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T Oba

Publications and source records attributed to T Oba.

At least 55 records · Page 3Linked to original sources
PubMed1991

CGG: an unassigned or nonsense codon in Mycoplasma capricolum.

CGG is an arginine codon in the universal genetic code. We previously reported that in Mycoplasma capricolum, a relative of Gram-positive eubacteria, codon CGG did not appear in coding frames, including termination sites, and tRNA(ArgCCG) pairing with codon CGG, was not detected. These facts suggest that CGG is a nonsense (unassigned and untranslatable) codon--i.e., not assigned to arginine or to any other amino acid. We have investigated whether CGG is really an unassigned codon by using a cell-free translation system prepared from M. capricolum. Translation of synthetic mRNA containing in-frame CGG codons does not result in "read-through" to codons beyond the CGG codons--i.e., translation ceases just before CGG. Sucrose-gradient centrifugation profiles of the reaction mixture have shown that the bulk of peptide that has been synthesized is attached to 70S ribosomes and is released upon further incubation with puromycin. The result suggests that the peptide is in the P site of ribosome in the form of peptidyl-tRNA, leaving the A site empty. When in-frame CGG codons are replaced by UAA codons in mRNA, no read-through occurs beyond UAA, just as in the case of CGG. However, the synthesized peptide is released from 70S ribosomes, presumably by release factor 1. These data suggest strongly that CGG is an unassigned codon and differs from UAA in that CGG is not used for termination.

Amino Acid Sequence↗
PubMed1990

Analysis of MRI and SPECT in patients with acute head injury.

Traumatic lesions defined by magnetic resonance (MR) imaging were divided into two groups according to findings on computed tomography (CT). This classification reflected difference in the regional cerebral blood flow (rCBF). In the contusional lesions which CT could demonstrate, rCBF varied from hyperperfusion to hypoperfusion, while it was almost always decreased in the lesions which CT could not detect. These results suggest that the former may include a mixture of brain oedema and hyperemia and the latter may imply brain oedema. MR imaging can reveal the minor oedema which CT fails to show in patients with acute head injury.

Acute Disease↗
PubMed1990

Sulfhydryls on frog skeletal muscle membrane participate in contraction.

To examine the molecular mechanism underlying contractile activation, we studied effects of a sulfhydryl reagent, N-(7-dimethylamino-4-methylcoumarinyl)maleimide (DACM), on twitch, Ag(+)-induced contraction, and K+ and caffeine contractures in single toe muscle fibers of frog. DACM suppressed twitch and Ag(+)-induced contraction, dose dependently, but not caffeine contracture. K+ contracture also was decreased appreciably by exposure to 40 microM DACM for 10 min. DACM elicited no shift of the mechanical threshold or inhibition of resting potential but slightly inhibited action potential. Increase of the fluorescence intensity produced by binding of 10 microM DACM to sulfhydryl groups was depressed by brief pretreatment with 100 microM Ag+. When exposed to 1 mM dithiothreitol (DTT) within 5 s of the rising phase of 5 microM Ag(+)-induced contraction, the fiber rapidly decreased the tension to the resting level. In this case, reapplication of 5 microM Ag+ after washing out DTT elicited a new contraction similar to the first Ag(+)-induced contraction. The second contraction amplitude depended on the time between the onset of the first Ag(+)-induced contraction and DTT application. If DTT was applied after more than 16 s, tension no longer developed on the second exposure to Ag+ or K+. The experiments provide evidence that crucial sulfhydryl groups participate in muscle activation. The possible role of the sulfhydryl group on the transverse tubular membrane in tension development is discussed.

Animals↗
PubMed1990

Effects of tetraphenylboron-induced increase in inner surface charge on Ca2+ release from sarcoplasmic reticulum.

We studied the relationship between surface charge and release of Ca2+ in the heavy sarcoplasmic reticulum (SR) of skeletal muscle. The inner and outer surface potentials and charge densities of the membrane treated with a lipophilic anion, tetraphenylboron (TPB-), were measured using 1-anilino-8-naphthalene-sulfonate fluorescence. Ca2+ was loaded passively or actively by the SR. Ca2+ release was estimated by the fluorescence of chlortetracycline, and protein conformational change was monitored by use of the sulfhydryl group fluorescent probe, N-(7-dimethylamino-4-methyl-3-coumarinyl) maleimide (DACM). Treatment of Ca2(+)-loaded SR vesicles with micromolar TPB- dose-dependently increased the local fixed negative charge on the inner surface, and changed the DACM fluorescence intensity in parallel with the Ca2+ release. The changes in surface charge and in DACM fluorescence intensity did not originate from the Ca2+ flux. A lipophilic cation, tetraphenylarsonium (TPA+), screened the negative inner surface charge which was increased by TPB-, and inhibited both TPB(-)-induced change in DACM fluorescence intensity and Ca2+ release. Millimolar Mg2+ reduced degrees of TPB(-)-induced Ca2+ release from the SR and of TPB(-)-induced contraction in mechanically skinned fibers. Mg2+ did not inhibit the increase in the negative inner surface charge and DACM fluorescence intensity produced by TPB-. Thus, the local increase in negative charge on the SR inner membrane leaflet seems to be causally related to the Ca2+ release. Mg2+ and TPA+ are suggested to inhibit TPB(-)-induced Ca2+ release by different mechanisms.

