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Biomedical subjects

M Fujiwara

Publications and source records attributed to M Fujiwara.

At least 487 records · Page 27Linked to original sources
PubMed1989

In situ Ia expression on brain cells in the rat: autoimmune encephalomyelitis-resistant strain (BN) and susceptible strain (Lewis) compared.

In order to examine in situ Ia expression on brain cells of various strains of rat, experimental autoimmune encephalomyelitis (EAE) was induced in both EAE-susceptible (LEW) and EAE-resistant (BN) strains. For induction of EAE in the resistant strain, two methods were applied: one was injection of guinea-pig myelin basic protein (GPBP) in complete Freund's adjuvant into LBNF1----BN chimeras; the other was transfer of GPBP-reactive T-line cells from BN rats into syngeneic rats. LBNF----BN chimeras developed clinical EAE, whereas BN rats that received T-line cells did not. However, histological EAE was apparent in both groups. Immunohistochemical examination using two different monoclonal antibodies (OX3 and OX6) against rat Ia antigens revealed that microglia of LEW, BN and chimera rats expressed Ia antigens in the central nervous system (CNS) with EAE. On the other hand, astrocytes were negative for Ia antigens in all the strains. Furthermore, quantitative analysis was undertaken in order to compare the density of Ia-positive microglia in the BN CNS with that in the LEW CNS. It was revealed that the density of Ia-positive microglia in the vicinity of perivascular inflammatory cell aggregates was essentially the same in both strains regardless of the difference in methods of EAE induction or histological severity of the disease. Ia-positive microglia remote from inflammatory cell aggregates were somewhat fewer in rats with mild histological EAE. However, no strain difference was noted in this analysis. Therefore, we concluded that in situ Ia-inducibility on the brain cells of EAE-resistant rats is not different from that of EAE-susceptible rats. Although Ia-positive microglia in both strains may be involved in the immune responses in the CNS, it is unlikely that the difference in Ia-inducibility on brain cells would contribute to strain-specific susceptibility to EAE.

Animals↗
PubMed1989

[Two cases of B-cell lymphoma occurring in a mother and her daughter at the same time].

Many cases of familial aggregation of lymphoproliferative malignancy have been reported. But familial aggregation of non-Hodgkin lymphoma is less frequent than Hodgkin's disease, Burkitt's lymphoma, and adult T-cell leukemia. Here we report familial B-cell lymphomas occurring in a mother and her daughter at the same time. The daughter (43 years old) was admitted because of fever. She died of a rapidly progressive neurological disturbance. The mother (75 years old) was admitted because of fever and pleural effusion, and then died with a complication of cryoglobulinemia. The histological findings of their lymph nodes revealed diffuse lymphoma, medium-sized cell type, with B-cell phenotype and mixture of cleaved and non-cleaved nuclei. They had no common chromosome abnormalities, while they shared HLA-A 2, B 35, and Cw 3 antigen. Viral infection was not identified, by routine serological test and electronmicroscopic study.

Adult↗
PubMed1989

Endothelium-dependent contractions induced by angiotensin I and angiotensin II in canine cerebral artery.

Whether angiotensin I and angiotensin II caused endothelium-dependent contraction was examined in canine cerebral arteries. In endothelium-intact preparations, angiotensin I and angiotensin II at 10(-8) to 10(-6) M caused dose-dependent contractions, whereas both angiotensins caused much less contractions in endothelium-removed preparations. The contractions induced by angiotensin I and angiotensin II were strongly attenuated by aspirin (cyclooxygenase inhibitor) (5 x 10(-5) M), OKY-046 [thromboxane (TX) A2 synthetase inhibitor] (10(-5) M) and ONO-3708 (TX)A2 antagonist) (5 X 10(-9) M). Captopril (10(-6) M) significantly attenuated the contractions induced by angiotensin I but not those induced by angiotensin II. Angiotensin I- and angiotensin II- induced contractions were inhibited markedly by Sar1, Ala8-angiotensin II (10(-9) and 10(-8) M). The present experiments demonstrate that angiotensin I and angiotensin II produce endothelium-dependent contraction in canine cerebral artery via a factor which appears to be TXA2. Angiotensin I may be converted by endothelial cells to angiotensin II, which may activate the cells to produce TXA2 in canine cerebral artery.

Angiotensin I↗
PubMed1988

Induction of 68,000-dalton heat shock proteins by cyclopentenone prostaglandins. Its association with prostaglandin-induced G1 block in cell cycle progression.

