Alteration of generations in peanut plants was examined by the culturing of seeds. Flowers were induced in more than 50% of the seed cultures in vitro. Benzyl aminopurine stimulated the rate of flower induction most among the factors examined. Development of pegs was stimulated by gibberellic acid given after flowering. Dark culture allowed the development of ovaries in pegs to immature seeds and then to mature seeds ; with this, the life cycle of the peanut plant was complete. Mature seeds excised from the original cultures germinated normally in vitro, and the cotyledonary nodes proliferated multiple shoots on shoot-forming medium. Alteration of generations and the in vitro omission of certain steps of the life cycle in plants are discussed.

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Five extracellular endo-xylanases were recognized in the culture broth of shochu koji mold (Aspergillus kawachii, IFO 4308), and three major xylanases (Xy1A, Xy1B, and Xy1C) were purified and characterized. The molecular masses of Xy1A, Xy1B, and Xy1C were 35, 000, 26, 000, and 29, 000, and isoelectric points were pH 6.7, 4.4, and 3.5, respectively. Amino acid compositions and other properties were studied and these three xylanases were found to be greatly different in their properties. These three xylanases, Xy1A, Xy1B, and Xy1C, were stable between pH 3-10, 3-10, and 1-9 and the optimum pHs were 5.5, 4.5, and 2.0, respectively. Consequently, these xylanases were acid stable xylanases, especially Xy1C was an acidophilic xylanase (acid xylanase). These xylanases produced various xylooligosaccharides including xylose from xylan and the main product was xylobiose in all xylanases. The production of acid xylanase (Xy1C) was enhanced with a low initial pH of the medium.

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Gobo dietary fiber (GDF) obtained from the roots of edible burdock (Arctium lappa L.) was examined for its protective role against amaranth (Food Red No. 2, Am) toxicity in the upper gastrointestinal tract (from the mouth to the ileal end) of rats. Ileorectostomized rats were examined for their growth response to feeding with a purified basal diet containing 4% Am with or without 7.5% GDF. The transit half-time (TT₅₀) through the upper gastrointestinal tract was also examined with ileostomized rats by recovering the small intestinal contents from the ileal end, using Cr-EDTA as a nonabsorbable water-soluble marker. Although feeding Am to the ileorectostomized rats resulted in a 50% mortality, concurrent feeding of Am and GDF not only protected the rats from death but also significantly promoted their growth rate when compared to the effect on the survivors fed only with the Am-containing diet. The results of TT₅₀ measurements on ileostomized rats showed that the TT₅₀ was decreased by half in the presence of Am, but was restored to the value for the control without Am when the Am-containing diet was supplemented with GDF. These and previous rsults imply that Am toxicity develops mainly in the upper gastrointestinal tract as a result of decreased availability of nutrients that is produced by the rapid transit and inhibitory effect of Am on the digestion-absorption process. The beneficial effect of GDF appears to be in normalizing the rapid transit through the upper gastrointestinal tract of chyme containing Am and not in the cecum or colon.

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α-, β-, and γ-cyclodextrins (CDs) completely inhibited raw starch digestion by glucoamylase I (GA I, MW 90, 000) from Aspergillus awamori var. kawachi, and inhibited by 85% the raw starch adsorption of GA I at the CD concentrations of 1-5 ImM. CDs at 1-5 mM did not inhibit gelatinized starch hydrolysis by GA I, but at the concentration of 50 mM, they inhibited such hydrolysis slightly. GA I was specifically adsorbed onto CD-Sepharose 6B, but glucoamylase I' (GA I', MW 73, 000), which does not adsorb onto or digest raw starch, from the same strain was not adsorbed onto that gel. The adsorption of the glucoamylases onto raw starch and CD-Sepharose 6B was correlated to their digestion of raw starch. The hydrophobic adsorption of GA I onto CDs and raw starch occurred competitively at the Cp region, which is on the C-terminal side of Gp-I in the site for raw starch affinity of GA I, and inclusion complexes were formed.

