Biomedical subjects
Publications and source records attributed to M Umeda.
We isolated a rice cDNA encoding nucleoside diphosphate kinase (NDK, EC 2.7.4.6). The deduced amino acid sequence of the rice NDK shows highest homology to spinach NDK-I. The rice NDK gene exhibits a strong codon bias (73.8% GC) in the third position of the codon. DNA blot analysis indicated that at least single NDK gene is present in rice genome.
We isolated a rice cDNA clone which encodes an open reading frame of 382 amino acids. Its deduced amino acid sequence corresponds to an ATP/ADP translocator protein. Its homology with a maize ATP/ADP translocator was 83.9% in nucleotide sequence, and 90.2% of the amino acid level. Expression of this gene is regulated by such external stresses as salinity and low temperature.
We isolated a rice cDNA clone which is similar to the bovine brain-specific 14-3-3 protein (an activator protein of tyrosine and tryptophan hydroxylase involved in the synthetic pathway of monoamine) gene. The deduced amino acid sequence of the cDNA indicated a surprising similarity to a potent inhibitor of Ca(2+)-phospholipid-dependent protein kinase C. DNA blot analysis indicated that this gene is located at more than a single locus in rice genome DNA. Expression of this gene is regulated by external stresses.
Explore the source record for details and available documents.
We established a cell line (STKM-1) from tumor cells obtained from carcinomatous pleural effusion of a gastric cancer patient. The lymphocytes separated from her peripheral blood or pleural effusion were cryopreserved and immunological experiments were performed after the establishment of the cell line. They were treated with IL-2 or with both IL-2 and mitomycin C (MMC)-treated autologous STKM-1 cells. The cytolytic activity against STKM-1 cells was elevated in lymphocytes cultured with IL-2, and was more prominently augmented in lymphocytes cultured with both IL-2 and MMC-treated STKM-1 cells. The elevation in cytolytic activity was more marked with pleural effusion lymphocytes than with the peripheral blood lymphocytes. The results suggest that the lymphocytes obtained from the pleural effusion would be an excellent source for adoptive immunotherapy.
Thirty-eight human cancer cell lines and subclones derived from 12 different organs were screened for vitronectin secretion in their culture media. By immunoblotting analysis we detected high secretion by three out of five hepatoma cell lines tested but no secretion by the others. In addition, significant secretion was observed in seven non-hepatic cancer cell lines and subclones derived from the cervix, lung, and pancreas. These vitronectin-secreting cells included PLC/PRF/5, HuH-6 #5, HuH-7, HeLa S3, HeLa.P3 #2, #3, #6, #8, A549, and MIAPaCa-2. The results were further confirmed by quantitative analysis using sandwich enzyme-linked immunoassay, and activity analysis of cell attachment promotion on Western blotted filters.
Using a synthetic peptide with the amino acid sequence corresponding to residues 78-109 of human TGF-beta 1 (A78/109 peptide) as an immunogen, we generated and characterized monoclonal antibodies (Mabs) against transforming growth factor-beta 1 (TGF-beta 1). Spleen cells from a mouse immunized with carrier-free A78/109 peptide were fused with a myeloma cell line, P3U1. Two hybridoma cell lines were selected with A78/109 peptide used as the antigen, and the Mabs were designated as 3H10 and 4C10. Mab 4C10 recognized TGF-beta 1 only in immunoblotting analysis, but not in ELISA. In contrast, Mab 3H10 recognized it in not only ELISA but also immunoblotting analysis. Neither Mab was capable of recognizing both porcine TGF-beta 2 and chicken TGF-beta 3, and, therefore, both Mabs were TGF-beta 1 specific. From the results of epitope-mapping analysis, both Mab 3H10 and Mab 4C10 recognized the 78-87 portion of TGF-beta 1. The epitope recognized by Mab 4C10 was mapped specifically to amino acids 85-87. These results suggest that the region within 78-87 is exposed in the dimeric TGF-beta 1 and is one of the antigenic determinants in this molecule.
Explore the source record for details and available documents.
