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Biomedical subjects

M Furuya

Publications and source records attributed to M Furuya.

At least 109 records · Page 6Linked to original sources

Structure-activity relationships of alpha-human atrial natriuretic peptide (alpha-hANP) analogs: role of charged groups in alpha-hANP.

To determine whether the addition of a methylene unit in the side chain of the Asp or Arg residue in alpha-human atrial natriuretic peptide (alpha-hANP) influences its biological activity, analogs of alpha-hANP, [Glu13]-alpha-hANP (7-28) (1), [Aad13]-alpha-hANP (7-28) (2), and [Harn]-alpha-hANP(7-28) (where n is any possible combination of 11, 14 and 27) (3-9), where the original Asp or Arg residue was replaced by a homo-amino acid, were synthesized by the solid-phase synthesis method. All the analogs were evaluated for their receptor binding, cyclic guanosine monophosphate (cGMP) accumulation activity in rat vascular smooth muscle cells (VSMC), and for vasorelaxant activity employing rat aorta. 1 and 2 were 0.9 and 0.03 times as potent as alpha-hANP (7-28), respectively, in binding. Har-containing analogs (3-9) were as potent as alpha-hANP (7-28) in binding. Among the Har-containing analogs, [Har11,14]-alpha-hANP (7-28) (6) and [Har11,27]-alpha-hANP (7-28) (7) were remarkably vasorelaxant active, being 4.2 and 5.3 times potent than alpha-hANP (7-28), respectively, in spite of relatively lower cGMP accumulation activity in the case of 7. The roles of the chargeable amino acid residues in biological activity are discussed.

Amino Acid Sequence↗

Andean leishmaniasis in Ecuador caused by infection with Leishmania mexicana and L. major-like parasites.

Between 1986 and 1988, epidemiologic studies were carried out in a small rural community in an Andean region of Ecuador, where cutaneous leishmaniasis is highly endemic. A total of 25 human cases, positive for Leishmania parasites by culture and/or smear, were examined. Fourteen of the cases were in infants less than one year of age, suggesting intradomiciliary transmission of the disease. Clinically, many of these cases were similar to descriptions of "uta," a form of cutaneous leishmaniasis which occurs in Andean regions of Peru and is reported caused by L. peruviana. Of the 11 positive cultures obtained from human cases in the present study, eight were identified by molecular characterization as L. mexicana and three were identified as L. major-like. Two additional isolates of L. mexicana were also made from an infected dog and from a sand fly, Lutzomyia ayacuchensis, living in the region, thus implicating the latter species as possible reservoir and vector, respectively, of L. mexicana in this highland community. The significance and validity of recent isolates of L. major-like parasites from the New World are also discussed.

Animals↗

Photomovement in Dunaliella salina: fluence rate-response curves and action spectra.

We determined the action spectra of the photophobic responses as well as the phototactic response in Dunaliella salina (Volvocales) using both single cells and populations. The action spectra of the photophobic responses have maxima at 510 nm, the spectrum for phototaxis has a maximum at 450-460 nm. These action spectra are not compatible with the hypothesis that flavo-proteins are the photoreceptor pigments, and we suggest that carotenoproteins or rhodopsins act as the photoreceptor pigments. We also conclude that the phototactic response in Dunaliella is an elementary response, quite independent of the step-up and step-down photophobic responses. We also determined the action spectra of the photoaccumulation response in populations of cells adapted to two different salt conditions. Both action spectra have a peak at 490 nm. The photoaccumulation response may be a complex response composed of the phototactic and photophobic responses. Blue or blue-green light does not elicit a photokinetic response in Dunaliella.

Cell Movement↗

A stimulating electrode easily attached to and detached from the phrenic nerve.

An electrode system is developed for temporary diaphragm pacing. It is divided into four parts: an electrode complex, a tube, a branch for a lead and a syringe. Stainless steel electrodes are put forward and pulled back by pressure produced by the syringe so that they can be easily attached to and detached from the phrenic nerve. Experiments show that the system works well, the nerve is successfully stimulated and no nerve damages are visually observed.

