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Biomedical subjects

S Ohno

Publications and source records attributed to S Ohno.

At least 541 records · Page 30Linked to original sources
PubMed1994

Rapid detection of human seminal plasma proteins by membrane aspiration test (MAT).

A new dot-immunobinding method, called the membrane aspiration test (MAT), was developed for the detection of human seminal plasma (HSP) antigens. Biological materials, such as macerated vaginal swabs or extracts from seminal stains, were dotted on a nitrocellulose membrane in a disposable aspiration kit. After the drying and aspiration of a quenching solution, a suspension of gold particles (GP) coated with an immune serum to HSP was aspirated through the nitrocellulose membrane. The dotted portion on the membrane turned red after several repeated aspirations, if HSP antigen was present. Results can be obtained within a few minutes. This test has broad applicability because of its rapidity and simplicity.

Animals↗
PubMed1994

Purification and characterization of 3 alpha/beta-hydroxysteroid dehydrogenase from mature porcine testicular cytosol.

NADPH-dependent 3 alpha/beta-hydroxysteroid dehydrogenase (3 alpha/beta-HSD) was purified to apparent homogeneity from testicular cytosol of mature pigs. The purified enzyme catalyzes the conversion of 5 alpha-dihydrotestosterone (5 alpha-DHT) to both 5 alpha-androstane-3 alpha,17 beta-diol and 5 alpha-androstane-3 beta,17 beta-diol. The molecular weight of the enzyme was estimated to be 31 kDa by SDS-polyacrylamide gel electrophoresis and 40 kDa by gel filtration chromatography indicating that the native 3 alpha/beta-HSD is a monomer. The isoelectric point of the purified enzyme was found to be 6.2 by density gradient isoelectric focusing and 6.4 by chromatofocusing. The enzyme reduced both 5 alpha- and 5 beta-DHT, 5 alpha- and 5 beta-dihydroprogesterone, 5 alpha- and 5 beta-dihydrocortisol, prostaglandin E2, 13,14-dihydro-15-keto-prostaglandin E2 and 13,14-dihydro-15-keto-prostaglandin F2 alpha. Moreover, the enzyme caused rapid reduction of other carbonyl compounds including aldehydes, ketones and quinones. The rates of reduction of these compounds are fast relative to the rates of reduction of steroids and prostaglandins. The purified enzyme was inhibited by AgNO3, SH-reagent, quercetin, hexesterol, stilbestrol, disulfiram and divalent cation such as Cu2+, Hg2+ and Cd2+. The two enzymes show certain similarities (e.g. molecular weight, cross-reactivity to a common antibody) and certain striking differences (e.g. pI, effects of various inhibitors and greater enzyme activity towards steroids (neonatal form) or prostaglandins (mature form). Reasons are give for suggesting that these enzymes are closely related to carbonyl reductase.

3-Hydroxysteroid Dehydrogenases↗
PubMed1994

The cardinal principle of like attracting like generates many ubiquitous oligopeptides shared by divergent proteins.

Actual protein amino acid sequences are very different from random assemblages of 20 varieties of amino acids. The separate survey of 20 unrelated proteins in two steps that included eight of the 18 discussed in this paper, revealed that at the level of 5000 total residues, one out of every 32 tetrapeptides appeared in two or more identical copies, whereas at the level of 10,000 total residues, the frequency was elevated to one out of every 29. It would thus appear that only 60,000 or so, out of the possible 160,000 (20(4)) varieties of tetrapeptides, are regularly used by all proteins. These shall be defined as ubiquitous tetrapeptides. Those tetrapeptides occasionally found to be stray which did not belong to the above group of 60,000 must have been generated by new mutations. Thus, they are expected to return to the group by subsequent mutations. The above ubiquity is due to the cardinal principle of protein construction which is like attracting like. On the average, 28% of each residue is devoted to the formation of homodipeptides such as Leu-Leu, Asn-Asn and Trp-Trp. Consequently, homo-oligopeptides, pentapeptidic and longer, are readily found in two or more proteins unrelated to each other. The next in line among the ubiquitous oligopeptides are those made of similar residues. They usually contain palindromic cores such as Leu-Val-Leu, Ala-Gly-Ala and Lys-Arg-Lys.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1994

Molecular genetic studies of HLA class II alleles in sarcoidosis.