Animals↗
PubMed1990

[MR imaging of traumatic cerebellar dysfunction].

Four cases of cerebellar dysfunction following head trauma are presented. Cerebellar signs revealed were those such as dysmetria, dysdiadochokinesis, horizontal nystagmus and ataxia. T2-weighted magnetic resonance (MR) imaging (0.15 tesla, spin-echo method; TR 2000 msec. and TE 100 msec.) revealed focal lesions in these patients, although CT scan failed to demonstrate any changes in the cerebellum. The cerebellar symptoms were maximal immediately after the trauma but improved gradually in two cases. These are compatible with the transient traumatic cerebellar dysfunction postulated by R. C. Cantu in 1969. The pathophysiology of this syndrome, whether it is due to cerebellar concussion or contusion, has not yet been determined. The abnormality of the cerebellum revealed by MR imaging seemed to be contusion rather than concussion. Therefore the authors presume that transient traumatic cerebellar dysfunction is caused by minor cerebellar contusion. In the other two cases, delayed epidural hemorrhage ensued and the symptoms disappeared rapidly after evacuation of the hematoma. In these patients, occurrence of delayed epidural hematoma in the posterior cranial fossa was predicted by MR imaging. The authors regard the lesion as an alarm signal indicating the probable occurrence of infratentorial hematoma.

Adult↗
PubMed1989

Calcium release from frog sarcoplasmic reticulum by an imidazolyl reagent.

Calcium is released from the isolated heavy sarcoplasmic reticulum (SR) of frog skeletal muscle upon application of 0.1-1 mM diethylpyrocarbonate (DEP, an imidazolyl reagent). The Ca-ATPase activity of SR was suppressed by 20% in the presence of 1 mM DEP. More than 1 mM of free magnesium ion or 5 microM ruthenium red eliminated the effect of DEP on calcium release but not on Ca-ATPase activity. A plausible site of DEP action is on the calcium channel.

Animals↗
PubMed1989

Caffeine treatment inhibits drug-induced calcium release from sarcoplasmic reticulum and caffeine contracture but not tetanus in frog skeletal muscle.

Effects of pretreatment with caffeine on Ca2+ release induced by caffeine, thymol, quercetin, or p-chloromercuriphenylsulfonic acid (pCMPS) from the heavy fraction of sarcoplasmic reticulum (SR) were studied and compared with those effects on caffeine contracture and tetanus tension in single fibers of frog skeletal muscle. Caffeine (1-5 mM) did induce transient Ca2+ release from SR vesicles, but subsequent further addition of caffeine (10 mM, final concentration) induced little Ca2+ release. Ca2+ release induced by thymol, quercetin, or pCMPS was also inhibited by pretreatment with caffeine. In single muscle fibers, pretreatment with caffeine (1-5 mM) partially reduced the contracture induced by 10 mM caffeine. However, tetanus tension was almost maximally induced by electrical stimulus in caffeine-treated fibers. These results indicate that SR, which becomes less sensitive to caffeine, thymol, quercetin, or pCMPS by pretreatment with caffeine, can still respond to a physiological signal transmitted from transverse tubules.

4-Chloromercuribenzenesulfonate↗
PubMed1989

Ruthenium red and magnesium ion partially inhibit silver ion-induced release of calcium from sarcoplasmic reticulum of frog skeletal muscles.

