Biomedical subjects

H Okada

Publications and source records attributed to H Okada.

At least 685 records · Page 38Linked to original sources

PubMed1992

Complement regulatory proteins at the feto-maternal interface during human placental development: distribution of CD59 by comparison with membrane cofactor protein (CD46) and decay accelerating factor (CD55).

The complement (C) regulatory proteins decay-accelerating factor (DAF, CD55) and membrane cofactor protein (MCP, CD46), which control C3 convertases, together with CD59, an inhibitor of the membrane attack complex (MAC), were found to be present in the developing human placenta from at least 6 weeks of gestation until term. Immunostaining revealed differences in the distribution of these proteins on the fetally derived trophoblast epithelium, especially in early placentae which contain trophoblast populations of diverse proliferative potential and differentiation status. Expression of all three proteins occurred on the terminally differentiated syncytiotrophoblast epithelium covering chorionic villi and which is in direct contact with maternal blood. CD59 was also expressed on the underlying villous cytotrophoblast cells and on their extra-villous derivatives. These two populations showed differential expression of the C3 convertase regulators. Villous cytotrophoblast cells expressed MCP but were largely devoid of DAF. Proliferation of this population to generate extra-villous cytotrophoblast cell columns was associated with both an increase in DAF expression and a decrease in MCP expression. Throughout placental development, expression of DAF appeared to be lower than that of MCP and CD59 as assessed by solid-phase binding assays on isolated trophoblast membranes. Early placentae were also found to contain both DAF+ and DAF- chorionic villi. Conversely, expression of CD59 appeared comparatively high and transcripts for CD59 were found to be much more abundant than those for DAF in purified trophoblast cells. C regulatory proteins appear to play an important role throughout gestation in protecting the fetally derived human conceptus from maternal C. The differential expression patterns of the proteins on trophoblast may reflect differences in requirement for specific functional activities at different locations within the placenta.

Antigens, CD↗

PubMed1992

Species-specific restriction of complement by HRF20 (CD59) generated by cDNA transfection.

The 20-kDa homologous restriction factor (HRF20, CD59) is a phosphatidyl inositol-anchored membrane glycoprotein that inhibits the formation of human complement membrane attack complexes. The cDNA of HRF20 was transfected into Chinese hamster ovary (CHO) cells resulting in expression of human HRF20 protein on the cell surface anchored via glycosylphosphatidyl inositol. The transfected CHO cells were resistant to human complement-mediated cell killing. However, the cells remained sensitive to rat and guinea pig complement. Therefore, species specificity between HRF20 and complement is maintained in HRF20 generated on the CHO cells following transfection with HRF20 cDNA.

Animals↗

PubMed1992

Paroxysmal nocturnal hemoglobinuria due to hereditary nucleotide deletion in the HRF20 (CD59) gene.

HRF20 (CD59) is a membrane glycoprotein which protects cells from the membrane attack reaction of homologous complement. A patient who is completely deficient in HRF20 expression and is suffering from paroxysmal nocturnal hemoglobinuria (PNH) was studied. His parents are cousins and both have decreased HRF20 expression, suggesting that the deficiency is genetic. We established a cultured cell line (NCU1) which is HRF20 deficient from the patient's lymphocytes by Epstein-Barr-virus (EBV) infection. Northern blot analysis revealed HRF20 mRNA signals, indicating that HRF20 mRNA were transcribed. HRF20 cDNA was amplified by the polymerase chain reaction (PCR) method. Sequencing of the cDNA from the NCU1 showed two single-base deletions at amino acid 16 and 96 from the N terminus of the mature protein. Deletion in the genomic DNA of peripheral blood lymphocytes was confirmed by the DNA sequence of an HRF20 open reading frame containing amino acid 16. Furthermore, the patient's parents and sister possessed both intact and deleted genomic HRF20 DNA while his brother's DNA was intact. These findings demonstrate that the HRF20 deficiency was genomic in origin, and that complete deletion was brought about by a homozygous abnormality in the HRF20 gene. The base deletion caused a codon frame shift resulting in failure to produce intact HRF20 protein in the patient.

Antigens, CD↗

PubMed1992

A novel mutation occurring in the PHO80 gene suppresses the PHO4c mutations of Saccharomyces cerevisiae.

