Degradation ofp-aminoazobenzene by means of cell-free extracts fromAeromonas hydrophila var. 24B andPseudomonas pseudomallei 13NA

Summary By means of cell-free extracts (supernatant III: crude enzyme, precipitate II: cell debris and precipitate III: microsomal fraction) fromAeromonas hydrophila var. 24B andPseudomonas pseudomallei 13NA,p-aminoazobenzene was effectively degraded. Aniline andp-phenylenediamine were detected in the degradation products. No degradation ofp-aminoazobenzene was found in the supernatant I (extra-cellular enzyme fraction). These results clearly show that thep-aminoazobenzene degradation activity was found in the supernatant III and the precipitate II, III.     

Developing a Multilayered Integrated Numerical Model of Surface Physics – Growing Plants Interaction (MINoSGI)

This paper describes a Multilayered Integrated Numerical Model of Surface Physics – Growing Plants Interaction (MINoSGI), which represents interactions between the dynamics of forest ecosystems and microclimate. Aiming at a large‐scale study in the future, we describe forest dynamics by using area‐averaged prognostic equations for thedistributions of plant density and plant weight with respect to plant height classes, instead of individual‐based treatments for small‐scale forest patches. Growth and mortality of plants are modelled based on the carbon balance of each plant height class. The area‐averaged microclimate (e.g., light, wind speed, temperature, humidity, CO₂ concentration) within the forest canopy is simulated by a one‐dimensional multilayer canopy model, which includes most of the physical and physiological processes that control the forest microclimate. Owing to its multilayered framework, a direct specification is possible for the difference in the growing environment among plants of different size and species. Given hourly meteorological conditions, the model outputs energy, water, CO₂ and momentum fluxes to and from a forest, of which the structure changes through competition among plants. The model's performance was tested by comparing its outputs with observed data on the development of plant size distribution taken over a 5‐year period in an evergreen coniferous (Cryptomeria japonica) forest. The model produced realistic estimates of the total biomass increments during the period. The ratio of net primary production to gross primary production (=0.45) was consistent with previous estimates for temperate forests. The bimodal seasonal pattern in net ecosystem production was similar to the seasonal trend in the CO₂ flux measured over a forest of the same species. Although some limitations due to the one‐dimensional representation of microclimate were noticeable, the model adequately simulated distributions of annual growth rate, plant weight and diameter across plant height classes. Since the basic equations can be extended to include the effect of spatial variability with marginal increase of computational costs, the present model framework is feasible for large‐scale studies.     

Mucin-specific bark lectin from elderberry Sambucus sieboldiana and its applications to the affinity chromatography of mucin

Three bark lectins were isolated from elderberry Sambucus sieboldiana using fetuin-Sepharose 4B and mucin-Sepharose 4B, and were studied comparatively for their binding to glycoprotein and to clarify various physicochemical features. For each, a unique pattern on isoelectric focusing was noted and their affinity toward various glycoproteins differed, indicating the structures of their carbohydrate binding sites possibly differ. One bark lectin showed specific binding toward porcine mucin. The purity of mucin from a crude porcine stomach mucin or an extract of porcine submaxillary glands could be improved by affinity chromatography on immobilized lectin having binding specificity toward mucin.     

Glutamate Inhibits Adenylate Cyclase Activity in Dispersed Rat Hippocampal Cells Directly via an N-Methyl-d-Aspartate-Like Metabotropic Receptor

Three major subtypes of glutamate receptors that are coupled to cation channels--N-methyl-D-aspartate (NMDA), kainate, and alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) receptors--are known as ionotropic receptors in the mammalian CNS. Recently, an additional subtype that is coupled to GTP binding proteins and stimulates (or inhibits) metabolism of phosphoinositides has been proposed as a metabotropic receptor. Incubation of dispersed hippocampal cells from adult rats with glutamate or NMDA decreased forskolin-stimulated cyclic AMP (cAMP) accumulation; half-maximal effects were obtained with 5.6 +/- 2.2 and 6.4 +/- 2.3 microM, respectively. Kainate and quisqualate were less potent. The effect of glutamate was antagonized by 2,3-diaminopropionate and 2-amino-5-phosphonovalerate, NMDA/glutamate receptor antagonists, but not by 0.5 microM Joro spider toxin, a specific blocker of the AMPA receptor. The inhibitory effect of glutamate on cAMP formation was not blocked by 2 microM tetrodotoxin or by the absence of Ca2+. In hippocampal membranes, glutamate, similar to carbachol, inhibited adenylate cyclase activity in a GTP-dependent manner. These findings suggest that the glutamate inhibition of adenylate cyclase is direct and is not due to a result of the release of other neurotransmitters. The effect of glutamate on cAMP accumulation was observed in an assay medium containing 0.7 mM MgCl2, which is known to inhibit both ionotropic NMDA receptor/channels in the hippocampus and metabotropic NMDA receptors in the cerebellum. The inhibitory effect of glutamate was abolished by pertussis toxin treatment.(ABSTRACT TRUNCATED AT 250 WORDS)     

DNA-dependent adenosinetriphosphatase C1 from mouse FM3A cells has DNA helicase activity.

