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Biomedical subjects

Y Okano

Publications and source records attributed to Y Okano.

At least 343 records · Page 19Linked to original sources

In vitro morphological study of cochlear epithelium.

1. The mammalian cochlear epithelium was successfully kept alive in long-term tissue cultures. The Na and K concentration of the culture medium was similar to that of perilymph. 2. The most vulnerable cells in the culture epithelium were the outer and inner hair cells followed by the inner zones cells and the interdental cells of the spiral limbus. The difference in disintegration time between the outer and inner hair cells was about 48 hrs under the same culture conditions. The same phenomenon was noted between the rows of outer hair cells. 3. When the outer hair cells were not supported by Deiters' cells or when the cochlear epithelium was separated between Deiters' cells and Hensens cells, the outer hair cells degenerated quickly. 4. The interdental cells appeared to be relatively highly differentiated and showed much activity. No evidence of secretory function of interdental cells could be obtained. 5. The inner sulcus cells, Hensens cells, Claudius' cells and Reissners membrane cells were flattened and arranged like covering epithelium.

Animals↗

Effect of the high potassium medium on cultured cochlear epithelial cells.

1. A high potassium and low sodium culture medium does not produce morphological change in cochlear epithelial cells such as sulcus cell, Deiters' cell, Hensen's cell, Claudius' cell and Raissner's membrane cell. 2. Interdental cells of spiral limbus are affected by a high potassium medium but the change is to a moderate degree. 3. Outer hair cells degenerate quickly in a high potassium and low sodium medium. The onset and the grade of the degeneration is in a parallel with the potassium concentration, and a concentration of more than 46.4 mEq/l produces an acute and severe degeneration of outer hair cells.

Animals↗

Molecular basis for phenotypic heterogeneity in galactosaemia: prediction of clinical phenotype from genotype in Japanese patients.

We identified 14 mutations in 15 Japanese subjects from 13 families with galactose-1-phosphate uridyltransferase (GALT) deficiency using denaturing gradient gel electrophoresis (DGGE) and direct sequence analysis. These mutations accounted for 22 (96%) of 23 mutant alleles in 15 Japanese subjects. The mutational spectrum included nine missense mutations (M142V, G179D, A199T, R231H, W249R, N314D, P325L, R333Q, and R333W), two deletions (L275fsdelT and Q317fsdelC), a nonsense mutation (W249X), and two splicing mutations (V85-N97fsdel38bp and IVS4nt+1). Ten of the 14 mutations have not been reported in Caucasians. Differences in frequency and spectrum of GALT mutations suggest that the mutations may have occurred after racial divergence of Caucasians and Asians. The Duarte variant in Japanese was associated with the N314D mutation, g.1105G > C, g.1323G > A, and g.1391G > A (SacI -) polymorphisms, as in Caucasians. The Duarte variant may have occurred before racial divergence, and was an ancient mutation. In vitro GALT activities of nine missense mutations were determined by a COS cell expression system, and indicated between 1.3% and 35% of wild-type control. Patients with R333Q (29% in vitro GALT activity) or A199T (35%) showed mild clinical phenotypes, i.e. no ovarian failure or neurological deterioration. Genotype determination is useful for predicting biochemical and clinical phenotypes in classic galactosaemia, and can be of further help in managing patients with this disorder.

Adolescent↗

Gonadal function in patients with testicular germ cell tumors.

The gonadal function of 18 patients with testicular germ cell tumors was evaluated. Seminal parameters after orchiectomy were examined in 15 patients. Six of them were available for follow-up observation after 2 or 3 courses of adjuvant chemotherapy. Serum gonadal hormones before and after orchiectomy were evaluated in 7 patients (testosterone and PRL were not examined in one patient). Five of 15 (33.3%), 8 of 15 (53.3%), 13 of 15 (86.7%), 7 of 13 (53.8%), and 9 of 12 (75.0%) had abnormal values in seminal volume, sperm concentration, motility, morphology, and vitality, respectively. The sperm concentration gradually improved after chemotherapy following orchiectomy in 5 of 6 (83.3%) patients. In all the patients examined, serum levels of follicular stimulating hormone (FSH), luteinizing hormone (LH), and prolactin (PRL) increased after orchiectomy. Serum levels of testosterone increased in 4 patients, but decreased in 2 after orchiectomy. These findings suggest that several factors, including preexisting intrinsic defect and disturbance of the hypothalamus-pituitary-gonadal axis, are involved in the deterioration of gonadal function in patients with testicular germ cell tumors.

Adult↗

Processing and secretion of lysosomal acid alpha-glucosidase in Tetrahymena wild type and secretion-deficient mutant cells.

The proteolytic processing and secretion of a lysosomal enzyme, acid alpha-glucosidase, was studied by pulse-chase labeling with [35S]methionine in Tetrahymena thermophila CU-399 cells treated with ammonium chloride. This cell secreted a large amount of acid alpha-glucosidase into the cultured medium during starvation. The secretion was found to be repressed by addition of ammonium chloride (NH4Cl). Acid alpha-glucosidase was produced as a precursor form (108 kDa) and then processed to a mature polypeptide (105 kDa) within 60 min. This mature enzyme was secreted into the media within 2-3 h after chase, whereas the precursor form was not secreted by either control cells or NH4Cl-treated cells. NH4Cl did not affect the processing of the precursor acid alpha-glucosidase. Processing profile of this enzyme was apparently indistinguishable from that of the mutant MS-1 defective in lysosomal enzyme secretion. Furthermore, the purified extracellular (CU-399) and intracellular (MS-1) acid alpha-glucosidases were the same in molecular mass (105 kDa) and enzymatic properties. They contained no mannose 6-phosphate residues in N-linked oligosaccharides. These results suggested that unlike mammalian cells, Tetrahymena acid alpha-glucosidase may be transferred to lysosomes by a mannose 6-phosphate receptor-independent mechanism, and also that low pH was not essential for the proteolytic processing of precursor polypeptide.

Ammonium Chloride↗