Showing papers by "Hiroshima University published in 1990"

IRA2, a second gene of Saccharomyces cerevisiae that encodes a protein with a domain homologous to mammalian ras GTPase-activating protein.

Abstract: The IRA1 gene is a negative regulator of the RAS-cyclic AMP pathway in Saccharomyces cerevisiae. To identify other genes involved in this pathway, we screened yeast genomic DNA libraries for genes that can suppress the heat shock sensitivity of the ira1 mutation on a multicopy vector. We identified IRA2, encoding a protein of 3,079 amino acids, that is 45% identical to the IRA1 protein. The region homologous between the IRA1 protein and ras GTPase-activating protein is also conserved in IRA2. IRA2 maps 11 centimorgans distal to the arg1 locus on the left arm of chromosome XV and was found to be allelic to glc4. Disruption of the IRA2 gene resulted in (i) increased sensitivity to heat shock and nitrogen starvation, (ii) sporulation defects, and (iii) suppression of the lethality of the cdc25 mutant. Analysis of disruption mutants of IRA1 and IRA2 indicated that IRA1 and IRA2 proteins additively regulate the RAS-cyclic AMP pathway in a negative fashion. Expression of the IRA2 domain homologous with GAP is sufficient for complementation of the heat shock sensitivity of ira2, suggesting that IRA down regulates RAS activity by stimulating the GTPase activity of RAS proteins.

Cell Culture on a Thermo-Responsive Polymer Surface

Abstract: We have used a thermo-responsive polymer, poly-N-isopropyl acrylamide (PNI-PAAm), as a substratum for the culture of human dermal fibroblasts by conjugating it with collagen. The cells attached well, spread, and grew on the substratum, indicating that the polymer has no toxicity towards the cells. PNIPAAm is insoluble in water over the lower critical solution temperature (LCST; about 32°C) and reversibly solubilized below the LCST. Taking advantage of this conversion, monolayered fibroblasts cultured on the substratum containing the PNIPAAm over the LCST, were completely detachable from the substratum by simply lowering the temperature below the LCST, without the use of conventional detaching agents such as trypsin and EDTA. The detached cell sheet gradually aggregated and finally formed a multicellular spheroid. This polymer may provide a convenient and potentially useful technology for cell culture.

Inhibition of the Fermentation of Propionate to Methane by Hydrogen, Acetate, and Propionate

Abstract: Inhibition of the fermentation of propionate to methane and carbon dioxide by hydrogen, acetate, and propionate was analyzed with a mesophilic propionate-acclimatized sludge that consisted of numerous flocs (size, 150 to 300 μm). The acclimatized sludge could convert propionate to methane and carbon dioxide stoichiometrically without accumulating hydrogen and acetate in a propionate-minimal medium. Inhibition of propionate utilization by propionate could be analyzed by a second-order substrate inhibition model (shown below) given that the substrate saturation constant, Ks, was 15.9 μM; the substrate inhibition constant, Ki, was 0.79 mM; and the maximum specific rate of propionate utilization, qm, was 2.15 mmol/g of mixed-liquor volatile suspended solids (MLVSS) per day: qs = qmS/[Ks + S + (S2/Ki)], where qs is the specific rate of propionate utilization and S is the initial concentration of undissociated propionic acid. For inhibition by hydrogen and acetate to propionate utilization, a noncompetitive product inhibition model was used: qs = qm/[1 + (P/Kp)n], where P is the initial concentration of hydrogen or undissociated acetic acid and Kp is the inhibition constant. Kinetic analysis gave, for hydrogen inhibition, Kp(H2) = 0.11 atm (= 11.1 kPa, 71.5 μM), qm = 2.40 mmol/g of MLVSS per day, and n = 1.51 and, for acetate inhibition, Kp(HAc) = 48.6 μM, qm = 1.85 mmol/g of MLVSS per day, and n = 0.96. It could be concluded that the increase in undissociated propionic acid concentration was a key factor in inhibition of propionate utilization and that hydrogen and acetate cooperatively inhibited propionate degradation, suggesting that hydrogenotrophic and acetoclastic methanogens might play an important role in enhancing propionate degradation to methane and carbon dioxide.

