Nucleotide sequence of a cDNA encoding a protein kinase homologue in Arabidopsis thaliana.
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Amino Acid Sequence↗
Biomedical subjects
H Harada
Publications and source records attributed to H Harada.
Isolation and characterization of a cDNA that encodes ECP31, an embryogenic-cell protein from carrot.
A full-length cDNA for ECP31, an embryogenic cell protein from carrot (Daucus carota L.) with a M(r) of 31,000 (Kiyosue T, Satoh S, Kamada H, Harada H (1991) Plant Physiol 95: 1077-1083), was isolated from a cDNA library prepared from embryogenic cells using PCR-amplified DNA as a probe. The genomic Southern blot analysis revealed that there are two or three genes for ECP31 in the carrot genome. The transcripts of ECP31 accumulated in the peripheral regions of clusters of embryogenic cells and disappeared in the course of somatic embryogenesis that was induced by transfer of the embryogenic cells to auxin-free media. The cDNA encodes a polypeptide of 256 amino acids, and the calculated molecular weight of this polypeptide is 26,111. The deduced amino acid sequence shows a high degree (62.2%) of similarity to that of a protein that is abundant during late embryogenesis of cotton (LEA D34; Baker JC, Steele C, Dure III (1988) Plant Mol Biol 11: 227-291). The mRNAs for ECP31 started to accumulate in zygotic embryos at a late stage of embryogenesis but were undetectable in mature embryos within 24 h after imbibition of seeds. In dry fruits (seeds), the transcripts were detected only in zygotic embryos by in situ hybridization. The level of ECP31 transcripts increased after treatment with abscisic acid (ABA) in torpedo-shaped somatic embryos but not in seven-day-old seedlings. These results suggest that both embryo-specific factor(s) and ABA are involved in the expression of the gene for ECP31.
Abscisic Acid↗
Primary and secondary culture of rat ameloblasts in serum-free medium.
Enamel is the hardest tissue in vertebrates. Ameloblasts are derived from epithelial cells and are responsible for enamel formation. They secrete enamel matrix components in which amelogenins are the major proteins, the biochemical properties of which are well known. However, little is known about the characteristics of ameloblasts themselves or about the functions of amelogenins. In this study, we developed a novel primary and secondary culture system for ameloblasts using a monoclonal antibody which recognized amelogenin (En3). The cell layer on dentine removed from rat mandibular incisors was isolated and cultured in low calcium, serum-free medium. Primary culture was performed on collagen-coated culture plates and typically, two types of cells appeared. One major type changed morphology after the addition of a high concentration of calcium to the medium. Expression of amelogenin was shown as cytoplasmic particles in these cells using En3. In the secondary culture, expression of amelogenins was also observed. In this system, the cells grew and maintained the expression of amelogenin for about 3 weeks.
Ameloblasts↗
Effects of L-threo- and erythro-3,4-dihydroxyphenylserine on learning performance and concentrations of brain noradrenaline and its metabolites in rats.
Effects of L-threo and L-erythro-3,4-dihydroxyphenylserine [DOPS, precursor amino acids for noradrenaline (NA)] on the learning performance in a maze paradigm designed to model on the water maze paradigm using a multicomputerized behavioral analysis system were studied. A marked facilitation of learning performance was observed in rats after an intraventricular injection of 5 micrograms L-threo-DOPS (the s-NA precursor), and this effect was inhibited by a simultaneous administration of 1 or 2 micrograms propranolol (a beta-adrenergic antagonist). As concentrations of brain NA, 3-methoxy-4-hydroxyphenylglycol, and normethanephrine were increased by the injection of 5 micrograms L-threo-DOPS, the effect seemed to be derived from activation of beta-adrenoceptors in the CNS by the formed s-NA. On the other hand, an intraventricular injection of 5 micrograms L-erythro-DOPS (the r-NA precursor) attenuated the learning performance, and this effect was probably caused by the formed r-NA from L-erythro-DOPS.
