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Biomedical subjects

M Toriyama

Publications and source records attributed to M Toriyama.

At least 37 records · Page 2Linked to original sources

Such hydrophobic peptides as dansylated mastoparan can elevate the fertilization membrane of sea urchin eggs.

Melittin is known to be a major hydrophobic peptide component in honeybee venom that can cause as much elevation of fertilization membrane of sea urchin eggs as normal fertilization. The action of melittin has been thought to be closely related with its ability to facilitate the phospholipase A2 activity on the eggs. However, another peptide "mastoparan" from wasp venom was not found here to cause any elevation of the membrane, although it can activate the enzyme as well as melittin. On the other hand, mastoparan was found to get the membrane-elevating activity only when its amino groups were modified with hydrophobic substituents. N epsilon-Substituted mastoparan with a dansyl group in Lys11 residue was most effective among the analogs examined here. Our findings indicate that the facilitation of phospholipase by the peptides have little relation with the membrane generation. Such hydrophobic moiety as the dansyl group in the peptides must cause the cortical reaction on the eggs in cooperation with peptide moiety. The dansylated peptide will be a useful tool to induce the artificial fertilization of sea urchin eggs.

Amino Acid Sequence↗

Purification of a low molecular weight microtubule binding protein from sea urchin eggs.

A low molecular weight microtubule binding protein(SU-MAP34) was purified from sea urchin eggs. This protein bound strongly to the microtubule formed from purified echinoderm tubulin but showed no cross-linking of microtubules. Monospecific antibody against SU-MAP34 was produced and an immunoblotting analysis showed that this protein was not a breakdown product of a protein of a higher molecular mass. Whole cell staining and confocal laser scanning microscope observation showed that SU-MAP34 localized on the filamentous structure of mitotic apparatus and this structure was identified as the microtubule with double staining using anti-SU-MAP34 and anti-tubulin. An immunoblotting experiment showed an enrichment of SU-MAP34 in a microtubule protein fraction prepared using taxol from a crude extract of the cell.

Animals↗

[Histopathological studies on paranasal mucosa from patients treated with erythromycin].

Low-dose and long-term administration of erythromycin (EM therapy) has been reported to be very effective for patients with intractable chronic sinusitis including sinobronchial syndrome. However, we sometimes encounter patients whose sinusitis is extremely resistant to EM therapy. Therefore, the present study was carried out to determine the correlation between the clinical efficacy of EM therapy and the histopathological features of the ethmoidal mucosa and nasal polyps of patients treated with erythromycin. Patients with significant lymphocytic infiltration in the submucosal area responded well to EM therapy. Furthermore, patients with neutrophilic infiltration within and beneath the ciliary epithelium tended to show improvement with this therapy. On the other hand, EM therapy was minimally effective in patients whose subepithelial layer showed marked eosinophilic infiltration. In patients showing excellent and good clinical results, histological changes in the paranasal mucosa after therapy were as follows: a decrease in the number of inflammatory cells, reduced interstitial edema, increased fibrosis and normalization of the secretory glands. From these results, we conclude that erythromycin may suppress chronic inflammation except for an allergic reaction characterized by marked eosinophilic infiltration.

Adolescent↗

Purification of tropomyosin from bovine adrenal medulla and its inhibitory effect on the actin severing activity of adseverin (adrenal medulla 74 kDa actin severing protein).

Tropomyosin was purified from a gelation product produced in the crude extract of bovine adrenal medulla using several column chromatographies mixed with a boiling treatment. Purified tropomyosin was a mixture of major isoforms with low molecular weights and minor isoforms of high molecular weights. Purified tropomyosin showed a dose-dependent protective effect on actin filaments against the severing activity of adseverin derived from adrenal medulla, suggesting its important role in the regulation of cell shape and structure in vivo.

Actin Cytoskeleton↗

Purification of a sea urchin calpactin like protein which is excluded from the mitotic apparatus region.

A calpactin like protein (SUAX34) was purified from fertilized sea urchin eggs. Ca2+ dependent binding to and EGTA induced elution from a phenyl column were used to detect and isolate the calpactin like protein. SUAX34 showed monomeric nature having a molecular mass of 34,000. Immunoblotting of the cell extract showed that closely positioned two bands were reactive to the antibody against SUAX34 and immunostaining experiments showed specific exclusion of these proteins from the mitotic apparatus of metaphase eggs.