Cyclopentenone prostaglandins (PGs) such as PGA2 and delta 12-PGJ2 act specifically on cells in the G1 phase and induce block of cell cycle progression (Ohno, K., Sakai, T., Fukushima, M., Narumiya, S., and Fujiwara, M. (1988) J. Pharmacol. Exp. Ther. 245, 294-298). In this study, we characterized proteins induced by these PGs in HeLa S3 cells of synchronized growth and examined its association with the cell cycle block. HeLa S3 cells transiently expressed two 68-kDa proteins of isoelectric points of 5.5 and 5.6 in the G1 phase of cell cycle. When G1-enriched cells were incubated with either PGA2 or delta 12-PGJ2, synthesis of these proteins was markedly enhanced. Enhancement by delta 12-PGJ2 was persistent and irreversible, whereas that by PGA2 was reversible. delta 12-PGJ2 also enhanced the synthesis of two additional 68-kDa proteins with isoelectric points of 5.8 and 5.9. On two-dimensional gel electrophoresis, these proteins overlapped exactly with the 68-kDa heat shock proteins induced in cells treated at 43 degrees C for 90 min. They were also indistinguishable from the heat shock proteins in limited proteolysis. When delta 12-PGJ2 was incubated with G2/M phase cells, it induced only a small and transient increase in the 68-kDa proteins. These results suggest that cyclopentenone PGs extensively induce 68-kDa heat shock proteins in the G1 phase HeLa S3 cells and this induction is closely associated with the G1 block of cell cycle progression caused by these PGs.

Cell Cycle↗
PubMed1988

Substrate for botulinum ADP-ribosyltransferase, Gb, has an amino acid sequence homologous to a putative rho gene product.

Gb (b for botulinum) is an Mr 21,000-22,000 guanine nucleotide binding protein which is specifically ADP-ribosylated by an ADP-ribosyltransferase produced by types C1 and D Clostridium botulinum (Morii, N., Sekine, A., Ohashi, Y., Nakao, K., Imura, H., Fujiwara, M., and Narumiya, S. (1988) J. Biol. Chem. 263, 12420-12426). In this study, we have identified the partial amino acid sequence of this protein. Purified Gb was digested with various proteases, and the proteolytic fragments were separated by high performance liquid chromatography. Isolated peptides were subjected to protein sequencing by an automated pulse-liquid phase protein sequenator. Eight partial amino acid sequences consisting of a total of 105 amino acid residues were obtained from 16 peptides. Computerized homology analysis revealed that they are homologous to the sequence deduced from cDNA of the rho genes. About 88% of the amino acid residues was identical between the sequenced Gb fragments and the corresponding regions of the Aplysia rho gene product. The result strongly suggests that Gb is a putative rho gene product.

Adrenal Glands↗
PubMed1988

Purification and properties of the cytosolic substrate for botulinum ADP-ribosyltransferase. Identification as an Mr 22,000 guanine nucleotide-binding protein.

The substrate for ADP-ribosyltransferase from Clostridium botulinum was purified from the cytosol of bovine adrenal gland. Purification procedures consisted of ammonium sulfate fractionation, chromatographies on columns of DEAE-Sepharose and phenyl-Sepharose, gel filtration on a TSK-gel G3000SW column, and Mono Q fast protein liquid chromatography. On DEAE-Sepharose chromatography, the substrate activity was eluted in two separate peaks, and electrophoretic analyses revealed that the substrates in the two peaks are of similar molecular weight but different isoelectric points. The major peak of the substrate was further purified. It was purified about 1,800-fold with a recovery of 2.2% by the above procedures. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the final preparation showed a single protein band at Mr 22,000. The purified protein served as a substrate for botulinum ADP-ribosyltransferase and was maximally ADP-ribosylated to the extent of about 0.7 mol of ADP-ribose/mol of protein. A guanosine 5'-(3-O-thio)triphosphate (GTP gamma S) binding activity was co-purified with the ADP-ribosylation substrate, and the purified protein maximally bound about 0.5 mol of GTP gamma S/mol. GTP gamma S binding was effectively competed by GTP and GDP but not by GMP, ATP, and ADP. Thus, the ADP-ribosylation substrate is a GTP-binding protein. This protein, designated Gb (b for botulinum), is widely distributed in various tissues. It was rich in brain, pituitary, and adrenal glands, and poor in heart, smooth, and skeletal muscles.

Adenosine Diphosphate Ribose↗
PubMed1988

Subcellular distribution and isoelectric heterogeneity of the substrate for ADP-ribosyl transferase from Clostridium botulinum.