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Fine droplets of sardine oil either embedded in or covered with egg white powder, which had been prepared from an oil-in-water emulsion containing 10% protein-oil (9 : 1) mixture by spray-drying and freeze-drying, were examined for their oxidative stability during storage under moderate conditions (RH=45-55%, 40°C). The antioxidative effect was estimated by measuring the peroxide value as well as the residual unsaturated fatty acid (mainly eicosapentaenoic and docosahexaenoic acids). Microencapsulation of the oil droplets with albumen by freeze-drying was not so effective in stabilizing against oxidation as that by spray-drying, probably because of the porosity of a protein coating on oil or leakage of oil through crevices. As a practical application of powdered sardine oil (i.e., entrapped in spray-dried albumen particles), plain cookies and those enriched with the oil were baked, and any difference in taste between them was evaluated by a paired preference test assessed by 32 amateur panelists. Sardine oil fortification of the cookies was judged not to affect their quality from the results of the sensory test. Spray-dried egg white powder inclusive of sardine oil was stable during prolonged storage, so that its use would be favorable for supplying an n-3 series of polyunsaturated fatty acids.

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A new gibberellin (GA) was identified from extracts of cotyledons of 7 day-old canola seedlings (Brassica campestris cv. Tobin). This GA is 12α-hydroxy-GA₁ and has been assigned the trivial name of GA₈₅ Isolation was monitored by the Tan-ginbozu dwarf rice micro-drop assay after each high-performance liquid chromatography (HPLC) step. Identification was based on Kovats retention index (KRI) and the mass spectrum of the methyl ester, trimethylsilyl ether (MeTMSi) derivative after analysis by gas chromatography-mass spectrometry (GC-MS) in comparison with an authentic sample of 12α-hydroxy-GA₁. Based on quantitation by the dwarf rice micro-drop assay, GA₈₅ is one of the major biologically active GAs in cotyledons of young canola seedlings.

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Casein was conjugated with dextran and galactomannan in a controlled dry state at a relative humidity of 79% and at 60°C for 24 hr. The covalent attachment of polysaccharides to casein was confirmed by SDS-PAGE and HPLC. The emulsifying activity of the casein-dextran and casein-galactomannan conjugates was 1.5 times higher than that of casein. The emulsion stability of the casein-dextran and casein-galactomannan conjugates was 10 times higher than that of casein. The improvement in these emulsifying properties reached a steady state when the conjugation of casein with polysaccharide was done for 24 hr in a controlled dry state, suggesting the rapid formation of conjugates through a Maillard reaction in the case of casein. Compared to commercial emulsifiers, the casein-polysaccharide conjugates showed better emulsifying properties in acidic and high-salt concentration systems.

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Lactic acid bacteria in four samples of sour dough sponges were studied quantitatively and qualitatively. In each sponge, there were one or two species of the genus Lactobacillus : L. reuteri and L. curvatus in San Francisco sour dough sponge, L. brevis and L. hilgardii in panettone sour dough sponge produced in Italy, L. sanfrancisco from a rye sour dough sponge produced in Germany, and L. casei and L. curvatus from a rye sour dough sponge produced in Switzerland. For all isolates except the L. reuteri strains oleic acid, a component of the Tween 80 added to the medium, was essential for growth. It was of interest that lactobacilli requiring oleic acid were the predominant flora of lactic acid bacteria in the microbial environment of sour dough sponges.

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The effects of one-time ethanol intoxication on ascorbic acid and lipid metabolism and on drugmetabolizing enzymes in liver of rats were investigated. Male Donryu rats that had been fed semi-purified feed were given 5 g/kg ethanol solution (25%, w/v) via a stomach tube and killed 16 h after intubation. The amount of ascorbic acid excreted in the urine after ethanol administration increased, but renal and adrenal concentrations of ascorbic acid decreased. The serum levels of total cholesterol, high-density-lipoprotein cholesterol, triglycerides, phospholipids, and non-esterified fatty acids were elevated in rats given ethanol, but hepatic level of total lipids, cholesterol, triglycerides, phospholipids were not. The hepatic concentrations of cytochrome P-450 and cytochrome b₅ did not increase, but this large dose of ethanol increased the activities of aminopyrine N-demethylase and cytochrome c reductase. These results indicated that the single dose of ethanol affected the ascorbic acid and lipid metabolism of rats, and induced drug-metabolizing enzymes in their liver.