Animals↗

Erythromycin therapy for otitis media with effusion in sinobronchial syndrome.

Chronic sinusitis is frequently associated with chronic lower respiratory tract diseases, and the association is referred to as sinobronchial syndrome (SBS). This study was carried out to determine the incidence of otitis media with effusion in the patients with sinobronchial syndrome and to investigate the efficacy of low-dose and long-term erythromycin therapy for otitis media with effusion associated with sinobronchial syndrome. We have found a high incidence of otitis media with effusion in patients with sinobronchial syndrome, the morbidity rate being 54% in 50 cases studied. This ear disease seems to be the major cause of hearing disturbance in patients with sinobronchial syndrome. Sixteen patients with both sinobronchial syndrome and otitis media with effusion were given low-dose and long-term erythromycin therapy (erythromycin base, 600 mg/d for more than 4 months); of these, 13 became effusion-free and most subjects showed improvement in the symptoms of sinobronchial syndrome. The erythromycin therapy thus seems to be exceedingly effective for the treatment of sinobronchial syndrome and associated otitis media with effusion.

Administration, Oral↗

Chronic tonsillitis and IgA nephropathy. Clinical study of patients with and without tonsillectomy.

To determine whether tonsillectomy is a significantly effective treatment in the clinical course of IgA nephropathy, we did a comparative study on 50 patients with IgA nephropathy and chronic tonsillitis. We divided the patients into two groups: 35 patients with and 15 without tonsillectomy (control group). With or without tonsillectomy, renal function became progressively worse during the follow-up period in most patients with a serum creatinine level of > 1.4 mg/dl at the time of renal biopsy. In patients with a serum creatinine level of < or = 1.4 mg/dl, renal function remained normal in all subjects with tonsillectomy, but worsened in 3 patients out of 13 without tonsillectomy. Improvement in proteinuria/hematuria was found more frequently in the tonsillectomized group than in the controls. Furthermore, the serum IgA level was significantly reduced after tonsillectomy, especially in patients showing improvement. From these results we conclude that tonsillectomy was effective for patients with IgA nephropathy complicated by tonsillitis when the operation was performed before deterioration of renal function.

Adult↗

Prognostic factors for persistent middle ear effusion after acute otitis media in children.

The present study was carried out to determine the clinical course of acute otitis media (AOM) in children and to analyze the risk factors that lead to persistent effusion. One hundred and twenty children aged 9 months to 10 years diagnosed as having AOM were included in this study. Sixty-two children (52%) recovered from AOM without middle ear effusions within a month. In 26 cases (22%), middle ear effusions resolved after 1 to 3 months, while 32 children (26%) had persistent effusions for more than 3 months after the onset of acute inflammation. Statistical analysis between the group showing quick recovery and that with persistent effusion was carried out in relation to various clinical factors at the onset. The significant risk factors were: younger age, bilateral AOM, presence of otorrhea, tympanogram type B or C2 on an AOM ear. Moreover, the most significant prognostic factor was a tympanogram type B or C2 on an opposite ear at the acute onset. On the other hand, factors such as sex, fever, history of otitis media, season at onset or complications did not correlate with the duration of persistent effusion. From these results we conclude that careful examination should be performed not only on the ear with AOM but also on the opposite ear in order to predict the course of otitis media.

Acoustic Impedance Tests↗

[Growth rate analysis of pulmonary metastases in giant cell tumor of bone].

We reviewed six cases with pulmonary metastases in giant cell tumor of bone. The growth rate and the doubling time of pulmonary metastases was measured and compared to the clinical course. The six cases were classified into two each of the following three types: 1) spontaneous remission, 2) continuously slow growing and 3) rapid growing. A favorable factor was a doubling time of more than 80 days in pulmonary metastases. A poor prognosis was indicated by a doubling time of around 30 days. Aggressive primary lesions should be excised widely.

Adolescent↗

A novel 24-kDa microtubule-associated protein purified from sea urchin eggs.

