A Simple Bioassay for Chemicals Active to the Photosynthetic or Respiratory Systems of Plants

After male rats of the Sprague Dawley strain, 5 weeks old, were fed a 20% casein diet with or without 0.5% nicotinamide for 13 days, 180 mg/kg body weight of alloxan was injected in- traperitoneally into the rats. The rats were kept for 18 days with the same diet. The level of blood glucose was increased 6-fold in the group on a 20% casein diet by the injection of alloxan, while there was only a 2-foid increase in the group on a nicotinamide-containing diet and the decreased body weight was also lower in the group on the nicotinamide diet than the group on the casein diet. The body weight was indirectly related to the concentration of blood glucose. A marked increase was observed in the activities of tryptophan oxygenase, aminocarboxymuconate-semialdehyde decarboxylase, and nicotinamide methyltransferase upon the injection of alloxan with both diets; on the other hand, the activities of kynureninase and NAD⁺ synthetase were decreased by the injection of alloxan. The activity of kynurenine aminotransferase increased in the group on the 20% casein diet by the injection of alloxan, while in the group on the nicotinamide-containing diet its activity was not increased by the injection. These changes in the above enzyme activities mean that the conversion ratio from tryptophan to niacin is lower in the alloxan diabetic rat than normal rat. It was found that the activities of tryptophan oxygenase, aminocarboxymuconate-semialdehyde decarboxylase, and nicotinamide methyltransferase were directly related to the concentration of blood glucose, and that the activities of kynureninase and NAD⁺ synthetase were inversely related. There was no difference in the activities of 3-hydroxyanthranilic acid oxygenase and nicotinamide mononucleotide adenylyltransferase upon the injection of alloxan with both diets.     

Synthesis of Japanese Boletus edulis ectomycorrhizae with Japanese red pine

Boletus edulis is a well-known ectomycorrhizal mushroom. Although cultivation has been widely attempted, no artificial fruiting has been achieved owing to difficulties associated with mycorrhizal synthesis and acclimatization in fields. We collected fifteen B. edulis basidiomata samples from locations in Japan and identified them microscopically and by phylogenetic analysis of their nuclear ribosomal internal transcribed spacer (ITS) regions. Pure culture isolates of B. edulis were established efficiently on malt extract agar medium, and one isolate, EN-63, was inoculated to axenic Pinus densiflora seedlings in vitro. Brownish ectomycorrhizal tips were observed on the pine lateral roots within four months of inoculation. Ten pine seedlings that formed ectomycorrhizae were acclimatized under laboratory and greenhouse conditions. At four months after transplant, mycorrhizal colonization by B. edulis was observed on newly grown root tips under laboratory conditions, but no B. edulis ectomycorrhiza survived under greenhouse conditions. These results suggest that B. edulis ectomycorrhizae synthesized in vitro with P. densiflora requires additional steps for acclimatization to greenhouse conditions.     

Purification and characterization of membrane-bound alcohol dehydrogenase from Acetobacter polyoxogenes sp. nov.

Summary Membrane-bound alcohol dehydrogenase (ADH) was purified from the membrane fraction of an industrial-vinegar-producing strain, A. polyoxogenes sp. nov. NBI1028 by solubilization using Triton X-100 and subsequent column chromatography on DEAE-Sepharose CL-6B and hydroxyapatite. The purified enzyme was homogeneous on polyacrylamide disc gel electrophoresis (PAGE). Upon sodium dodecyl sulphate-PAGE, the enzyme showed the presence of two subunits with a molecular mass of 72 000 daltons and 44 000 daltons, respectively. The small subunit was identified as cytochrome c. In addition, absorption and fluorescence spectra showed the the presence of pyrroloquinoline quinone in the purified ADH. The ADH preferentially oxidized aliphatic alcohols with a straight carbon chain except for methanol. Formaldehyde and acetaldehyde were also oxidizable substrates. The apparent K _m for ethanol was 1.2 mM. The optimum pH and temperature were 5.0–6.0 and 40°C, respectively. p-Chloromercuribenzoic acid and heavy metals such as CuSO₄ were inhibitory to the enzyme activity. Ferricyanide was effective as an electron acceptor.Offprint requests to: M. Fukaya     

Adsorption of polyhedra of a cytoplasmic-polyhedrosis virus by soil particles

Tests on the quantitative adsorption of polyhedra of a cytoplasmic-polyhedrosis virus of Bombyx mori to soil showed an increase in the adsorption of polyhedra with an increase in the amount of soil, but the number of polyhedra adsorbed per unit weight of soil decreased. The number of polyhedra adsorbed to a fixed quantity of soil was in direct proportion to the polyhedron concentration, and the amount of adsorption increased with acidity but decreased with the addition of reagents which masked the polyvalent cations exposed on the surfaces of soil particles. The polyhedra applied to the top of a soil column were detected microscopically within 4 cm from the surface after the equivalent of 1,120 cm depth of water was passed through the column. The polyhedra occurred on the upper surface of the soil column and on the walls of voids formed by random packing of the soil particles.     

