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Biomedical subjects

H Yamane

Publications and source records attributed to H Yamane.

At least 73 records · Page 4Linked to original sources

Induction of macrophage exudation in the inner ear by OK432 treatment.

OK432, a heat- and penicillin-treated lyophilized powder of a low virulent Su strain of Streptococcus pyogenes, was injected into the perilymphatic space of guinea pigs in order to determine the kinetics of exudation of inflammatory cells into the cochlea over a 7-day period. OK432 induced exudation of many neutrophils and Asialo GM1-positive cells into the scala tympani. The numbers of these cells peaked on the first day after OK432 treatment, and then gradually decreased. Four days after treatment very few of either of these types of cells were observed in the scala tympani. Asialo GM1-positive cells were confirmed to be activated macrophages. In ears treated with physiological saline or with the additive included in OK432 in control studies, no notable changes in the inner ear were recognized. These findings suggest that the inner ear can undergo induction of macrophage rapidly exudation by OK432 treatment as has previously been observed for other organs.

Animals↗

Local substances regulating cochlear blood flow.

The regulation of cochlear lateral wall blood flow was investigated in rats using histochemistry, immunohistochemistry and transmission electron microscopy. The contractile protein, tropomyosin (TM) was localized in the pericyte around the vessels of spiral ligament but little was seen around strial capillaries. Prostaglandin I2 (PGI2) is a vasodilatory substance produced by PGI2 synthase (PGI2S), which is present in the endothelium of the vessels of spiral ligament but not much is present around strial capillaries. Endothelial nitric oxide synthase (eNOS) was present in the endothelium of both the vessels of spiral ligament and strial capillaries. These findings indicate that the blood flow of the cochlear lateral wall is regulated mainly in the spiral ligament by locally produced substances and that strial blood flow is not significantly regulated by pericytes responsible for enlargement or constriction of capillaries.

Animals↗

Surgical treatment of cancer extending from the retromolar fossa to the masticator space.

Cancer extending from the retromolar fossa to the masticator space with neck metastasis (T4N2M0, Stage IV) was surgically removed en bloc by maxillectomy using a mandibular swing technique. This approach allowed a wide surgical view and was useful as handling the external and internal pterygoid muscles and upper neck dissection could be combined. Though there have been several surgical approaches to the masticator space, in the case of tumors which are commonly malignant there, this approach is useful and satisfactory.

Carcinoma, Squamous Cell↗

Study of tuberculosis in the field of otorhinolaryngology in the past 10 years.

Over the 10 years from 1988 to 1997, a total of 18 cases of tuberculosis of the head and neck (8 of the cervical lymph node, 5 of the larynx, 2 of the salivary glands, and 1 each of the hypopharynx, tongue and middle ear) were diagnosed in our department. Four cases were diagnosed in 1996 and 8 in 1997, indicating a rapid increase over these 2 years. Definitive diagnosis was based on histopathological examination in 14 cases, detection of M. tuberculosis in 1 case and exclusion diagnosis, mainly using the tuberculin skin test, in 3 cases. The length of time from the first examination to obtaining a definitive diagnosis was 14.2 +/- 6.5 days in five subjects in whom tuberculosis was suspected from the beginning or in whom tuberculosis was suggested by the physician in making a differential diagnosis, 25.3 +/- 17.2 days in the group in whom malignant tumour was suspected and 64.3 +/- 57.3 days in other cases. Although no significant difference was found (p = 0.077), the length of time until diagnosis tended to be shorter in the cases that were suspected of having tuberculosis from the beginning. Family history and past history of tuberculosis, contact with a tuberculous patient and abnormal findings at chest X-ray were examined as background factors, and some of these were present in nine out of the 18 cases. In the five cases diagnosed in the early stage, some of the background factors were found in four cases. However, no significant difference in the time until definitive diagnosis was found between the groups with or without background factors in all cases (p = 0.675). These results suggest the importance of considering tuberculosis when performing differential diagnoses.