We have previously shown that mouse L.P3 cells secrete fibronectin and a novel protein, gelatin-non-binding and heparin-binding cell-adhesive protein (GNCP). Here, we screened and characterized cell-adhesive proteins in the conditioned media (CM's) of a series of cell lines growing in a protein- and lipid-free synthetic medium (P3 cell lines). Although cell-attachment activity of the CM's ranged from undetectable to highly significant, fractionation with affinity columns revealed the presence of significant cell-attachment activity in all CM's. Using cell-attachment assay and immunoassay on blotted filters, various cell-adhesive proteins were detected in the CM's, and most of them were identified as fibronectin, laminin, vitronectin, and collagen. GNCP-like proteins were detected in the CM's of HeLa.P3, JTC-16.P3, L.P3, and JTC-12.P3. There was no relationship between the origin of the cell lines and the cell-adhesive proteins secreted. GNCP purified from L.P3-CM was separated into 120-, 140-, 150-, and 160-kDa proteins on SDS-PAGE, which were judged to be a type of mouse type I procollagen from the following results: (1) they were digested by collagenase, (2) pepsin treatment converted the 150- and 160-kDa proteins into 120- and 140-kDa proteins, (3) they were recognized by anti-type I collagen antiserum, and (4) amino-terminal sequence of the pepsin-digested 140-kDa protein had significant homology with type I collagen. GNCP showed half-maximum activity of cell attachment at 0.03 micrograms/ml, indicating that GNCP was a cell-adhesive protein with an extremely high specific activity compared to other known cell-adhesive proteins.
Quantitative evaluation of the effectiveness of taurine against ocular surface damage caused by hypochlorous acid (HOCl) was investigated using albino rabbits. The activity of lactate dehydrogenase (LDH) released from ocular tissues into meniscus tears at eye irritation was used as an index of ocular surface damage. Instead of collecting meniscus tears directly with a glass micropipette, a new sampling method, where 150 microliters of saline was instilled into the cul-de-sac of rabbit eyes and collected all of the diluted tears within 10 s, was developed. The LDH activity after serial instillations of HOCl increased dose-dependently with increasing HOCl concentration. After serial instillation of taurine, HOCl was instilled in the same way. Pre-application of taurine effectively suppressed (p < 0.01, n = 11) the HOCl-induced LDH release as compared to saline, suggesting that the residual taurine in ocular surface tissues was still effective in protecting the tissues against HOCl by scavenging HOCl. LDH activity at 30 in after post-application of taurine was significantly lower (p < 0.01, n = 10) than that in the case of saline. This result indicates that taurine is effective in protecting the ocular surface after it has been attacked by HOCl. LDH activity in meniscus tears became a good index of quantitatively estimating ocular surface damage due to HOCl by devising the new sampling method. By using this method, we were able to prove objectively and quantitatively that taurine is effective in protecting the ocular surface against HOCl. It was suggested that taurine is clinically useful in the treatment of ocular surface damage caused by oxidants, such as HOCl.
We examined the cardioprotective effect of nisoldipine against myocardial dysfunction during ischemia and reperfusion in comparison with those of diltiazem and nifedipine in rabbit hearts perfused at constant pressure. These calcium antagonists were administered to the hearts before 60 min of ischemia. They inhibited the increase of end-diastolic pressure during ischemia in a dose-dependent manner. Diltiazem at 1.0 microM, nifedipine at 3.0 microM and nisoldipine at 0.01 microM produced the maximal cardioprotective effect. Nisoldipine had a beneficial effect with less negative inotropic effect than those of diltiazem and nifedipine and it produced a significant increase of coronary flow during reperfusion. When the vascular component was eliminated under constant flow perfusion, nisoldipine also showed the cardioprotective effect. Nisoldipine did not produce any beneficial effect without the inhibition of the increase in end-diastolic pressure during ischemia nor did it do so without the increase of reperfusion flow. Therefore, the nisoldipine-increased coronary flow during reperfusion as well as the inhibition of ischemic contracture by nisoldipine seems to play a crucial role in improving the myocardial dysfunction of ischemic-reperfused hearts.