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5-Methyluridine (5MU) was synthesized efficiently from adenosine, thymine, and phosphate by a combination of adenosine deaminase (ADA), purine nucleoside phosphorylase (PUNP), pyrimidine nucleoside phosphorylase (PYNP), and xanthine oxidase (XOD). Adenosine was converted into inosine first by ADA. 5MU and hypoxanthine were synthesized from inosine and thymine by PUNP and PYNP. The hypoxanthine formed was converted into urate via xanthine by XOD. After inosine was completely consumed, an equilibrium state, in which 5MU, thymine, ribose-1-phosphate, and phosphate were involved, was achieved. At the equilibrium state, the maximum yield of 5MU was obtained. The yield of 5MU was 74%, when the initial concentrations of adenosine, thymine, and phosphate were 5 mM each. On the other hand, in the absence of ADA or XOD the yield of 5MU was 1.8%. Several kinds of nucleosides were also synthesized with high yield by the same method.

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A partial amino acid sequence of acid phosphatase-1¹ (apase-1¹), one of acid phosphatase isozymes of tomato, was identified. This information enabled us to synthesize degenerated primer pools of oligonucleotides for polymerase chain reactions (PCR) using cDNA for poly(A)⁺ RNA of tomato leaves as a template. As a result, a 135-bp, then a 467-hp PCR product were obtained. Nucleotide sequencing of these two PCR products gave a total of 522-bp sequence that was identified as a part of the Asp-1¹ gene judging from the amino acid sequence deduced from it. Using the 135-bp PCR product as a probe, we detected the restriction fragment length polymorphism (RFLP) in two different lines of tomato by genomic Southern blot analysis. We also did pulsed-field gel electrophoresis (PFGE) and Southern blot analysis to search for suitable fragments to clone into a YAC vector. As a result, a single band with a size that could be cloned into a YAC vector was detected when the genomic DNA was digested with some kinds of restriction enzymes.

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Effects of tea polyphenol intake (0.4 g/volunteer, 3 times per day, for four weeks) on fecal microflora, bacterial metabolites, and pH were investigated using eight healthy human volunteers. Counts for Clostridium perfringens and other Clostridium spp. were significantly decreased during the tea polyphenol intake periods. Percentage of volunteers having C. perfringens in their feces decreased significantly, but not for other Clostridium spp. Percentage of Bifidobacterium spp. (the acid forming bacteria) in total counts and the content of volatile fatty acids including acetic and propionic acids increased significantly, which might have reduced the fecal pH. However, the tea polyphenols had no effect on fecal enzyme activities, ammonia, or putrefactive products. Two weeks after discontinuing the intake, the microflora counts and their biological parameters appeared to have returned to normal.

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The effects of green tea polyphenols, inhibitors of various biological activities of cariogenic bacteria in vitro, on caries development were examined using conventional rats. A total of 96 male rats were divided into 8 groups and the rats in the test groups were given tea polyphenols ranging from 0.1% to 0.5% in their cariogenic diet or drinking water for 40 days. Total fissure caries lesions was significantly reduced by the addition of tea polyphenols to the diet or in the drinking water. Diet containing 0.1% tea polyphenols demonstrated about 40% reduction of total fissure caries lesions. No toxic effect of tea polyphenols on rats were observed under these experimental conditions.

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The exothermic and endothermic peaks in cooling and heating curves of differential scanning calorimetry(DSC) for gellan gum gels without and with potassium chloride and sodium chloride were analyzed. The gelling and melting temperatures shifted to higher temperatures with increasing gellan and salt concentration in the concentration range of gellan from 0.3 to 2.0% (w/w). The exothermic and endothermic enthalpy increased with increasing gellan and salt concentrations. Cooling DSC curves showed one exothermic peak for samples with salts and at low gellan concentration. Heating DSC curves showed many peaks for all samples except 0.3% (w/w) gellan gum gels. The sol-gel transition of samples was examined numerically by using a zipper model approach. The introduction of cations increases the number of junction zones or zippers and decreases the rotational freedom of parallel links. This makes the structure of junction zones more heat resistant, and increases the elastic modulus of the gel.