Previous HLA serological studies showed positive associations of the DR52 antigen, the DR52-associated antigens (DR3, DR5 and DR6) and the DR8 antigen with sarcoidosis. To investigate the HLA alleles that may contribute to the genetic susceptibility to sarcoidosis at the DNA level, HLA-DRB1, -DRB3, -DQA1 and DQB1 genotyping using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was performed in 63 Japanese patients with sarcoidosis. The frequencies of the DR52-associated DRB1 alleles (DRB1*11, DRB1*12 and DRB1*14 except DRB1*1302), DRB1*08, DRB3*0101, DQA1*0501 and DQB1*0301 were significantly increased in patients compared with healthy controls. The significant increase of DRB3*0101, DQA1*0501 and DQB1*0301 could be explained by linkage disequilibrium with the DR52-associated DRB1 alleles. It must be noted that the DR8 haplotype, which does not possess the DRB3 gene, also showed a significant increase in sarcoidosis. These results suggest that the HLA-alleles responsible for the susceptibility to sarcoidosis are located at the HLA-DRB1 locus rather than the HLA-DRB3, -DQA1 and -DQB1 loci. In contrast, DRB1*1302 may confer resistance to the disease.

HLA-DQ Antigens↗
PubMed1994

The eta isoform of protein kinase C is localized on rough endoplasmic reticulum.

The eta isoform of protein kinase C, isolated from a cDNA library of mouse skin, has unique tissue and cellular distributions. It is predominantly expressed in epithelia of the skin, digestive tract, and respiratory tract in close association with epithelial differentiation. We report here that this isoform is localized on the rough endoplasmic reticulum in transiently expressing COS1 cells and constitutively expressing keratinocytes. By the use of polyclonal antibodies raised against peptides of the diverse D1 and D2/D3 regions, we found that immunofluorescent signals were strongest in the cytoplasm around the nucleus and became weaker toward the peripheral cytoplasm. Under immunoelectron microscopic examination, electron-dense signals were located on the rough endoplasmic reticulum and on the outer nuclear membrane which is continuous with the endoplasmic reticulum membrane. However, no signals were detected in the nucleus, inner nuclear membrane, smooth endoplasmic reticulum, Golgi apparatus, mitochondria, or plasma membrane. Treatment of the cells in situ with detergents suggested association of the isoform of protein kinase C with intracellular structures. By immunoblotting, a distinct single band with an M(r) of 80,000 was detected in whole-cell lysate and in rough microsomal and crude nuclear fractions, all of which contain outer nuclear membrane and/or rough endoplasmic reticulum. We further demonstrated the absence of a nuclear localization signal in the pseudosubstrate sequence. The present observation is not consistent with the report of Greif et al. (H. Greif, J. Ben-Chaim, T. Shimon, E. Bechor, H. Eldar, and E. Livneh, Mol. Cell. Biol. 12:1304-1311, 1992).

Amino Acid Sequence↗
PubMed1994

HLA-DRB1 typing of Vogt-Koyanagi-Harada's disease by PCR-RFLP and the strong association with DRB1*0405 and DRB1*0410.

Vogt-Koyanagi-Harada's (VKH) disease is reported to be closely associated with the HLA class II antigen, HLA-DR4. Serologically defined DR4 is further divided into 11 alleles by molecular HLA genotyping. However, no study of HLA-DNA typing of VKH patients has been reported. To clarify molecular genetic mechanism underlying the susceptibility/resistance to VKH disease, HLA-DNA typing of DR antigens (DRB1 genotyping) by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was performed. It was found that DRB1*0405 showed a significant association with VKH disease compared with the healthy controls (corrected p value < 1 x 10(-5)) and that all the patients had DRB1*0405 and/or DRB1*0410. The specific amino acid residue shared only by these two alleles is Ser at position 57 which is located in the antigen binding groove and may influence the immunological function as an antigen-presenting molecule, suggesting that Ser at position 57 plays an important role in the susceptibility to VKH disease, although the possibility that the involvement of the HLA-DQ molecule, DQ4, in strong linkage disequilibrium with DRB1*0405 and DRB1*0410, cannot be excluded.

Gene Amplification↗
PubMed1994

Erythrocytosis complicated by multiple paraganglioma.

We report the case of a 22-year-old woman with onset of erythrocytosis at the age of 9 years. Endocrinological and radiological examinations revealed an elevated catecholamine level and the presence of multiple abdominal tumors. After the removal of the tumors, the catecholamine level normalized, whereas erythropoietin remained at the same level and erythrocytosis persisted. The tumor lysate contained considerable amounts of catecholamine but not erythropoietin. Moreover, no erythropoietin mRNA was detected in the tumor by in situ hybridization. These data suggest that this paraganglioma did not produce erythropoietin. A review of the literature showed the existence of patients with early-onset erythrocytosis complicated with paraganglioma, whose erythrocytosis was not relieved even after the resection of paraganglioma.

