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Biomedical subjects

S Takemori

Publications and source records attributed to S Takemori.

At least 73 records · Page 4Linked to original sources

Myosin heads contact with thin filaments in compressed relaxed skinned fibres of frog skeletal muscle.

When skinned skeletal muscle fibres with rest sarcomere length (L = 2.5 microns) are compressed by the addition of various concentrations ([PVP]) of polyvinylpyrrolidine, the relation between the 1,0 spacing (d) of thick filament lattice and [PVP] has been known to break at d of around 35 nm, resulting in a steeper slope of the relationship at d greater than 35 nm. To clarify the cause of this, X-ray diffraction and crosslinking experiments were carried out. The d versus [PVP] relationship of stretched fibres (L = 3.5 microns) breaks at a d of around 29 nm. The difference between these characteristic d values, 35-29 = 6 nm, is close to the diameter of thin filaments (8 nm). The crosslinking efficiency of formaldehyde between myosin heads and thin filament surface, measured by radial stiffness increase, was found to begin to markedly increase when the relaxed fibre with rest L was compressed to a d of nearly 35 nm. In addition to these results, the d versus [PVP] relationship obtained in rigor and in high [Mg2+] (30 mM) relaxing solutions, and the crosslinking efficiency seen in high [Mg2+] solutions supported our previous hypothesis that in normal relaxing solution (containing 1 mM Mg2+) the probability of myosin heads coming into contact with the thin filament surface abruptly increases at d near 35 nm in fibres with rest L.

Adenosine Triphosphate↗

The clinical pharmacokinetics of two different preparations of intrarectal ketoprofen following spinal or local anesthesia for anal surgery.

Two different preparations of commercially available suppositories containing Ketoprofen (KP) were administered to 49 patients immediately following anal surgery. The KP was prepared as either fatty suppositories (FS) or gelatin capsulated suppositories (GCS) and surgery was performed under either spinal (n = 37) or local anesthesia (n = 12). Similar results were observed in the kinetics of KP after both FS and GCS administration. The extent of bioavailability of the two dosage forms in the patient groups and control subjects (n = 10) were essentially equal. When the pharmacokinetic parameters of KP were compared between patient groups under spinal and local anesthesia, significant differences were found in the values of the peak level (C max), peak time (T max), and terminal phase half-life (t 1/2). The C max decreased by one-half, while the T max and t 1/2 increased twice and four times, respectively, in patient operated on under spinal anesthesia compared to those operated on under local anesthesia. The absorption rate constant (Ka) following spinal anesthesia was significantly less than that following local anesthesia or that of the healthy subjects (p less than 0.01). A "flip-flop" phenomena could be seen in the time profiles of plasma KP concentration following spinal anesthesia.

Administration, Rectal↗

Immunoreactive cytochrome P-450(17 alpha) in rat and guinea-pig gonads, adrenal glands and brain.

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.

Adrenal Cortex↗

Role of the steroid 17 alpha-hydroxylase in spironolactone-mediated destruction of adrenal cytochrome P-450.

Previous investigations have established that spironolactone (SL) is converted to a reactive metabolite by adrenal microsomal enzymes, resulting in the degradation of cytochrome P-450 (P-450). Deacetylation of SL to 7 alpha-thiospironolactone (7 alpha-thio-SL) is the first step in the activation pathway, but further NADPH-dependent metabolism of 7 alpha-thio-SL is required for P-450 destruction. Studies were done to evaluate the role of the steroid 17 alpha-hydroxylase in the activation of 7 alpha-thio-SL by adrenal microsomes. Incubation of guinea pig adrenal microsomes with 7 alpha-thio-SL in the presence of NADPH effected greater than 50% declines in P-450 content and in 17 alpha-hydroxylase activity but no change in the rate of 21-hydroxylation. Preincubation of the microsomes with antisera to the 17 alpha-hydroxylase P-450 isozyme (P-450(17 alpha,lyase)) decreased 17 alpha-hydroxylase but not 21-hydroxylase activity and prevented the degradation of P-450 by 7 alpha-thio-SL. Control IgG had no effect on 17 alpha-hydroxylase activity or on the 7 alpha-thio-SL-mediated destruction of P-450. When added to a purified P-450(17 alpha,lyase) preparation, 7 alpha-thio-SL and the endogenous substrate progesterone caused typical type I spectral changes, but SL did not. Incubation of a purified and reconstituted 17 alpha-hydroxylase system, consisting of P-450(17 alpha,lyase), NADPH-P-450 reductase, cytochrome b5, and dilauroylphosphatidylcholine, with 7 alpha-thio-SL plus NADPH effected the complete degradation of the P-450(17 alpha,lyase). Neither progesterone nor SL caused P-450 destruction with the reconstituted enzyme preparation. The results provide direct evidence for the activation of 7 alpha-thio-SL by the 17 alpha-hydroxylase and support the hypothesis that a mechanism-based inhibition of the enzyme occurs. The data also provide additional evidence that 7 alpha-thio-SL is an obligatory intermediate in the degradation of P-450 by SL.