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A simple method was developed to establish serum-independent Chinese hamster ovary (CHO) cells that grew and secreted high level of human antithrombin-III (AT-III). First, human AT-III and mouse dihydrofolate reductase (DHFR) cDNAs were transfected into DHFR-deficient CHO cells. Transfected cells were treated with increasing concentrations of methotrexate (MTX) and clones secreting high levels of AT-III (10-20 μg/ml/3 day) in a serum-containing medium were obtained. Serum-independent clones were derived from the serum-dependent clones by simply culturing the cells for a few weeks in a serum-free medium. In a serum-free medium the established serum-independent clones grew at normal rate and produced almost equivalent amount of AT-III to that of the serum-dependent, parent clones. In addition, AT-III from the serum-independent clones has specific activity similar to that of plasma-derived AT-III.

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A fluorometry of hydroperoxides in oil and food samples was developed using diphenyl-1-pyrenylphosphine (DPPP) with a sample size of less than 20 mg by a batch method. The sensitivity was more than 10, 000 times that of the conventional iodometry. A good accordance was obtained between peroxide values (POV) measured this way and by iodometry (r=0.9995, n=41, POV=0.04-240).

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A levanase from Bacillus sp. was purified to a homogeneous state. The enzyme had a molecular weight of 135, 000 and an isoelectric point of pH 4.7. The enzyme was most active at pH 6.0 and 40°C, stable from pH 6.0 to 10.0 for 20hr of incubation at 4°C and up to 30°C for 30 min of incubation at pH 6.0. The enzyme activity was inhibited by Ag⁺, Hg²⁺, Cu²⁺, Fe³⁺, Pb²⁺, and p-chloromercuribenzoic acid. The enzyme hydrolyzed levan and phlein endowise to produce levanheptaose as a main product. The limit of hydrolysis of levan and phlein were 71% and 96%, respectively.

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(-)-Perillaldehyde (1) was photoirradiated by a 400W high-pressure mercury lamp under various conditions. In MeOH, 1 was degraded to give two new MeOH adducts (2 and 3) and a new methyl ether (4) under an N₂ stream, while 1 was converted to a dimethyl acetal (5) and oxidation product (6) in addition to 2 and 3 under an O₂ stream. In EtOAc, 1 disappeared to afford two new dimers (7 and 8) under an N₂ stream, while 1 was changed to two oxygenated products (9 and 10) including a new compound under an O₂ steam. In the presence of rose bengal (RB), 1 was converted to 5 by visible light in MeOH under an N₂ stream. Photosensitizabon of RB was observed under an O₂ steam, and two new oxygenated products (11 and 12) were formed. Perillaldehyde was relatively stable in n-hexane under an N₂ stream or in the presence of RB under an O₂ stream.

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New derivatives of pyridinyloxyphenoxypropionic acid were synthesizd, which have a keto group in their ester-alkyl structures. Some acylmethyl esters, e.g., acetylmethyl ester, hydroxyacetylmethyl ester and diazoacetylmethyl ester, were found to show outstanding herbicidal activity against grass weeds. A study of the hydrolysis rate of esters with regard to their pro-herbicide action is also discussed.

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In a solution, cis-abscisic acid (ABA) isomerizes into the trans-isomer which is physiologically inactive, and this structural instability is regarded as a reason for insuitability of ABA in agricultural applications. The side chain of ABA, the 2-cis-4-trans-3-methyl-2, 4-pentadienoic acid moiety, was replaced with various substituted phenyl groups to examine biological effects of the inflexibility of the part. Construction of the phenyl moiety was achieved by cyclizing the ionone derivatives and subsequent aromatization. Some of those new compounds showed ABA-like activity in both seed germination and transpiration assays.

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Five 2-substituted 6/7-methoxy-1, 4-benzodioxan-7/6-carbaldehydes and 6-methoxy-1, 4-benzodioxan-7-carbaldehyde available for the syntheses of insecticidal neolignan analogs were prepared from 4/3-benzyloxy-3/4-hydroxybenzaldehydes and 1, 4-benzodioxan-6-carbaldehyde, respectively.