Adolescent↗

Apoptotic cell death in Kikuchi's disease: a TEM study.

The pathogenesis of Kikuchi's disease (Kikuchi-Fujimoto disease, histiocytic necrotizing lymphadenitis) remains unclear. However, some previous studies have suggested that a disorder in cellular immunity is responsible for this disease, and apoptotic cell death appears to be the principal finding in the histogenesis of this disease. In the present study, a lymph node from a female patient with Kikuchi's disease was examined by transmission electron microscopy (TEM). TEM revealed specific morphological features of apoptotic cells, such as nuclear chromatin condensation and fragmentation along the nuclear membrane with intact organelles, and the presence of histiocytes phagocytosing karyorrhectic debris (apoptotic bodies) in areas affected by Kikuchi's disease. Although neither the role nor the trigger of apoptosis in Kikuchi's disease has been clearly determined, our findings show that apoptotic cell death clearly plays a role in the pathogenesis of Kikuchi's disease.

Adult↗

Differential effects of LPS and CD40 ligand stimulations on the induction of IL-12 production by dendritic cells and macrophages.

In this experiment, we have examined the IL-12 production of murine macrophages (Mphi) and dendritic cells (DC) in response to LPS and CD40 ligand (CD40L) stimulations. Splenic Mphi and DC were purified by sorting on the basis of Mac-1 and CD11c expression and stimulated with LPS or Chinese hamster ovary cells expressing CD40L. The results showed that the ligation of CD40 induced the enhancement of IL-12 p40 mRNA accumulation and IL-12 production in DC as well as in Mphi; however, neither the accumulation of IL-12 p40 mRNA nor the production of bioactive IL-12 was detected in DC stimulated with various concentrations of LPS, although Mphi produced IL-12 on LPS stimulation. There was a remarkable difference in the expression of LPS receptor CD14 between Mphi and DC. Mphi evidently expressed CD14 but the CD14 expression of DC was quite low. Possible mechanisms of the failure in IL-12 production of DC are discussed.

Animals↗

Cloning and characterization of genes encoding an enzyme which oxidizes dimethyl sulfide in Acinetobacter sp. strain 20B.

Acinetobacter sp. strain 20B was isolated based on the ability to utilize dimethyl sulfide as the sole sulfur source. Since strain 20B oxidized indole as well as dimethyl sulfide, indigo production by recombinant Escherichia coli clones carrying Acinetobacter DNA was used as a selection for cloning genes encoding dimethyl sulfide oxidation genes. The gene encoding an indole-oxidizing enzyme was also found to oxidize dimethyl sulfide. The dimethyl sulfide-oxidizing enzyme genes consisted of six open reading flames designated dsoABCDEF. The deduced amino acid sequences of dsoABCDEF were homologous with those of the multicomponent phenol hydroxylases. DsoABCDEF oxidized dimethyl sulfide to dimethyl sulfoxide, and dimethyl sulfoxide to dimethyl sulfone.

Acinetobacter↗

The product of a gas6 splice variant allows the release of the domain responsible for Axl tyrosine kinase receptor activation.

The product of gas6 (Gas6) is a growth factor with high level of similarity to protein S and was identified as the ligand for Axl family of tyrosine kinase receptors. Gas6 contains an N-terminal gamma-carboxylated domain (Gla), four epidermal growth factor like domains and a large C-terminal D region. An alternative Gas6 spliced form (Gas6SV) having an additional 43 amino acids between fourth EFG like and D domain was characterised. Here we show data indicating that Gas6SV is specifically cleaved within the inserted sequence, thereby splitting the D domain from the remaining part of the protein. The resulting two proteolytic products of 36 kDa and 50 kDa were separated and the 50 kDa fragment corresponding to region D was shown to be responsible for Axl receptor activation. Furthermore a deletion mutant of Gas6 containing only the D domain was shown to similarly activate Axl receptor phosphorylation unequivocally demonstrating that D domain can act as a signalling molecule. The possible roles of the proteolytic processing of Gas6SV in the regulation of growth factor availability are discussed.