Adrenal Glands↗

[Interstitial hyperthermia (MINERVE) for perineal local recurrence of rectal cancer].

Interstitial hyperthermia (MINERVE) was performed for 2 patients with perineal local recurrence of rectal cancer. The MINERVE was a 20 W generator of 550-610 kHz with 4 pairs of probes creating an electromagnetic field (Odam Co., France). The basic regimen was five irradiation treatments of 2 Gy per week, with two hyperthermia treatments (for 60 minutes at more than 43.5 degrees C) a week, 500 mg of 5-FU or 25 mg of CDDP per hyperthermia treatment, and 3 g/day of PSK. The radiation therapy was performed prior to hyperthermia, the chemotherapy at the same time. Perineal tumors and symptoms disappeared completely in 1 patient and were reduced in 1 patient. This treatment is expected to be clinically as a part of multidisciplinary treatment for superficial malignant tumors.

Adult↗

Immunocytochemical and biochemical studies on the localization and changes of 17 alpha-hydroxylase/C17-C20 lyase activity in immature rat ovary treated with PMSG and hCG.

The localization of cytochrome P-450 of 17 alpha-hydroxylase/C17-C20 lyase (P-450(17 alpha, lyase] and the changes of the enzyme activity were studied immunocytochemically and biochemically in the ovaries of immature rats treated with PMSG (pregnant mare serum gonadotropin) and hCG (human chorionic gonadotropin). Immunocytochemically, P-450(17 alpha, lyase) was localized in both the theca interna cells and interstitial gland cells of the ovaries of immature rats treated with PMSG for 48 h. After hCG administration, the immunoreactive cells rapidly decreased in number in the PMSG-pretreated rat ovary. Namely, 6 h after the hCG injection, positive staining for P-450(17 alpha, lyase) was recognized only in a few theca interna cells, while 12 h after the injection to immunostained cells were detected in the ovary. Forty-eight hours after the hGC treatment (96 h after the PMSG injection), most of the theca interna cells and the interstitial gland cells became immunopositive for P-450(17 alpha, lyase) again. The 17 alpha-hydroxylating activity of P-450(17 alpha, lyase) was 0.5, 0.22 and 0.03 nmol/min/mg protein in the ovarian microsomes of PMSG-treated, PMSG + hCG(3 h)-treated and PMSG + hCG(6 h)-treated rats, respectively. Changes of the hydroxylase activities in all the experimental groups are almost parallel to those of P-450 contents in the microsomes.(ABSTRACT TRUNCATED AT 250 WORDS)

Aldehyde-Lyases↗

Influence of osmotic swelling on cross section and resting tension in isolated skeletal muscle fibers.

The causes of change in resting tension with hypotonic swelling of isolated living frog skeletal muscle fibers were studied by observing their cross-sectional shape. The cross-sectional area steadily increased, in an almost osmometer-like manner, to one-third the standard tonicity. When the cross-sectional shape became almost circular and the circumferential length began to extend, the resting tension began to decrease/increase prominently at sarcomere lengths shorter/longer than 2.8 microns. The coincidence of the occurrence of prominent changes in tension and in the cross-sectional shape indicates that tension changes are closely related to the circumferential extension of the sarcolemma. For reference purposes, we studied the effects of reduction in ionic strength and in osmotic compressive force on the resting tension of mechanically and chemically skinned fibers. We concluded that (1) the decrease in resting tension with hypotonic swelling was mediated by an elevation in intracellular hydrostatic pressure due to the circumferential extension of sarcolemma, and that (2) the increase in tension was due to longitudinal contraction of the two-dimensionally tense sarcolemma, arising from its circumferential extension.

Animals↗

Steroidogenesis in liposomal system containing adrenal microsomal cytochrome P-450 electron transfer components.