Cell membrane stability, an indicator of drought tolerance, as affected by applied nitrogen in soyabean.

Abstract: Four soyabean cultivars were grown with two N application rates (50 and 300 kg N/ha) in the field at Hiroshima University, Japan, from June to August 1988. Cell membrane stability (CMS) by the polyethylene glycol (PEG) test, leaf water relations and nutrient concentrations in cell sap and leaf tissues were measured when the plants were 50 days old, in the uppermost fully expanded leaves.Cell membrane stability was higher at the higher N rate, the increase over the lower rate being greater in the cultivars Lee+ and Lee–than in Tamahomare and T201. Leaf water potential was not affected by the higher rate of N application. Osmotic adjustment, which was independent of water stress, was observed with the higher rate of N and it was higher in Lee + and Lee–than in Tamahomare and T201. It is suggested that osmotic potential in leaf tissues may influence CMS measured by the PEG test. Solute concentrations in cell sap and leaf tissues were higher at the higher N rate. Sugar and K were the major contributors to osmotic potential.

Aggressive natural killer cell leukaemia/lymphoma: report of four cases and review of the literature POSSIBLE EXISTENCE OF A NEW CLINICAL ENTITY ORIGINATING FROM THE THIRD LINEAGE OF LYMPHOID CELLS

Abstract: The morphologic, immunologic, genotypic and functional properties of peripheral blood and bone marrow cells or cultured cells from four patients with a clinically aggressive non-T, non-B natural killer cell leukaemia/lymphoma (ANKL/L) are described. The leukaemic cells possessed medium to large granules in the cytoplasm, antigens against CD38, CD2, OKIa 1 and NKH-1 CD56) monoclonal antibodies on their cell-surface, and also showed natural killer (NK) activity. In addition, these ANKL/L belonged to neither T- nor B-cell lineage, proved by studying clonal gene rearrangement for the T beta, T gamma and T delta receptors, and immunoglobulin. After comparing them with the seven cases of ANKL/L reported in other institutions, with regard to immunophenotype, genotype and function, we conclude that ANKL/L originating from a third lineage of lymphoid cells is a distinct clinical entity.

Formation of an anisotropic energy gap in the valence-fluctuating system of CeNiSn.

Abstract: Measurements of susceptibility \ensuremath{\chi}, resistivity \ensuremath{\rho}, and thermoelectric power S have been performed on single-crystal CeNiSn. Only along the a axis of the orthorhombic structure do \ensuremath{\chi}(T) and \ensuremath{\rho}(T) exhibit pronounced peaks at 12 K, whereas no anomaly was found in the specific heat. The gap energies estimated from \ensuremath{\rho}(T) are 2.4, 5.5, and 5.0 K along the a, b, and c axes, respectively. Near 3 K, ${\mathit{S}}_{\mathit{a}}$(T) and ${\mathit{S}}_{\mathit{c}}$(T) exhibit extremely sharp peaks, which indicate the presence of a density of states within the gap. The magnetic contribution to the specific heat divided by temperature, ${\mathit{C}}_{\mathit{m}}$/T versus T reveals a maximum of 0.19 J/${\mathrm{K}}^{2}$ mol near 6.7 K. These results suggest that an antiferromagnetic correlation develops near 12 K, which induces the formation of the pseudogap in the narrow band of heavy quasiparticles.