Animals↗
Sequencing and population analysis of four novel HLA-DPA1 alleles.
We determined the base sequences of the HLA-DPA1 gene from four B-lymphoblastoid cell lines (CB6B, LKT3, AMAI, and T7526) that showed distinct electrophoretic patterns of single-stranded polymerase chain reaction products of the HLA-DPA1 gene. The novel HLA-DPA1 alleles of CB6B, LKT3, AMAI, and T7526 were designated DPA1*02021, DPA1*02022, DPA1*0301, and DPA1*0401, respectively. Although there was only one base substitution between DPA1*02021 and DPA1*02022, the single-strand conformation polymorphism of these two alleles was clearly demonstrated by electrophoresis in a nondenaturing polyacrylamide gel containing 10% glycerol. In addition, we genotyped for the HLA-DPA1 gene of healthy unrelated Oriental individuals--i.e., 227 Japanese, 88 Papua New Guineans, and 41 Buyi-Chinese--to demonstrate the ethnic distribution of the HLA-DPA1 alleles.
Alleles↗
Light requirement for shoot regeneration in horseradish hairy roots.
Hairy roots of horseradish (Armoracia rusticana) were induced by inoculation with Agrobacterium rhizogenes harboring Ri plasmid and cultured on phytohormone-free Murashige and Skoog medium after eliminating the bacteria. Hairy roots grew vigorously and sometimes formed yellowish calli under dark conditions. On the other hand, growth of hairy roots stopped after several weeks of culture with light, then shoots were regenerated. Frequency of shoot formation from hairy roots increased as the culture period in light lengthened and the light intensity increased. The shoot regeneration was induced by treatment with white or red light, but not with far-red light. Shoot regeneration by red light was inhibited by following treatment with far-red light. Red and far-red light reversibly affected shoot regeneration. Excised roots of nontransformed plants grew quite slowly on phytohormone-free Murashige and Skoog medium and occasionally formed shoots under white light conditions.
Journal Article↗
DNA typing of HLA class II genes in B-lymphoblastoid cell lines homozygous for HLA.
The HLA class II genotypes were determined in the B-lymphoblastoid cell lines selected for the Tenth International Histocompatibility Workshop. The HLA class II genes were determined by the PCR-SSOP method using the reagents provided by the Eleventh Histocompatibility Workshop. Additional studies have been performed for further characterization of HLA class II polymorphism on these cell lines. It is observed that several cell lines have HLA class II haplotypes with the same DRB1, DQA1 and DQB1 alleles on both haplotypes but different alleles at the other class II loci, confirming that these cell lines are not truly HLA class II-homozygous. Other cell lines carried HLA class II haplotypes which were only different at the DRB1 gene. These results suggest double recombination events or gene conversion-like events in generation of HLA DR, DQ haplotypes. These cell lines provide an important tool as references for HLA DNA typing.
Alleles↗
Expression of MUC1 and MUC2 mucins by human tumor cell lines.
The secretion and nature of mucins produced from a panel of recently available new gastric and colon carcinoma cell lines (LIM1839, LIM1215, LIM1863, LIM1899, LIM2099, LIM2405, LIM2408, LIM2412, LIM2463), as well as other colon (LS174T, HT29, HT29-SB, COLO533, COLO206), breast (T47D, MCF-7, BT20, ZR75-1) and ovarian (COLO316) tumor cell lines, was investigated. ELISA and Western blotting of the culture supernatants with novel anti-MUC1 and anti-MUC2 monoclonal antibodies (MAbs) specific for mucin core proteins showed their secretion by most of these cell lines. In addition, mucins produced by these cell lines expressed the tumor-associated carbohydrate detected by MAb 3E1.2 (glycolylsialyl-Tn, mammary serum antigen or MSA) and the Tn or T antigens reactive with lectin SSA-M. SSA-M detected MUC1 or MUC2 captured by MAbs BC2 or CCP58, while 3E1.2 only detected MUC1-associated carbohydrate, indicating that the MAb may react with a conformationally dependent epitope, or that the sialyl/glycolyl-transferases involved in MSA production may be sequence specific. In addition, the BC2/SSA-M and CCP58/SSA-M assays detected mucins in some samples which were not detected by BC2/BC2 or CCP58/CCP58 dual determinant assays, indicating that this format may be more appropriate for the detection of tumor-associated mucins in body fluids. These new cell lines and assays should be of use in the investigation of mucin core proteins, particularly LIM2463 and LIM1839 which express significant quantities of both MUC1 and MUC2.