We have isolated suppressors of a PHO4c (a positive regulator) mutant which normally confers weak constitutivity for acid phosphatase production on the Saccharomyces cell. One dominant suppressor (PHO80-2) was found to be an allele of PHO80 (a negative regulator) that changes G to A, resulting in substitution of isoleucine for methionine 42 of the Pho80 protein. Substitution of valine (PHO80-3) or leucine (PHO80-4) for the same methionine by site-directed mutagenesis also suppressed PHOc. Suppression by PHO80-2) did show some allele specificity. From these results we were able to delimit the region of PHo80 which may interact with the Pho4 protein.

Alleles↗

PubMed1992

Keratin profiles in normal/hyperplastic prostates and prostate carcinoma.

Immunoreactivities in 25 cases of prostatic adenocarcinoma and 10 normal/hyperplastic prostates were investigated in methacarn-fixed, paraffin-embedded serial sections using a panel of nine anti-keratin monoclonal antibodies (mAbs); 34 beta E12, CK8.12, 312C8-1, CK4.62, RPN1165, RPN1162, 35 beta H11, CK5, M20, and one of anti-actin mAb, HHF35. In normal/hyperplastic prostates, RPN1162, 35 beta H11, CK5 and M20 stained luminal cells without staining basal cells, and 34 beta E12, CK8.12 and 312C8-1 stained basal cells but not luminal cells. Other mAbs, CK4.62 and RPN1165, stained basal cells as well as luminal cells. All of the mAbs labelling luminal cells stained cancer cells with variable frequencies in a manner unrelated to the grade of tumour differentiation. Of the prostate cancer cases 92% were scored positive with M20, 84% with 35 beta H11, 80% with CK5, 68% with CK4.62, 60% with RPN1165 and 4% with RPN1162. However, basal cell-specific keratins labelled with 34 beta E12, CK8.12 and 312C8-1 were totally negative in the cancer cells. HHF35 showed no labelling in normal, hyperplastic or neoplastic epithelial cells of the prostate. Our findings indicate that the major part of the cells of prostatic adenocarcinomas have keratin phenotypes similar to luminal cells but not basal cells, and that no myoepithelial differentiation can be detected in epithelial cell of the prostate. Thus, mAbs for keratins facilitate the identification of epithelial cell phenotypes in normal, benign and malignant conditions of the prostate.

Adolescent↗

PubMed1992

Flow cytometric evaluation of phagocytosis by peripheral blood polymorphonuclear leucocytes in human periodontal diseases.

The complement-dependent phagocytic functions of polymorphonuclear leucocytes (PMNL) in peripheral blood from 15 patients with localized juvenile periodontitis (LJP), 13 with generalized juvenile periodontitis (GJP) and 52 with adult periodontitis (AP), and from 30 normal subjects as controls were measured by flow cytometry. Heparinized blood was collected and incubated with fluorescent microspheres, and erythrocytes were removed. By means of single-cell analysis the percentage of phagocytosing cells (% phagocytosis) and the mean number of microspheres phagocytosed by one PMNL (degree of phagocytosis; d-phagocytosis) were measured. Some but not all patients with LJP (53%) and GJP (46%) showed consistently low % phagocytosis and d-phagocytosis. On the other hand, only 6% of AP patients and no healthy subjects showed a reduction of PMNL phagocytosis. Phagocytosis was unchanged after initial periodontal treatment in all subjects, suggesting the depression of PMNL phagocytosis may not be a transient phenomenon associated with periodontal status. Furthermore, PMNLs from the LJP patients that showed depressed phagocytic function exhibited depressed phagocytic responses with either autologous or normal plasma, while control PMNLs with either normal or the patients' plasma showed normal responses. These results suggested that the depressed phagocytic responses in LJP patients could be due to cell-associated defect(s) on the PMNL.

Adolescent↗

PubMed1992

CD59 (homologous restriction factor 20), a plasma membrane protein that protects against complement C5b-9 attack, in human atherosclerotic lesions.

Blood cells express a cell membrane protein, termed homologous restriction factor 20 (HRF20) and identical to CD59, that can inhibit complement C5b-9 insertion into their membranes. In this report, we investigated by immunohistochemistry whether CD59 was present on cells in human atherosclerotic lesions since membranous C5b-9(m) has been found in lesions. Using a monoclonal anti-CD59 antibody, a cellular CD59 staining pattern was apparent in nearly all lesion specimens. CD59 stain co-localised with macrophage (CD14), T lymphocyte (CD7), endothelial cell (anti-factor VIII related antigen) and smooth muscle cell cytoskeletal-specific antigens (anti-alpha actin and muscle myosin). Endothelial cells always exhibited a more intense stain than the other cell types. CD59 antigen was not localised to any one area of the lesions. Usually CD59-positive cells occurred in clusters rather than as randomly spaced individual cells. CD59 did not stain all cells of the lesion and in particular did not appear to stain all smooth muscle cells. Areas of CD59-negative cells were sometimes observed to exhibit a cellular C5b-9 staining pattern. C5b-9 deposits were also observed in CD59-positive regions. Normal saphenous vein stained strongly for CD59 at the endothelial lining and weakly in the media. Capillaries in atherosclerotic intima always stained strongly for CD59. We conclude that HRF20 is constitutively expressed on endothelium and is under regulatory control in smooth muscle cells. Cellular C5b-9 attack in atherosclerotic lesions is therefore most likely to occur on smooth muscle cells.