Growth factors and oncogenes in human gastrointestinal carcinomas

Abstract: Multi-autocrine loops of the epidermal growth factor (EGF), transforming growth factorα (TGFα), platelet-derived growth factor (PDGF) and TGFβ system are expressed in human gastrointestinal carcinomas. In esophageal and gastric carcinomas, they evidently play an important role in tumor progression. Gastrin, one of the major gut hormones, may also act as an autocrine growth factor for gastric and colonic carcinomas. TheHST1 andINT-2 genes, belonging to the fibroblast growth factor gene family, are coamplified in approximately 50% of primary tumors and in all the metastatic tumors of esophageal carcinoma. TGFα and EGF are the ligands of the tumor cells that overexpress EGF receptor in esophageal carcinomas. The synchronous expression of EGF and its receptor, as well as TGFα andras p21, is evidently correlated with the depth of tumor invasion, metastasis and prognosis of gastric carcinomas. Amplification of c-erbB-2 and EGF receptor genes has been observed in many metastatic sites of gastric carcinomas regardless of histological type. In addition to TGFα and EGF, TGFβ and PDGF A chain produced by tumor cells may stimulate collagen synthesis not only by fibroblasts but also by tumor cells themselves, resulting in extensive progression and diffuse fibrosis of scirrhous gastric carcinomas. Moreover, TGFα or EGF and estrogen may also play a cooperative role in the development of scirrhous gastric carcinoma. In colorectal carcinoma, it has been shown that the accumulation of several alterations inras genes and p53 genes is most important for the conversion of adenoma to carcinoma. Critical genetic changes, including activation of oncogenes, mutation and deletion of tumor suppressor genes and disturbances in transcriptional regulatory sequences, may bring about aberrant expression of growth factors and their receptors in gastrointestinal carcinomas. The understanding of the significance of EGF-related growth factors in tumor progression provides a framework for a biological approach to the therapy of human gastrointestinal carcinomas. 8-Cl-cAMP, which inhibits expression of oncogenes and TGFα, may be useful not only for cancer therapy but also for the study of cell differentiation.

Expression of ERBB2 in Human Gastric Carcinomas: Relationship between p185ERBB2 Expression and the Gene Amplification

Abstract: The expression of p185ERBB2 in a total of 34 human gastric carcinoma tissues as well as in corresponding normal mucosa was examined by Western blotting. More than 70% of both tumor tissues and normal mucosa showed p185ERBB2 expression at various levels. Eighteen (55%) cases revealed higher levels of p185ERBB2 in the tumor than in normal mucosa, while 13 (38%) cases showed lower levels in the tumor tissues. Higher expression of p185ERBB2 was frequently observed in well differentiated adenocarcinomas, with the incidence between well differentiated type and poorly differentiated type being significantly different (P less than 0.05). Comparative immunohistochemical analysis revealed the consistent results with p185ERBB2 expression obtained by Western blotting in well differentiated adenocarcinomas. Of the 34 cases, three well differentiated adenocarcinomas had extremely high levels of p185ERBB2. ERBB2 gene was amplified in two of the three tumors, but the amplification differed by the tumor site from where the sample was obtained. Another tumor which showed an extremely high level of p185ERBB2 but no gene amplification demonstrated a high level of binding protein to the TATA box that is located in the promoter of the ERBB2 gene. A high level of TATA-binding protein was also detected in gastric carcinoma cell lines which contain a single copy of ERBB2 gene and a high expression of p185ERBB2.

Kinetics of the Methanogenic Fermentation of Acetate

Abstract: Inhibition of the fermentation of acetate to methane and carbon dioxide by acetate was analyzed with an acetate-acclimatized sludge and with Methanosarcina barkeri Fusaro under mesophilic conditions. A second-order substrate inhibition model, q ch 4 = q m S /[ K s + S + ( S 2 / K i )], where S was the concentration of undissociated acetic acid, not ionized acetic acid, could be applicable in both cases. The analysis resulted in substrate saturation constants, K s , of 4.0 μM for the acclimatized sludge and 104 μM for M. barkeri. The threshold concentrations of undissociated acetic acid when no further acetate utilization was observed were 0.078 μM (pH 7.50) for the acclimatized sludge and 4.43 μM (pH 7.45) for M. barkeri. These kinetic results suggested that the concentration of undissociated acetic acid became a key factor governing the actual threshold acetate concentration for acetate utilization and that the acclimatized sludge in which Methanothrix spp. appeared dominant could utilize acetate better and survive at a lower concentration of undissociated acetic acid than could M. barkeri. Images