In our previous study, we identified four chromatographically distinct DNA-dependent ATPases, B, C1, C2, and C3, in mouse FM3A cells (Tawaragi, Y., Enomoto, T., Watanabe, Y., Hanaoka, F., and Yamada, M. (1984) Biochemistry 23, 529-533). The DNA-dependent ATPase C1 has been purified and characterized in detail. A divalent cation and a polynucleotide cofactor were required for the ATPase activity. Poly(dT), single-stranded circular DNA, and heat-denatured DNA were very effective. Almost no ATPase activity was observed with S1 nuclease-treated native DNA. ATPase C1 hydrolyzed ATP only among the ribo- and deoxyribonucleoside triphosphates tested, and this fact distinguished ATPase C1 from ATPases B, C2, and C3, because the latter enzymes are capable of hydrolyzing both ATP and dATP. The purified DNA-dependent ATPase C1 fraction was shown to have a DNA helicase activity that was dependent on hydrolysis of ATP. The helicase activity and DNA-dependent ATPase activity cosedimented at 5.2 S on glycerol gradient centrifugation. Both activities showed similar preferences for nucleoside 5'-triphosphates and similar requirements for divalent cations. The DNA helicase activity was inhibited by the addition of single-stranded DNAs that served as cofactor for the ATPase activity. The efficiency of a single-stranded DNA to inhibit DNA helicase activity correlated well with the capacity of the DNA to serve as cofactor for DNA-dependent ATPase activity. The helicase was shown to migrate along the DNA strand in the 5' to 3' direction, which is the same direction of migration of the mouse DNA helicase B (Seki, M., Enomoto, T., Yanagisawa, J., Hanaoka, F., and Ui, M. (1988) Biochemistry 27, 1766-1771).     

Effects of the brachyury (T) mutation on mitotic activity in the neural tube.

Mitotic activity in the neural tube was examined in +/+, $\text{T}\text{+}$ and $\text{T}\text{T}$ mouse embryos. The number of mitotic figures per unit area of ependymal layer (area index) was higher in $\text{T}\text{+}$ embryos than in +/+ embryos. In +/+ embryos, area index was higher in the dorsal part of the neural tube than in the ventral part, whereas this pattern is disordered in $\text{T}\text{+}$ embryos. In $\text{T}\text{T}$ embryos, these values fluctuate. The process of nuclear migration accompanying cell division seems to be normal in the neural tube of $\text{T}\text{+}$ and $\text{T}\text{T}$ embryos.     

Activity of synthetic gibberellin A15 and (±)-gibberellin A15isolactone

The activities of (±)-gibberellin A₁₅ ((±)-GA₁₅) and (±)-gibberellin A₁₅-isolactone ((±)-iso-GA₁₅) which were obtained by stereocontrolled total synthesis and gibberellin A₁₅ (E-GA₁₅) synthesized by interconversion of enmein were assayed by the rice seedling test. As expected, (±)-GA₁₅ showed half the activity of natural gibberellin A₁₅ (GA₁₅). E-GA₁₅ which has a natural configuration showed the same activity as natural gibberellin A₁₅ while (±)-iso-GA₁₅ was almost inactive. These samples were also submitted to the cucumber hypocotyl assay. Contrary to what has already been reported, they were almost inactive.     

Serological identification of sperm antigens specified by lethalt-alleles in the mouse

Cytotoxic antisera were prepared by immunizing wild-type recipient mice with sperm from donors carrying different recessive lethal alleles at theT locus (T/t ⁰,T/t ^{w 1} ,T/t ^{w 5} , andT/t ^{w 32} ). After removal of sperm autoantibody by absorption with sperm of recipient type, each antiserum reacted only with sperm from males whose genotype contained at least one of the immunizing alleles. Cytotoxicity was high against sperm populations in which both immunizing alleles were represented and was lower when only one was present. Thus, each allele at theT locus which has so far been tested serologically is recognizable as a discrete antigen on the surface of sperm.This paper is dedicated to Professor Hans Grüneberg, F.R.S. on the occasion of his retirement from the chair of Animal Genetics at University College London, in appreciation of the fact that it was he who first suggested that surface properties played an important role in the origin of abnormalities associated with mutants at theT-locus.     

Suppression of mitochondrial response to hepatectomy in diabetic rats

In non-diabetic rats, the mitochondrial phosphorylative activity per unit of cytochrome a(+a₃) in the remnant liver 3 hours after hepatectomy increased to approximately 140% of sham-operated controls without significant decrease in the energy charge. In diabetic rats the enhancement in mitochondrial phosphorylative activity following hepatectomy was inhibited in proportion to the severity of impaired insulin secretion, with a concomitant decrease in hepatic energy charge. It is suggested that insulin plays an important role in initiating an enhancement of mitochondrial oxidative phosphorylation.