Purified adrenal microsomal P-450C21 and/or P-45017 alpha,lyase were incorporated with purified NADPH-cytochrome-P-450 reductase into liposome membranes composed of phosphatidylcholine, phosphatidylethanolamine and phosphatidylserine at a molar ratio of 5:3:1. The rate dependences of reduction of liposomal P-450C21 in the fast phase as well as progesterone hydroxylation activities of P-450C21 and P-45017 alpha,lyase on the reductase concentration in the liposome membranes suggested that electrons were delivered through random collisions between the reductase and cytochrome P-450s in the liposome membranes. A rapid exchange of the steroid metabolic intermediate between vesicles was observed in a reaction system consisting of P-450C21-proteoliposomes and P-45017 alpha,lyase-proteoliposomes. Using the combined liposomal system, it was definitely proved that androstenedione was formed from progesterone mainly by a successive hydroxylation reaction without the intermediate 17 alpha-hydroxyprogesterone leaving from P-45017 alpha,lyase. It was also found that 21-hydroxylation of 17 alpha-hydroxyprogesterone into 11-deoxycortisol was inhibited by a physiological concentration of progesterone.

Adrenal Glands↗

Adrenal cytochrome P-45011 beta-proteoliposomes catalyzing aldosterone synthesis: preparation and characterization.

Purified cytochrome P-45011 beta from bovine adrenocortical mitochondria was successfully incorporated into the liposome membranes composed of phosphatidylcholine, phosphatidylethanolamine and cardiolipin at a molar ratio of 2:2:1. The incorporation of P-45011 beta into the liposome membranes was ascertained by the Ficoll density gradient centrifugation and the protein refractoriness to trypsin digestion. The prepared proteoliposomes containing P-45011 beta and phospholipid at a molar ratio of 1:3000 were unilamellar vesicles of about 40 nm in average diameter. The P-45011 beta embedded in the liposome membranes was found to be more stable than the detergent-solubilized form. The reconstituted system containing the P-45011 beta-proteoliposomes, adrenodoxin and NADPH-adrenodoxin reductase showed catalytic activities not only for the hydroxylation of 11-deoxycorticosterone at 11 beta- and 18-positions but also for its conversion into aldosterone with a turnover number of 2.3 nmol/min per nmol of P-45011 beta. A successive reaction without the intermediates leaving from the enzyme was suggested for the P-45011 beta-mediated conversion of 11-deoxycorticosterone to aldosterone following the result that the formation of aldosterone was linear with respect to time without the lag phase; this was confirmed by the result that radioactivity in aldosterone from 3H-labeled 11-deoxycorticosterone was scarcely decreased by the addition of unlabeled intermediates to the reactions system.

Adrenal Cortex↗

Butanedione monoxime suppresses contraction and ATPase activity of rabbit skeletal muscle.

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.

Animals↗

An analysis of ocular counter-rolling measured with search coils.

Ocular counter-rolling (OCR) was studied by using a scleral search coil magnetic system in normal subjects and in pathological cases. Normal ocular counter-rolling was 2.7 degrees-7 degrees when the head was tilted 10 to 30 degrees. In most cases of benign paroxysmal positional vertigo, the OCR to the ipsilateral side was reduced, while that to the contralateral side was normal or only slightly reduced. Soon after unilateral labyrinthectomy, the OCR to the ipsilateral side was reduced or was 0, whereas OCR to the contralateral side was normal or slightly reduced. Some 3-5 years after the operation, however, the OCR seemed to depend on the compensation achieved. In cases of acoustic neurinoma, OCR to both sides was reduced, that to the ipsilateral side being more strongly impaired than the OCR to the contralateral side.

Ear, Inner↗

High-resolution MR imaging of the inner ear.

The magnetic resonance (MR) images of the temporal bones have been analyzed, these MR images taken with head coils utilizing a 1.5-Tesla magnet whole-body imaging system. The MR images were acquired by means of spin-echo pulse sequences, and the thickness of the sections was 2.5 mm. By this modality, the T2-weighted images could clearly delineate the details of the liquid-containing labyrinthine structures and facial nerve. MR imaging can provide information that is unobtainable from a CT scan in the diagnosis of inner ear disorders, particularly as to whether the membranous labyrinth is filled with the lymph fluid or is fibrosed. This point is one of the greatest advantages of MR imaging over a high-resolution CT scan.

Ear, Inner↗

[Unilateral palatal myoclonus with peculiar ocular movements--neurotological studies and MRI].

