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Biomedical subjects

S Ohno

Publications and source records attributed to S Ohno.

At least 757 records · Page 42Linked to original sources
PubMed1989

Purification and characterization of pig adrenal 20 alpha-hydroxysteroid dehydrogenase.

Two distinct molecules of 20 alpha-hydroxysteroid dehydrogenase (20 alpha-HSD) from pig adrenal cytosol have been purified to homogeneity and characterized. Purification was achieved by ammonium sulphate precipitation followed by DEAE-cellulose column chromatography. 20 alpha-HSD activity was separated into two fractions including 20 alpha-HSD-I and 20 alpha-HSD-II, and these were further purified by affinity chromatography on 2',5'ADP-Sepharose, reactive red 120-agarose and some conventional column chromatography. Both enzymes catalyzed the reduction of 17 alpha-hydroxyprogesterone to 17 alpha, 20 alpha-dihydroxy-4-pregnen-3-one in the presence of NADPH as the preferred cofactor. The apparent Km and Vmax values against 17 alpha-hydroxyprogesterone were 26.2 microM and 1.3 nmol/min/mg for 20 alpha-HSD-I, and 118 microM and 19.4 nmol/min/mg for 20 alpha-HSD-II, respectively. Furthermore, remarkable differences between 20 alpha-HSD-I and 20 alpha-HSD-II were demonstrated under the influence of ionic strength, heat treatment and divalent cations. The molecular weight was estimated to be 39 kDa for 20 alpha-HSD-I and 40 kDa for 20 alpha-HSD-II by sodium dodecyl sulphate-polyacrylamide gel electrophoresis, and 30 kDa for both enzymes by gel filtration on Sephadex G-100. These were glycoproteins. There was a large difference in their isoelectric points (8.5 for 20 alpha-HSD-I and 5.5 for 20 alpha-HSD-II). The peptide mapping of the two distinct enzymes was greatly different although there was a slight difference in the amino acid composition.

20-Hydroxysteroid Dehydrogenases↗
PubMed1989

IgG1 plasmacytosis in interleukin 6 transgenic mice.

Interleukin 6 (IL-6) has been suggested to be involved in the pathogenesis of polyclonal and monoclonal plasma cell abnormalities. To address this possibility, transgenic mice carrying the human IL-6 genomic gene fused with a human immunoglobulin heavy chain enhancer were generated. High concentrations of human IL-6 and polyclonal increase in IgG1 (120- to 400-fold) in sera of all transgenic mice were observed. A massive plasmacytosis in thymus, lymph node, and spleen and an infiltration of plasma cells in lung, liver, and kidney were observed. However, the plasma cells were not transplantable to syngeneic mice and were found not to contain chromosomal aberrations including c-myc gene rearrangements. The evidence indicates that deregulated gene expression of IL-6 can trigger polyclonal plasmacytosis but cannot induce plasmacytoma. It is suggested that additional genetic changes may be required for the generation of plasma cell neoplasia. Other interesting findings in these transgenic mice were the development of mesangio-proliferative glomerulonephritis and an increase in megakaryocytes in bone marrow.

Animals↗
PubMed1989

Concordant evolution of coding and noncoding regions of DNA made possible by the universal rule of TA/CG deficiency-TG/CT excess.

The universal rule of TA/CG deficiency-TG/CT excess previously proposed as the construction principle of coding sequences applies to noncoding regions of the gene as well. Analysis of a 1989-base-long gene sequence for mouse immunoglobulin gamma 2a heavy-chain constant region as well as the 19,002-base-long gene sequence for human serum albumin revealed deficiency and overabundance of very similar sets of base trimers and tetramers in the coding and noncoding regions of the same gene, in spite of the fact that noncoding regions were considerably richer in A + T. Inasmuch as this universal rule does not discriminate one strand of DNA double helix from another, two complementary DNA strands of the entire gene maintained nearly perfect symmetry. That is to say, the degrees of excesses, deficiencies of the 64-base trimers remained nearly identical between two complementary strands, and this symmetry was only slightly disturbed in the coding region. It would thus appear that the universal rule as an intrinsic force has been exerting far greater influence than natural selection in the evolution of genes.

Animals↗
PubMed1989

Enzymatic properties of a novel phorbol ester receptor/protein kinase, nPKC.

