Effects of Tween 80 and sodium fluoride on extracellular glucosyltransferase production and membrane lipids of Streptococcus mutans

1. Both Tween 80 and sodium fluoride significantly enhanced total extracellular glucosyltransferase activities of Streptococcus mutans. 2. Water-insoluble and water-soluble glucan formation were uniformly increased by Tween 80, whereas fluoride stimulated only water-soluble glucan formation. 3. Elevated glucan formation was due to an increase in enzymes secreted from bacterial cells. 4. Fatty acid composition and phospholipid content in bacterial membrane were changed by Tween 80, although sodium fluoride scarcely showed these changes. 5. Comparative results suggest that modulation of membrane lipids participates in mutansucrase production but not in dextransucrase production of S. mutans.     

A real time learning algorithm for recurrent analog neural networks

A new learning algorithm is described for a general class of recurrent analog neural networks which ultimately settle down to a steady state. Recently, Pineda (Pineda 1987; Almeida 1987; Ikeda et al. 1988) has introduced a learning rule for the recurrent net in which the connection weights are adjusted so that the distance between the stable outputs of the current system and the desired outputs will be maximally decreased. In this method, many cycles are needed in order to get a target system. In each cycle, the recurrent net is run until it reaches a stable state. After that, the weight change is calculated by using a linearized recurrent net which receives the current error of the system as a bias input. In the new algorithm the weights are changed so that the total error of neuron outputs over the entire trajectory is minimized. The weights are adjusted in real time when the network is running. In this method, the trajectory to the target system can be controlled, whereas Pineda's algorithm only controls the position of the fixed point. The relation to the back propagation method (Hinton et al. 1986) is also discussed.     

A quantitative comparison of foliage development among allopatric ferns,Dryopteris crassirhizoma,D. coreano-montana andD. filix-mas

A quantitative comparison was conducted on the foliage development during sporophyte development of three allopatric ferns in cool temperate and subalpine regions of Hokkaido and Tirol, European Alps. The foliage development ofDryopteris crassirhizoma, D. coreano-montana andD. filix-mas was quantitatively described by the leaf development (NV, number of veins); NV correlates the leaf-shape complexity from a circle (DI, L/2(3.14×S)^1/2). Nearly similar patterns were detected on frequency distribution of fertile leaves, fertility increase and number of leaves in threeDryopteris ferns which exhibit funnel-shaped foliage arrangements in mature sporophyte. No difference was found in number of leaves, maximum NV, fertility rate and leaf-shape parameters among three ferns. A positive difference was found only on changes in order of pinnae with maximum number of costa branches (NVP) and the DI of outline of pinnae betweenD. crassirhizoma andD. filix-mas. These allopatricDryopteris ferns seem to have a similar foliage structure, in spite of some sympatricDryopteris ferns capable of producing putative hybrids (D. austriaca andD. amurensis; D. tokyoensis andD. monticola) having different foliage structures in Hokkaido. Contribution No. 3346 from the Institute of Low Temperature Science, Hokkaido University.     

Expression of human lymphotoxin in Namalwa KJM-1 cells adapted to serum-free medium

A Namalwa cell line, KJM-1, which was adapted to serum-free medium is thought to be a good host cell line for recombinant DNA technology. We previously reported the expression of human -interferon (-IFN) in Namalwa KJM-1 (Miyaji, 1989a). The utility of Namalwa KJM-1 for expression of foreign genes was further examined. As a target gene to be expressed, human lymphotoxin (hLT) cDNA was used. It was engineered for expression in Namalwa KJM-1 using a simian virus 40 (SV40)-based expression vector pAGE107 (Miyaji, 1989a). It contains all components necessary for the expression of cDNA in mammalian cells. The expression vector was introduced into Namalwa KJM-1 by electroporation. Among the transformants, clone 7 was further examined for the expression of hLT in serum-free medium. The production level of hLT was augmented with the increase of the cell density. Thus it was further indicated that Namalwa KJM-1 is useful for production of foreign gene products.Abbreviation HEPES 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid     