Amino Acid Sequence↗
Synthesis of 3-substituted 2-oxo-1,4-thiazines and evaluation of their protective effect against endotoxin shock in mouse.
5-Methyl-2,3-dioxo-2H,4H-1,4-thiazine (7) was obtained by the oxidation of 5-methyl-2H-1,4-thiazin-3(4H)-one (5) with m-chloroperbenzoic acid in MeOH, followed by acid hydrolysis of the resulting 2,2-dimethoxy-1,4-thiazine (6). 3-Chloro-2-oxo-1,4-thiazine (10), which was obtained from 7 by heating with phosphorous oxychloride, reacted with various nucleophiles to give 3-substituted 2-oxo-1,4-thiazines (11a--y). Some of these 2-oxo-1,4-thiazines, 11a--b, e, o and r--s, showed a protective effect against endotoxin shock in D-galactosamine-sensitized mice.
Animals↗
Novel benzamides as selective and potent gastrokinetic agents. III. Synthesis and structure-activity relationships of 4-amino-5-chloro-2-methoxy- and 2-ethoxy-N-[(4-substituted 2-morpholinyl)methyl]-benzamides.
A series of 4-amino-5-chloro-2-methoxy- and 2-ethoxy-N-[(4-substituted 2-morpholinyl)methyl]benzamides (11-64) were prepared and evaluated for gastrokinetic activity by determining their effects on the gastric emptying of phenol red semisolid meal in rats. The N-4 substituent includes alkyl, phenoxyalkyl, (4-fluorobenzoyl)alkyl, and heteroarylmethyl groups. The benzamide derivatives, having an isopropyl, isoamyl, neopentyl, 3-(4-chlorophenoxy)-propyl, or pyridylmethyl group at N-4, showed potent in vivo gastric emptying activity. In particular, 4-amino-5-chloro-2-ethoxy-N-[[4-(3-pyridylmethyl)-2- morpholinyl]methyl]benzamide (57b) was equipotent to the 4-fluorobenzyl analogue 1b (AS-4370 as its citrate) in the gastrokinetic activity on phenol red semisolid meal in rats and mice, and on resin pellet solid meal in rats. Moreover, compound 57b was free from dopamine D2 receptor antagonistic activity in both in vitro ([3H]spiperone binding) and in vivo (apomorphine-induced emesis in dogs) tests. Structure-activity relationships of compounds with various substituents at N-4 are also discussed.
Animals↗
Synthesis and structure-activity relationships of substituted 2-[(2-imidazolylsulfinyl)methyl]anilines as a new class of gastric H+/K(+)-ATPase inhibitors. II.
A series of 2-[(2-imidazolylsulfinyl)methyl]anilines (2) having various substituents on their imidazole and aniline rings was synthesized and examined for their H+/K(+)-ATPase (adenosine triphosphatase) inhibitory effects and antisecretory activity against histamine-stimulated gastric acid secretions in Heidenhain pouch dogs. Although substitutions on the imidazole ring did not enhance biological activity, substitutions on the aniline ring by electron-donating substituents potently enhanced the enzyme inhibitory activity and also showed an inhibitory effect on histamine-stimulated gastric acid secretion after oral administration. In particular, the in vitro activity of the dimethyl (2u--w) and trimethyl (2ac) derivatives was about 10 times that of omeprazole. Also, 4-methyl (2k), 4-methoxy-5-methyl (2y) and 3,5-dimethyl-4-methoxy (2ab) derivatives showed a potent antisecretory effect of more than 80% after oral administration at 6 mg/kg. Although these aniline derivatives have relatively low stabilities in aqueous solution, replacement of the isobutyl group at the aniline nitrogen atom with N-(2-methoxyethyl) group enhanced the stability.