Animals↗

Rapid confirmation and revision of the primary structure of bovine serum albumin by ESIMS and Frit-FAB LC/MS.

Incorrectness of the amino acid sequence of bovine serum albumin (BSA) was suggested from the observed molecular weight of BSA obtained by electrospray ionization mass spectrometry (ESIMS). Lack of a tyrosine residue in the position of 156th was found rapidly, by the combination of frit-fast atom bombardment mass spectrometry/liquid chromatography (Frit-FAB LC/MS), automated Edman degradation and tandem mass spectrometry (MS). Then it turned out that BSA is composed of 583 amino acid residues, and that its average molecular weight is not 66267.1, and it is corrected to 66430.3. Moreover the amino acid sequence of the positions of 94th and 95th was corrected to -QE- by using automated Edman degradation method.

Amino Acid Sequence↗

Importance of hydrophobic residues in alpha-human atrial natriuretic peptide (alpha-hANP) for vasorelaxant activity.

To investigate the roles of the hydrophobic residues in the ANP molecule on biological activities, we synthesized a series of analogs containing various phenylalanine-homologs in position 8 or methionine-homologs in position 12. Among the analogs [pCl-Phe8]-alpha-hANP(7-28) was 4.8 times as potent as alpha-hANP(7-28) in cGMP accumulation and 3.5 times as potent in vasorelaxant activity. All the analogs showed nanomolar affinity to the receptor. In contrast, vasorelaxation and cGMP accumulation activity of the analogs ranged widely. These results suggest that these hydrophobic residues in the cyclic core are critical for vasorelaxant activity rather than for the "apparent receptor binding", and that these residues may possibly discriminate the "bioactive receptor" which is coupled to guanylate cyclase from the non-coupled receptor.

Amino Acid Sequence↗

Novel natriuretic peptide, CNP, potently stimulates cyclic GMP production in rat cultured vascular smooth muscle cells.

The newly identified peptide C-type natriuretic peptide (CNP) caused only a slight elevation of cGMP in rat renal glomeruli. In contrast, CNP potently increased cGMP levels in cultured rat vascular smooth muscle cells (VSMC) and stimulated guanylate cyclase activity in the particulate fraction of the cells. The extent of maximum activation of the enzyme induced by CNP was 4-fold higher than that by human atrial natriuretic peptide (alpha-hANP) while CNP was 4- and 16-fold weaker than alpha-hANP in binding affinity for the putative receptors on VSMC and vasorelaxant activity for rat aorta, respectively. These results indicate that CNP is a potent stimulator of cGMP formation in VSMC but not in glomeruli and pharmacological feature of CNP is distinct from that of ANP.

Amino Acid Sequence↗

Cooperative regulation of cytoplasmic streaming and ca fluxes by pfr and photosynthesis in vallisneria mesophyll cells.

In mesophyll cells of Vallisneria gigantea Graebner, Ca(2+) regulates the induction and cessation of cytoplasmic streaming. Streaming is induced when the level of calcium in the cytoplasm is lowered through light-accelerated release of Ca(2+) from the cells (S Takagi, R Nagai [1988] Plant Physiol 88: 228-232). We have now initiated an investigation on the nature of the photoreceptor(s) that are involved in the regulation of Ca(2+) movements across the cell membrane and of streaming. Streaming is induced only when phytochrome exists in the phytochrome-far redabsorbing form (Pfr)-and photosynthesis is allowed to take place for at least 4 minutes. The former effect is typically photoreversible by red and far-red light, and phytochrome is spectro-photometrically detectable in the crude extract from the leaves. The latter effect is assessed in terms of the wavelength dependency and the effects of diuron and atrazine, two inhibitors of photosynthesis. A similar requirement for Pfr and photosynthesis is found to be associated with the acceleration of Ca(2+) efflux in the protoplasts. The results suggest that phytochrome and photosynthetic pigment(s) cooperatively regulate cytoplasmic streaming via modulation of the Ca(2+) transport in the cell membrane.

Journal Article↗

Synthesis and biological property of alpha-human atrial natriuretic peptide analogs with a constrained or stereochemically modified cyclic moiety.

