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Biomedical subjects

T Asano

Publications and source records attributed to T Asano.

At least 703 records · Page 39Linked to original sources
PubMed1988

Immunoassay for the beta gamma subunits of GTP-binding proteins and their regional distribution in bovine brain.

Antibodies were raised in rabbits against the beta gamma subunits of bovine brain GTP-binding proteins, and were purified with a beta gamma-coupled Sepharose column. Purified antibodies reacted strongly with 36,000-dalton beta subunit and slightly with 35,000-dalton beta and gamma subunits, but not with other proteins in an immunoblot assay. Using these purified antibodies, a sensitive enzyme immunoassay method for the quantification of brain beta gamma was developed. The assay system consisted of polystyrene balls with immobilized antibody F(ab')2 fragments and the same antibody Fab' fragments labeled with beta-D-galactosidase from Escherichia coli. The minimum detection limit of the assay was 3 fmol, or 130 pg. Samples from various regions of bovine brain were solubilized with 2% sodium cholate and 1 M NaCl, and the concentrations of beta gamma were determined. The beta gamma were detected in all the regions, and the highest concentrations were observed in the cerebral cortex and nucleus caudatus. The concentrations of beta gamma were higher than those of alpha subunit of GTP-binding protein, Go, in all the regions.

Animals↗
PubMed1988

Ontogeny of the GTP-binding protein Go in rat brain and heart.

We determined the ontogeny of the GTP-binding protein Go in rat brain and heart by employing highly sensitive enzyme immunoassay methods. In the brain, the alpha subunit of Go (Go alpha) gradually increased and reached adult levels approximately 20 and 30 days after birth in cerebral cortex and cerebellum, respectively. Concentrations of beta subunits, which were also quantified by the immunoassay, were almost equal to those of Go alpha in the brain of rats younger than 10 days, but were higher than those of Go alpha after 10 days. These results suggest that late development of GTP-binding proteins other than Go. Go alpha was immunohistochemically positive in neuropils and negative in cell bodies at any age tested. In the heart, the concentrations of Go alpha increased up to several times of the adult level just after birth, and then gradually decreased after the 20th postnatal day. The level of Go alpha in the liver, however, was very low and constant throughout ontogenic development. An immunohistochemical study indicated that Go alpha was positive in the cardiac muscle of young rat, but negative in that of adult rat. These results indicate that Go alpha exists in cells other than those of nervous tissues and neuroendocrine cells in some periods of ontogenic development.

Aging↗
PubMed1988

Activation of the arachidonate 5-lipoxygenase pathway in the canine basilar artery after experimental subarachnoidal hemorrhage.

Severe cerebral vasospasm as confirmed by angiography was induced in dogs by injection of autologous blood into the cisterna magna, and the resultant leukotriene formation in the isolated basilar artery was examined. When stimulated with calcium ionophore (A 23187), the arteries of the treated animals produced a significant amount of leukotrienes B4 (85 +/- 12 pmol/mg protein, n = 3) and C4 (72 +/- 14 pmol/mg), in addition to 5(S)-hydroxy-6,8,11,14-eicosatetraenoic acid. Structural elucidations of these metabolites were performed by radioimmunoassays or gas chromatography-mass spectrometry, following purification with HPLC. The artery of the untreated dog produced none of these compounds from either exogenous or endogenous arachidonic acid, under stimulation with the calcium ionophore. However, the homogenates from both animals converted exogenous leukotriene A4 to leukotrienes B4 and C4. These observations suggest that the normal basilar artery contains no detectable amount of 5-lipoxygenase, and that a prominent activation of this enzyme occurred (2.1 nmol 5-HETE/5 min/mg of protein) after subarachnoidal hemorrhage. The observation that fatty acid hydroperoxides stimulated the 5-lipoxygenase activity indicates a possible role of lipid peroxides in the development of vasospasm.

Animals↗
PubMed1988

Participation of lipoxygenase products from arachidonic acid in the pathogenesis of cerebral vasospasm.

To examine the possible involvement of lipoxygenase products from arachidonic acid in the pathogenesis of delayed vasospasm after subarachnoid hemorrhage (SAH), we measured the contents of hydroxyeicosatetraenoic acids (HETEs) in the subarachnoid clot, the cerebrospinal fluid, and the basilar artery, using the canine "two-hemorrhage" model. Lipoxygenase activity in the subarachnoid clot and the basilar artery was measured, ex vivo, using samples obtained 7 days after SAH. For a quantitative analysis of HETEs, each sample was homogenized with either ice-cold saline or methanol. The lipid extract was then submitted to reverse-phase HPLC. The identity of each HETE was further confirmed using straight-phase HPLC and gas chromatography-mass spectrometry. When the basilar artery was homogenized with ice-cold saline, a significant increase in the 5-HETE content was observed on SAH day 8. However, when the artery was homogenized with methanol, HETEs were not detected. In the case of incubation in the presence of arachidonic acid and calcium ionophore A23187, the 5-lipoxygenase activity was remarkably increased in the basilar artery exposed to SAH, compared to that of normal dogs. The subarachnoid clot contained a significant amount of 12-HETE (average 1.8 nmol/g wet weight) from day 2 to day 8. The administration of 1,2-bis(nicotinamido)propane significantly ameliorated vasospasm in the two-hemorrhage model, simultaneously inhibiting the 5-lipoxygenase activity of the basilar artery. Our observations show that the activities of 12- and 5-lipoxygenases are significantly increased after SAH in the subarachnoid clot and the basilar artery, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

12-Hydroxy-5,8,10,14-eicosatetraenoic Acid↗
PubMed1988

Go, a GTP-binding protein: immunochemical and immunohistochemical localization in the rat.