Adenosine Triphosphatases↗
Asthma classification by a score calculated from clinical findings and examinations in subjects sensitive to inhalant allergens.
Twenty-one patients with atopic asthma were classified into three types according to their symptoms (clinical diagnosis): Ia, simple bronchoconstriction; Ib, bronchoconstriction + hypersecretion; and II, bronchiolar obstruction, and this classification was compared with a classification made according to clinical findings and examinations (score diagnosis). Type Ib asthma was characterized by the increased incidence of eosinophils in bronchoalveolar lavage fluid (BALF), while type II was characterized by ventilatory dysfunction in small airways and the increased incidence of neutrophils in BALF. Four patients, whose expectoration was between 50 and 99ml/day, of the 12 with type Ia assessed by clinical diagnosis were evaluated as type Ib by score diagnosis. One patient with type II by clinical diagnosis was assessed as questionable type II by score diagnosis. In the other 16 patients, the clinical and score diagnoses were the same.
Administration, Inhalation↗
Spa therapy improves ventilatory function in the small airways of patients with steroid-dependent intractable asthma (SDIA).
The improvement of ventilatory function by spa therapy was examined in 37 patients with steroid-dependent intractable asthma (SDIA) in relation to clinical asthma types. All subjects had been on long-term corticosteroid therapy before spa therapy. Spa therapy was found to improve the values of ventilatory parameters. The percent improvement in all subjects was +4.1% in %FVC, +8.9% in FEV1.0%, +9.8% in %PEFR, +22.0% in %MMF, +19.7% in %V50 and +28.2% in %V25, respectively. Relating to clinical asthma types, moderate improvement in %MMF and %V25 from 16.7% to 16.9% was observed in type Ia cases, and marked increase in %MMF, %V50 and %V25 was observed from 42.2% to 43.2% in type Ib cases. However, no significant increase was found in these parameters of types Ia or Ib after spa therapy. In patients with type II, a significant increase was shown in %V50 (p less than 0.05) and %V25 (p less than 0.01) after spa therapy. The results show that spa therapy improves the condition of small airways disorder in patients with SDIA.
Adrenal Cortex Hormones↗
Cellular composition of fluid in the airways of patients with house dust sensitive asthma, classified by clinical symptoms.
The cellular composition of bronchoalveolar lavage (BAL) fluid was observed in 28 patients with house dust mite-sensitive asthma. The proportion of neutrophils in the BAL fluid was significantly higher in the patients between the ages of 50 and 59 and those over 60 than in the patients younger than age 39. There was a significant difference in the number of neutrophils in patients aged 30-39 years and in those over 40 at the onset of disease. The cellular composition varied among the three types of asthma; the mean percentage of cells in the BAL fluid was 1.1% neutrophils and 21.3% eosinophils in type Ia (simple bronchoconstriction), 9.1% neutrophils and 23.0% eosinophils in type Ib (bronchoconstriction + hypersecretion), and 34.4% neutrophils and 12.0% eosinophils in type II (bronchiolar obstruction). The results suggested that an increased frequency of both neutrophils and eosinophils correlates with the mechanism of hypersecretion, and that a marked increase in the proportion of neutrophils is involved in bronchiolar obstruction.
Adult↗
Rheumatoid nodules of the lung in a patient with palindromic rheumatism.