Chromatographic fractionation of a crude extract of sea urchin eggs on a hydrophobic column enabled us to find a new 24-kDa microtubule-associated protein (SU-MAP24) that bound tightly to the column and was eluted under alkaline conditions. Biochemical studies using the purified protein showed its direct binding to microtubules reconstituted from tubulin purified from starfish sperm outer fibers. SU-MAP24 promoted tubulin polymerization in a dose-dependent manner. Immunoblotting analysis showed that SU-MAP24 is present in a microtubule protein fraction obtained from a crude extract using taxol, and immunostaining of paraffin-sectioned metaphase eggs showed its localization in the mitotic apparatus. These results show that SU-MAP24 is a newly identified microtubule-associated protein.

Alkaloids↗

Erythromycin inhibition of lipopolysaccharide-stimulated tumor necrosis factor alpha production by human monocytes in vitro.

The mechanism of clinical effectiveness of low-dose and long-term erythromycin (EM) treatment for diffuse panbronchiolitis, sinobronchial syndrome, and associated otitis media with effusion was investigated by studying the effects of EM on tumor necrosis factor alpha (TNF-alpha) production by cultured human monocytes stimulated with lipopolysaccharide. At concentrations of 0.1 microgram/mL or more, EM inhibited TNF-alpha release from human monocytes stimulated by lipopolysaccharide in a dose-dependent manner. Of the other macrolides tested, roxithromycin, an EM derivative, also showed significant inhibition of TNF-alpha production, whereas josamycin failed to inhibit TNF-alpha release from monocytes. Nonmacrolidic drugs such as minocycline hydrochloride, ofloxacin, or penicillin G had no significant effect on TNF-alpha production. These results suggest that the clinical improvement of chronic respiratory diseases by EM may depend on the suppression of production of inflammatory cytokines such as TNF-alpha.

Anti-Bacterial Agents↗

[Human monocytes show chemotaxis in response to cholesteatoma debris].

Using a microchamber technique, we tested cholesteatoma debris and certain of its constituents for effects on the migration of human peripheral blood monocytes and polymorphonuclear leukocytes. Cholesteatoma debris induced significant migration of monocytes. When the individual constituents of cholesteatoma debris, i.e., alpha-keratin, cholesterol, lauric acid and lipopolysaccharides, were tested for monocyte chemotaxis, only alpha-keratin induced significant monocyte migration. alpha-keratin extracted from the cholesteatoma debris with 8 M urea also induced migration of monocytes with a bell-shaped dose-response curve, which is frequently encountered with chemoattractants. Therefore, cholesteatoma debris and one of its components, alpha-keratin, are potent chemoattractants for human monocytes. On the other hand, cholesteatoma debris showed no significant chemotactic effect on polymorphonuclear leukocytes. Based on the present and our previous results, cholesteatoma debris acts on monocytes/macrophages as a strong chemotactant, a potent activating (priming) factor, and an inducer of production of tumor necrosis factor, which is a bone-resorbing cytokine. Therefore, we concluded that macrophages induced by cholesteatoma debris may play an important role in the pathogenesis of bone resorption in cholesteatoma otitis.

Chemotaxis, Leukocyte↗

Two Ca2(+)-binding proteins in the mitotic apparatus of sea urchin eggs.

Two Ca2(+)-binding proteins of sea urchin eggs were purified and partially characterized. They showed Ca2(+)-dependent binding to actin filaments and Ca2(+)-dependent changes of fluorescence intensity which was used to estimate the affinity constant of these proteins to Ca2+ ions. Ca2+ ions did not increase phospholipid binding ability of these proteins. Therefore these proteins are distinguished from the calpactin family. Staining of sections of metaphase eggs embedded in paraffin showed their localization in the mitotic apparatus. Furthermore, staining of whole mount eggs with anti-tubulin and antibodies against these proteins, followed by observations with confocal laser-scanning microscopy showed their co-localization with microtubules more clearly. In vitro co-sedimentation assay of microtubules with these proteins, however, showed no interaction between them. This suggested that some structures surrounding the mitotic apparatus microtubules are responsible for their localization.

Actin Cytoskeleton↗

The mitotic apparatus-associated 51-kDa protein from sea urchin eggs is a GTP-binding protein and is immunologically related to yeast polypeptide elongation factor 1 alpha.