The scent gland of Syrian hamsters is named the flank gland and thought to play an important role in the communication among individuals of the same species as in a few other rodents. In the present paper, the flank gland of Syrian hamsters of both sexes was macroscopically and histologically studied with special reference to the aging. This gland reached the plateau in size at 21 days of age in females, while it continued to develop until 70 days of age in males. Its shape was circular in adult females, and elliptical in adult males, respectively. Histologically, it was identified as the developing sebaceous glands with hair follicles in both sexes at 21 days of age. Thereafter, it gradually increased in size and number of lobules. It was always better developed in males than in females at each day examined in the present study, although there were no sex differences in its histological structure.
The musk gland of the adult house musk shrews (Suncus murinus) of both sexes was studied lectin histochemically. The musk gland was a kind of scent gland, consisted of congregation of branched or unbranched simple tubuloalveolar gland holocrine in nature and was attached by an apocrine gland-like structure (sweat gland) in the deeper layer of its periphery. Acinar cells of the musk gland were distinguishable into three type from basal to luminal parts of the acinus; immature cells, mature cells and degenerating cells. There was no histological difference between both sexes. Lectin-binding pattern of the musk gland was examined in comparison with that of the sweat gland and ordinary sebaceous gland by histochemical staining techniques using seven lectins: ConA, RCA I, PNA, SBA, UEA-I, DBA, WGA, WGA and PNA labelled the duct of the musk gland more intense than the acinus. Several lectins showed a tendency to label the cells situated near the luminal surface more intense than those near the basement membrane in both the acinus and duct of the musk gland. In the sweat gland and ordinary sebaceous gland, the lectin-binding pattern was different with each other and from that in the musk gland. These findings suggest that the musk gland, sweat gland, and ordinary sebaceous gland are different to each other in nature of cells and the secretion.
The presence of the scent glands were examined macroscopically and histologically in the Japanese vole, Microtus montebelli and the Hungarian vole, Microtus arvalis. A pair of scent glands were observed on the buttocks of the Japanese vole as the oval nodules with yellowish colored skin. The size of the glands was larger in males than in females. Histologically, the glands were composed of the aggregation of many acini of sebaceous glands. The epidermis above the scent glands were thicker than that in the other areas. In the Hungarian vole, no such gland could be detected with macroscopical and histological examinations.
Hypophosphatasia is a congenital disease characterized by defective bone mineralization, deficiency of alkaline phosphatase (ALP) activity, increased excretion of the phosphoethanolamine (PEA) in the urine, and premature loss of the deciduous teeth. A male hypophosphatasia patient (aged 15 years 6 months) with premature exfoliation of the deciduous teeth was referred to our hospital because of severe periodontal destruction in the permanent dentition. Blood and urine tests as well as oral and periodontal examinations were performed. Serum antibody titers against 7 periodontopathic bacteria by the enzyme-linked immunosorbent assay (ELISA), monocyte and neutrophil chemotaxis measurements, and cellular immunity tests were also performed. Low levels of ALP in serum and PEA in the urine were found. Monocyte and neutrophil chemotaxis exhibited normal values. Slightly depressed CD2+, CD3+, and CD4+ and slightly elevated activity of NK cells were found. An elevated level of serum antibody to Porphyromonas gingivalis was observed. Oral radiographic examination showed a mirror pattern of alveolar bone loss which is similar to that seen in localized juvenile periodontitis. Periodontal treatment of this patient was carried out for 4 years. The severely affected sites, the lower right and left first molars and the upper right first molar, had to be extracted. However, the other sites were well maintained. The serum IgG level against P. gingivalis was decreased after 4 years of periodontal treatment. Infection with P. gingivalis was suspected to be associated with the destruction of this patient's hypophosphatasia, but other dental abnormalities such as abnormal enamel, dentin, and cementum formation may also have contributed to the periodontal pathology.