The tissue and cellular distribution of a GTP-binding protein, Go, was investigated in the rat by immunochemical and immunohistochemical methods. Because the specific antibody for the alpha subunit of bovine Go (Go alpha) cross-reacted with rat Go alpha, an enzyme immunoassay method developed for bovine Go alpha was applied for measuring the tissue concentration of Go alpha in the rat. Go alpha was detected in all tissues examined except blood cells. The concentration of Go alpha was highest in the CNS (approximately 7.7 and 4.4 nmol/g in the cerebrum and cerebellum, respectively), followed by the pituitary gland and sciatic nerve. Among the other peripheral tissues, relatively high concentrations of Go alpha were observed in the urinary bladder, stomach, and intestines; however, these values were less than 2% of the concentration in the cerebrum. Go alpha in the intestine was located mostly in the muscle layer. Immunohistochemical study showed that Go alpha was associated mostly with the neural elements but not with cells particular to each peripheral organ. Go alpha was also present in the membranes of neuroendocrine cells, including glandular cells in the anterior lobe of the pituitary gland, chromaffin cells in the medulla of the adrenal gland, islets cells in the pancreas, and parafollicular cells in the thyroid. These results indicate that Go is localized exclusively in the nervous tissues and neuroendocrine cells.

Animals↗
PubMed1988

Protein kinase activity in canine basilar arteries after subarachnoid hemorrhage.

To investigate the intracellular physiology of cerebral arterial vasospasm after subarachnoid hemorrhage (SAH), we measured the activity of three protein kinases related to vascular smooth muscle function. Myosin light chain kinase (MLCK), cyclic adenosine monophosphate (cAMP)-dependent protein kinase and cyclic guanosine monophosphate (cGMP)-dependent protein kinase activity was determined in canine basilar arteries using the SAH model created by two intracisternal injections of autologous blood. Moderate chronic cerebral vasospasm was confirmed angiographically 7 days after the first injection of blood, with a mean decrease in the diameter of the basilar artery to 61.2 +/- 2.3% (SE) of preinjection controls. Enzymatic analysis of basilar arteries excised from control dogs and from SAH model dogs 2 to 10 days after an intracisternal injection of blood demonstrated detectable MLCK and cyclic nucleotide-dependent protein kinase activity. The MLCK activity was approximately 3 times higher than the cyclic nucleotide-dependent protein kinase activity. There was no significant difference in basilar artery protein kinase activity between control animals and SAH model animals at any time for up to 10 days after the intracisternal injection of blood. As MLCK is involved with smooth muscle contraction and cAMP- and cGMP-dependent protein kinases are involved in smooth muscle relaxation, our results suggest that the metabolic machinery of vascular smooth muscle in the cerebral arterial wall is intact after SAH and that prolonged vasospasm represents an appropriate physiological response by the vascular smooth muscle to external stimuli. If this is correct, the successful pharmacological manipulation of vasospasm after SAH is more likely than if profound disruption had occurred, and further investigative efforts in this area should be encouraged.

Animals↗
PubMed1988

Maximizing present value: A model to explain why moderate response rates obtain on variable-interval schedules.

In Phases 1 and 3, two Japanese monkeys responded on a multiple variable-ratio 80 variable-interval X schedule, where the value of X was adjusted to ensure equal between-schedule reinforcement rates. Components strictly alternated following the delivery of a food pellet, and each session ended following 50 components. Phase 2 differed from the others only in that the 50 pellets previously earned during the session were delivered together at session's end. Variable-ratio response rates did not decrease across phases, but variable-interval response rates decreased substantially during the Phase 2 procedure. This rate decrease was attributed to the food-at-session's-end manipulation removing the greater immediacy of reinforcement provided by short interresponse times relative to long interresponse times. Without this time preference for short interresponse times, the variable-interval interresponse-time reinforcement feedback function largely controlled response emission, dictating a response-rate reduction. This result was explained in terms of the economic notion of "maximizing present value."

Journal Article↗
PubMed1988

Choice in the repeated-gambles experiment.