We investigated the biochemical characteristics of the 51-kDa protein that is a major mitotic apparatus-associated basic protein of sea urchin eggs (Toriyama, M., Ohta, K., Endo, S., and Sakai, H. (1988) Cell Motil. Cytoskeleton 9, 117-128). The amino acid composition of the 51-kDa protein was apparently different from those of tubulin, actin, histones, and myelin basic protein; yet it was similar to those of polypeptide elongation factors 1 alpha (EF-1 alpha). In addition, antibody to EF-1 alpha from yeast cross-reacted with the 51-kDa protein. [3H] GTP binding activity was detected in the phosphocellulose-purified fraction (PC fraction) which predominantly contained the 51-kDa protein and was shown to be specific to GTP, GDP, guanylyl imidodiphosphate, and ITP. Photo-affinity labeling using [alpha-32P]8-azidoguanosine triphosphate (8-azido-GTP) demonstrated that a 51-kDa polypeptide in the PC fraction specifically bound 8-azido-GTP. This GTP-binding polypeptide was bound to a GTP affinity column, could be eluted by the addition of GTP, and was immunoreactive with anti-51-kDa protein antibodies. When the PC fraction was applied to a gel filtration chromatography column, GTP binding activity was completely coeluted with the 51-kDa protein. Furthermore, the PC fraction and the gel filtration-purified fraction had EF-1 alpha activity: [14C]Phe-tRNA transferring activity to ribosomes in the presence of poly(U) and ribosome-dependent GTPase activity. The results indicate that the mitotic apparatus-associated 51-kDa protein is a GTP-binding protein and suggest that it is structurally and functionally related to yeast EF-1 alpha.

Affinity Labels↗

Purification and characterization of a Ca2+-dependent actin filament severing protein from bovine adrenal medulla.

We describe the purification of an actin regulatory protein from bovine adrenal medulla. This protein caused a dose-dependent decrease of the specific viscosity of actin solution within 30 s of its addition in a Ca2+-sensitive way. Sedimentation assays and the observation by electron microscopy showed that this effect was ascribable to the fragmentation of actin filaments. This protein apparently promoted nucleation of actin polymerization and increased the critical concentration of actin for polymerization nearly 5-fold, suggesting its binding to the barbed end of actin filaments. The inhibitory effect of this protein on the elongation of actin from the barbed end of the myosin subfragment S1-labeled actin seeds confirmed this suggestion. These properties are similar to those of gelsolin. However, the physicochemical properties of this protein having a single polypeptide chain with a molecular weight of 74,000, a Stokes radius of 3.9 nm, a sedimentation coefficient (s0(20),w) of 4.5 S, and an immunological characterization showed that this protein is different from gelsolin.

Actin Cytoskeleton↗

Tropomyosin in the sea urchin egg cortex.

Tropomyosin was purified from the Triton-treated cortex fraction of fertilized sea urchin egg. Egg tropomyosin showed characteristics typical of nonmuscle tropomyosins such as low molecular mass, short periodicity of Mg2+-paracrystals, low lysine/arginine ratio, high Mg2+ requirement in binding to F-actin, in addition to the properties of all tropomyosins, namely, stability to high temperature, anomalous migration of SDS/urea gel, dissociation from F-actin under high ionic conditions and very acidic isoelectric point. Co-sedimentation assay of egg tropomyosin with actin in the presence of the previously purified high-molecular-mass actin binding protein (260-kDa protein) showed that these two proteins bind to actin filaments in a non-competitive manner. This suggested that both the proteins play a cooperative role in the formation of actin-filament-based cytoskeletal structure in the cortex.

Actins↗

Human leukocyte antigen in childhood unilateral deafness.

Human leukocyte antigen frequencies were examined in 40 Japanese patients with severe unilateral sensorineural hearing loss found in childhood. Human leukocyte antigen-Bw54 was significantly increased in frequency. No statistically significant deviation in the Bw54 frequency was observed among several possible subgroups studied. Although this group of patients may have various causes, and underlying mechanism is still unclear, this preliminary study suggests that childhood unilateral deafness may be associated with a genetically determined factor(s).

Child↗