A protein kinase C-related cDNA encodes a novel phorbol ester receptor/protein kinase, nPKC epsilon, clearly distinct from the four "conventional" PKCs [Ohno, S., Akita, Y., Konno, Y., Imajoh, S., & Suzuki, K. (1988) Cell 53, 731-741]. We purified nPKC epsilon from COS cells transfected with nPKC cDNA and compared its enzymatic properties with a conventional PKC, PKC alpha. nPKC epsilon was eluted from a hydroxyapatite column at a position coincident with type II PKC and thus was separated from type III PKC (PKC alpha), the only PKC expressed in COS cells. The protein kinase activity of nPKC epsilon is activated by phospholipids and diacylglycerols (or phorbol esters) in a manner similar to conventional PKCs. However, the cofactor dependencies and substrate specificities were clearly different from PKC alpha. A phospholipid, cardiolipin, enhances the kinase activity three- to fourfold compared with phosphatidylserine. The optimum Mg2+ concentration (3 mM) is clearly different from those of conventional PKCs (10-20 mM). The activation of nPKC epsilon by these cofactors is totally independent of Ca2+. Similar to conventional PKCs, nPKC epsilon autophosphorylates serine and threonine residues, indicating the specificity of the kinase to these amino acid residues. However, it shows a clearly different substrate specificity against exogenous substrates in that myelin basic proteins rather than histone are good substrates. These properties of nPKC epsilon permit clear discrimination of nPKC epsilon from conventional PKCs.

Amino Acids↗
PubMed1989

Erythroderma with generalized lymphadenopathy induced by phenytoin.

A 40-year-old man developed a generalized erythematous rash, fever, and systemic lymphadenopathy after treatment for two months with anticonvulsants including phenytoin. Laboratory examinations revealed increased white blood cell counts with eosinophilia, abnormal liver function, and increased gammaglobulin levels. A lymphocyte stimulation test showed a significant stimulation index for phenytoin. Skin biopsy findings were nonspecific cellular infiltrates in the upper dermis whereas in lymph nodes the normal architecture was replaced by massive cellular infiltrates consisting largely of lymphocytes, immunoblasts, and eosinophils suggesting a pattern of phenytoin lymphadenopathy. No malignant changes were noted. Phenytoin was discontinued and after a two-month treatment with oral corticosteroids, all manifestations had nearly subsided. However, the patient should be carefully followed up since phenytoin occasionally induces lymphoproliferative states including malignant lymphoma even after cessation of the medication.

Adult↗
PubMed1989

Effects of hyperthermochemoradiotherapy on KSE-1 cells, a newly established human squamous cell line derived from esophageal carcinoma.

Hyperthermia alone inhibits growth of the human esophageal cancer cell line (KSE-1). Even temporary exposure to hyperthermia results in substantial decreases in growth in a time- and temperature-dependent manner. A precipitous decrease in growth occurred at a temperature of approximately 42.5 degrees C. Concomitant application of hyperthermia, bleomycin and irradiation (hyperthermochemoradiotherapy, HCR) had a maximal effect on cell growth, compared to findings in cells treated with only one or two of these modalities. Fractionated hyperthermia (42 degrees C, 60 min) at a time interval of under 12 h was less effective than a single application (42 degrees C, 120 min). However, over a 12-hour time interval, the effects of both treatments were similar. Fractionated HCR (hyperthermia: 42 degrees C, 60 min; chemotherapy: 2 micrograms/ml of bleomycin, 60 min; radiotherapy: 2 Gy) at a time interval of under 21 h was less effective than a single HCR (hyperthermia: 42 degrees C, 120 min; chemotherapy: 2 micrograms/ml of bleomycin, 120 min; radiotherapy: 4 Gy). We conclude that care should be taken regarding development of thermotolerance when combined modality therapy is given.

Animals↗
PubMed1989

Hypotensive effect of carteolol on intraocular pressure elevation and secondary glaucoma associated with endogenous uveitis.