Conformationally restricted analogs of alpha-human atrial natriuretic peptide (alpha-hANP) containing L- or D-penicillamine, or D-cysteine in place of cysteine residues at positions 7 and 23 were synthesized by the liquid phase procedure. Their biological properties in the assays of receptor binding and cyclic guanosine monophosphate (cGMP) accumulation employing rat vascular smooth muscle cells (VSMC), vasorelaxant activity using rat isolated aorta were evaluated. We found that the constrained and/or stereochemically altered ring moiety generally did not influence the receptor binding activity, however, cGMP accumulation and vasorelaxant activities were quite sensitive to conformational perturbation. Furthermore, a lack of correlation between cGMP accumulation activity and vasorelaxant activity was observed. Dissociation between these activities was typical in the case of [DPen7,23]-alpha-hANP(7-28), which showed quite weak vasorelaxant activity in spite of its full cGMP accumulation and receptor binding potencies. This result suggests that cGMP accumulation alone is not sufficient to promote ANP-induced vasorelaxation, and that the other second messenger(s) may mediate this activity.

Amino Acid Sequence↗

[A clinicopharmacological study of serum sodium valproate free level in children with epilepsy].

The VPA total level (T value), the free level (F value), and the free fraction (FF) were measured in 74 epileptic children under valproic acid (VPA) monotherapy. The relationship between the T value, blood collection time, therapy duration, serum free fatty acid, clinical features, F value and FF were then studied. Blood was taken either before the morning (pre-breakfast) administration of medicine (Cmin), 2-3 hours after the post-breakfast administration (Cmax), or both. The subjects were divided into one Cmin. and one Cmax. group. The results of serum VPA measurement revealed that T values in the Cmin. group ranged from 23.0 micrograms/ml to 113.0 micrograms/ml (average: 50.0 +/- 16.2 micrograms/ml), F values from 2.0 micrograms/ml to 16.0 micrograms/ml (6.0 +/- 2.7 micrograms/ml) and FF from 5.9% to 21.7% (11.8 +/- 3.8%). In the Cmax. group, T values ranged from 41.0 micrograms/ml to 163.0 micrograms/ml (80.5 +/- 24.3 micrograms/ml), F values from 3.7 micrograms/ml to 22.8 micrograms/ml (10.0 +/- 4.2 micrograms/ml) and FF from 7.1% to 22.2% (12.2 +/- 3.3%). There was no difference in FF between the two groups. In both groups, T and F values significantly and positively correlated and FF was not affected by age or VPA therapy duration. However, FF varied in the early stage of medication. In individual cases with no seizures and receiving constant doses, the longer the period of medication, the greater the decrease in FF. Although free fatty acid was concurrently measured in some cases, it did not correlate with the F value or the FF. In 45 cases, changes in the FF were followed in both groups on the same day, but no general tendencies were noted. Diurnal fluctuation was studied in 4 cases. Significance of the clinical features was evaluated. The subjects were then classified by seizure type for group comparison and no differences in the FF among the different types were observed. During the follow-up period, 5 cases had seizures, but when their serum levels were compared with those of members of both groups, the T values did not differ. The F values and the FF in the 5 cases were below the mean values of the two groups. These findings suggest that when factors affecting VPA protein binding are expected to be present or when seizures cannot be controlled despite a sufficient T value in the blood, the F value measurement is of particular importance.

Adolescent↗

Linear alpha-human atrial natriuretic peptide analogs display receptor binding activity and inhibit alpha-hANP-induced cGMP accumulation.

We have synthesized a series of [Cys(R)7,23]alpha-hANP analogs, in which the two Cys residues were modified with various alkyl groups(R); i.e., R=Acm, Pe, Qe, Cam, Me, Ae, Bzl, Cm, Ocam and sulfo. The Acm-, Cam-, and Me-analogs exhibited binding activity as potent as alpha-hANP in rat vascular smooth muscle cells (VSMC). Binding activity of the analogs decreased progressively as the bulkiness of the R group increased. None of the analogs caused accumulation of cGMP in VSMC and vasorelaxant activity in rat aorta. Acm-, Cam- and Me-analogs substantially antagonized alpha-hANP-induced cGMP accumulation, but did not antagonize vasorelaxation induced by alpha-hANP in vitro.