When the homogenate of bovine adrenal gland was subjected to subcellular fractionation, an Mr 21,000 substrate for botulinum ADP-ribosyl transferase was found not only in the membrane fractions but also in the cytosol; the amounts in the 10,000 x g precipitates and the 100,000 x g supernatant were about 21 and 56% of the total amount, respectively. Each fraction gave a single ADP-ribosylated protein band on SDS-polyacrylamide gel electrophoresis, but yielded on isoelectric focusing at least three bands between pH 5.5 and 6.0, suggesting the presence of multiple forms of the substrate of a similar molecular weight but different isoelectric points. ADP-ribosylated protein bands from the membrane and cytosol overlapped each other on both electrophoreses. After ammonium sulfate fractionation, the substrate from the cytosol showed requirement of divalent cations or guanine nucleotides for the reaction. Among cations tested, calcium, magnesium and manganese stimulated, whereas cadmium and lanthanum inhibited the reaction. Guanine nucleotides such as GTP, GDP and GTP-gamma-S also stimulated the substrate activity in the cytosol as that in the membrane fraction. However, no additive effects were observed when the nucleotides and cations were added together.

ADP Ribose Transferases↗
PubMed1988

Nicotinic acetylcholine receptors in the rat stomach: I. (-)-[3H]nicotine binding.

The nicotinic acetylcholine receptors in the rat stomach were characterized by means of a radioligand binding assay with (-)-[3H]nicotine as ligand. Saturation binding studies on the gastric fundus membranes revealed the presence of two binding sites with dissociation constant (KD) values of 3.1 and 289 nM, and maximum binding capacity (Bmax) values of 3.6 and 76 fmol/mg protein, respectively. The Bmax of the high affinity binding site was greatest in the cardia, followed by fundal mucosa, fundal muscle, and, finally antrum. The IC50 values of cholinergic drugs to inhibit (-)-[3H]nicotine binding in fundus membranes were as follows: (-)nicotine, 0.12 nM; cytosine, 9.3 nM; acetylcholine, 17.7 nM; carbachol, 700 nM; hexamethonium, 2270 nM. The IC50 values of alpha-bungarotoxin, d-tubocurarine and atropine were greater than 100 microM. The muscarinic acetylcholine receptors were also characterized with [3H]quinuclidinyl benzilate and the choline acetyltransferase activity was measured. These results suggest that nicotinic acetylcholine receptors as well as muscarinic acetylcholine receptors are present in the rat stomach and that the regional distribution of these receptors is uneven.

Animals↗
PubMed1988

Nicotinic acetylcholine receptors in the rat stomach: II. Alterations after vagotomy.

Nicotinic and muscarinic acetylcholine receptors were studied in the stomach of vagotomized rats, with ligand binding techniques, and with the concomitant measurement of choline acetyltransferase (ChAT). The maximum binding capacity (Bmax) of the high affinity sites of (-)-[3H]nicotine binding was significantly increased in all the regions of the stomach of subdiaphragmatically vagotomized animals while the values for the low affinity sites remained the same as those in sham-operated animals. On the other hand, [3H]quinuclidinyl benzilate binding to the stomach was unchanged after vagotomy. ChAT activity was significantly reduced in the vagotomized rats. These results suggest that surgical vagal denervation causes an increase in the density of nicotinic acetylcholine receptors presumably located on the parasympathetic ganglion cells. There is also a reduction of ChAT activity, probably due to a degenerative loss of preganglionic cholinergic neurons. Preganglionic denervation (decentralization) has little influence on the muscarinic receptors of postsynaptic effector organs.

Animals↗
PubMed1988

Tetraploidy in a 15-month-old girl.

We present a 15-month-old girl with tetraploidy and compare the manifestations with those of 3 previously reported liveborn infants with the same type of polyploidy. Common anomalies noted included micro-turricephaly, a prominent but narrow forehead, microphalmia or anophthalmia, limb anomaly, sacral meningomyelocele, and mental retardation.

Chromosome Banding↗
PubMed1988

Changes in benzodiazepine receptors in Alzheimer-type dementia.

We studied benzodiazepine receptors in brains obtained at autopsy from 7 normal controls and 7 patients with Alzheimer-type dementia, using 3H-flunitrazepam as a ligand. Compared with controls, 3H-flunitrazepam binding sites were significantly reduced in number in frontal and temporal cortex and hippocampus in the Alzheimer-type dementia brains. These results suggest that there are significant changes in the number of benzodiazepine receptors in selected brain regions of patients with Alzheimer-type dementia.

Aged↗
PubMed1988

Effects of goniopora toxin on the action potential and membrane currents of guinea-pig single ventricular cells.

