Oxygen-dependent radiosensitivity of Escherichia coli W3623 his^- was confirmed. Regarding cellular superoxide dismutase (SOD), cells grown oxically gained higher activity than those grown anoxically, however, the reinforced enzyme level could not compensate the oxygen effect, i.e., the enhanced lethal effect of oxic γ-irradiation. Rather, the enhancement of oxygen effect was found in cells grown oxically compared with those anoxically. Oxygen enhanced lethality was mitigated to the extent by the amount of added SOD into the cell suspension to be irradiated. The results supported a proposal that superoxide anion, O^-₂, is involved in the oxygen effect, with the most likely site of the damage in the outer structure of cell but not in the cell matrix. Reverse oxygen effect could be found with λ phage DNA in transfecting ability. Added SOD protected phage DNA somewhat in oxic irradiation. While considerable protections were found in anoxic one with the added SOD even autoclaved but their function was still unknown.

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Formation and accumulation of protein components and their subunits in developing soybean seeds were investigated ultracentrifugally, electrophoretically and immunologically from flowering until maturity. Glycine max var. Ogura daizu took 80 to 90 days after flowering to mature its seeds. Only smaller molecular weight components were observed until 26 days after flowering in ultracentrifugal and electrophoretical patterns. Components of about 7S and 12S appeared 33 days after flowering. Immunological studies and sodium dodecyl sulfate electrophoresis, however, indicated that the specificities as antigens and subunit con-stituents of these components were different from those of the matured 7S and 11S globulins. Immunological specificity of mature 7S globulin was obviously detected 40 days after flowering, and that of 11S globulin was seen 50 days after flowering at least. These storage protein components were intensively accumulated from 40 days after flowering until maturity.

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The synergistic effect of trimethylamine oxide (TMAO) on the antioxidative activity of h-tocopherol (δ-Toc) to inhibit the autoxidation of methyl linoleate was investigated.
The peroxide value of methyl linoleate incubated with δ-Toc and TMAO at 50°C was maintained at constant values of 25_??_30, while a drastic increase in the peroxide value was observed in the absence of TMAO.
An oxidation product of methyl linoleate, which was formed only in the presence of δ-Toc and TMAO, was isolated and identified to be a mixture of methyl 9-keto-10, 12-octadeca-dienoate and methyl 13-keto-9, 11-octadecadienoate.
δ-Toc diphenyl ether dimer and two δ-Toc biphenyl dimers were identified, and the latters were thought to be atropisomers with each other. The amounts of reducing dimers of δ-Toc formed were larger in the presence of TMAO than in the absence of TMAO, and it was found that these dimers were likely to play an important role in synergism between δ-Toc and TMAO.

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The synergistic effect of trimethylamine oxide (TMAO) on the antioxidant activity of γ-tocopherol (γ-Toc) to inhibit the autoxidation of methyl linoleate was investigated.
Formation of the reducing dimers of γ-Toc was found to be closely related to the synergism of TMAO with γ-Toc. γ-Toc diphenyl ether dimer and two γ-Toc biphenyl dimers were identified, and the tatters were considered to be atropisomers with each other.
From the reaction of the hydroperoxide (LOON) of methyl linoleate with TMAO, TMAO was found to act as a peroxide decomposer. TMAO produced methyl keto-octadecadienoate, which was formed only in the presence of γ-Toc and TMAO, from LOOH, and markedly decreased the POV of LOOH.
A possible mechanism of synergism between Toc and TMAO in the inhibition of the autoxidation of methyl linoleate was proposed.

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Activities of AChE and ChE per g wet weight of the cerebrum were higher in the cerebrum of the MAM-induced microencephalic rats that were the offspring of the mother rats injected on the 15th day of pregnancy with MAM-acetate than in the normal rats cerebrum. They increased for 30 days after birth and were maintained constant thereafter. The activity of AChE more increased in the cerebral subcortex than in other regions. With the increase in dose of the drug administered to mother rats, the cerebral weight of their offspring decreased gradually whereas AChE activity per g wet weight of their cerebrum increased. These findings support existence of plasticity in the cerebrum.

