Biomedical subjects
Publications and source records attributed to R Sasaki.
The cDNA for the rat erythropoietin (EPO) has been cloned and sequenced. The deduced amino acid sequence consists of 166 amino acid residues, which has a 79% and 95% homology with human and mouse EPOs, respectively. Many short stretches, highly conserved in primate and rodent EPOs, are found in the 3'-noncoding region when insertions and deletions are taken into consideration.
A recombinant soluble form (sEPO-R) of erythropoietin (EPO) receptor (EPO-R) was produced by Chinese hamster ovary cells and isolated in high yield with the EPO-fixed gel. Ligand binding assays were done using three methods; precipitation of sEPO-R radiolabeled EPO complex and competition of sEPO-R for the binding of radiolabeled EPO with the cellular EPO-R. The results showed a Kd of 17 nM which was much lower than those for cellular EPO-R. One N-glycosylation site exists in sEPO-R but the glycosylation did not affect the binding affinity to EPO. A complex with a molecular size that corresponded to a 1:1 complex of EPO and sEPO-R was detected.
Erythropoietin (EPO) stimulates proliferation and differentiation of late erythroid precursor cells (CFU-E) and thereby determines the rate of erythropoiesis. Liver is the major erythropoietic site in a fetus. We dealt with developmental changes in CFU-E and EPO receptor (EPO-R) of fetal mouse liver. The affinity of the EPO-R to EPO was unchanged during fetal development. The population size of CFU-E, the number of EPO-R per liver cell, and EPO-R mRNA decreased as gestation proceeded, in a pattern indicating that the expression of EPO-R on erythroid precursor cells in fetal mouse liver is governed mostly by the process of mRNA production.
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Erythropoietin (EPO), a glycoprotein produced mainly by the kidney, is the major physiological regulator of erythropoiesis. We developed a sensitive and rapid ELISA for measurement of rat serum EPO with two monoclonal antibodies that recognize different epitopes. To understand the mechanism by which erythropoiesis is impaired in rats deficient in dietary protein, we investigated the levels of the immunoreactive EPO (iEPO) in serums and erythroid precursor cells in hemopoietic tissues during protein deprivation. The iEPO level of 32-d-old rats fed a protein-free diet was lowered to one-third that of rats fed 20% casein at 6 h after protein deprivation began. Protein deprivation decreased the number of EPO-responsive cells in spleen. These results indicate that the impairment of erythropoiesis during protein deficiency is caused by the decrease in serum EPO and the subsequent reduction of the population size of erythroid precursor cells in spleen.
We conducted a statistical review of 114 cases of dermatomyositis (DMS) treated primarily at the Department of Dermatology at Nagoya University Hospital over 27 years from 1965 to 1991 in order to determine the primary characteristics of juvenile DMS with the following results. 1) Juvenile DMS was found slightly more often in males than females; the male-to-female ratio was 1.4:1. Therefore, unlike adult DMS with its preponderance of females, there was no clear gender predominance. 2) Muscular manifestations tended to follow the appearance of cutaneous manifestations, but the frequency of minor muscular manifestations was high over the entire course of the disease. 3) Laboratory findings showed increases in serum aldolase and serum creatinine kinase with significant frequency when compared with adult patients (p < 0.01 and p < 0.05, respectively). Elevated serum aldolase most often occurred prior to or at the time of the appearance of muscular manifestations, suggesting its usefulness in early diagnosis. The positive rates for the antinuclear antibody on HEp-2 cells and anti-DNA antibody were significantly lower in children than in adults (p < 0.001 and p < 0.05, respectively). 4) There were no cases of juvenile DMS complicated by malignant tumors, interstitial pneumonia, or pulmonary fibrosis. There were also no deaths, and the rate of "remission or improvement" was significantly higher than in adult DMS cases (p < 0.05). Adult cases which remained the same or worsened usually presented with intractable muscular manifestations. In children, however, the cutaneous manifestations were more difficult to treat.(ABSTRACT TRUNCATED AT 250 WORDS)
In order to study the effect of omeprazole on the K/[H+] value of ascorbic acid (K = the ascorbic acid-ascorbic free radical equilibrium constant), changes of the concentration of ascorbate free radical in guinea pig sera were examined after intraperitoneal administration of omeprazole. Furthermore, to see the in vitro effect of omeprazole on ascorbate free radical, changes were examined in ascorbate solutions after addition of omeprazole. It was found that the K/[H+] value in the serum increased significantly after administration of omeprazole and also that the drug amplified the electron spin resonance (ESR) signal intensity of ascorbate free radical in vitro. These results suggested that omeprazole acts like oxygen radicals.
