Chemotaxis and phagocytosis of the neutrophils mobilized by granulocyte colony-stimulating factor in healthy donors for granulocyte transfusions.
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Blood Donors↗
Biomedical subjects
M Umeda
Publications and source records attributed to M Umeda.
Infection-induced transient remission and increase in the platelet count in idiopathic thrombocytopenic purpura.
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Communicable Diseases↗
Expression of functional proteins of cDNA encoding rice nucleoside diphosphate kinase (NDK) in Escherichia coli and organ-related alteration of NDK activities during rice seed germination (Oryza sativa L.).
The GST (glutathione S-transferase)-NDK (nucleoside diphosphate kinase) fusion protein was expressed in Escherichia coli. The GST-NDK protein was capable of transferring gamma-phosphate from ATP to nucleoside diphosphates such as GDP, CDP, TDP and UDP. Western blot analysis using anti-NDK antibody indicated that NDK in endosperm gradually decreased during 36 h of imbibition. On the contrary, NDK in embryo increased during the same period. NDK activities in both tissues were in accord with these observations. Whereas the NDK protein in roots of rice seedlings during 7 days of imbibition remained constant, in shoots it declined after 5 days of imbibition. Thus, NDK may play a significant role in the cellular event modulated by adenylate energy charge level.
Cloning, Molecular↗
Analysis of randomly isolated cDNAs from developing endosperm of rice (Oryza sativa L.): evaluation of expressed sequence tags, and expression levels of mRNAs.
Using a cDNA library prepared from poly(A)+ RNA from 10-day-old rice endosperm, partial nucleotide sequences of randomly isolated clones were analyzed. A total of 153 (30.6%) out of 500 cDNA clones showed high amino acid identity to previously identified genes. There was significant redundancy in cDNAs encoding prolamine and glutelin. About 21.0% of the cDNA clones were found to code for seed storage protein genes. Consequently, 37 independent genes were identified. Using cDNA clones encoding glutelin, prolamine, seed allergen, alpha-1,4-glucan branching enzyme, glycine-rich RNA binding protein, metallothionein, non-specific lipid-transfer protein and ubiquitin conjugating enzyme the accumulation of mRNA during rice seed development was compared. Genes associated with seed storage protein and starch biosynthesis were expressed according to expected developmental stages. Glycine-rich RNA binding protein genes as well as metallothionein-like protein genes were highly expressed in developing seeds, but low in leaves of whole plants.
Blotting, Northern↗
Marked induction of gelatinases, especially type B, in host fibroblasts by human ovarian cancer cells in athymic mice.
Two human ovarian adenocarcinoma cell lines, MCAS-3 and OVISE-3 were found to secrete little of any type of gelatinase in tissue culture. However, when these cell lines were implanted subcutaneously into nude mice the cyst fluids from the resultant tumors contained gelatinase A and/or B. The enzyme activities, especially of gelatinase B, were much higher in the malignant MCAS-3 tumors than in those of the less malignant OVISE-3 tumor cells. To elucidate the origin of gelatinase B in cyst fluids of the MCAS-3 tumors, murine skin fibroblasts (MSF) were isolated from a subcutaneous tumor in a nude mouse and tested for their proteinase secretion in culture. MSF cells, which secreted some gelatinase A and gelatinase B, were induced to secrete high levels of both enzymes, especially gelatinase B, by co-cultivation with MCAS-3 cells. In addition, gelatinase A activity was induced by incubation of MSF cells with the conditioned medium of either MCAS-3 or OVISE-3 cells, whereas gelatinase B was induced only with that of MCAS-3. Although cytokines or growth factors such as IL-1 beta, TGF-beta 1, TNF-alpha or EGF stimulated the secretion of gelatinases A and B from MSF cells, their effects on gelatinase B activity were far less than that of the MCAS-3 conditioned medium. These results indicate that the major part of gelatinase B activity in the cyst fluids of the ovarian tumors is secreted by host interstitial cells stimulated by tumor-derived humoral factors. Similar tumor cell-host cell interactions may be important in the production of various proteinases in other tumor types.