Actins↗

PubMed1992

Histochemistry of glandular metaplasia at the trigone of the urinary bladder in cows.

The epithelium at the trigone of the urinary bladder showed intestinal metaplasia of a colonic type in three Holstein-Friesian cows affected with chronic polypoid cystitis. Except for Ricinus communis (RCA-I), almost all goblet cells in the whole crypt were positive to periodic acid-Schiff (PAS), alcian blue (AB) pH 2.5, AB pH 1.0, periodate borohydride-potassium hydroxide-PAS (PB-KOH-PAS), Ulex europaeus (UEA-I), Triticum vulgare (WGA) and Arachis hypogaea (PNA) after neuraminidase digestion. This result indicated that most goblet cells contained acidic and neutral glycoconjugates as O-acetylated sialomucin, L-fucose, N-acetyl-glucosamine, neuraminic acid residues and sialic acid-galactose dimers and were devoid of beta-D-galactose. The goblet cells at the surface in the upper half of the crypt contained both sulpho- and sialo-mucins with N-acetyl-galactosamine residues by AB (pH 2.5)-PAS, high iron diamine (HID)-AB (pH 2.5), Dolichos biflorus (DBA) and Glycine maximus (SBA) reactions. On the other hand, the lower goblet cells were found to contain predominantly sulphated mucins with D-mannose and D-glucose residues by AB-PAS, HID-AB and Concanavalia ensiformis (Con A) reactivities. This suggested that mucin secreted from these cells was similar to that secreted from the goblet cells of the large intestine in cattle.

Alcian Blue↗

PubMed1992

A pathological study of bovine alimentary mycosis.

A 10-year study showed that 45 of 692 cattle aged > 6 months had systemic mycosis and, of these, 38 had alimentary mycosis. The percentages of these 38 cattle with lesions in various sites were: rumen (73.7), omasum (71.1), abomasum (34.2), reticulum (21.1), intestine (15.8), omasoabomasal orifice (7.9) and tongue (2.6). Infections in both the forestomachs and abomasum occurred in 31.6 per cent; mucormycosis was found in 94.7 per cent and aspergillosis in 31.6 per cent; 26.3 per cent (10 cattle) had complicated infections with both Mucorales and Aspergillus and eight of these animals had mixed lesions of both mucormycosis and aspergillosis. Macroscopically, focal haemorrhagic necrosis was seen in all 38 cattle. Histologically, thrombosis, coagulative necrosis and hyphae typical of the order Mucorales or of Aspergillus spp. were found. Many of the cattle were also affected by ruminal acidosis-inducing factors (ruminal atony 81.6 per cent, mastitis 63.2 per cent, "downer cow syndrome" 57.9 per cent and parturition 50.0 per cent), which may have predisposed to alimentary mycosis.

Animals↗

PubMed1992

New method for analysis of biodegradable polyesters by high-performance liquid chromatography after alkali hydrolysis.

A new method was developed for analysis of biodegradable polyesters, which involves alkali hydrolysis of polyesters to the corresponding hydroxyacids and determination of the hydroxyacids by high-performance liquid chromatography. It can be used to monitor weight change in polyesters and change in molecular ratio of the hydroxyacid constituents of polyesters during in vitro and in vivo degradation.

Animals↗

PubMed1992

Kinetics of inter-heavy chain disulfide bond formation of liganded and unliganded human immunoglobulin G by radioimmunoassay.