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Chemical and rheological properties of a highly viscous and acidic polysaccharide produced by a soil bacterium idendfied as Bacillus circulans are described. The molecular weight of the native polysaccharide was about 116×10⁴ by gel filtration with HPLC. The molar ratio of D-galactose, D-mannose, and L-rhamnose contained in the polysaccharide as neutral sugar components was 1.3 : 1 : 2. The viscosity of 1% solution of the polysaccharide was about 5000cp, which was higher than that of guar gum, a seed polysaccharide, used as a reference standard. The viscosity was affected by the pH of the polysaccharide solution, a maximum viscosity being observed at pH 5.5 for native and decationized polysaccharide solution. The viscosity of 0.3% solution was decreased to about 70% by heating at 100°C and 22% at 120°C for 30 min as compared to the viscosity before heating. The addition of sugars, especially sucrose and glucose, at 10-30% concentration brought about a 130-180% increase of the viscosity. The addition of CaCl₂ at a low concentration markedly increased the viscosity, and a maximum viscosity was attained at 2% concentration of the salt.

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When Escherichia coli cells were stressed by NaCl, cellular levels of glutathione (GSH) increased, and potassium ions (K⁺) accumulated. To identify the role of the stress-induced synthesis of GSH in the accumulation of K⁺ a GSH-deficient mutant of E.coli was used. The mutant grew even in a medium with a low concentration of K⁺ after a lag of about 32 hr. In such cells grown under osmotic stress in this medium, K⁺ did not accumulate in the lag phase. In the logarithmic phase, however, intracellular levels of K⁺ increased in spite of the lack of intracellular GSH. In a medium with the usual K⁺ concentration, K⁺ accumulated in a pattern similar to that of the parent strain in both phases. These results showed that GSH participates in the accumulation of K⁺ in cells under osmotic stress only in the lag phase in medium with little K⁺.

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The metabolic fate of α- and β-L-aspartyl-[U-¹⁴C] glycine was investigated in normal young rats in vivo and in vitro. The radioactive dipeptides were synthesized from L-aspartic acid and [U-¹⁴C] glycine in our laboratory. When labeled β-aspartylglycine was given intraperitoneally, about 66% of the dose was excreted in the urine and 8% was recovered in the expired carbon dioxide over a 24-hr period. More than 90% of the urinary radioactivity was present in the β-aspartylglycine fraction of the urine. When the labeled α-aspartylglycine was given, 3% and 22% of the dose were recovered, respectively, in the urine and expired carbon dioxide. In slice experiments with kidney, liver and small intestine from normal rats, α-aspartylglycine was rapidly and almost completely hydrolyzed, and a large amount of free ¹⁴C-glycine was released. In contrast, β-aspartylglycine was hardly hydrolyzed to the corresponding amino acids in liver and small intestine, and only slightly in kidney. These results suggest that in normal young rats most β-aspartylglycine, which may originate from endogenous tissue proteins, is hardly hydrolyzed and rapidly excreted into the urine.

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Lactobacillus acidophilus TK8912 produces an antibacterial substance, designated acidocin 8912, which is active against strains of Lactobacillus and Lactococcus. Of all conditions tested, the production of acidocin 8912 was maximum at 30°C in MRS broth. Acidocin 8912 was stable to heat treatment (120°C for 20 min), but completely inactivated by protease treatment. Curing a plasmid pLA103 resulted in the loss of both acidocin 8912 production (Acd⁺) and host immunity (Acd^r). A plasmid-cured stain, TK1-4 (Acd^- Acd^s), was transformed to Acd⁺ Acd^r with the pLA103 plasmid. These results provided the first direct evidence in lactobacilli for involvement of this plasmid in bacteriocin production and immunity.

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Cellobiose phosphorylase was purified from Cellvibrio gilvus cells by the method reported previously with some modifications, and its kinetic properties were studied in detail. The initial velocity of the synthetic reaction was 1.4 times as fast as that of the phosphorolytic one. The equilibrium constant of the phosphorolysis was 0.32 at 37°C and pH 7.0. No D-[U-¹⁴C] glucose exchange reaction was observed in the absence of Pi. Kinetic studies on the phosphorolytic reaction showed that the reaction follows an ordered bi bi mechanism. These results make a sharp contrast to those of sucrose phosphorylase, which catalyzes fructose exchange reaction and follows a ping pong bi bi mechanism. Kinetic parameters were calculated as K_mA=2.6mM, K_mB=0.61 mM, and K_iA=6.8 mM (A, D-cellobiose ; B, Pi).