Apparent gene conversion in an Escherichia coli rec+ strain is explained by multiple rounds of reciprocal crossing-over

Summary Gene conversion, the non-reciprocal transfer of sequence information between homologous DNA sequences, has been reported in lower eukaryotes, mammals and in Escherichia coli. In an E. coli rec ⁺ strain, we established a plasmid carrying two different deleted neo genes (neoDL and neoDR) in an inverted orientation and then selected for homologous recombination events that had reconstructed an intact neo ⁺ gene. We found some plasmids that had apparently experienced intramolecular gene conversion. Further evidence, however, suggests that they are products of multiple rounds of reciprocal crossing-over,apparently involving two plasmid molecules. First, most of the Neo⁺ clones contained multiple types of Neo⁺ plasmids, although the frequency of producing the neo ⁺ clones was low. Second, all the neo ⁺ clones also contained, as a minority, one particular form of dimer, which can be formed by reciprocal crossing-over between neoDL of one plasmid molecule and neoDR of another plasmid molecule. Third, in reconstruction experiments, we cloned and purified this dimer and transferred it back into the rec ⁺ cells. The dimer gave rise to clones containing multiple types of neo ⁺ recombinant monomers, including those apparent gene conversion types, and containing only few molecules of this dimer plasmid.     

A rapid bioassay method of gibberellin activity using pale malt

The availability of brewery pale malt as a substrate for gibberellinbioassay was investigated. GA₃ at the concentration of 0.001to 1 µg/ml caused an increase in a-amylase activity inpale malt under aerobic incubation, while no increase was observedunder anaerobic conditions. Pale malt heated at 130°C for2 hr showed no increase in a-amylase activity in the presenceof GA₃. Although the mechanism for the enhancement of a-amylaseactivity in pale malt by GA₃ is not clear, it is evident thatthis phenomena can be used in bioassay of gibberellins. Experimentalconditions for the bioassay using pale malt are described. Withthis method, the enhancement of a-amylase activity by differentgibberellins was: GA₃>GA₄>GA₂₀ (inactive). (Received October 16, 1975; )     

The neurobiology of pair bond formation,bond disruption,and social buffering

Enduring social bonds play an essential role in human society. These bonds positively affect psychological, physiological, and behavioral functions. Here, we review the recent literature on the neurobiology, particularly the role of oxytocin and dopamine, of pair bond formation, bond disruption, and social buffering effects on stress responses, from studies utilizing the socially monogamous prairie vole (Microtus ochrogaster).     

Chromosome constitution and its bearing on the chromosomal radiosensitivity in man

Blood samples obtained from patients with various types of inborn chromosome abnormalities were exposed to γ-rays and the relationship between the chromosome constitution and chromosomal radiosensitivity of lymphocytes was studied by analysing types and frequencies of radiation-induced chromosome aberrations. The results showed that the chromosomal radiosensitivity was consistently higher in the cells which were trisomic for the whole or a part of a chromosome than in the cells with normal karyotype, but it was not significantly influenced by the monosomic conditions, reciprocal translocation and inversion. Age of the subjects also affected the chromosomal radiosensitivity, which was elevated in the neonates.

The analysis of chromosome aberrations showed that the high frequency of radiation-induced chromosome aberrations was due to the increased production of exchange aberrations and that the level of deletions was not affected either by factors of the chromosome constitution or of the age of the subject. A hypothesis to explain the increased chromosomal radiosensitivity of the trisomic cells was given in line with the effects of altered enzyme activity on the production of exchange aberrations.

The parallelism between the increased chromosomal radiosensitivity in the trisomic cells and the susceptibility of the affected persons to neoplasia allowed us to recognize that the trisomic cells are particularly cancer-prone and that the illegitimate repair of chromosome damage, which is intrinsic to the trisomic cells, may play an important role in the development of cancer.     

Glycoprotein Ib distribution on the surface of platelets in resting and activation states: An electron microscope study

Summary Using a monoclonal antibody (TM60) against glycoprotein (GP) Ib, we determined immunocytochemically how GPIb is distributed on the platelet surface. When glutaraldehyde-fixed platelets were incubated with TM60, a uniform distribution of ferritin particles which represent the localization of GPIb was observed on the surface membrane of platelets. The particles were distributed at intervals of about 100 nm. The number of ferritin particles on the surface of one side were 2070–4150 (2940 ± 790; mean ±s.d.,n = 10) under the scanning electron microscope. The distribution of ferritin particles was somewhat disarranged on the surface of unfixed platelets incubated with TM60 compared to that in the fixed platelets. Cluster-like structures of ferritin particles were observed in several places. When platelets were activated with ristocetin or thrombin, the distribution of ferritin particles was disturbed and cluster formation was observed in several places on the surface. These findings suggest that GPIb is uniformly distributed on the surface of platelets in the resting state, and that cluster formation occurs during activation of platelets.     

Acrosomal ATPase in starfish and bivalve mollusk spermatozoa

ATPase activity was found in acrosomes of starfish and bivalve mollusk spermatozoa, using a cytochemical method with electron microscopy. The activity was located in central material of the starfish acrosome and in material lining the acrosomal membrane of the Mytilus acrosome, as well as in the basal part of the starfish acrosome. The ATPase activity in the former material was preferably activated by Ca²⁺, while that in the starfish basal material was preferably activated by Mg²⁺. Both types of activity persisted during and after the acrosome reaction. ATPase activity was also observed in the region of the axial filament complex of the flagella, in centrioles and in a basal matrix. ATPase in the acrosome also hydrolysed other nucleoside triphosphates. However, there was no detectable phosphatase activity, and little pyrophosphatase or 5′-nucleotidase activity. Evidence was obtained that adenylate kinase may be included in the acrosome. A possible role of the ATPase activity in the acrosome reaction is discussed.