Diurnal patterns of heat production and heart rate under thermoneutral conditions in Holstein Friesian cows differing in milk production

Abstract: Ten dairy cows were allocated into three groups according to milk productivity (four high, four intermediate and two dry cows, respectively). Heat production and heart rate, but not rectal temperature, were significantly different (P < 0·05) between groups. Heat production increased during feeding in the morning and in the afternoon and reached a peak 3 h later. Minimum heat production was observed in the early morning before feeding. The diurnal pattern for heart rate reflected that of heat production. These results suggest that cooling dairy cows during hot summer days is most effective at feeding time and 3 h afterwards.

New approximate optimization method for distribution system planning

Abstract: An algorithm to obtain an approximate optimal solution to the problem of large-scale radial distribution system planning is proposed. The distribution planning problem is formulated as a MIP (mixed integer programming) problem. The set of constraints is reduced to a set of continuous variable linear equations by using the fact that the basis of the simplex tableau consists of the power flow variables of radial branch. This linear problem is solved by pivot operations which correspond to a branch-exchange of the radial network. Numerical examples are presented to demonstrate the validity and effectiveness of the algorithm. >

Egf and tgf alpha the ligands of hyperproduced egfr in human esophageal carcinoma cells act as autocrine growth factors

Abstract: In order to ascertain autocrine growth factors in esophageal carcinomas, we analysed expression of mRNAs and proteins for epidermal growth factor (EGF), transforming growth factor-alpha (TGF-alpha) and epidermal growth factor receptor (EGFR) in 6 esophageal carcinoma cell lines. Gene alterations were also examined. All of the esophageal carcinoma cell lines expressed mRNA for EGFR and TGF-alpha genes. Interestingly, EGF mRNA of about 5.0 kb was also detected in TE-1, TE-2, and TE-8 cells. Production of protein was also confirmed by binding assay and ELISA on 3 of the 6 cell lines. The cells had a relatively high number of EGFRs and produced TGF-alpha and EGF protein at the same time. Furthermore, anti-EGF (KEM-10) and anti-TGF-alpha (WA-3) monoclonal antibodies (MAbs) inhibited spontaneous uptake of tritiated thymidine (3H-TdR) by TE-1 cells which expressed EGF, TGF-alpha and EGFR mRNA and protein. These results strongly suggest that EGF and/or TGF-alpha produced by carcinoma cells function as autocrine growth factors for human esophageal carcinomas.

Corrections to the expression for gain in GaAs

Abstract: When the expressions for gain derived in various papers are compared, a number of inconsistencies are found to exist. These inconsistencies have propagated through the literature and continue to do so. This study is specifically devoted to explaining how these inconsistencies originated so that they will not be repeated in future work on the subject. A general discussion of the matrix element is given, including spin degeneracy considerations, the procedure typically used in estimating the magnitude of the momentum matrix element, as well as the enhancement of the matrix element in quantum well structures. The correct expression for gain and spontaneous emission in semiconductors is given for comparison purposes within the framework of Fermi's golden rule. >

Characterization of sodium dodecyl sulfate-stable Staphylococcus aureus bacteriolytic enzymes by polyacrylamide gel electrophoresis.