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Hexose 1-phosphate uridylyltransferase (EC 2.7.7.12) was present constitutively in Bifidobacterium bifzdum. The enzyme was purified to a homogeneous state from B. bifzdum grown on a glucose medium and characterized. The molecular weight of the enzyme is about 110, 000. The pH optimum of the enzyme was 7.5. The enzyme was very labile on the acidic side below pH 4.5. Thymidine diphosphate glucose could serve as a substrate with about 60 efficiency of UDP-glucose. The Km values for UDP-glucose, galactose 1-phosphate (Gal-1-P), UDP-galactose and glucose 1-phosphate (Glc-1-P) were estimated to be 2.3×10^-5M, 5.0×10^-4M, 3.1×10^-5M and 1.4×10^-4M, respectively. From these results the physiological roles of the enzyme were considered in relation to galactose metabolism in B. bifidum.

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UDP-galactose 4-epimerase (EC 5.1.3.2) was purified to a homogeneous state from Bifzdobacterium bifzdum grown on a glucose medium. The molecular weight was estimated to be about 90, 000. The purified enzyme was very stable and 60% of its initial activity survived three months of storage at 4°C even at a low protein concentration (0.2mg/ml). The optimum pH was 9.0, and the Km values for UDP-galactose and UDP-glucose were 5.4×10-4M and 1.4×10^-4M. UDP was a competitive inhibitor. The enzyme activity was stimulated by various sugar phosphates, but was slightly inhibited by fructose 1, 6-diphosphate (FDP). A high concentration of galactose or glucose, which had no effect by itself, inhibited the activity in combination with UMP. The inhibition by FDP was also enhanced by combi-nation with UMP.

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As one of the applications of a technique for providing W/O/W-type multiple-phase emulsions, an attempt was made to prepare water-in-olive oil-in-water systems with soy lecithin, Span 80 and sucrose-fatty acid ester as a model for estable multiple-phase emulsions. The procedure tested was divided into two operations, as follows; (1) preparation of a water-in-olive oil emulsion stabilized by a mixture of soy lecithin and Span 80, and (2) mixing of the above emulsion with an aqueous solution of sucrose-fatty acid ester so as to prepare a W/O/W dispersion composed of aqueous compartments surrounded by the oil-phase layer. The results obtained suggest that the concentrations of soy lecithin and Span 80 in the oil phase are of critical importance to the formation of the W/O/W systems. Both components seem to cooperate in constituting an incorporated film at the olive oil/water interface in a mechanically stable form.

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The regulation of aspartokinase (EC 2.7.2.4) in Brevibacterium lactofermentum was studied. One mM of lysine or threonine or threonine plus lysine inhibited the activity of aspartokinase to the levels of 45, 41 or 81%, respectively, as compared with no addition.
In order to obtain mutants with aspartokinase which is genetically desensitized to the feedback inhibition, we tried to derive mutant resistant to S-(2-aminoethyl) L-cysteine (AEC) or α-amino-β-hydroxyvarelic acids (AHV) from B. lactofermentum.
Most of AEC resistant mutants produced more than 5mg/ml of L-lysine. These aspar-tokinase were not inhibited by threonine or threonine plus lysine, but inhibited by lysine.
On the other hand, about 30% of AHV resistant mutants produced more than 5mg/ml of L-lysine and these aspartokinases were partially desensitized to the feedback inhibition by lysine.
Furthermore, the homoserine dehydrogenase level and the degree of inhibition by threo-nine in ARV resistant mutants were almost the same as those in parental strain. Both AEC and AHV resistant mutants produced L-lysine twofold as compared with that of either AEC or AHV resistant mutants.
The data provide significant evidence that aspartokinase in Brevibacterium lactofermentum was subject to the single or concerted regulation by threonine and lysine.

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A cell free protein synthesizing system was established from alkalophilic Bacillus. Amino acid incorporation was dependent on ribosomes, supernatant, poly uridilic acid (poly U) and magnesium. The optimum pH for protein synthesis by poly U was about 0.5 higher than that of Bacillus subtilis, In protein synthesis directed by endogeneous mRNA, the optimum pH was also about 0.5 higher. The properties of ribosomes and aminoacyl-tRNA synthetase were not remarkably different. A comparative study of alkalophilic Bacillus and B. subtilis showed that the protein synthesizing system of alkalophilic Bacillus was similar to that of mesophilic bacteria.