Myeloperoxidase (MPO)- and Sudan Black B-not more than 3%-positive, esterase staining-negative, lymphoid, megakaryocyte lineage and erythroid surface marker-negative and electron microscopic platelet peroxidase-negative acute leukemia (AL) was diagnosed as acute undifferentiated leukemia (AUL), and myeloid marker (CD13, CD33), electron microscopic MPO (EMMPO), and DNA analysis of immunoglobulin heavy chain and T cell receptor as well as chemotherapy and its reactivity were examined. Of 239 cases of AL, 10 (4.2%) were AUL, and of these 10 cases, 9 were CD13 or CD33-positive AML-MO (MO) cases. Of 9 cases examined for EMMPO, 4 (44%) were positive, and of 3 cases of MO subjected to DNA analysis, 1 and 1 showed rearrangements of immunoglobulin heavy chain and T cell receptor beta chain, respectively. Of 6 cases of MO on myeloid induction therapy, 1 and 1 showed complete remission (CR) and partial remission (PR), respectively, each having lymphoid genotype, and 4 showed no remission (NR), being 3 of them EMMPO-positive. Of 2 cases on lymphoid induction therapy, 1 and 1 showed CR and NR, respectively, the former being EMMPO-positive MO. BHAC-EM therapy with behenoyl cytosine arabinoside, VP-16 and mitoxantrone performed on 2 cases refractory to any one of both these myeloid and lymphoid induction therapies led to CR in all these 2 cases.
To clarify the factors relating to the prognosis of spinocerebellar degenerations (SCD), we performed a follow-up study on the survival of patients with Friedreich disease, familial spastic paraparasis, sporadic olivopontocerebellar atrophy (OPCA), hereditary OPCA of Menzel type, sporadic late cortical cerebellar atrophy (LCCA), cerebellar atrophy of Holmes type, Shy-Drager syndrome, striatonigral degeneration, dentatorubropallidoluysian atrophy, or Joseph disease. One hundred and forty-eight patients admitted to the Nagoya University Hospital during the period of 1976 to 1990 were dealt with. They had been followed-up for 15 years at longest through the medical records, or through the direct informations from the patients or their family members or both. In order to find factors influencing the survival of the patients, Kaplan-Meier method was used as the 1st step to construct the survival curve for each factor. These were calculated by generalized Wilcoxon test. Employing Cox's proportional hazard model, a multivariate analysis was then performed based on the factors which were shown significant in the 1st step. The multivariate analysis showed that the following four factors are related to the prognosis of SCD patients, i.e., lack of hereditary trait, existence of orthostatic hypotension, lack of walking disorder at onset, and lack of nystagmus on admission. Of these factors, the lack of hereditary trait had the most intimate relation to their poor prognosis. Therefore, these four factors are statistically informative to predict prognosis of each patient with SCD.
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Erythropoietin (Epo) has three N-linked sugar chains. Codons for asparagine at N-glycosylation sites in genomic human Epo DNA were replaced with those for glutamine. The wild-type Epo gene and seven mutants that lacked N-glycosylation sites in every possible combination were introduced into baby hamster-kidney cells. To study the role of the N-linked sugars in Epo biosynthesis, Epo protein expressed transiently was measured by an enzyme-linked immunoassay. The elimination of all three N-glycosylation sites decreased Epo production to 10% of that of the wild-type Epo. Wild-type and mutant Epos produced by stably transfected cells were partially purified to investigate their properties. Removal of N-glycosylation sites changed affinity of Epo to the receptor. The in vitro activity of Epo that lost all N-glycosylation sites was comparable with that of the wild-type Epo, while the in vivo activity severely decreased. These results indicate that N-linked sugars of Epo have two major functions; N-linked sugars are important for 1) proper biosynthesis and/or secretion and 2) expression of the in vivo activity probably by enhancing survival in the circulation. N-Linked sugars of Epo affect binding affinity of the ligand to the receptor but do not play a key role in expression of the in vitro activity.
The exposure of K-562 cells to ethidium bromide or acridine orange as DNA-intercalating agents induced their differentiation to hemoglobin-producing cells. The addition of L-ethionine or dimethylsulfoxide induced neither such differentiation nor changes in poly(A) polymerase activity of K-562 cells, though the addition of ethidium bromide, acridine orange, hydroxyurea, or 1-beta-arabinofuranosylcytosine induced erythroid differentiation and decreases in poly(A) polymerase activity. These results suggest that DNA-intercalation is one of the pathways triggering differentiation of K-562 cells and the decrease in poly(A) polymerase activity may be an important change associated with differentiation of K-562 cells.