Abstract: Profiles of the bacteriolytic activities of Staphylococcus aureus culture supernatants, sodium dodecyl sulfate cell extracts, LiCl cell extracts, cell wall extracts, and cell membranes were analyzed in sodium dodecyl sulfate-polyacrylamide gels containing Micrococcus luteus or S. aureus. A total of 20 distinct bands of bacteriolytic activity could be detected in gels containing M. luteus, 8 of these bands were found in culture supernatants. The sodium dodecyl sulfate cell extracts, the LiCl cell extracts, and the cell membranes each contained 20 bands (P1 to P20), but no activity was found in cell wall extracts. Less bacteriolytic activity could be detected in gels containing S. aureus, although three bands were found in culture supernatants and LiCl extracts and cell membranes contained one major band, P13. Crude cell extracts showed five bacteriolytic bands of which the major bacteriolytic bands were distributed in an identical manner in all 10 strains of S. aureus studied. The effects of chemical and physical factors were determined, and it was shown that iodoacetic acid, Hg2+, and Cibacron Blue 3G-A reduced activity, and an optimum pH for enzyme detection was between 7 and 8. Preincubation at 100 degrees C for 30 min reduced the activity of P1 and P2 bands. Images

Optimal allocation and control problems for software-testing resources

Abstract: Two kinds of software-testing management problems are considered: testing-resource allocation to best use specified testing resources during module testing, and a testing-resource control problem concerning how to spend the allocated amount of testing-resource expenditures during it. A software reliability growth model based on a nonhomogeneous Poisson process is introduced. The model describes the time-dependent behavior of software errors detected and testing-resource expenditures spent during the testing. The optimal allocation and control of testing resources among software modules can improve reliability and shorten the testing stage. Based on the model, numerical examples of these two software testing management problems are presented. >

The cisA cistron of Bacillus subtilis sporulation gene spoIVC encodes a protein homologous to a site-specific recombinase.

Abstract: The nucleotide sequence of the sporulation gene spoIVC cisA in Bacillus subtilis was determined and found to encode a protein of 500 amino acid residues with a calculated molecular weight of 57,481, which is in good agreement with the size of the gene product estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The amino acid sequence of the N-terminal region of this protein is homologous to the site-specific DNA recombinases. Hybridization of a 3.6-kilobase EcoRI fragment carrying the spoIVC cisA gene with the EcoRI-restricted chromosomal DNA prepared from cells of various stages showed that DNA rearrangement occurs only in the mother cell in the region adjacent to spoIVC cisA 3 h after the initiation of sporulation. This result coincides with that of Stragier et al. (P. Stragier, B. Kunkel, L. Kroos, and R. Losick, Science 243:507-512, 1989). The timing of the DNA rearrangement coincides very well with the timing of spoIVC cisA gene expression. The DNA rearrangement was not observed in spoIVC cisA mutants. These results strongly suggest that the spoIVC cisA gene encodes a site-specific DNA recombinase having a very important role in sporulation.

A lighting model aiming at drive simulators

Abstract: Many techniques for rendering natural objects such as the sea, terrains, and trees have been developed; they are indispensable for flight simulators. In this paper, techniques for rendering road surfaces under various conditions are discussed. Rendering road surfaces is quite useful for the evaluation of driving safety, and it will play an important part in the development of drive simulators. Light sources with high intensity often disturb drivers especially under wet road surface conditions.This paper proposes two models, a reflection model for road surfaces taking into account weather conditions, and a model on streaks of light taking into account both refraction and diffraction of light. Some examples demonstrate the possibility of applications for drive simulators in the future.

Initiation of meiosis and sporulation in Saccharomyces cerevisiae requires a novel protein kinase homologue.

Abstract: SME1 was cloned due to its high copy number effect: it enabled MATα/MATα diploid cells to undergo meiosis and sporulation in a vegetative medium. Disruption of SME1 resulted in a recessive Spo− phenotype. These results suggest that SME1 is a positive regulator for meiosis. DNA sequencing analysis revealed an open reading frame of 645 amino acids. An amino terminal peptide of ca 400 amino acids in the deduced protein was similar to known protein kinases. Transcription of SME1 was regulated negatively by nitrogen and glucose and positively by MATα/MATα and IME1, another positive regulator gene of meiosis. By complementation analysis, SME1 was found to be identical to IME2, which had been shown to be important in meiosis. These results suggest that IME1 product stimulates meiosis by activating transcription of SME1 (IME2) and that protein phosphorylation is required for initiation of meiosis.