Animals↗

The primary structure of human EGF produced by genetic engineering, studied by high-performance tandem mass spectrometry.

The primary structure of human epidermal growth factor (hEGF), which was produced by Escherichia coli using recombinant DNA technique, has been studied by tandem mass spectrometry. The molecular weight of hEGF (about 6200 amu) was determined by fast atom bombardment mass spectrometry. Then reduced and carboxymethylated hEGF was digested by chymotrypsin into seven peptides which could cover the whole sequence of hEGF. The amino acid sequences of five of these seven peptides could be confirmed by tandem mass spectrometry with or without isolation by high-performance liquid chromatography (HPLC). After isolation by HPLC, the other two peptides were digested with trypsin or thermolysin into small peptides, and sequenced by tandem mass spectrometry.

Amino Acid Sequence↗

Primary photoprocesses of phytochrome. Picosecond fluorescence kinetics of oat and pea phytochromes.

The primary photoprocesses of etiolated oat and pea phytochromes (Pr forms) are diffusion-modulated by the microscopic viscosity within the chromophore pocket. The chromophore pocket is preferentially accessible to glycerol but not to Ficoll. Glycerol preferentially retarded the rate (rate constant ca. 1-2 X 10(10) s-1) of the initial reaction from the Qy excited state of phytochrome, whereas it increased the long fluorescence lifetime (nanosecond) component that can be attributed to either an emitting intermediate or to modified/conformationally heterogeneous phytochrome populations. The picosecond time-resolved fluorescence spectra of different phytochrome preparations (i.e., full-length vs 6/10-kDa NH2-terminus truncated forms of phytochromes from monocot and dicot plants) revealed no significant differences. The spectra in the picosecond time scale showed no spectral shifts, but at longer time scales of up to approximately 1.90 ns, significant blue spectral shifts were observed. The shifts were more in the truncated than in the full-length pea phytochrome. Comparison of the fluorescence decay data and the picosecond time-resolved fluorescence spectra suggests differences in conformational flexibility/heterogeneity among the preparations of the monocot vs dicot phytochromes and the full-length native vs the amino terminus truncated phytochromes.

Kinetics↗

Molecular properties and biogenesis of phytochrome I and II.

Previously, phytochrome was thought to consist of a single molecular species. However, physiological and spectrophotometric evidence has accumulated to indicate that there are two phytochrome pools in tissues, one of which is predominant in dark-grown tissues and rather unstable in the light, and the other present in very low concentrations but stable, even in the Pfr form, irrespective of light condition. Recently, two immunochemically distinct phytochromes I and II, PI and PII, were found in both dark- and light-grown tissues, and their comparative amino acid sequences shown to be 64% homologous. This is crucial evidence for the presence of chemically different phytochrome apoproteins in a single plant species. However, it is still an open question as to which phytochrome, PI or PII, is a component of the photolabile and photostable pools of phytochrome. Our understanding of the molecular structure of phytochrome has been greatly improved by recent, rapid progress in the cloning and characterization of phytochrome genes. The expression of PI genes is photoreversibly inhibited by the photostable Pfr pool, while that of several other genes, like Cab, appears to be induced by PI in the Pfr form. It is suggested that autoregulation of phytochrome gene expression is not so simply governed in plants as thought earlier. If there are two different phytochromes in a plant cell, the most important physiological problem to be solved is which phytochrome triggers the numerous red/far-red reversible reactions reported in the literature. Photomorphogenetic mutants and transgenic plants with engineered phytochrome genes will probably help to solve this problem in the future, and preliminary work along this line has already introduced in this article. A model of the molecular structure of pea PI dimer was proposed on the basis of small angle X-ray scattering analysis, and the model then confirmed by rotary shadowing electron microscopy. Important questions are still open, such as: what is the nature of phytochrome's partner compounds in cells (phytochrome receptor)? How is/are the phytochrome-induced signal(s) transmitted in the signal transduction chain?

Amino Acid Sequence↗

Evidence for active interactions between microfilaments and microtubules in myxomycete flagellates.