Alternative Splicing↗

Subunit Ya-specific glutathione peroxidase activity toward cholesterol 7-hydroperoxides of glutathione S-transferases in cytosols from rat liver and skin.

Dermal 7alpha- and 7beta-hydroperoxycholest-5-en-3beta-ols (cholesterol 7alpha- and 7beta-hydroperoxides), regarded as good aging markers in the rat (Ozawa, N., Yamazaki, S., Chiba, K., Aoyama, H., Tomisawa, H., Tateishi, M., and Watabe, T. (1991) Biochem. Biophys. Res. Commun. 178, 242-247), were reduced in the presence of glutathione (GSH) with concomitant formation of GSSG by cytosol from rat liver in which no detectable level of the hydroperoxides had been demonstrated to occur. The GSH peroxidase (GSH Px) activity toward the toxic steroid hydroperoxides was exerted to almost the same extent by both Alpha-class GSH S-transferases (GSTs), Ya-Ya and Ya-Yc, and by selenium-containing GSH Px (Se-GSH Px) in rat liver cytosol. None of three Mu-class GSTs, Yb1-Yb1, Yb1-Yb2, and Yb2-Yb2, and a Theta-class GST, Yrs-Yrs, from rat liver and a Pi-class GST, Yp-Yp, from rat kidney showed any appreciable GSH Px activity toward the hydroperoxides. The subunit Ya-bearing GSTs and Se-GSH Px purified from rat liver cytosol showed marked differences in apparent specific activity toward the cholesterol hydroperoxides (GSTs Ya-Ya > Ya-Yc >> Se-GSH Px). However, a kinetic study indicated that Se-GSH Px had a higher affinity for steroid hydroperoxides than did the GSTs, so that Se-GSH Px could catalyze the reduction of lower concentrations of cholesterol 7-hydroperoxides with approximately equal Vmax/Km values to those by the GSTs. Rat skin had no GST bearing the subunit Ya but contained only a very low concentration of Se-GSH Px, possibly resulting in the accumulation of cholesterol 7-hydroperoxides in the skin but not in the liver. From rat skin cytosol, GSTs Yc-Yc, Yb1-Yb1, Yb1-Yb2, Yb2-Yb2, and Yp-Yp were isolated, purified to homogeneity, and identified with the corresponding GSTs from liver and kidney. The GSTs accounted for 0.23% of total skin cytosolic protein, and the most abundant isoform of skin GSTs was Yb2-Yb2, followed by Yc-Yc, Yp-Yp, Yb1-Yb1, and Yb1-Yb2 in decreasing order.

Animals↗

Cell death caused by the acute effects of aminoglycoside and zinc in the ampullary cristae of guinea pigs.

We reported that apoptosis occurred in the guinea pig vestibular hair cells after chronic aminoglycoside treatments. In the present study, we used in situ nick-end labeling to determine whether apoptosis was also induced by the acute effects of aminoglycosides in guinea pig ampullar cristae. In addition, we evaluated the effect of zinc supplements upon these ototoxic treatments. After a local application of streptomycin directly to the round window, we found labeled bodies in the vestibular hair cells. The zinc supplement increased the number of labeled bodies resulting in severe hair cell loss. These findings indicate that the acute effects of aminoglycosides also induce apoptosis of the vestibular hair cells, and that zinc enhances aminoglycoside ototoxicity. Consequently, we propose that an interaction with ion channels may play a key mechanism in the processes of apoptosis affecting the vestibular hair cells.

Administration, Topical↗

Gentamicin ototoxicity induced apoptosis of the vestibular hair cells of guinea pigs.

To clarify mechanisms of inner ear cell death induced by aminoglycosides, we used an in situ nick-end labelling method to examine guinea pig vestibular epithelia after chronic systemic treatments with gentamicin to produce apoptosis. Such changes occurred in damaged hair cells, suggesting that this process may be crucial for subsequent repair and cell regeneration.