Animals↗
Effect of Clostridium perfringens-derived wound healing substance as compared with epidermal growth factor on the growth and morphological transformation of BALB/3T3 A31-1-1 cells.
Clostridium perfringens-derived wound-healing substance (WHS), having growth-stimulating activity, was examined to determine its effect on the growth and morphological transformation of BALB/3T3 A31-1-1 cells. WHS accelerated the cell growth at the exponential growth phase, shortening the doubling time by 8-18%. The maximum cell density of the treated cultures was slightly higher than that of the control culture, and the cell number decreased in the same way as the control cells did. On the other hand, the cells treated with epidermal growth factor (EGF) or insulin showed growth rates similar to that of the control cells during the exponential growth phase, and after the control cells attained the maximum cell number, the number of the treated cells continued to increase gradually for more than 4 days and then decreased. Under the experimental conditions of the two-stage transformation assay, application of WHS at the tumor-initiation or promotion stage did not accelerate the formation of transformed foci. Although treatment with EGF at the initiation stage induced no enhancement, marked enhancement of morphological transformation was observed in the treatment at the promotion stage. These results indicate that the mode of action between WHS and EGF or insulin is different on the growth-stimulating activity and morphological transformation of BALB/3T3 A31-1-1 cells.
3T3 Cells↗
Possible anti-tumour-promoting activity of components in Japanese soybean fermented food, Natto: effect on gap junctional intercellular communication.
In order to detect any protective agent against tumor formation, we examined the anti-tumor-promoting effect of a Japanese traditional soybean fermented food, Natto. Dye transfer was employed as an assay method. When fluorescent dye was microinjected into cultured BALB/3T3 cells, the dye was transformed into the neighboring cells through the gap junction. This dye transfer was blocked by the treatment with the tumor promoters 12-O-tetra-decanoylphorbol-13-acetate (TPA), a high concentration of NaCl and lithocholic acid (LCA). This reduction of the dye transfer by TPA treatment was not observed when the cells were pretreated with retinoic acid, an anti-tumor promoter. Thus, the recovery of the dye transfer in TPA-treated BALB/3T3 cells was proven to ge a good indicator for detecting some possible anti-tumor promoters. After extraction and fractionation of Natto, we obtained an active fraction (H1) which showed recovery of the dye transfer in TPA-treated cells. The fraction contained straight-chain saturated hydrocarbons. A comparison of the fraction and the authentic samples by GC analysis suggests that the H1 fraction contained straight-chain saturated hydrocarbons from around C30 to C32. Among these hydrocarbons, hentriacontane (C31) was found at the highest concentrations, and was shown to have the highest activity. Hentriacontane at a very low concentration of 0.65 ng/ml was shown to recover the dye transfer inhibited by the treatment with TPA as well as with NaCl and LCA.
3T3 Cells↗
Improvement in the efficiency of the in vitro transformation assay method using BALB/3T3 A31-1-1 cells.
In order to improve the in vitro transformation assay using BALB/3T3 cells, which is routinely conducted with minimal essential medium (MEM) containing 10% fetal calf serum (FCS), we examined the effect of a new medium after the cells had been treated with carcinogens. Preliminary experiments suggested that the use of T medium (modified DME/F-12) supplemented with insulin, transferrin, ethanolamine and sodium selenite plus 2% FCS resulted in a high transformation frequency. The present study confirmed that this medium was very efficient at inducing transformation foci. Transformation frequency was highest when 12-O-tetradecanoylphorbol-13-acetate (TPA) was started from 1 week after the carcinogen treatment. According to the new protocol using this medium, the transformation frequency was five times higher and transformation foci appeared much earlier than with the protocol using usual MEM plus 10% FCS medium. In addition, we tested several typical carcinogens and a promoter in order to confirm the applicability of the new protocol to the two-stage transformation assay. N-Methyl-N'-nitro-N-nitro-soguanidine (MNNG) and benzo[a]pyrene, but not noncarcinogenic benzo[e]pyrene, induced transformation foci. In the metabolic activation system dimethylnitrosamine gave a highly positive transformation. Okadaic acid, a non-TPA-type tumor promoter, enhanced transformation of MNNG-initiated cells. These studies demonstrate that the use of the proposed medium drastically improves transformation frequency in the two-stage in vitro transformation assay.