The relationship between serum concentrations of beta-cryptoxanthin and zeaxanthin and the consumption of cigarettes and alcohol or the intake frequencies of foods was investigated in 775 Japanese inhabitants. Serum concentrations of beta-cryptoxanthin and beta-carotene in healthy Japanese aged 40-70 years were higher in females than in males. Serum beta-cryptoxanthin levels were inversely associated with the consumption of cigarettes and alcohol, as were the serum beta-carotene levels. Serum zeaxanthin (including lutein) levels were associated with cigarette consumption. The levels of serum beta-cryptoxanthin were positively correlated with the intake frequency of some fruits, and those of zeaxanthin, with the intake frequency of green vegetables, milk and eggs. These xanthophylls may play a role with beta-carotene in smoking-related cancer prevention.
The effect of human recombinant erythropoietin (Epo) on B cell responses was studied in a serum-free medium. Epo enhanced IgM production and thymidine uptake by a human IgM-producing lymphoblastoid cell line, CBL. This effect was specific to Epo since enhancement was blocked by anti-Epo antibody but not by control antibody. Among the various cytokines, interleukin-4 (IL-4) enhanced IgM production and thymidine uptake while IL-6 enhanced IgM production without affecting thymidine uptake. In contrast, other cytokines including IL-1 beta, IL-2, IL-5, interferon-alpha (IFN-alpha), interferon-gamma (IFN-gamma), or granulocyte/macrophage colony-stimulating factor (GM-CSF) were without effect. However, the enhancing effect of Epo is different from that of IL-4 or IL-6, since Epo effect was not blocked by anti-IL-4 antibody or anti-IL-6 antibody. Moreover, specific binding of Epo was detected on CBL cells. Epo also enhanced immunoglobulin (IgG, IgM and IgA) production and thymidine uptake by purified tonsil small resting B cells stimulated by Staphylococcus aureus Cowan strain I (SAC) or by large activated B cells. In contrast, Epo had no effect on unstimulated small resting B cells. These results indicate that Epo could directly stimulate activated and differentiated B cells and could enhance B cell immunoglobulin production and proliferation.
The relationship between serum concentrations of carotenoids, tocopherols and retinol and the consumption of cigarettes, alcohol and foods was investigated in 835 healthy inhabitants (aged 35-79 years) of a rural area of Hokkaido. Serum concentrations of beta-carotene (BC), alpha-carotene (AC), lycopene (LY), beta-cryptoxanthin (BX), zeaxanthin (including lutein, ZX), canthaxanthin (CX) and beta-tocopherol (including gamma-tocopherol, BT) were higher in females than in males, while serum retinol (RE) concentrations were lower. Serum alpha-tocopherol (AT) concentrations were the same for both sexes. The serum values of BC, BX, AC, LY and ZX for males were inversely associated with the consumption of cigarettes in multiple-regression analysis. The serum values of BC, AC and BX for males were inversely associated with alcohol consumption, while serum RE values were positively related to alcohol consumption for both sexes. There were significant relations between serum values of BC, AC, and ZX and the intake frequencies of carrots, tomatoes, milk and/or green-leaf vegetables, especially for females, and between serum BX values and the intake of oranges, juices and fruits. Serum RE and AT concentrations were not associated with any food intake, and BT concentrations were associated with the intake of tomatoes and juices for males.
A method for estimating the intervals for the number of patients with intractable diseases from nationwide epidemiological surveys was developed under the assumption that response is independent of frequency of patients. This method is based on data utilizing the number of response hospitals classified by their reported number of patients. The approximate 95% confidence intervals of numbers of several intractable diseases patients were estimated using this method.
A factor with burst-promoting activity (BPA) stimulates the formation of erythroid bursts in the presence of erythropoietin, acting on early erythroid progenitor cells (erythroid burst-forming units, or BFU-E). Here we investigated the biological properties of this factor partially purified from the urine of anemic patients. The human urinary factor did not cause the formation of late erythroid progenitor cells (erythroid colony-forming units, or CFU-E) or enhance such colony formation in the presence of erythropoietin. Thus, the urinary factor was a different substance from erythroid potentiating activity and from activin, which act on both BFU-E and CFU-E. The urinary factor promoted the colony formation of BFU-E from both humans and mice, but the human hematopoietic growth factors such as recombinant interleukin-3, interleukin-6, granulocyte-macrophage colony-stimulating factor, and macrophage colony-stimulating factor did not stimulate the formation of BFU-E derived colonies from mice. The results suggested that the factor in the urine of anemic patients was different from the hematopoietic growth factors identified so far.
Terminal deoxynucleotidyl transferase (TdT) is an important marker for the diagnosis and the therapy of leukemia and lymphoma patients. In this study, we developed the quantitative method for the assay of TdT antigen in leukemic cells using a combination of monoclonal and polyclonal antibodies prepared for this enzyme. High correlation was obtained between the values measured by biochemical assay and enzyme-linked immunosorbent assay (ELISA). This method (ELISA), using antibodies to TdT, permits the rapid and quantitative estimation of TdT antigen in leukemic cells.