We report a case of rheumatoid nodules of the lung seen in a patient with palindromic rheumatism. A 54-year-old man with palindromic rheumatism was admitted for evaluation of three nodules in the right upper lobe on chest roentgenogram. Wedge resection was performed for the purpose of confirmative diagnosis and treatment. Histology of these lung lesions revealed palisaded histiocytic cells surrounding a layer of central necrosis, which were considered to be characteristic findings of rheumatoid nodule. Such a case is extremely rare. To our knowledge, only one other case has been reported before in the literature.
Arthritis, Rheumatoid↗
A new alternative hepatectomy method for resection of segments 3 and 4 of the liver.
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Blood Loss, Surgical↗
A retrospective comparative study of reconstructive methods following pancreaticoduodenectomy--pancreaticojejunostomy vs. pancreaticogastrostomy.
Reconstructive methods following pancreaticoduodenectomy in our department are discussed and evaluated in this study. Between January 1980 and November 1990 fifty-two consecutive patients underwent pancreaticoduodenectomy because of pancreas head disease. Thirty-one patients underwent pancreaticojejunostomy and twenty-one had pancreaticogastrostomy as reconstructive procedures. Mortality rate was 6% in pancreaticojejunstomy versus zero in pancreaticogastrostomy. Six patients had leakage from the pancreaticojejunostomy, but only one patient had necrosis of the gastric stump and leakage from the pancreaticogastrostomy. This case had previous distal gastrectomy done for gastric ulcer. The residual stomach might not have been large enough, and the blood supply of the gastric stump might not have been adequate for pancreaticogastrostomy. Except for this case, none was observed with leakage from the pancreatic anastomosis in the pancreaticogastrostomy group. No statistical significance in operating time or blood loss was observed between the two methods. The pancreaticogastrostomy cases without complications had significantly less loss of body weight than those with pancreaticojejunostomy at the date of discharge (p < 0.05). It is concluded that pancreaticogastrostomy is the safer reconstructive method following pancreaticoduodenectomy, although it may not be indicated in patients with prior gastrectomy.
Adult↗
[Analysis of acute graft-vs-host like reaction in [B6-lpr----B6] spleen chimera mice].
The lpr autoimmune mice develop massive lymphadenopathy and autoimmune disease. Transfer of lpr autoimmune disease to normal mice by bone marrow transplantation did not succeed, but caused a severe wasting syndrome like graft-vs-host (GvH) disease (lpr-GvH). We previously demonstrated that the transfer of B6-lpr spleen cells to B6 mice ([B6-lpr----B6] chimera) caused acute GvH-like disease accompanied by remarkable CD8+ T cell expansion, and that chimera spleen cells had suppressive activity on mitogen responses of spleen cells from normal mice. In this paper, I studied strain difference of acute lpr-GvH disease and mechanism of suppressive activity further. [B6-lpr----B6] chimera spleen contained more than 70% of CD8+ cells. However, [MRL-lpr----MRL-(+)] chimera and [C3H-lpr----C3H] chimera contained less than 20% of CD8+ cells. Both B220+ DN T cells and CD 8+ T cells were requisite for expansion of CD8+ T cell in the chimera spleen. Mixed chimera experiments using CD8+ T cells from B6-lpr-Thy1.1 mice and B220+ DN T cells from B6-lpr mice showed that CD8+ but not B220+ DN T cells were direct precursors of expanding CD8+ T cells. The mechanism of suppressive activity on Con A responses was different from that on LPS responses. A cell-to-cell interaction was essential for suppression of Con A response without involvement of CD8- cells, while suppression of LPS responses was mainly exhibited by soluble factors. Suppression of LPS responses was abrogated by the presence of anti-IFN-gamma monoclonal antibody, suggesting that those factors contain IFN-gamma. These data suggest that CD8+ T cells from B6-lpr mice were activated by unidentified antigen(s) which were expressed by B6 but not B6-lpr cells, and that IFN-gamma and/or unknown inflammatory factors secreted by the CD8+ T cells induced pathological outcome indistinguishable from GvH reaction in B6 recipient mice.
Animals↗