The therapeutic effects of carteolol hydrochloride were evaluated in the treatment of 44 uveitis patients (51 eyes) with intraocular pressure elevation or secondary glaucoma. Carteolol ophthalmic solutions, either 1% or 2%, were given twice day for more than 4 weeks in glaucomatocyclitic crisis and for more than 8 weeks in other forms of uveitis. Intraocular pressure significantly decreased from week 1 of treatment and persisted within normal limits until week 8 in glaucomatocyclitic crisis. Similarly, intraocular pressure decreased significantly from week 2 in other forms of uveitis. Intraocular pressure was well controlled in patients with open-angle glaucoma, but the control was insufficient in patients with angle-closure glaucoma. No adverse reactions such as systemic hemodynamic effects or exacerbations of intraocular inflammation were observed during this study. Carteolol therefore seems to be effective for the treatment of intraocular pressure elevation and secondary glaucoma associated with endogenous uveitis.

Adolescent↗
PubMed1989

20 Beta-hydroxysteroid dehydrogenase of neonatal pig testis: cofactor requirement and stereospecificity of hydrogen transfer from nicotinamide adenine dinucleotide phosphate, reduced form.

The cofactor requirement of purified 20 beta-hydroxysteroid dehydrogenase from cytosol fraction of neonatal pig testis, in the reduction of 17 alpha-hydroxyprogesterone was investigated. The enzyme required beta-nicotinamide adenine dinucleotide phosphate, reduced form (beta-NADPH) as the preferred cofactor, with an apparent Km value of 17 microM. Furthermore, alpha-nicotinamide adenine dinucleotide phosphate, reduced form (alpha-NADPH), beta-3'-NADPH and beta-nicotinamide adenine dinucleotide (beta-NADH) were also utilized as hydrogen donors in the reduction at relatively high concentration with apparent Km values of 85.2 microM, 179.2 microM and 1.00 mM, respectively. The optimum pH was 5.5 when beta-NADPH was used as the cofactor, while it was 6.0 when beta-NADH was used. The hydrogen transfer from the beta-NADPH to the product, 17 alpha,20 beta-dihydroxypregn-4-en-3-one catalyzed by 20 beta-hydroxysteroid dehydrogenase was stereospecific, and the 4-pro-S-hydrogen of the nicotinamide moiety was transferred to the product.

20-Hydroxysteroid Dehydrogenases↗
PubMed1989

[Effectiveness of local delivery of ofloxacin using controlled-release strips (PT-01) in periodontal patients. Part 2. Successive delivery system and combination with scaling].

Four different sites were randomly selected in each periodontal patient and divided into four group: PT-01-treated site (A), PT-01 treatment combined with scaling (S + A), placebo-treated site (P) and placebo treatment combined with scaling (S + P). PT-01 and the placebo were applied to the periodontal pocket weekly on days 0 to 28, and clinical and microbiological evaluations were made. The results obtained were as follows: 1. Weekly changes in plaque index, plus discharge and mobility of the tooth were not observed in any group. 2. In gingival index, bleeding on probing, pocket depth and gingival crevicular fluid, significant improvement was found in A in comparison with P and in S + A in comparison with S + P. 3. In total number of subgingival bacteria and in the ratio of motile rods and spirochetes to the total number of bacteria, significant reduction was found in A in comparison with P and in S + A in comparison with S + P.

Delayed-Action Preparations↗
PubMed1989

Relationship among iodine, bromine and chlorine concentrations in cow's milk in Japan.

In order to know the relationship among some elements in biological materials, iodine, bromine and chlorine concentrations in cow's milk samples in Japan were determined by the thermal neutron activation analysis using a low power research reactor and a Van de Graaff accelerator. The iodine contents in cow's milk samples ranged from 0.041 to 0.316 ppm with an average of 0.096 ppm. The bromine and chlorine in these samples ranged from 2.3 to 11.1 ppm and from 475 to 1650 ppm, respectively. The average concentration of the bromine was calculated to be 5.6 ppm and that of the chlorine was 853 ppm. The relationship among iodine, bromine and chlorine concentrations in cow's milk samples in Japan was studied with a regression analysis. It was suggested that the correlation has a power function as follows; Y = K(Z)-A where, Y is elemental concentration in ppm, Z is atomic weight [corrected] of element, A (= 7.4) is exponent and K (= 14.7) is a constant.

Animals↗
PubMed1989

Molecular cloning and expression of human leukotriene A4 hydrolase cDNA.