Humans chose 10 times between two roulette wheels projected on a monitor. During the first trial, the left wheel provided a hypothetical $100 with p = .94, and the right wheel provided $250 with p = .39. A titration procedure adjusted the probability of a $250 win across trials to permit estimation of an indifference point between alternatives. In Experiment 1, intertrial-interval duration (25 vs. 90 s) and whether sessions began with an intertrial interval or a trial were varied in a 2 x 2 design in this risky-choice procedure. Risk aversion (preference for the $100 wheel) increased with intertrial interval but was unaffected by whether sessions began with a trial or an intertrial interval. In Experiment 2, all sessions began with a trial, and subjects were informed that the experiment ended after 10 trials. Intertrial-interval duration had no effect on choice. In Experiment 3, intertrial-interval duration and whether subjects were given $10 or $10,000 before beginning were varied among four groups in a 2 x 2 design. In all other ways, the procedure was unchanged from Experiment 2. Intertrial interval had no effect on choice, but the $10,000 groups showed less risk aversion than the $10 groups. These results can be explained more readily in terms of Kahneman and Tversky's (1984) notion of "framing of the prospect" than in terms of Rachlin, Logue, Gibbon, and Frankel's (1986) behavioral account of risky choice.

Journal Article↗
PubMed1988

Reduced function of the stimulatory GTP-binding protein in beta adrenoceptor-adenylate cyclase system of femoral arteries isolated from spontaneously hypertensive rats.

Arterial relaxant responses to beta adrenoceptor agonists are decreased in spontaneously hypertensive rats (SHR) when compared with normotensive Wistar-Kyoto rats (WKY). To determine which component of the beta adrenoceptor-adenylate cyclase (AC) system is involved in the decreased beta adrenoceptor responses, effects of two activators of AC-forskolin and cholera toxin and of dibutyryl cyclic AMP (DBcAMP) were compared between the strips of femoral arteries isolated from 13-week-old SHR and age-matched WKY. Arterial relaxant responses to either forskolin, an activator of AC or DBcAMP were not significantly different between the SHR and WKY, whereas the relaxant responses to norepinephrine (NE) via beta adrenoceptors were significantly weaker in the SHR than in the WKY. In the absence of timolol, a beta adrenoceptor antagonist, contractile responses to NE were significantly greater in the SHR than in the WKY. Timolol augmented the contractile responses to NE to a greater extent in the WKY than in the SHR. After the blockade by timolol of beta adrenoceptors, contractile responses to alpha adrenoceptor stimulation with NE were not significantly different between the two strains. The pretreatment of the strips with cholera toxin, an activator of stimulatory GTP-binding protein (Gs), antagonized the alpha adrenoceptor-mediated contractions much greater in the WKY than in the SHR. The alpha adrenoceptor-mediated contractions after the pretreatment with cholera toxin were comparable to the contractile responses to NE determined in the absence of timolol in either the SHR or the WKY. Forskolin and DBcAMP also antagonized the alpha adrenoceptor-mediated contractions. However, these antagonisms were not significantly different between the two strains. The cellular cAMP content in arterial strips after the stimulation with NE was significantly less in the SHR than in the WKY, whereas the cAMP contents were similar in arterial strips from both strains which were stimulated with forskolin. These results suggest that the reduced function of Gs is involved in the abnormality of beta adrenoceptor-AC system in the SHR femoral artery.

1-Methyl-3-isobutylxanthine↗
PubMed1988

A new T cell subset expressing B220 and CD4 in lpr mice: defects in the response to mitogens and in the production of IL-2.

Autoimmune-prone mice homozygous for the lpr gene develop prominent lymphadenopathy composed mainly of Thy-1+ CD8- CD4- B220+ cells. Expression patterns of B220 vs CD4 on lymph node cells from lpr mice were analysed using two-colour flow microfluorometry. B220+CD4+ cells, which were hardly seen in lymph nodes of B6-+/+ mice, increased significantly in B6-lpr mice with ageing. Functional analysis of purified B220+ CD4+ cells from lpr mice revealed that these cells scarcely responded to T cell mitogens with or without rIL-2. Furthermore, B220+ CD4+ cells were defective in IL-2 production when cultured with Con A. On the other hand, B220-CD4+ cells from B6-lpr mice showed an ability to respond to T cell mitogens similar to that of B220- CD4+ cells from B6-+/+ mice. These results indicate that an unusual T cell subset expressing both B220 and CD4 in lpr mice is functionally defective, but the intrinsic ability of B220-CD4+ cells is almost intact as compared with the counterpart in normal mice.

Aging↗
PubMed1988

[A fatal case of skin cancer 18 years after cadaveric donor renal transplantation].

A 39 years old man was admitted with chronic renal failure, and in June, 1968, he received renal transplantation from a cadaveric donor, 30 years old male, died from cerebral contusion. The graft had been well functioning for 18 years, and he had been engaged in his occupation. At the age of fifty-seven skin cancer (squamous cell carcinoma) occurred in his face, and surgical treatment was carried out. But it metastasized to left parotid, bilateral lungs and other areas of skin, and he died in October 1986, complicated by cerebral infarction. Some reports mention that the incidence of malignancies in posttransplant patients is greater than in general population. Our study reveals that immunological surveillance is weakened in posttransplant patients because of decrease of NK activity, but that the incidence of malignancies (male 0.78%, female 0.45%) is not so great as that reported. There is need of further investigation in this problem.

Adult↗