Monoclonal antibodies (mAbs) to the hinge region of human IgG1 immunoglobulins were prepared by immunization with a proteolytic fragment of hinge segment coupled to keyhole limpet hemocyanin. A mAb, 4G3, was obtained capable of binding to intact IgG but not to partially reduced IgG. Using this mAb, inter-heavy (H) chain disulfide bond formation from partially reduced anti-tetanus antibodies (Abs) in the presence or absence of antigens (Ags) was studied by solid phase radioimmunoassay. When the Abs were partially reduced in solution and then coated to plastic plates, only 10% regeneration of inter-H chain disulfide bonds occurred after reoxidation, although 100% formation occurred in solution [Kishida et al., J. Biochem. (Tokyo) 79, 91-105 (1976)]. This difference in the extent of disulfide bond formation can be explained by the fact that there are two convertible isomers in solution, Conformer I and II, one of which (Conformer I) can form disulfide bonds but is present as a minor component. Since the motion of IgG molecules on plates is restricted by hydrophobic interactions, the two conformers are not convertible as in solution. Therefore only Conformer I which existed before coating formed inter-H chain disulfide bonds. Similar kinetic measurements were performed using plates coated with tetanus toxoid. Abs, partially reduced in solution and then allowed to react with Ags on the plate were able to completely regenerate their inter-H chain disulfide bonds, although the rate of reaction was slow. These results can also be explained by the fact that the two isomers are convertible since Ag-Ab complexes are dissociable. Ag-binding therefore did not significantly perturb conformation of the hinge segments. In addition, no difference between liganded and unliganded Abs was observed in the binding of anti-hinge mAbs. These results imply little or no contribution of the hinge region to transmission of the signal produced by Ag-binding to Fc.

Amino Acid Sequence↗

PubMed1992

Enhanced muscle sympathetic nerve activity during sleep apnea in the elderly.

To examine how muscle sympathetic nerve activity (MSNA) becomes modified during sleep apnea in the elderly, we analyzed polysomnographic recording simultaneously with microneurographically recorded MSNA. Subjects were three healthy elderly males aged 72, 75, and 76. MSNA was suppressed with deeper non-REM sleep stages in these elderly subjects. In all three subjects, sleep apnea for 10 s or longer was observed during sleep of 00:00-06:00. During sleep apnea, MSNA was enhanced concomitantly with a blood pressure fall and a reduction in saturation rate of oxyhemoglobin. With the termination of sleep apnea, MSNA was maximally enhanced with a transient elevation of blood pressure. We conclude that sleep apnea induces an enhancement of MSNA, which may be responsible for hypertensive episodes during sleep.

Aged↗

PubMed1992

Isolation and partial characterization of a growth factor from human cementum.

Cementum is the mineralized interface through which collagen fibers of periodontal connective tissues are anchored onto the tooth surface. We have isolated and partially characterized a mitogenic factor from human cementum which has properties different from other growth factors. Cementum was harvested from healthy human teeth, extracted in 1.0 M CH3COOH and mitogenic activities were fractionated by heparin-affinity chromatography. Proteins eluted by 0.4-0.6 M NaCl, which contained most of the cementum mitogenic activity, were precipitated by trichloroacetic acid and resolved by HPLC through ion-exchange and reverse-phase columns. NaDodSO4-polyacrylamide gel electrophoresis revealed that the purified preparation contained a M(r) 23,000 protein and this protein was associated with mitogenic activity. The purified cementum-derived growth factor (CGF) was active alone, but at suboptimal concentrations its activity was potentiated by small quantities of plasma-derived serum and epidermal growth factor (EGF). The activity was resistant to heat, but it was destroyed by trypsin digestion. Reduction and alkylation destroyed the mitogenic activity, however electrophoretic mobility was not affected. Binding of EGF to fibroblast membranes was not affected by the CGF and assays to detect platelet-derived growth factor were negative. These characteristics indicated that CGF is a distinct molecular species. Our data show that cementum contains several mitogenic factors and that CGF is the major cementum mitogen.

Cell Division↗

PubMed1992

Protein binding of four antiepileptic drugs in maternal and umbilical cord serum.

The total and protein free levels of 4 antiepileptic drugs (AEDs) in serum from 35 maternity patients who had been treated with AED monotherapy throughout pregnancy were studied. Results were compared with those in the umbilical cord serum at the time of delivery, and the placental transfer of AEDs was evaluated from the viewpoint of the protein binding capacity of the drug. The materials consisted of 35 samples of maternal and umbilical cord serum in total and included 13 patients on phenobarbital (PB), 7 on phenytoin (PHT), 7 on carbamazepine (CBZ) and 8 on valproic acid (VPA). The mean fetal/maternal total concentration ratios were 0.86, 0.91, 0.73 and 1.59 for PB, PHT, CBZ and VPA, respectively, only the VPA ratio being above 1. On the other hand, the mean fetal/maternal free fraction ratios were 1.13, 1.10, 1.42 and 0.50 for PB, PHT, CBZ and VPA, respectively, only the VPA ratio being less than 1. Correlation of the 2 ratios showed a reciprocal proportion with a correlation coefficient of -0.90 (P < 0.005). It was considered that the fetal/maternal total concentration ratio of 4 AEDs was regulated by the fetal/maternal free fraction ratio of the corresponding AEDs and that the difference in fetal/maternal free fraction ratio depended on the type of drug being administered.