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The effects on liver and serum lipids of rats of excess cystine added to their casein or soy protein isolate diets were studied. When excess cystine was added to the casein diet, liver lipids were not changed. Using soy protein isolate as a protein source, both food consumption and body weight gain were depressed with increasing levels of dietary cystine. Liver total lipids, cholesterol, triacylglycerols, and phospholipids were increased in rats with 2.5% or 3.5% cystine diets. Serum cholesterol of rats on the 3.5% cystine diet was significantly higher than those of the other groups, which yielded similar values. Serum triacylglycerols were increased by the addition of 0.5% or 1.5'% cystine, and decreased by the addition of 2.5% or 3.5% cystine to the diet. Serum free fatty acids increased or tended to increase when liver lipid accumulation was observed. The addition of either methionine or choline to the cystine-excess diet containing soy protein isolate prevented the accumulation of hiacylglycerols and phospholipids in the liver although serum triacylglycerols did not return to the levels of rats on the diet without cystine, and serum free fatty acid levels were not changed. Thus, it is surmised that this fatty liver might be due to the choline deficiency induced by the excess cystine.

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An extracellular lipase was isolated from a culture filtrate of Fusarium oxysporum f.sp. lini SUF 402 by hydrophobic chromatography. Purity of the preparation was 38-fold and recovery yield was 32%. The molecular mass of the isolated enzyme was 30 kDa. The lipase had a sugar chain, and the N-terminal amino acid was modified. The optimun pH at 37°C was 7.0. The enzyme had higher activity toward p-nitrophenyl esters with long and middle chain fatty acids (C₈-C₁₈) than with a short chain fatty acid (C₄). The enzyme was not inhibited by phenylmethylsulfonyl fluoride or 2-mercaptoethanol. In the case of the lipase, both the hydrolysis rate of tristearin and final concentration of fatty acid content were higher than those of triolein. The lipase was examined with respect to its ability of concentrating the n-3 polyunsaturated fatty acid (n-3 PUFA) content of partially hydrolyzed glycerides (TG and DG) obtained from two kinds of fish oil (cod liver oil and refined sardine oil). The lipase gave increases in n-3 PUFA contents as the hydrolysis progressed. The lipase concentrated only docosahexaenoic acid with little increase in icosapentaenoic acid. Maximal total n-3 PUFA contents were about 30% in both fish oils.

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We investigated whether the fate of tryptophan (Trp) to nicotinamide (Nam) is changeable according to nitrogen sources or not. Male rats were fed with a nicotinic icid-free diet (Trp content was arranged at about 234mg/100 g of diet) containing casein, casein hydrolysates, or mixtures of amino acids (simulating the amino acid pattern of casein) for 12 days. From the urinary excretion of Nam and its metabolites, the conversion ratio was calculated. The conversion ratio was lower in the groups fed with the casein hydrolysate diets than in the groups fed with the casein and amino acid diets. In order to find the reason, the urinary excretion of kynurenic acid, xanthurenic acid, anthranilic acid, and 3-hydroxyanthranilic acid from the upper part of the Trp-Nam conversion pathway, the contents of free Trp in whole blood, and eight kinds of enzyme activities involved in the Trp-Nam pathway were measured. From these results, it is considered that the reason why the conversion ratio was lower in the casein hydrolysate group is due to the lower concentration of free Trp in whole blood and the higher activity of aminocarboxymuconatesemialdehyde decarboxylase. As is known that this enzyme activity is induced by adrenal cortical hormone, it was suggested that the release of this hormone is stimulated by feeding the casein hydrolysate diet.

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To analyze the genetic system of paralytic shellfish poisoning (PSP) toxin production in the dinoflagellate Alexandrium catenella, we examined toxin compositions and mating type of F1 progenies from crosses between algal stains having different toxin compositions. In all strains used, the mole percentage of their toxin composition did not significantly change in any growth phase, although total toxin levels increased rapidly in the early to middle exponential growth phase and then decreased by 95% in the stationary phase. One parental strain produced gonyautoxin (GTX) 4, and C4, while the other produced neosaxitoxin (neoSTX) and saxitoxin (STX) during all growth phases. F1 progenies showed one parental toxin composition and segregated independently with the mating type. These data suggest that A. catenella is a toxin producer and that Mendelian inheritance of toxin profiles occurs in the heterothallic dinoflagellate A. catenella.

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