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The active groups of the lipase from rice bran were examined by chemical modification techniques. Diisopropylphosphofluoridate (DFP), paraoxon and diazonium-l-H-tetrazole inhibited completely the activity of the lipase in contrast to the partial inactivation with p-chloromercribenzoic acid and N-ethylmaleimide. Photooxidation as well caused strong inactivation of the lipase. The rate of inactivation of the lipase with paraoxon was much faster than that with DFP. The pH-dependence of the inactivation by DFP was observed over the pH range from 7 to 8, which was similar to that of enzymatic hydrolysis of tributyrin. These experimental results suggest that the lipase is likely to have a serine residue involved in active site, although the role of a histidine residue in catalytic function is not made clear yet. However, the possibility that the sulfhydryl group plays a vital role in the catalytic function of the rice bran lipase was eliminated.

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Amadori compounds were isolated from soy sauce by ion exchange chromatography, gel filtration and paper chromatography. Five Amadori compounds, fructose-glycine, fructose-alanine, fructose-valine, fructose-isoleucine and fructose-leucine, were identified and their concentration in soy sauce was estimated to be about 0.2, 0.3, 1.2, 1.3 and 1.5mM, re-spectively. The Amadori compounds exhibited remarkable browning in the presence of oxygen and iron, but darkened very little without oxygen or iron. Amino acids promoted the oxidative browning of the Amadori compounds.

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A fluorescent reagent, N-(9-acridinyl)maleimide, was studied for its application to the determination of GSH and the total thiol content (crude GSH) in mammalian tissues. The fluorescence was reproducible and stable at pH 8.8. The fluorescence intensity was linear in the range of the GSH concentration from 0.001 nmole/ml to 100 nmole/ml. The total thiol content of the mammalian tissue determined by this method agreed well with that ob-tained by the DTNB colorimetric method. The sensitivity of the present method was more than fifty times higher than the DTNB method; determination with 0.03_??_15mg of tissue was adequate.

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Applicability of PMR spectroscopy to the discrimination of the SCP-related yeast was investigated because the yeast was inactivated by heat treatment. When Candida subtropicalis was cultured in a medium containing glucose as a sole source of carbon, its cell wall mannan (mannan A) showed a PMR spectral pattern characterized by three intense peaks at δ:4.97, 5.10, 5.29 and two small peaks at δ:4.90, 5.19. Mannan (mannan B) from the yeast cultured in a medium containing n-pentadecane and triton X-100 showed a different spectral pattern in which the signals were observed at δ: 4.97, 5.10, 5.29, the intensity ratios of the signals were also different from those of mannan A. Acetolysis mannan was analyzed to compare the differ-ence between two specified structures by using a gel elution method, methylation analysis and PMR spectra. Mannan B contained a less amount of the a (1→3) linkage than mannan A did, and differed from mannan A in its distribution pattern of side chain units. Our previous results together with the present ones proved the PMR method to be effective for the dis-crimination of the yeast.

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Soybean 11S globulin was polymerized by incubating with peroxidizing linoleic acid. The molar ratio of the acidic subunits to the basic subunits of IiS globulin decreased with the elapse of the incubation time. The acidic subunits were lost faster and formed polymers more easily than the basic subunits. The acidic and basic subunits in 1tS globulin were fractionated by DEAE-Sephadex gel chromatography. Each of the acidic and basic subunits was allowed to react with peroxidizing linoleic acid individually. The results also showed that the acidic subunits formed polymers faster than the basic subunits. Both succinylated and acetylated 11S globulins were also submitted to the incubation with peroxidizing linoleic acid. The polymerization of the modified protein was suppressed by masking r-amino groups.

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Four analogues of the intermediates of gibberellin biosynthesis, 1-geranylimidazole (V) and three 17-nor-16-azakauranes (VIII), (IX) and (X) were found to inhibit gibberellin production of Gibberella fujikuroi by the procedure of bioassay which the author established. These compounds showed also growth retarding effects on rice seedlings.

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The authors have reported) a new maleimide type fluorescent thiol reagent, N-(9-acridinyl)-maleimide (NAM). In this paper the syntheses of NAM and its coupling products with thiol compounds are presented. NAM was synthesized from 9-aminoacridine and maleic an-hydride through dehydratic cyclization in polyphosphoric acid. NAM showed no sub-stantial fluorescence but its coupling products with thiol compounds exhibited strong blue fluorescence. Application of NAM for the fluorometrical analysis of cysteine and glutathione are suggested.

