second myosin meridional reflection (at 1/21.5 nm-1) at 81%. In overstretched semitendinosus muscle, 3 mM BDM did not significantly reduce the intensity increase of the second actin layer-line during activation, suggesting that enough calcium is released to activate the regulatory system and the regulatory proteins are intact. These results indicate that BDM suppresses tetanic tension by mainly inhibiting actin-myosin interaction. It has a smaller effect on the equatorial reflections and myosin layer-lines than on the actin layer-lines, suggesting that BDM-influenced myosin heads may bind to actin without following the symmetry of the actin helix.

The possibility of dextran magnetite complex (DM) as a new hyperthermic material was examined in this study. DM suspension of 56 mg ml(-1) iron concentration was locally injected into the normal tongue of golden hamster. DM injected tongues were heated by 500 kHz alternating current (AC) magnetic field and its serial changes in temperature were recorded at 30-s intervals. The temperature of DM injected tongue was maintained at about 43.0-45.0 degrees C for 30 min by changing the AC magnetic field intensity. While temperature elevations of the contralateral tongue and the rectum were only of minor degree. In experiment on the extent of heating area, there was correlation between volume of black stain area and amount of the injected DM suspension (Y = - 18.1 + 1.94X, r = 0.931, P < 0.0001, n = 9 ). Histological examination after heating revealed brown uniform DM accumulation in the connective tissue between fibers of the tongue muscle. Except for vascular dilatations, no tissue damage was seen in the heated tongue. Thus, DM which has the possibility of selective and uniform heating in local hyperthermia might be useful for oral cancer therapy.

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