3T3 Cells↗
Specific binding of a synthetic peptide derived from an antibody complementarity determining region to phosphatidylserine.
We have established a series of monoclonal antibodies that bind to phosphatidylserine (PS). One mAb, PS4A7, showed a strict specificity for PS and distinguished the stereospecific configuration of its serine moiety. We determined the amino acid sequences of the heavy and light chain variable regions of PS4A7, and examined the reactivity of the synthetic peptides corresponding to the complementarity determining region (CDR) of the mAb with phospholipids. We found that a 12-amino acid synthetic peptide corresponding to the third CDR of the heavy chain (amino acid residues 93-102, referred to as CDR3-H) bound specifically to PS. Although the affinity of the peptide to PS was markedly lower, the peptide was shown to bind to 1,2-diacyl-sn-glycero-3-phospho-L-serine (PS), but not to 1,2-diacyl-sn-glycero-3-phospho-D-serine, showing a similar specificity to that of PS4A7. The specific binding of the CDR3-H peptide to PS was confirmed by ELISA and TLC-immunostaining assay. The interaction between the CDR3-H peptide and water-soluble PS-derivatives was investigated by inhibition of the ELISA. PS effectively inhibited the binding and phosphoserine showed a weak but significant inhibition, but no appreciable inhibition was observed with serine. These observations suggest that the CDR3-H peptide plays a major role in the interaction of PS4A7 with the phosphoserine residue of the PS molecule.
Amino Acid Sequence↗
Characterization of matrix-degrading proteinases and their inhibitors secreted by human gynecological carcinoma cells.
Matrix-degrading proteinases secreted by tumor cells play crucial roles in tumor cell invasion and metastasis. Serum-free conditioned media of 7 human gynecological carcinoma cell lines were examined for proteinases and their inhibitors by using gelatin zymography, reverse zymography and immunoblotting. All of three ovarian adenocarcinoma cell lines secreted urokinase-type plasminogen activator. Among them, a mucinous cystadenocarcinoma cell line also secreted tissue-type plasminogen activator, plasmin-like enzyme and trypsinogen. On the other hand, two ovarian undifferentiated carcinoma cell lines mainly secreted glatinase A or B. A choriocarcinoma cell line secreted multiple metalloproteinases in the highest amount, whereas an endometrial adenocarcinoma cell line (HEC-1) derived from an early clinical stage hardly secreted any gelatinolytic enzyme. The five high proteinases producers hardly secreted the corresponding inhibitors, such as tissue inhibitor of metalloproteinases (TIMP)-1, -2 or plasminogen activator inhibitor-1. In contrast to these highly malignant cell lines, a poor proteinase producer, HEC-1, secreted a large amount of TIMPs. Therefore, an enhanced proteolytic tendency appears to be associated with gynecological cancer cells established from highly malignant tumors.
Adenocarcinoma, Clear Cell↗
Characterization of oral treponemes isolated from human periodontal pockets.
A total of 90 clinical strains of oral treponemes was isolated from subgingival plaque in patients with periodontal disease. They were characterized by biochemical means as well as cell enzyme, protein analysis using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and DNA dot blot hybridization. Sixty strains were isolated on Medium 10 (M10), which was fundamentally serum-free. The remainder were isolated on serum-containing media. Isolates were divided into 6 groups according to their biochemical characteristics. Three of the 6 groups were asaccharolytic, and 2 of these 3 groups were Treponema denticola and "Treponema vincentii". The other 3 groups were saccharolytic and further divided into 9 subgroups. The analysis of cell enzyme, cell protein and dot blot hybridization with the DNA probe of Treponema socranskii indicated that all the saccharolytic groups were T. socranskii or closely related species. This study indicated that the newly characterized saccharolytic oral treponemes could be identified using M10 from the human periodontal pocket.