This is the first report of unilateral palatal myoclonus with which two different ocular movements were synchronized. A 55-year-old woman was admitted to our hospital due to intubation and dysarthria of sudden onset after three similar attacks for these four years. On admission right ptosis, hypalgesia in the right face, right facial nerve palsy, dysarthria, bilaterally increased deep tendon reflexes and trunkal ataxia were noted. Continuous repetitive contractions at 110/min frequencies were observed in the left soft palate, pharynx and larynx. On electronystagmography and electro-magnetic search coil system (Skala system), two different ocular movements, i.e., rotary oscillation with torsion and nystagmus to the right, successively alternated each other at random during eyelids closure. They were synchronized with palatal myoclonus. True nystagmus synchronized with palatal myoclonus has not been reported. When she calculated, rotary oscillation disappeared. In sleep polygraphy, rotary oscillation reduced in amplitude in stage 1 and disappeared in stages 2, 3, 4 and REM. On the other hand, the nystagmus reduced in amplitude in stage 1 and 2 and disappeared in stages 3, 4 and REM. The direction of nystagmus was converted to the left in stages 1 and 2. Similarly, in a drowsy state induced by intravenous injection of 7 mg diazepam, the direction of the nystagmus was converted to the left. On brain magnetic resonance imaging (MRI) right inferior olive was identified as a well circumscribed, enlarged increased signal area on T2-weighted and proton density-weighted images in addition to the lesions of infarcts in left corona radiata, posterior limb of right internal capsule and tegmentum pontis.(ABSTRACT TRUNCATED AT 250 WORDS)

Brain Stem↗

Interaction of steroids with adrenal cytochrome P-450 (P-450(17)alpha,lyase) in liposome membranes.

Purified cytochrome P-450(17)alpha,lyase from guinea-pig adrenal microsomes, which catalyzes progesterone 17 alpha-hydroxylation and sequentially C17-C20 bond cleavage of the 17 alpha-hydroxyprogesterone, was successfully incorporated into liposomal membranes composed of only phosphatidylcholine or of a phospholipid mixture of phosphatidylcholine, phosphatidylethanolamine and phosphatidylserine at a molar ratio of 5:3:1. Although the purified P-450(17)alpha,lyase was readily converted into P-420 in the detergent-solubilized system without substrates, the P-450 embedded in the liposomal membranes was found to be quite stable without the substrates. Using the P-450(17)alpha,lyase-proteoliposomes, the interaction of steroids with P-450(17)alpha,lyase was studied for progesterone, 17 alpha-hydroxyprogesterone and androstenedione in the liposomal system by optical difference spectroscopy and by equilibrium dialysis. The partition coefficients of steroids between the aqueous phase and the liposomal membranes were determined by the equilibrium dialysis. They were about 1.4-1.6-times higher in phosphatidylcholine liposomes than in the liposomes of the lipid mixture. The dissociation constants of the P-450-steroid complexes were calculated from the apparent dissociation constants using the partition coefficients for the situation where the substrate-binding site faces the lipid phase of the membranes or where it faces the aqueous phase. The dissociation constant in the former case was not affected by the lipid composition. These results suggest that P-450(17)alpha,lyase might interact only with the substrates in the lipid phase of the liposomal membranes.

Adrenal Glands↗

Examination of differences between benzo[a]pyrene and steroid hydroxylases in guinea pig adrenal microsomes.

The effects of antibodies to cytochromes P-45017 alpha,lyase and P-450C21 on benzo[a]pyrene hydroxylase activity were measured in microsomes from guinea pig adrenals. Anti-cytochrome P-45017 alpha,lyase IgG inhibited about 30% of the benzo[a]pyrene hydroxylase activity of the microsomes in the presence of excess amounts of the IgG, but anti-cytochrome P-450C21 IgG did not affect the activity. In a reconstituted system, consisting of cytochrome P-450, NADPH-cytochrome-P-450 reductase and dilauroylphosphatidylcholine, cytochrome P-45017 alpha,lyase catalyzed, in addition to steroid hydroxylation, benzo[a]pyrene hydroxylation, but cytochrome P-450C21 did not hydroxylate benzo[a]pyrene. Since anti-cytochrome P-45017 alpha,lyase IgG inhibited benzo[a]pyrene hydroxylase activity completely in the reconstituted system with cytochrome P-45017 alpha,lyase, the presence of non-inhibited benzo[a]pyrene hydroxylase in the microsomes suggests that the residual activity in the microsomes may be due to some enzyme other than cytochrome P-45017 alpha,lyase. Benzo[a]pyrene hydroxylase activity was detected in detergent-solubilized microsomes from which cytochrome P-45017 alpha,lyase was removed by using an immobilized anti-cytochrome P-45017 alpha,lyase IgG-Sepharose column. This shows the existence in microsomes of a benzo[a]pyrene hydroxylase other than cytochrome P-45017 alpha,lyase. Benzo[a]pyrene hydroxylase in the solubilized microsomes required O2, NADPH and NADPH-cytochrome-P-450 reductase for its activity and was inhibited by CO. This suggests that benzo[a]pyrene hydroxylase is a cytochrome P-450-dependent monooxygenase. This novel enzyme was also active in xenobiotic metabolism, such as 2-nitropropane denitrification and aminopyrine demethylation.

Adrenal Glands↗