Heterocyclic amine-DNA adducts analyzed by 32P-postlabeling method

DNA adducts formed by 12 heterocyclic amines were analyzed by 32P-postlabeling method. Several DNA adducts were detected in rat liver by administration of each heterocyclic amine. Total adduct levels ranged from 0.5 for 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) to more than 250 for 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1) per 10(7) nucleotides 24 hr after intragastric administration of these compounds. The N-hydroxy derivative of 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) was reactive toward DNA in vitro to form adducts. Addition of acetic anhydride to N-OH-MeIQx greatly enhanced its reactivity to DNA. 32P-Postlabeling analysis revealed that the MeIQx-DNA adducts formed in vivo and in vitro were identical. Thus, MeIQx would be metabolized in vivo to N-hydroxy form and further esterified to produce more reactive species, such as N-acetoxy form, which modify DNA to form adducts.     

Processing, secretion, and biological properties of a novel growth factor of the fibroblast growth factor family with oncogenic potential.

We recently reported that the protein encoded in a novel human oncogene isolated from Kaposi sarcoma DNA was a growth factor with significant homology to basic and acidic fibroblast growth factors (FGFs). To study the properties of this growth factor (referred to as K-FGF) and the mechanism by which the K-fgf oncogene transforms cells, we have studied the production and processing of K-FGF in COS-1 cells transfected with a plasmid encoding the K-fgf cDNA. The results show that, unlike basic and acidic FGFs, the K-FGF protein is cleaved after a signal peptide, glycosylated, and efficiently secreted as a mature protein of 176 or 175 amino acids. Inhibition of glycosylation impaired secretion, and the stability of the secreted K-FGF was greatly enhanced by the presence of heparin in the cultured medium. We have used the conditioned medium from transfected COS-1 cells to test K-FGF biological activity. Similar to basic FGF, the K-FGF protein was mitogenic for fibroblasts and endothelial cells and induced the growth of NIH 3T3 mouse cells in serum-free medium. Accordingly, K-fgf-transformed NIH 3T3 cells grew in serum-free medium, consistent with an autocrine mechanism of growth. We have also expressed the protein encoded in the K-fgf protooncogene in COS-1 cells, and it was indistinguishable in its molecular weight, glycosylation, secretion, and biological activity from K-FGF. Taken together, these results suggest that the mechanism of activation of this oncogene is due to overexpression rather than to mutations in the coding sequences.     

Role of pili in the pathogenesis of Pseudomonas aeruginosa burn infection

The present study using three isogenic mutants (F+P-, F-P+, F-P-) of Pseudomonas aeruginosa indicates that the presence of pili enhances the virulence of the organisms in experimental P. aeruginosa burn infection of mice. The 50% lethal dose (LD50) value for burned mice inoculated with non-piliated (P-) mutant was at least ten times higher than those inoculated with piliated (P+) bacteria. Meanwhile the LD50 value for burned mice inoculated with non-flagellated (F-) mutant was at least 10(5) times higher than those inoculated with flagellated (F+) bacteria. At 24 hr after inoculation, the bacterial counts in burned skin of mice inoculated with P+ bacteria were ten times higher than those inoculated with P- bacteria; and at 48 hr the bacterial counts became a hundred times higher in the former mice than the latter. At 24 hr after inoculation, P+ bacteria were isolated from blood, liver (F+P+), lung (F+P+), and kidney, while P- bacteria were not present in these tissues. And at 48 hr after inoculation, P+ bacteria were isolated from all tissues, while P- bacteria were isolated from some sites only. These results suggested that pili and flagella each play an important role as virulence factors independently, and that pili-mediated enhancement of virulence of P. aeruginosa was attributed to pili-mediated enhanced colonization of the organisms at the burned skin surfaces.     

Androgenic regulation of epidermal growth factor in the mouse ventral prostate

Castration of adult male mice caused a marked reduction in the amount of immunoreactive epidermal growth factor (EGF) in the ventral prostate, and the treatment of such castrated mice with testosterone increased the EGF level significantly. Gel filtration of prostate extract showed that the immunoreactive EGF in the prostate had the same molecular weight (6,045) as the submandibular gland EGF. Moreover, its isoelectric point (pH 4.5) was almost similar to that (pH 4.55) of the submandibular gland peptide. These results suggest that under the control of androgens, mouse ventral prostate synthesizes EGF structurally and functionally identical to the submandibular gland EGF.