A 66-year-old woman visited a local hospital complaining of left brachial tumor in June 1988. She was diagnosed as having non-Hodgkin's lymphoma (diffuse medium-sized type) by biopsy of the tumor. No other lesions were detected at any site. The disease was classified as stage IA and radiotherapy was carried out resulting in complete remission (CR). Because a tumor was observed at the same site again she was admitted to our hospital. Under a diagnosis of recurrence, she received 2 courses of COP-BLAM therapy, and achieved CR. Subsequently she was treated with chemotherapy as an outpatient. As another tumor developed at the same site in June 1992, the patient was readmitted. The re-evaluation of clinical stage revealed no other lesions. Under a diagnosis of second recurrence, still in stage I, she underwent both chemotherapy (IMV-triple P therapy) and radiotherapy, and CR was achieved again. She is now being treated as an outpatient. Malignant lymphoma originating from soft tissue, particularly the skeletal muscle, is rare. The present case was considered to be rare because the tumor was a non-Hodgkin's lymphoma originating from the left brachial soft tissue, in which the tumor recurred twice at the same site.
Although the third generation regimens for the treatment of malignant lymphoma are used more predominantly than those of other generations in recent years, a number of elderly patients die due to the toxicity of those treatments. We treated elderly patients with non-Hodgkin's lymphoma (NHL) with COP-BLAM III, and compared its therapeutic and adverse effects on elderly NHL patients (more than 65 years of age) with those in non-elderly patients. Nine previously untreated elderly NHL patients (4 males and 5 females, with a median age of 71 years) and 13 previously untreated NHL non-elderly patients (6 males and 7 females, with a median age of 43 years) were treated with COP-BLAM III. According to the Ann Arbor staging rules for malignant lymphoma, 4 and 6 elderly patients were classified in stage III and IV, respectively, while 6 and 7 non-elderly patients were in stage III and IV, respectively. There were 5 elderly patients with diffuse large cell type lymphoma (D. large), 3 with diffuse medium-sized cell type (D. medium) and one with diffuse mixed-cell type (D. mixed), while there were 6 non-elderly patients with D. large, 6 with D. medium and one with D. mixed. The COP-BLAM III regimen consisting of portion A with continuous infusion of the drugs and portion B with intravenous bolus administration of drugs was identical to that used by Boyd et al. Portion A and portion B were repeated every 3 weeks alternatingly. The administration of granulocyte-colony stimulating factor (G-CSF) was started, when the granulocyte count fell to below 1,000/microliters.(ABSTRACT TRUNCATED AT 250 WORDS)
We examined neutrophil functions in seven elderly patients with non-Hodgkin's lymphoma before and during treatment with granulocyte colony-stimulating factor (G-CSF) at the neutropenic stage after combination chemotherapy. Subcutaneous injection of 75 micrograms/d of G-CSF produced by E. coli was started when the neutrophil count decreased less than 1,500/microliter, and continued until the neutrophil count increased to about 10,000/microliter. The phagocytic activity of neutrophils from the elderly on day 3 of G-CSF treatment was markedly enhanced; 1,129.9 +/- 403 ps/100 PMNs, which was 185.7 +/- 31.4% (p < 0.001) as compared with that before G-CSF treatment. The neutrophil alkaline phosphatase (NAP) activity was also enhanced on day 3; 398.3 +/- 48 score, which was 135.2 +/- 5.1% (p < 0.001) as compared with that before G-CSF treatment. Two patients developed interstitial pneumonitis during or shortly after the treatment with G-CSF. Interstitial pneumonitis suddenly developed when their neutrophil count was increased, and the phagocytic activity and NAP activity recovered. The phagocytic activity of neutrophils from them was enhanced to 1,090 +/- 26 ps/100 PMNs and 772 ps/100 PMNs during the treatment with G-CSF, as compared with that before G-CSF treatment of 644 +/- 29 ps/100 PMNs and 465 +/- 69 ps/100 PMNs, respectively. The NAP activity was also enhanced to 372 from 264. One patient suffered from transient pulmonary dysfunction during the treatment with G-CSF. His neutrophil count was more than 13,000/microliter, and the phagocytic activity enhanced to 949 +/- 105 ps/100 PMNs. Dyspnea with suppressed PaO2 recovered reversibly after cessation of G-CSF.(ABSTRACT TRUNCATED AT 250 WORDS)