Bacterial Proteins↗
COP-BLAM regimen combined with granulocyte colony-stimulating factor and high-grade non-Hodgkin's lymphoma.
The clinical efficacy of COP-BLAM chemotherapy combined with human recombinant granulocyte colony-stimulating factor (G-CSF) was evaluated in 104 previously untreated patients with non-Hodgkin's lymphoma (NHL). According to the method of Laurence et al., a modified COP-BLAM regimen was administered every 21 days. G-CSF was added when the granulocyte count fell below 1000 x 10(9)/l. Ninety-eight of 104 (94.2%) patients achieved a complete remission; the 4-year survival rate was 82.4% with a median duration of observation of 26 months. Survival was significantly longer in patients with low serum LDH levels, B-cell type or low CRP or in earlier clinical stages, than in patients with high serum LDH levels, T-cell type of higher CRP levels or in advanced clinical stages. The mean duration of administration of G-CSF was 5.4 days. Twelve patients developed infections during treatment. The adverse effects of G-CSF included interstitial pneumonia, bone pain and fever. Patients administered COP-BLAM combined with G-CSF achieved a high rate of remission and had a low incidence of infection. Nearly all the patients could be treated in 21-day cycles. The results suggest that G-CSF combined with COP-BLAM was effective in treating NHL, because the patients can tolerate a higher dose of the anticancer agents.
Adolescent↗
Non-invasive measurement of brain activity using functional MRI: toward the study of brain response to acupuncture stimulation.
We studied functional MRI in 15 male and 13 female normal volunteers on a clinical MRI system using gradient echo sequence. During the experiments, brain activation was induced by grasping the unilateral hand once or twice a second for motor tasks. A localized increase of MRI signal in the contralateral motor cortex was observed in 17 out of 21 cases (81%) under right hand motor task and 11 out of 21 cases (52%) under left hand motor task. The application of this method may be useful to evaluate brain response to acupuncture.
Acupuncture Therapy↗
Regression of hypertrophic osteopathy following removal of intrathoracic neoplasia derived from vagus nerve in a dog.
Surgical removal of an intrathoracic tumor derived from a vagus nerve was undergone in a dog with hypertrophic osteopathy. The tumor was pathologically diagnosed as malignant schwanoma. Soft tissue swelling, lameness, and itchiness in four limbs disappeared within 7 days after surgery. The proliferated periosteal osteophytes of the four limbs was progressively reduced with time by follow-up radiography on the 58th day after surgery. On the 710th day after surgery, these osteophytes were greatly decreased as osteopathy, malignant schwanoma.
Animals↗
[Evaluation of respiratory tract disease accompanying malignant lymphoma in the elderly].
We studied the usefulness of bronchoscopy in the diagnosis of pulmonary complications in elderly patients with non-Hodgkin's lymphoma (NHL). The subjects were 12 patients 65 or older with NHL. Two of these patients underwent transbronchial lung biopsies (TBLB), all of them underwent bronchiolavage (BAL) and 2 with tumors underwent transbronchial biopsies of the tumors. Diagnosis was obtained in 8 out of 12 patients (66.7%). Two patients showed infiltration of lymphoma cells, 2 had cytomegalovirus (CMV) infection, 1 had pneumocystosis (P. carinii), 2 had fungal infections and 1 had a bacterial infection. TBLB was performed in 2 patients with platelet counts of over 5 x 10(4)/microliters. Lymphoma cell infiltration was diagnosed in one of these patients and a fungal infection in the other. BAL was performed on all patients. CMV was detected in 2 cases by the PCR method, P. carinii was detected in 1 case and aspergillus in 1 case. Klebsiella pneumonia was diagnosed in 1 patient. Since respiratory tract complications are often serious in NHL and the prognosis depends on early diagnosis and treatment, it is important to perform bronchoscopy on elderly patients and obtain a definitive diagnosis.