The effects of Goniopora toxin (GPT), a polypeptide isolated from a coral, Goniopora spp., on action potential and membrane currents were studied in single ventricular cells of the guinea-pig using the whole-cell clamp technique with a single patch electrode. GPT at a concentration of 10 nmol/l prolonged the duration of the action potential without significant change in the resting membrane potential and action potential amplitudes. This prolongation became more evident at lower stimulus frequencies and persisted after washing with toxin-free solution. Tetrodotoxin (TTX, 1 mumol/l), but not Co2+ (2 mmol/l), abolished the prolonged action potential. Under voltage-clamp conditions, a sustained inward current, not present in the control, followed the transient inward current during depolarizing pulses in the GPT-treated cells. The current-voltage relationship for the sustained inward current was much the same as that for the fast sodium current reported in rat single ventricular cells (Brown et al. 1981). Both the sustained and transient currents were abolished by the shift of holding potential in the direction of depolarization and reappeared after repolarization; the reappearance of the sustained current was much slower than that of the transient current. TTX but not Co2+ abolished both the sustained and transient inward currents. Calcium current and time-independent current were not affected by GPT. Time-dependent outward current induced by large depolarizing pulses was attenuated by GPT.(ABSTRACT TRUNCATED AT 250 WORDS)

Action Potentials↗
PubMed1988

Effects of hypoxia on contractile responses of rabbit aortic strips to transmural electrical stimulation.

To clarify the effects of hypoxia on adrenergic transmission, we examined the contractile responses of isolated rabbit aortic strips to electrical stimulation, the concentration-response relationships for noradrenaline and KCl, and the electrical stimulation-evoked overflows of total [3H] and [3H]noradrenaline from strips preloaded with [3H]noradrenaline in media equilibrated with gas mixtures containing various concentrations of O2. Contractile responses to electrical stimulation were completely inhibited by tetrodotoxin and alpha-adrenoceptor antagonists such as phentolamine and phenoxybenzamine, but were not affected by indomethacin. When the concentration of O2 in the gas mixture was decreased from 95% to 20%, the contractile responses to electrical stimulation remained unchanged, but as the concentration of O2 was further decreased, the responses were inhibited concentration-dependently. At 0% O2, the response was inhibited by about 80% when compared with control values obtained at 95% O2, and the electrical stimulation-evoked overflows of total [3H] and [3H]noradrenaline into the superfusates were decreased by about 55%. At 0% O2, the concentration-response curve for exogenous noradrenaline was shifted to the right about 50-fold and the maximum response was decreased by 25%. The maximum contractile responses of aortic strips from animals pretreated with reserpine or 6-hydroxydopamine to high KCl were decreased slightly (about 15%). These results suggest that inhibition of adrenergic transmission under hypoxic conditions is mainly the result of a decrease in the stimulus-evoked release of noradrenaline and of a decrease in the affinity of alpha-adrenoceptor for noradrenaline and/or inhibition of signal transduction mechanisms, although hypoxia also causes a slight decrease in the contractility of vascular smooth muscle.

Animals↗
PubMed1988

Possible activation of intracellular beta-adrenoceptors by extraneuronally accumulated isoprenaline in perfused rat heart.

Two rat hearts were perfused in series by a modified Loewi's method. The recipient heart was perfused with the perfusate collected from the donor heart. 1. After perfusion with 3H-isoprenaline in the presence of tropolone, an inhibitor of catechol-O-methyltransferase, the donor heart was washed out with amine-free medium containing tropolone and corticosterone. The heart rate of the recipient heart increased after the change to the perfusate from the donor heart during the wash-out. After wash-out the heart rate of the donor heart (which had accumulated 43.4 pmol.g-1 3H-isoprenaline) was higher than that of the recipient heart (which had accumulated 0.44 pmol.g-1 3H-isoprenaline), and the rates of efflux of 3H-isoprenaline from both hearts were similar. 2. After perfusion with 3H-isoprenaline and corticosterone in the absence of tropolone, the enhanced heart rate of the donor heart decreased during wash-out with amine-free medium in the presence of corticosterone. The heart rate of the recipient heart increased after the medium change to the perfusate from the donor heart, and the heart rates in both hearts were similar after wash-out. Only small amounts of 3H-isoprenaline remained in both hearts after wash-out, and the rates of efflux of 3H-isoprenaline from both hearts were similar. 3. After perfusion with 3H-isoprenaline in the presence of tropolone, the effects of propranolol and atenolol on the heart rate during wash-out with amine-free medium containing tropolone and corticosterone were compared. The inhibitory effect of propranolol on the heart rate was significantly greater than that of atenolol.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1988

MR imaging of syringobulbia with giant syrinx.

Syringobulbia with a large syrinx was revealed by MR imaging. The syrinx extended from the medulla to vertebral level T10. We suspect that this giant syrinx may be produced by a congenital meningioma at C7, generating pulsatile flow in the original syrinx and causing a giant syrinx.

Aged↗