Effects of Ca2+-induced Ca2+ release blockers, ruthenium red (RR) and Mg2+, on Ag+-induced Ca2+ release were studied using skinned muscle fibers or fragmented heavy SR (HSR) prepared from frog muscle, and compared with those on caffeine-induced one. Exposure of the skinned fibers to 5 microM Ag+ produced a rapid and large contraction in the presence of 0.043 mM free Mg2+. When Mg2+ concentration was increased to 0.86 mM, Ag+ led to a large transient contraction, combined with a small tonic one. The transient component was completely blocked by high Mg2+ (3.64 mM), but the tonic one was not. Ca2+-ATPase activity was not stimulated by increase of Mg2+ from 0.86 to 3.64 mM. Ag+ and caffeine induced a rapid Ca2+ efflux from HSR in a dose-dependent manner. RR over a range from 1 to 10 microM dose-dependently inhibited the Ca2+ efflux induced by 10 microM Ag+. Despite increase of RR to 30 microM, however, further inhibition of the Ca2+ efflux was not produced any more (77.8 +/- 12.2% inhibition). A 10 mM caffeine-induced efflux of Ca2+ was blocked slightly by only 0.5 microM RR and almost completely by 3 microM. A slight inhibition (about 28%) of the Ca2+-ATPase activity was observed in the presence of 10 microM Ag+ in 0.5 mg SR protein/ml of medium. RR and caffeine did not affect the enzyme activity. These results indicate that frog SR could induce a rapid release of Ca2+ upon Ag+ and caffeine, suggesting that Ag+ may have two different binding sites to release Ca2+; one is on Ca2+-induced Ca2+ release channel and the other on RR-insensitive site.

Animals↗
PubMed1989

Change in surface charge of sarcoplasmic reticulum membrane may elicit conformational change in sulfhydryl groups of membrane proteins to release calcium.

A lipophilic anion, tetraphenylboron (TPB-)-induced Ca2+ release from fragmented sarcoplasmic reticulum (SR) of frog skeletal muscle was monitored by chlortetracycline fluorescence. TPB- caused change in surface charge of the membrane and in the protein conformation with a time course similar to that of the Ca2+ release. Tetraphenylarsonium (TPA+) inhibited these effects of TPB-. Change in surface charge of SR is suggested to cause conformational change in SR membrane proteins, and then result in Ca2+ release from the SR.

Animals↗
PubMed1987

Similar inhibitory effects of dantrolene sodium on twitch tension and on silver ion-induced contracture in skeletal muscle.

To determine the mechanism by which Ag+ induces a transient contracture in skeletal muscle, the effect of dantrolene sodium on the Ag+ contracture was examined and the findings compared with those for the twitch, tetanus and caffeine contracture. The inhibition of twitch by dantrolene was equivalent to that of the Ag+ contracture at concentrations of 1, 2 or 5 microM of dantrolene. The tetanus tension was slightly inhibited by dantrolene, but not the caffeine contracture. These observations suggest that the Ag+ contracture may be governed by the same mechanism as that involved in the development of twitch tension.

Animals↗
PubMed1987

Effects of extracellular calcium and calcium channel blocker on silver-induced contractures in frog skeletal muscle fibers.

The mechanism by which Ag+ induces muscle contracture was elucidated by investigating the effect of external Ca2+ concentration and Ca2+ channel blocker on the maximum tension amplitude in single fibers from frog toe skeletal muscle. Five microM Ag+ induced two different types of contracture in the presence of external Ca2+ more than 0.1 mM, i.e., a phasic and a subsequent tonic contracture. The phasic contracture appeared only in fibers with intact T-tubules immersed in a solution with or without Ca2+ after a lag time of 5.7 +/- 0.9 s (N = 5). The maximum amplitude was 58% of the tetanus tension observed in the same fiber immediately before Ag+ exposure. Diltiazem at high-concentration (100 microM) inhibited the Ag+-induced phasic contracture only to a small extent (17%). The contracture was not affected by 1 microM TTX or 1 mM DAP at all. These results indicate that Na+, K+, and Ca2+ channels on the T-tubular membrane would not be attributed to the phasic tension development induced by Ag+. On the contrary, a tonic contracture did not require intact T-tubules. The amplitude and the rate of rise of the contracture depended on external Ca2+ concentrations and were inhibited by a high concentration of diltiazem. Neither 1 microM TTX nor 1 mM DAP affected them. Therefore, the tonic contracture seems to be triggered by Ca2+ which entered the muscle fiber through the surface but not T-tubular membranes.

4-Aminopyridine↗
PubMed1987

A local anesthetic, tetracaine, similarly inhibits Ag+ and K+ contractures in frog skeletal muscle.

To evaluate usefulness of Ag+ contracture as a tool for elucidating the mechanism underlying the excitation-contraction coupling, the effects of tetracaine on Ag+ contracture were compared with those on K+ and caffeine contractures in frog skeletal muscle. Tetracaine less than 100 microM shortened the duration of 120 mM K+ contracture, without affecting tension amplitude. At higher concentrations of tetracaine, K+ contracture was inhibited dose-dependently and the duration shortened. Treatment of the fibers with 20-500 microM tetracaine for 3 min did not block the contracture induced by 25 mM caffeine. Effects of tetracaine on Ag+ contracture were similar to those on K+ contracture. In the presence of 200 microM tetracaine, 41% inhibition was observed in 120 mM K+ contracture, while 43% in 100 microM Ag+ contracture. Also, 200 microM tetracaine completely inhibited the contractures induced by 40 mM K+ or 5 microM Ag+. These findings suggest that the Ag+ may induce contractures via its action on the T/SR junction, not a direct action on the SR. Therefore, understanding the mechanism involved in the development of Ag+ contracture would be helpful to elucidate the mechanism of excitation-contraction coupling.