Abdominal Neoplasms↗
PubMed1994

Differential expression of protein kinase C genes in cultured mast cells derived from normal and mast-cell-deficient mice and mast cell lines.

We investigated the expression of mRNA of protein kinase C (PKC) isozymes (alpha, beta, gamma, delta, epsilon, xi, and eta) in cultured mast cells (CMC) derived from normal (+/+) mice, CMC derived from genetically mast-cell-deficient (W/W, Wv/Wv, and mi/mi) mice, and murine mast cell lines (IC2, MC9, and P-815) by Northern blotting. In +/+ CMC, abundant expression of PKC delta and moderate expression of PKC alpha and beta was observed, while other PKCs (types gamma, epsilon, xi, and eta) were not detected. In vivo expression of PKC delta was demonstrated in the skin by in situ hybridization. In mast cell lines, the expression pattern of PKC isozymes was similar to that of +/+ CMC, except that the expression of PKC eta was detected in the IC2 cell line. The expression levels of PKC delta in CMC derived from c-kit-deficient mutants, W/W, Wv/Wv, and mi/mi, were lower than that of +/+ mice. These results indicate that PKC delta is the main isozyme in various types of murine mast cells and also suggest that the reduced level of PKC delta expression in mutant mice may be caused by a deficit in the signal transduction system through c-kit receptor.

Animals↗
PubMed1994

Synthesis and structure-activity relationships of new (5R,8R,10R)-ergoline derivatives with antihypertensive or dopaminergic activity.

A series of new (5R,8R,10R)-ergoline derivatives was synthesized, and their antihypertensive and dopaminergic activities were tested in conscious spontaneously hypertensive rats and in rats with unilateral 6-hydroxydopamine-induced lesions of the substantia nigra. (5R,8R,10R)-6-Alkyl-8-ergolinemethanols, prepared from the corresponding ergolinecarboxylates, were converted to the tosylates, which were treated with various five-membered heterocycles containing nitrogen atoms to afford the new ergolines. (5R,8R,10R)-8-(1,2,4-Triazol-1-ylmethyl)-6-methylergoline (4s, maleate: BAM-1110) exhibited potent dopaminergic activity, about 18-fold greater than that of bromocriptine mesylate. (5R,8R,10R)-8-(1,2,4-Triazol-1-ylmethyl)-6-propylergoline (8b, fumarate: BAM-1602) showed extremely potent dopaminergic activity, being about 220 and 1.15 times more active than bromocriptine mesylate and pergolide mesylate, respectively. Several compounds exhibited potent antihypertensive activity. Structure-activity relationships for antihypertensive and dopaminergic activities are discussed.

Animals↗
PubMed1994

Synthesis and structure-activity relationships of new (5R,8S,10R)-ergoline derivatives with antihypertensive or dopaminergic activity.

A series of new (5R,8S,10R)-ergoline derivatives was synthesized, and their antihypertensive and dopaminergic activities were evaluated in conscious spontaneously hypertensive rats and in rats with unilateral 6-hydroxydopamine-induced lesions of the substantia nigra, respectively. (5R,8S,10R)-6-Methyl-8- ergolinemethanols, prepared from the corresponding ergolinecarboxylates, were converted to the tosylates, which were treated with various five-membered heterocycles containing nitrogen atoms to afford the new ergolines. (5R,8S,10R)-8-(1-Imidazolylmethyl)-6-methylergoline (5a, BAM-2101) and (5R,8S,10R)-2-bromo-6-methyl-8-(1,2,4-triazol-1-ylmethyl) ergoline (7c, BAM-2202) exhibited potent antihypertensive activities. The maximum falls of systolic blood pressure after oral administration of 5a and 7c at 3 mg/kg were 95 and 132 mmHg, respectively, while those of cianergoline, bromocriptine mesylate, hydralazine, and nifedipine at the same dose were 40, 37, 47, and 49 mmHg, respectively. The durations of significant antihypertensive effects of these compounds except nifedipine were more than 7 h. None of the ergolines exhibited potent dopaminergic activity. Structure-activity relationships are discussed.

Animals↗
PubMed1994

Peripheral intrapulmonary hamartoma accompanied by a similar endotracheal lesion.