Animals↗

Nitric oxide synthase and contractile protein in the rat cochlear lateral wall: possible role of nitric oxide in regulation of strial blood flow.

The present study demonstrated by histochemical and immunohistochemical methods that NADPH diaphorase reactivity, endothelial nitric oxide synthase (eNOS)-like immunoreactivity, and tropomyosin-like immunoreactivity, were located within the rat cochlear lateral wall. Both NADPH diaphorase reactivity and eNOS-like immunoreactivity were found mainly in the endothelium of the strial capillaries (ESC) and that of the vessels of the spiral ligament (ESL). These reaction products appeared to be somewhat more common in the ESC than in the ESL. On the other hand, tropomyosin-like immunoreactivity was localized in tissues outside the endothelium and its intensity was greater in the ESL than in the ESC. These findings suggest that nitric oxide (NO) produced by eNOS may play a role in regulating the blood flow of the cochlear lateral wall. In addition, NADPH diaphorase reactivity, eNOS-like immunoreactivity, and tropomyosin-like immunoreactivity showed different patterns of distribution between ESC and ESL. This suggests that in these two sites blood circulation is controlled by NO through two different mechanisms that are suitable for regulating strial blood flow.

Animals↗

Triggers of hair cell regeneration in the avian inner ear.

We performed an in vitro study in order to determine possible triggers of hair cell regeneration in the chick basilar papilla following degeneration. We compared the response of sensory epithelium damaged by collagenase treatment with that damaged by acoustic trauma. The former exhibited no proliferative activity, but the latter did. The basilar papillae damaged by acoustic trauma could have proliferating activity in medium containing fetal bovine serum (FBS) or epidermal growth factor (EGF) but not in the medium without FBS or EGF. These findings indicate that regeneration of basilar papillae depends on the manner of cell death and that FBS or EGF is required for regeneration.

Animals↗

Identification of stylar RNases associated with gametophytic self-incompatibility in almond (Prunus dulcis).

Stylar proteins of 13 almond (Prunus dulcis) cultivars with known S-genotypes were surveyed by IEF and 2D-PAGE combined with immunoblot and N-terminal amino acid sequence analyses to identify S-RNases associated with gametophytic self-incompatibility (SI) in this plant species. RNase activities corresponding to Sa and Sb, two of the four S-alleles tested, were identified by IEF and RNase activity staining. The Sa-RNase band reacted with the anti-S4-serum prepared from Japanese pear (Pyrus serotina); no reaction with the antiserum was observed with the Sb-RNase band. When the Sa-RNase band was excised from an IEF gel stained for RNase activity, subjected to SDS-PAGE, and detected by immunoblotting, it appeared that this band consisted of a single protein that reacted with the anti-S4-serum with M(r) of about 28 kDa. With 2D-PAGE and silver staining of the stylar extracts, all four S-proteins could be successfully distinguished from each other in the highly basic zone of the gel. Although Sb-, Sc-, and Sd-proteins had roughly the same M(r) of about 30 kDa, the Sc-protein seemed to be slightly smaller than the Sb-protein and slightly larger than the Sd-protein. In 2D-PAGE profiles as well, the Sa-protein had M(r) of about 28 kDa, apparently smaller than the other three proteins. A bud sport, in which one of the two S-alleles of the original cultivar is impaired, was visualized as a loss of Sc-protein, which is consistent with the previous pollination study. All four S-proteins reacted with the anti-S4-serum, probably because of the differing conformations of these S-proteins in the IEF and 2D-PAGE gels. The Sa-protein in 2D-PAGE appeared to be identical to Sa-RNase in IEF; both had the same M(r) and were reactive with the anti-S4-serum. N-terminal amino acid sequence analysis of the four S-proteins revealed that they were highly homologous to each other and similar to the S-RNases of Malus, Pyrus, Scrophulariaceae, and Solanaceae. Taken together, RNases in the style are strongly suggested to be associated with the gametophytic SI of almond. This is the first report identifying and characterizing S-RNase in almond.