Aged↗
[Usefulness of COP-BLAM therapy with concomitant G-CSF in elderly patients with non-Hodgkin's lymphoma in comparison with patients not given G-CSF].
COP-BLAM therapy with concomitant G-CSF was performed in patients 65 years of age of older with non-Hodgkin's lymphoma (NHL) and the therapeutic effects and its adverse effects were compared with those in a group not given G-CSF concomitantly. The subjects were 64 patients with NHL, divided into 36 in the G-CSF group and 28 in the non-G-CSF group. In the G-CSF group, complete remission (CR) was achieved in 88.9% and the efficacy rate was 94.5%. In the non-G-CSF group, 89.3% achieved CR, and there was no significant difference between the G-CSF and non-G-CSF groups. The survival time and duration of remission also showed no significant differences between the G-CSF and non-G-CSF groups. The frequency of granulocytopenia as an adverse effects was significantly reduced in the G-CSF group, but the other adverse effects showed no intergroup differences. The occurrence rates of fever of at least 37.5 degrees C and documented infection were significantly less in the G-CSF group. These results indicate that COP-BLAM therapy with concomitant G-CSF achieved a high remission rate, showed few severe adverse effects, especially infections, and can be safely performed in elderly patients.
Aged↗
[Subgingival distribution of periodontopathic bacteria in periodontic patients and susceptibility of these bacteria to minocycline-HCl].
The present study was carried out to examine the distribution of six periodontopathic bacteria in deep periodontal pockets and to reconfirm the effect of Periocline on these periodontopathic bacteria. Samples from sixty-two periodontal pockets were collected at pocket depths of over 4 mm in twenty-one periodontitis patients aged 43 to 75 years. After sampling, Periocline was applied topically to the selected pockets once a week for four weeks and reexamined. The detected rates of the periodontopathic bacteria were Capnocytophaga sputigena (37.1%), Prevotella intermedia (22.6%), Porphyromonas gingivalis (22.6%), Fusobacterium nucleatum (20.1%), Actinobacillus actinomycetemcomitans (9.7%) and Eikenella corrodens (4.8%). The distribution of the bacteria was compound because two or three bacterial species were found to coexist. In view of the MIC of minocycline-HCI for these bacteria, increase of most of the measured bacteria was suppressed by the concentration of drugs, including Periocline. However, clinical strains of P. i. were considered to have low susceptibility to minocycline-HCl. In view of the effect of topical application of drugs, no significant differences were found. From these results, it was suggested that Periocline contained effective concentration of minocycline-HCl.
Adult↗
[Usefulness of 123I-MIBG myocardial SPECT in patients with hematologic malignancies with chemotherapeutic agent-induced cardiomyopathy].
In 59 patients with hematologic malignancy undergoing chemotherapy regimens including adriamycin (ADR), myocardial imaging was performed by SPECT with 123I-metaiodobenzylguanidine (MIBG) to detect cardiac sympathetic nerve abnormalities. SPECT imaging was performed 20 min and 4 hr after intravenous injection of 123I-MIBG, and the washout rate (WR) for the entire left ventricle was calculated. Then the relationship of the WR to the total dose of ADR, the left ventricular ejection fraction, and the frequency of arrhythmias was evaluated. It was found that the WR was related to the total dose of ADR, suggesting that it is a useful parameter for cardiac sympathetic nerve damage. As the WR increased, the frequency of ventricular arrhythmias also increased. The WR was usually normalized 3-6 months after the discontinuation of ADR. In patients with a WR > or = 50%, however, prolonged follow-up was required. In conclusion, the WR on 123I-MIBG myocardial SPECT scans may be useful as a parameter for predicting cardiac toxicity requiring the discontinuation of ADR or dose reduction.
3-Iodobenzylguanidine↗