We describe a case of peripheral intrapulmonary hamartoma accompanied by a similar endotracheal lesion. A 70-year-old asymptomatic man had a polypoid lesion in the trachea and a nodular shadow in a peripheral lung area. A biopsy specimen from the trachea revealed a chondroid hamartoma, and the surgically resected tissue of the intrapulmonary tumor showed the same histologic findings. This is the second case of multiple hamartomas showing coexistence of tracheobronchial and parenchymal tumors.

Aged↗
PubMed1994

[A tentative plan of measuring the amount of the stretch of the skin using origin on body surface].

The present study was made to search for the origin on human dorsal surface, which is available for a correct grasp of a change of body surface (skin surface) with human activities and to try to re-evaluate, using the above origin, quantitative skin deformation of human dorsal surface on raising arms. The following results were obtained: 1) X. examination in standing (static) and arm-raising (dynamic) positions revealed a site with inter-positional difference, as zero, in slippage on dorsal median line. This site was supposed to be the origin and was confirmed to be Th10 and Th12. The palpation of the site of, Th10 and Th12 on dorsal median line was thought to be simple and effective in searching for a site with the least bone and skin slippage, thus in presuming the origin. 2) Based upon the origins on dorsal median line and scapular line parallel thereto in sitting position, 5 cm phi-correct circles were stamped up and down, then arm raising action was taken. The utilization of the origin enabled us to determine the length, directional angle, theta, and area, S, of the slippage between correct and distorted circles as obtained by these actions. 3) For arm raising, the skin slipped up and down on median line for lower and higher sites than the origin, respectively, with the angle theta on scapular line indicating the + direction, while and theta appeared greater and smaller for a nearer site to the origin, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Arm↗
PubMed1994

Diffuse panbronchiolitis and rheumatoid arthritis: a possible correlation with HLA-B54.

We analyzed the HLA antigens in two patients with rheumatoid arthritis accompanied by diffuse panbronchiolitis in whom HLA typing could be done and found the presence of B54 and DR4 alleles. We previously reported that the frequency of HLA-B54 is significantly increased in patients with diffuse panbronchiolitis. This allele demonstrates a linkage disequilibrium with DR4 (or DR4.1) in Japanese individuals. The association of rheumatoid arthritis with HLA-DR4 is well established in various ethnic groups including Japanese people. Since both diseases show the same pattern of HLA correlation, more Japanese patients with these two diseases are likely to be encountered.

Arthritis, Rheumatoid↗
PubMed1994

Lyme disease spirochetes in a wild fox (Vulpes vulpes schrencki) and in ticks.

Lyme disease spirochetes were demonstrated in a wild female fox (Vulpes vulpes schrencki) and in Ixodes persulcatus ticks collected from the fox on Sapporo, Hokkaido, Japan. Spirochetes were detected in I. persulcatus, as well as skin lesions, brain, heart, kidney, and liver of the fox. Five of seven isolates reacted with a monoclonal antibody against Borrelia afzelii specific Osp B. Deoxyribonucleic acid (DNA) relatedness of a brain isolate was 89% to B. afzelii, and ranged from 50 to 67% to three other species. Immunoglobulin G antibodies to B. afzelii, B. garinii and B. burgdorferi sensulato, when tested in an enzyme-linked immunosorbent assay (ELISA), were negative in the fox. There were no antibodies against seven serovars of Leptospira interrogans.

Animals↗
PubMed1994

Cholesterol sulfate, a novel activator for the eta isoform of protein kinase C.

Activity of protein kinase C depends on the interaction with polar head-groups of two membrane lipids, i.e., phosphatidylserine and diacylglycerol. In the present study, we demonstrated a novel activation mechanism of the eta isoform of protein kinase C (nPKC eta), which is predominantly expressed in epithelial tissues in close association with epithelial differentiation. We found that the nPKC eta was activated by cholesterol sulfate, a metabolite of cholesterol formed during squamous differentiation. This activation was greater than that by phosphatidylserine plus phorbol ester; the Vmax for the activation by cholesterol sulfate was 3.6 times that by phosphatidylserine plus phorbol ester, while Kms were almost equal. In the presence of cholesterol sulfate, phorbol ester only weakly enhanced the activity of nPKC eta. Activation of nPKC eta by cholesterol sulfate was further demonstrated by autophosphorylation of the kinase molecule. However, the alpha and delta isoforms of protein kinase C were not activated by cholesterol sulfate. The present observation affords a new insight into a signal transduction pathway of squamous differentiation.

Amino Acid Sequence↗