Amino Acid Sequence↗

Cloning of genes involved in carbazole degradation of Pseudomonas sp. strain CA10: nucleotide sequences of genes and characterization of meta-cleavage enzymes and hydrolase.

The DNA fragment encoding meta-cleavage enzymes and the meta-cleavage compound hydrolase, involved in carbazole degradation, was cloned from the carbazole-utilizing bacterium Pseudomonas sp. strain CA10. DNA sequence analysis of this 2.6-kb SmaI-SphI fragment revealed that there were three open reading frames (ORF1, ORF2, and ORF3, in this gene order). ORF1 and ORF2 were indispensable for meta-cleavage activity for 2'-aminobiphenyl-2,3-diol and its easily available analog, 2,3-dihydroxybiphenyl, and were designated carBa and carBb, respectively. The alignment of CarBb with other meta-cleavage enzymes indicated that CarBb may have a non-heme iron cofactor coordinating site. On the basis of the phylogenetic tree, CarBb was classified as a member of the protocatechuate 4,5-dioxygenase family. This unique extradiol dioxygenase, CarB, had significantly higher affinity and about 20-times-higher meta-cleavage activity for 2,3-dihydroxybiphenyl than for catechol derivatives. The putative polypeptide encoded by ORF3 was homologous with meta-cleavage compound hydrolases in other bacteria, and ORF3 was designated carC. The hydrolase activity of CarC for 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid, the meta-cleavage compound of 2,3-dihydroxybiphenyl, was 40 times higher than that for 2-hydroxy-6-oxohepta-2,4-dienoic acid, the meta-cleavage compound of 3-methylcatechol. Alignment analysis and the phylogenetic tree indicate that CarC has greatest homologies with hydrolases involved in the monoaromatic compound degradation pathway. These results suggest the possibility that CarC is a novel type of hydrolase.

Amino Acid Sequence↗

Identification and characterization of genes encoding carbazole 1,9a-dioxygenase in Pseudomonas sp. strain CA10.

Nucleotide sequence analysis of the flanking regions of the carBC genes of Pseudomonas sp. strain CA10 revealed that there were two open reading frames (ORFs) ORF4 and ORF5, in the upstream region of carBC. Similarly, three ORFs, ORF6 to ORF8, were found in the downstream region of carBC. The deduced amino acid sequences of ORF6 and ORF8 showed homologies with ferredoxin and ferredoxin reductase components of bacterial multicomponent dioxygenase systems, respectively. ORF4 and ORF5 had the same sequence and were tandemly linked. Their deduced amino acid sequences showed about 30% homology with large (alpha) subunits of other terminal oxygenase components. Functional analysis using resting cells harboring the deleted plasmids revealed that the products of ORF4 and -5, ORF6, and ORF8 were terminal dioxygenase, ferredoxin, and ferredoxin reductase, respectively, of carbazole 1,9a-dioxygenase (CARDO), which attacks the angular position adjacent to the nitrogen atom of carbazole, and that the product of ORF7 is not indispensable for CARDO activity. Based on the results, ORF4, ORF5, ORF6, and ORF8 were designated carAa, carAa, carAc, and carAd, respectively. The products of carAa, carAd, and ORF7 were shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to be polypeptides with molecular masses of 43, 36, and 11 kDa, respectively. However, the product of carAc was not detected in Escherichia coli. CARDO has the ability to oxidize a wide variety of polyaromatic compounds, including dibenzo-p-dioxin, dibenzofuran, biphenyl, and polycyclic aromatic hydrocarbons such as naphthalene and phenanthrene. Since 2,2',3-trihydroxydiphenyl ether and 2,2',3-trihydroxybiphenyl were identified as metabolites of dibenzo-p-dioxin and dibenzofuran, respectively, it was considered that CARDO attacked at the angular position adjacent to the oxygen atom of dibenzo-p-dioxin and dibenzofuran as in the case with carbazole.

Amino Acid Sequence↗