Anticonvulsants↗

PubMed1992

Contribution of aromatase to the deoxyribonucleic acid synthesis of MCF-7 human breast cancer cells and its suppression by aromatase inhibitors.

We have studied the effects of various steroids on DNA synthesis in MCF-7 human breast carcinoma cells, which have aromatase activity and which exert an oestrogen receptor-mediated growth, to assess the significance of intracellular aromatase on growth stimulation as well as inhibition by aromatase inhibitors. The cells were cultured for 96 h in phenol red-free medium containing 10% charcoal-treated fetal bovine serum and test reagents and pulse-labelled with [3H]thymidine. Physiological concentrations of oestradiol, oestrone, testosterone (T) and androstenedione (AD) stimulated thymidine incorporation. However, oestrone-sulphate and dihydrotestosterone (DHT) only stimulated at concentrations greater than the physiological levels. T and DHT stimulation was blocked by tamoxifen, but not by cyproterone acetate, suggesting that the stimulation was mediated via the oestrogen receptor but not by the androgen receptor. Stimulation by T and AD was reduced by aminoglutethimide and 14 alpha-hydroxy-4-androstene-3,6,17-trione, both of which inhibit aromatase activity, however, stimulation by nonaromatizable DHT was not reduced by the inhibitors, suggesting that androgens were converted by the intracellular aromatase to oestrogens which stimulated the thymidine incorporation. It is suggested that intracellular aromatase significantly contributes to the stimulation of DNA synthesis and that aromatase inhibitors suppress the stimulation.

Androgens↗

PubMed1992

Estrogen as a growth factor to central nervous cells. Estrogen treatment promotes development of acetylcholinesterase-positive basal forebrain neurons transplanted in the anterior eye chamber.

In a previous report, we demonstrated in vivo ameliorating effects of conjugated estrogen in women suffering from senile dementia-Alzheimer's type. To investigate the effects of estrogen on the growth of cholinergic neurons, the present study was performed using rat cholinergic tissue implanted into the anterior chamber of the eye. Fetal diagonal band tissue containing cholinergic neurons was grafted into the anterior eye chamber of adult female rats that had either been treated or not with 2 mg estradiol valerate injected every 3 days after oophorectomy. Two and four weeks after transplantation, the axonal and/or dendritic growth of cholinergic neurons in the graft was studied using acetylcholinesterase histochemistry. At both times, acetylcholinesterase positive processes were densely distributed in the grafts of estradiol valerate treated rats, while in rats without estradiol valerate treatment acetylcholinesterase positive reaction was essentially localized only on the cell bodies. These findings were more obvious at 2 weeks after transplantation than at 4 weeks. These results suggest that estrogen acts on cholinergic neurons as a growth factor.

Acetylcholinesterase↗

PubMed1992

Two cases of eosinophilic cystitis induced by tranilast.

We report 2 cases of eosinophilic cystitis induced by Tranilast, which was used for the treatment of bronchial asthma. In case 2 Tranilast itself and its metabolic derivative proved to be inciting agents by a drug-induced lymphocyte stimulation test. The literature on this association is reviewed.

Aged↗

PubMed1992

Quantitative analysis of immunocompetent cells in human periapical lesions: correlations with clinical findings of the involved teeth.

Human periapical lesions develop as a result of a pathological immune response to continuous stimuli from infected root canals. This study identified the immunocompetent cells in such lesions immunohistochemically and quantified them to examine their interrelationships and correlations with clinical findings. The number of IgG-containing cells in CD4+ cell (Th/i)-rich lesions (> or = 55 CD4+ cells/2 x 10(4) microns 2) was significantly larger than in CD4+ cell-poor lesions (< 55 CD4+ cells/2 x 10(4) microns 2). This indicated that the CD4+ cells and the IgG-containing cells acted together against antigenic stimuli. The proportion of T cells in the mononuclear infiltrates varied with the size of the periapical lesions. The proportion of CD11+ cells (monocytes/macrophages) was significantly larger in lesions which showed a positive reaction to percussion or were tender on palpation than in the lesions without these symptoms. These observations suggest that T cells may play an important role in the development of periapical lesions and that CD11+ cells may be involved in the development of symptoms.

Antigens, CD↗