We have isolated a near full-length cDNA encoding human leukotriene A4 (LTA4) hydrolase from a human spleen cDNA library. The mature form of the enzyme consists of 610 amino acid residues and its molecular weight is calculated to be 69,153. The cDNA was inserted into an expression vector (pUC 9) and expressed in Escherichia coli. The fusion protein (EX85) possessed the LTA4 hydrolase activity with kinetic properties similar to the native enzyme.

Amino Acid Sequence↗
PubMed1989

Malignant mixed mesodermal tumour of the peritoneum with a complete response to cyclophosphamide.

We treated a 66-year-old Japanese woman with malignant mixed mesodermal tumour of the heterologous type arising from the paracolic peritoneum. The tumour contained heterologous elements in the form of cartilage in addition to carcinosarcomatous areas. This is the third case of malignant mixed mesodermal tumours originating from the peritoneum to be documented in the literature. A complete resection of the tumour was carried out but multiple lymph nodes metastases in the left neck occurred 5 months later. Cyclophosphamide in doses of 150 mg daily by mouth was initiated, with the result of a complete regression of the malignant lesions. She died of myocardial infarction 21 months after surgery.

Aged↗
PubMed1989

Three-dimensional studies of cytoskeletal organizations in cultured thyroid cells by quick-freezing and deep-etching method.

Isolated porcine thyroid cells were cultured on collagen gels (control group, TSH-stimulated group, and double-layered culture). They were split or cut to remove cytoplasmic soluble proteins for replica preparations. Some specimens were immunostained with anti-actin antibody or decorated with S1 myosin fragments to identify actin filaments. The basal cell membranes of thyroid cells of monolayer culture were in contact with collagen gels and the apical cell membranes faced the culture medium. Networks of actin filaments were attached to the cytoplasmic sides of the apical cell membranes, while intermediate filaments were localized along the basal ones. The thyroid-stimulating hormone (TSH) treatment induced the formation of microvilli only on the apical cell membranes and the accumulation of actin filaments under the apical cell membranes, indicating the apical-basal polarity of the cells. In double-layered culture, the primitive follicular lumens with microvilli appeared between two adjacent cells. The interaction of cell membranes with collagen gels is a determinant factor in the orientation of apical-basal polarity. Moreover, the TSH treatment and cell-cell contact further intensify the polarization through reorganizing the cytoskeletons.

Animals↗
PubMed1989

[Clinical evaluation of systematic lymph node dissection for the intrathoracic esophageal carcinoma].

We evaluated the effectiveness and complication of systematic lymph node dissection for the intrathoracic esophageal carcinoma, which includes cervical, intrathoracic and abdominal lymph node dissection. Two hundred and thirteen individuals with intrathoracic esophageal carcinoma underwent esophageal resection in the Department of Surgery II, Kyushu University from 1979 to 1988. Of these 213, systematic lymph node dissection in addition to esophageal resection was performed on 19 patients. Lymph node recurrence has been reduced with this procedure and survival rate was more favorable in the cases with systematic lymph node dissection than those without it at present. On the other hand, although the occurrence of postoperative recurrent nerve palsy in the cases with systematic lymph node dissection and in those without it were 47.4 and 11.9%, rates of postoperative pulmonary complications were 5.2 and 16.0%, respectively. Operative death was none in those with systematic lymph node dissection. Therefore, this procedure has been performed in safety with intensive perioperative cares and it would contribute more favorable prognosis.

Aged↗
PubMed1989

C-myc-S mu rearrangement in a murine plasmacytoma without visible chromosomal translocations.

We report here that one murine plasmacytoma (DCPC 21) arising in a Balb/c mouse implanted i.p. with a Millipore diffusion chamber lacks any known chromosomal translocations. In situ molecular hybridization analysis of the metaphase spreads of DCPC 21, however, revealed that the c-myc gene has moved to the IgH locus on the chromosome 12. Further, an Ig switch region-mediated recombination was detected. The DNA sequencing analysis demonstrated that the c-myc gene has juxtaposed directly to S mu 5'-flanking sequence in a 'head-to-tail' orientation. The results strongly support our notion that the c-myc rearrangement is an essential step in the genesis of murine plasmacytomas, whether or not they have visible chromosomal translocations.

Animals↗