We have previously observed the apparent displacement of microfilaments over microtubules in the backbone structure of permeabilized flagellates of Physarum polycephalum upon addition of ATP (Uyeda, T. Q. P., and M. Furuya. 1987. Protoplasma. 140:190-192). We now report that disrupting the microtubular cytoskeleton by treatment with 0.2 mM Ca2+ for 3-30 s inhibits the movement of the microfilaments induced by subsequent treatment with 1 mM Mg-ATP and 10 mM EGTA. Stabilization of microtubules by pretreatment with 50 microM taxol retarded both the disintegrative effect of Ca2+ on the microtubules and the inhibitory effect of Ca2+ on the subsequent, ATP-induced movement of the microfilaments. These results suggest that the movement of the microfilaments depends on the integrity of the microtubular cytoskeleton. EM observation showed that the backbone structure in control permeabilized flagellates consists of two arrays of microtubules closely aligned with bundles of microfilaments of uniform polarity. The microtubular arrays after ATP treatment were no longer associated with microfilaments, yet their alignment was not affected by the ATP treatment. These results imply that the ATP treatment induces reciprocal sliding between the microfilaments and the microtubules, rather than between the microfilaments themselves or between the microtubules themselves. While sliding was best stimulated by ATP, the movement was partially induced by GTP or ATP gamma S, but not by ADP or adenylyl-imidodiphosphate (AMP-PNP). AMP-PNP added in excess to ATP, 50 microM vanadate, or 2 mM erythro-9-[3-(2-hydroxynonyl)]adenine (EHNA) inhibited the sliding. Thus, the pharmacological characteristics of this motility were partly similar to, although not the same as, those of the known microtubule-dependent motilities.

Actin Cytoskeleton↗

Differential stimulation of rat lung particulate guanylate cyclase activity by atrial natriuretic peptide and sodium nitroprusside.

The effects of alpha-rat atrial natriuretic peptide (alpha-rANP) and sodium nitroprusside on the activity of rat lung particulate guanylate cyclase were examined. The particulate guanylate cyclase in partially purified rat lung membranes was stimulated by both alpha-rANP and nitroprusside. The effects of alpha-rANP and nitroprusside were, however, not additive. Diamide and N-ethylmaleimide almost completely abolished the nitroprusside-mediated stimulation, while they had only moderate effects on the alpha-rANP-mediated stimulation of the enzyme activity. ATP potentiated the enzyme stimulation by alpha-rANP, whereas it had no effect on the nitroprusside-mediated stimulation. These findings suggest that the stimulation of lung particulate guanylate cyclase activity by alpha-rANP and nitroprusside is mediated by different mechanisms.

Animals↗

[Efficacy of UFT in advanced gastric carcinoma under comparative study of MMC + UFT and MMC + tegafur therapies].

Antitumor effect of MMC + UFT(A) and MMC + tegafur(B) therapies for advanced gastric carcinoma was compared from February 1985 to March 1988. UFT and tegafur were orally given at dose of 400 mg/m2 daily, and MMC was intravenously administered at dose of 6-8 mg/m2 every two weeks. The following results were obtained. 1. Twenty-nine cases entered in this study were divided into A or B therapies at random. All cases entered were eligible. Fourteen of 29 cases were randomized into A therapy and 15 cases into B therapy. One case treated with B was evaluated as incomplete. 2. There were no differences in the characteristics of patients between A and B therapies. 3. Among 29 eligible cases, a partial response was obtained in 3 out of 14 cases (21.4%) treated with A and in 3 out of 15 cases (20.0%) treated with B. Among 28 cases evaluated completely, a partial response was obtained in 3 out of 14 cases (21.4%) treated with each A and B. 4. Response rate with every ps or for every lesion did not differ between A and B therapies. 5. Median survival day treated with A and B therapies was 223 and 181 days, respectively. The survival curve did not differ significantly between the two therapies. 6. The frequency of adverse reactions within eight weeks after beginning the therapy was 64.2 and 73.3% with A and B therapies, respectively. From the results mentioned, it was concluded that the antitumor effect of UFT was not superior to that of tegafur for advanced gastric carcinoma.

Adult↗