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Bovine trypsin (EC 3.4.4.4) was bound to RNA in the presence of water-soluble carbo-diimide [1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide], and the enzymic properties of this complex were compared with those of free trypsin. RNA bound trypsin does not lose its activity in the presence of 0.5% SDS, and this is consistent with the fact that the intensity of the fluorescence spectrum of this complex does not change when SDS is added. The RNA-enzyme complex seems suitable for studies on the conformation of immobilized enzyme.

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Structure-activity relationships were studied on 25 analogs of 3-pyridylguanidines toward powdery mildew. Only the di-sec-alkyl analogs were active among the 1-alkyl or 1, 1-dialkyl-2-(3-pyridyl)-3 p-ethoxyphenylguanidines on available data. The activity was linearly cor-related with Hammett's σ constant of the substituents on the phenyl group, among which cyano, benzyloxy, alkoxy and diethylamino groups showed higher activity than chloro, nitro, methyl, bromo and phenyl groups. Fungitoxic spectrum and other fungicidal natures were also shown and compared with those of S-1358 and its derivatives.

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Reactions of DHA with aromatic amines in ethanol produced fairly stable esr signals with characteristic hyperfine structures that indicated the presence of the aromatic moiety in the radical molecules. Several intermediates of ninhydrin reactions, i.e., carbinolamines and Schiff bases, were isolated and these two were found to give rise to similar esr signals when subjected to reaction with ascorbic acid. This also provided basis for the elucidation of the structures of the radical species from the reaction of DHA and aromatic amines.

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Factors contributing to the heat-induced aggregation of ovalbumin were studied. The first half of this work was about the aggregation behavior of heat-denatured ovalbumin, and the last half, about the heat-induced aggregation of ovalbumin solution. Factors studied in both experiments were protein concentration, pH, addition of salts and succinylation of amino groups. From the results of the two experiments, it was concluded that the main factor contributing to the heat-induced aggregation of ovalbumin was the degree of electrostatic repulsion on the packing of the denatured protein molecules.

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The cell wall fraction prepared from the yeast-like cells of Tremella fuciformis was frac-tionated by proteinase digestion, hot-water and alkaline extractions. The acidic polysac-charide, which may originate from the outer layer of the cell wall, was purified from the hot-water extract. It was composed of D-glucuronic acid, D-mannose and n-xylose (molar ratio, 0.5:3.8:1.0). The methylated polysaccharide yielded on acid hydrolysis 2, 3, 4-tri-O-methyl-D-xylose, 2, 3, 4, 6-tetra-O-methyl-, 2, 4, 6-tri-O-methyl-and 4, 6-di-O-methyl-n-mannose, and 2, 3, 4-tri-O-methyl-n-glucuronic acid (molar ratio, 1.0:0.8:2.7:2.3:0.5) together with a trace of 3, 4-di-O-methyl-D-xylose, suggesting that the polysaccharide consists of a backbone of (1→3)-linked D-mannose residues, some of which are substituted at the C-2 positions with single or short side chains of D-xylose, D-mannose and D-glucuronic acid residues.
The alkali-insoluble residue of the cell wall was composed of D-glucose, D-glucuronic acid, D-mannose and D-xylose (molar ratio, 4.3:0.6:2.5:1.0). Methylation and periodate oxidation studies suggested it to comprise two polysaccharide moieties, i.e., β-D-glucan and glucurono-xylo-mannan, the structure of the latter resembling that of the cell surface acidic polysaccharide. The glucan moiety was shown by methylation and enzymatic degradation to have a branched structure consisting of β-(1→3)- and (1→6)-linkages (approx. ratio, 2:3). The controlled Smith degradation of the alkali-insoluble polysaccharide yielded an insoluble, degraded, predominantly (1→3)-linked gluco-mannan which may represent the backbone of the cell wall polysaccharide. On the basis of these findings, the constitution of the cell wall is discussed.

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The O-glycosidically linked carbohydrate units of ovomucin were released from serine and threonine in peptide as oligosaccharide chains by alkali treatment with and without borohydride. Two sulfated oligosaccharides were fractionated by using gel filtration and ion-exchange chromatography. The yield of sulfated oligosaccharides released by alkali treatment was higher in the presence of borohydride than in the absence of borohydride. The sulfated oligosaccharides released by alkali treatment with borohydride were as follows: an oligosaccharide composed of N-acetylgalactosaminitol, galactose, N-acetylneuraminic acid and sulfate in a molar ratio of about 1:1:1:1 and another oligosaccharide in a molar ratio of about 1:1:0.6:0.5.