Action Potentials↗
PubMed1986

Inhibition of Zn2+-induced potentiation of twitch tension by Ni2+ in frog muscle.

Effect of Ni2+ on Zn2+-induced potentiation of twitch tension was studied electrophysiologically in the toe muscle fibers of Rana catesbeiana. The major findings of this investigation are as follows. When 2 mM Ni2+ was applied to fibers in a normal Ringer's solution containing 50 microM Zn2+ (Zn2+ solution), the Zn2+-potentiated twitch tension decreased remarkably to about one-third of that before Ni2+ treatment. This concentration of Ni2+ caused a 23% decrease in the duration of action potential which had been prolonged by Zn2+ (6.61-5.09 ms). Ni2+ (2 mM) added to normal Ringer's solution led to increases of about 30 and 42% in twitch tension and in the duration of action potential, respectively. A slight increase in the mechanical threshold was induced by 2 mM Ni2+. The inhibitory action of Ni2+ on the twitch tension in Zn2+ solution was larger than that in the case of tetanus tension. Diltiazem (40 microM), a Ca2+ channel blocker, did not inhibit the twitch tension potentiated in Zn2+ solution. These results suggest that the decrease in Zn2+-potentiated twitch tension by Ni2+ may possibly derive from impairment of the propagation of action potential along the T tubules.

Action Potentials↗
PubMed1986

Chemical modification of calcium release from the sarcoplasmic reticulum of mechanically skinned skeletal bullfrog muscle fibers.

Several types of reagents that react with amino acid side chains induced repetitive phasic contracture of skinned skeletal muscle from frogs. The presence of 10 mM procaine or 5 mM magnesium in the medium or disruption of the sarcoplasmic reticulum (SR) eliminated this contracture, indicating that the calcium-induced calcium-release mechanism of SR is involved in the contraction. Dithiothreitol inhibited the contracture induced by chloramine T, N-acetylimidazole, or p-chloromercuriphenylsulfonic acid (pCMPS) but not in the case of carbodiimide, phenylglyoxal, trinitrobenzenesulfonic acid, diethylpyrocarbonate (DEP), or N-chlorosuccinimide (NCS). Therefore, modification of groups other than the sulfhydryl ones seems to induce contractures under such conditions. The amplitude of the caffeine-induced contracture decreased after treatment with pCMPS, DEP, or NCS. NCS shifted the pCa-tension curve toward low pCa in the SR-disrupted fibers. This shift would explain the decrease in the caffeine contracture. It is tentatively concluded that pCMPS and DEP release a large amount of calcium from SR.

Animals↗
PubMed1985

Silver ion-induced tension development and membrane depolarization in frog skeletal muscle fibres.

Silver ions elicit dose-dependently a transient contracture in single fibres of bull-frog toe muscle placed in 0-Ca2+, Cl- -free MOPS solution containing 3 mM Mg2+ and NO3-. To elucidate the mechanisms involved, changes in membrane potential and in tension development were continuously measured following exposure to Ag+. The effect of Ag+ on contraction in fibres in which the membrane had been depolarized by elevating the external K+ concentration was also examined. The major findings of this investigation are as follows. (1) The mechanical threshold was shifted towards more negative potentials by 5 mV (-51 to -56 mV), when Ca2+ and Cl- in the Ringer's solution were replaced with Mg2+ and NO3-, respectively. (2) On the exposure of the fibres to 5 microM Ag+, the membrane potential decreased by 1.6 mV from -87.8 mV and tension was developed. (3) In fibres soaked in a solution containing 10 mM K+ (corresponding to a membrane potential of -69.5 mV), 5 microM Ag+ produced a large contracture similar to that seen in the control solution. (4) The Ag+-induced contracture was inactivated when more than 20 mM K+ was used. (5) The membrane depolarization evoked by either 20 or 50 microM Hg2+ did not produce contraction. (6) Muscle fibres which had been exposed to 20 microM Hg2+ for 5 min responded to 5 microM Ag+ by a transient tension development. These findings strongly suggest that Ag+-induced tension development is not associated with depolarization of the surface membrane but rather is caused by specific actions of Ag+ on membrane proteins in the T-tubules.

Animals↗