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N-Acetyl-L-glutamic γ-semialdehyde was enzymatically prepared in 51% yield from Na-acetyl-L-ornithine. By the reaction with 4-benzyloxy(or methoxy)-phenylhydrazine this aldehyde was converted into N-acetyl-5-benzyloxy(or methoxy)-L-tryptophan which is known to be transformed into 5-hydroxy-L-tryptophan, a biogenetic precursor of serotonin. By this work an amino acid of L-glutamic acid family was first chemically derived to that of L-trypto-phan series.

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The insecticidal activity of methyl substituted cyclohexenylmethyl and cyclohexadienyl-methyl chrysanthemate was investigated from the view point that these compounds have partial structure of the hypothetically cyclized rethrolones. 4-Methyl-l-cyclohexenylmethyl-and 4, 5-dimethyl-1, 4-cyclohexadienylmethyl chrysanthemate showed the equivalent insecticidal activity to houseflies as those of natural pyrethrins or α-dl allethrins. The insecticidal activity of other related compounds is also discussed.

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Two constituent polypeptide chains (lle and Ala chains) of ricin E were separated by an affinity chromatography on acid-treated Sepharose 4B in the presence of β-mercaptoethanol, and the Ala chain was purified by CM-cellulose column chromatography at pH 7.0 and crystal-lized.
From the analyses of isoelectric points, amino acid compositions and tryptic peptides, it was revealed that the Ile chain will be common in ricin E and ricin D while the Ala chain of ricin E is homologous protein having several amino acid replacements to that of ricin D.
Furthermore, the toxicity of the hybrid molecule consisting of the Ile chain of ricin E and the Ala chain of ricin D to malignant cultured cells was determined to be similar to that of ricin D. It was concluded that the difference in the toxicity toward the cultured cells between ricin E and ricin D was due to the difference in the function of Ala chain.

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The mutagenic activities of quinoline, isoquinoline, phenanthridine, benzo(f)quinoline, benzo(h)quinoline and their a-amino derivatives were compared in relation to the effect of structural changes using the Salmonella typhimurium test system. All mutagenic compounds tested require the liver microsomal fraction for their mutagenic activity. Phenanthridine, two benzoquinolines and quinoline were mutagenic, α-Amination of two benzoquinolines and quinoline resulted to increase their mutagenic activity intensively. Addition of a benzene ring to the benzene moiety of 2-aminoquinoline, so that two carbon atoms are shared, affected distinctly the increase in the mutagenic activity. The co-existence of benzoquinoline series with 2-aminobenzo (f) quinoline showed the clear synergistic action.

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(+)-Biotin was synthesized from D-glucosamine in a sequence of 13 reactions.

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Single and double asymmetric inductions were achieved by the sodium borohydride reduction of α-ketoesters in the presence of chiral phase transfer catalysts. The quaternary ammonium salts with hydroxyl group in the substituent moiety showed the effective catalytic activity. The reduction of (-)-(3R)-menthyl benzoylformate gave (3R)-menthyl (S)-mandelate in contrast to the prediction by the Prelog rule.

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The effect of light on nicotine production in cultured tobacco callus tissues was investi-gated. Illumination strikingly inhibited the nicotine production even though the growth of the tissues was slightly stimulated by the light. The inhibitory effect of light increased as the intensity and the length of the illumination increased. No decisive difference in nicotine production was observed between the effects of blue light and red light. The depression of nicotine production by light was restored completely when the tissue was transferred to the dark. Therefore, nicotine production is concluded to be regulated by the light just as in the case of plant growth regulators. The inhibitory effect of light was assumed to be caused rather by the inhibition of nicotine biosynthesis than by the promotion of nicotine decomposition.

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It was found that trans-2-octenoic acid was hydrated to 3-hydroxyoctanoic acid by the resting cells of Mucor species. By application of the hydrating activity of the fungi, we prepared seven L 3-hydroxyalkanoic acids (C₆_??_C₁₂) in moderate yields. The method is of value for the preparation of dextrorotatory 3-hydroxy acids.

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