Reproduction site preference and performance by sexuparae,and mating behavior of their sexual generation on the primary host plant in a heteroecious aphid,Neothoracaphis yanonis (Homoptera: Aphididae)

We studied reproduction site preference and performance by the sexuparae (autumnal migrants) of Neothoracaphis yanonis and mating behavior of their sexual generation on its primary host plant, Distylium racemosum. The sexuparae preferred younger leaves of D. racemosum for settlement and imbibing leaf sap, and they produced more offspring there than on older leaves. Thus, it is suggested that the sexuparae selected more nutritious younger leaves to increase their own fecundity. The offspring consist of yellowish dark-grey and creamy yellow type nymphs, which develop into small males and large oviparae, respectively. Yellowish dark-grey nymphs were deposited gregariously on the basal part of the abaxial surface of a leaf blade, while creamy yellow nymphs were deposited evenly over the abaxial surface. Such a localized distribution pattern probably resulted from sexuparae's strategy to enhance a possibility for their male offspring to mate with oviparae at the base of leaf blade, over which oviparae crawl to twigs. During mating and sometimes afterwards, the ovipara incidentally carried a male on her back and crawled downward on twig to find an axillary bud for oviposition. This behavior may be advantageous to males, who can guard their mate during this period against other conspecific males. Such mate-guarding behavior seems to be related to the development of dual mate-seeking strategies, in which males try to copulate, first at the basal part of midrib, and second on the axillary bud.     

DSC Measurement of Frozen Water in Liquid Foods

We investigated the state of water in frozen liquid foods, supposed to influence the performance of freeze-drying, by DSC. The amount of unfreezable water in milk was constant irrespective of the initial water content. As expected from the phase equilibrium, the water content of the non-ice part of CAS was constant among the samples with different initial water contents. The volume fraction of ice crystals, which should be known in the analysis of the freeze-drying rate, was significantly smaller at the low initial water content than the one usually estimated with the assumption that the water was all freezable.     

Adaptation to a Starch Environment and Regulation of alpha-Amylase in Drosophila

The adaptation to glucose and starch foods insix species, D. melanogaster, D.virilis, D. saltans, D. funebris,D. levanonensis and D. americana, wasstudied by measuring productivity. D.melanogaster and D. virilis adapted more to thestarch environment than to the glucose environment,while D. saltans adapted more to the glucoseenvironment than to the starch environment. D.funebris, D. levanonensis, and D. americana did not distinctlyadapt to either environment. In addition, the regulationof amylase in the six species was investigated bymeasuring the levels of amylase activity with glucoseand starch food environments. The levels of amylaseactivity in D. levanonensis and D.saltans were substantially low, indicating thatthese species cannot utilize starch as a carbon source.The starch-adapted species, D. melanogaster and D.virilis, showed higher levels of amylase activitywith the starch environment and higher inducibility.These results suggest that changing the regulation ofamylase is important for the adaptation to a starch environment inDrosophila.     

Determination of Norepinephrine in Microdialysis Samples by Microbore Column Liquid Chromatography with Fluorescence Detection Following Derivatization with Benzylamine

A microbore column liquid chromatographic method is described for the determination of norepinephrine (NE) in microdialysis samples from rat brain. The method is based on precolumn derivatization of NE with benzylamine in the presence of potassium hexacyanoferrate(III) resulting in a highly fluorescent and stable benzoxazole derivative. Typically, a 10-microl sample was mixed with 10 microl derivatization reagent containing 0.45 M Caps buffer (pH 12.0), 0.2 M benzylamine, 10 mM potassium hexacyanoferrate(III), and N,N-dimethylformamide (1:1:1:15, v/v). The derivatization was carried out at 50 degrees C for 20 min. Under these conditions only NE and epinephrine produced high fluorescence yields at excitation and emission wavelengths of 345 and 480 nm, respectively, while fluorescence of other catechols and 5-hydroxyindoles was quenched by 10-100 times. The NE derivative was separated on a reversed-phase column (100 x 1.0 mm i.d., packed with C18 silica, 5 microm) within 10 min with no late eluting peaks. The mobile phase consisted of 40 mM Britton-Robinson buffer (pH 7.5) containing 1 mM didodecyldimethylammonium bromide and acetonitrile (34%, v/v), the flow rate was 40 microl/min. The limit of detection (signal-to-noise ratio of 3) for NE was 90 amol in 10 microl sample injected. Microdialysis samples were collected in 5-min intervals from the probes implanted in the hippocampus, frontal cortex, or hypothalamus of awake rats. The basal extracellular NE levels in the respective areas were 4.7 +/- 0.9, 1.8 +/- 0.3, and 0.8 +/- 0.2 fmol/10 microl (mean +/- SE, n = 7). Perfusion with a Ringer solution containing 100 mM K+ increased hippocampal NE levels by 700%, while NE uptake inhibitors maprotiline and amitriptyline administered orally or subcutaneously increased extracellular NE in the frontal cortex by about 300%. On the other hand, reserpine (5 mg/kg) reduced cortical NE levels by 40% 3 h after the administration. This new fluorescence derivatization method provides better selectivity, sensitivity, and speed for NE determination than the electrochemical detection since no late-eluting compounds such as dopamine, serotonin, and their metabolites are detectable in the chromatograms of the microdialysis samples.     

Volatile Flavor Components of Watermelon (Citrullus vulgaris)

Volatile flavor components of watermelon fruit (Citrullus vulgaris) were obtained by distilling juice under reduced pressure and extracting the resulting distillate with Freon-11. The volatiles were separated into acidic and neutral fractions and then analyzed by gas chromatography and gas chromatography-mass spectrometry.

Fifty-two compounds were found for the first time as flavor components of watermelon. Among them, the following compounds have not been previously reported as naturally occurring flavor components: 4-oxononanal and 2-hydroxy-, 2-pentyloxy-, 2-hexyloxy-, 2-octyloxy-, 2-nonyloxy-, 2-[(Z)-3-noneyloxy]-, 2-[(Z)-2-nonenyloxy]-, 2-[(Z,Z)-3,6-nonadienyloxy]-, 2-benzyl-oxy-, and 2-phenetyloxy-5-pentyltetrahydrofurans.     

A Component of Wheat Flour Globulin Polymerized at Alkaline Sides and Depolymerized by Reduction Reversibly

1. The SDS-disc electrophoretical analysis of wheat flour globulin revealed that upon incubation with “Kansui,” a mixture of alkali carbonates, one (No. 9) of the components disappeared and another component (No. 11) increased significantly whose molecular weight was estimated to be more than one million. Incubation of flour globulin with “Kansui” resulted in decrease of SH groups of the globulin.

2. However, a mild reduction of the globulin incubated with “Kansui” caused reappearance of component No. 9 and a decrease of component No. 11.

3. Previous treatment of flour globulin with PCMB or AN completely inhibited such effects of “Kansui” as described above.

4. A fraction containing component No. 9 of flour globulin which was the main component responsible to polymerization by “Kansui,” was isolated from “Kansui”-treated globulin by chromatography on columns of Sephadex G200 and Sepharose 4B.

5. The component protein thus isolated showed a typical phenomenon of reversibility in depolymerization to the original component corresponding to No. 9 component by chemical reduction and polymerization to a component corresponding to No. 9 at alkaline sides.

     

γ-Glutamylcysteine synthetase and γ-glutamyl transferase as differential enzymatic sources of γ-glutamylpeptides in mice

Some γ-glutamylpeptides in blood plasma are putative biomarkers for pathological conditions of the liver. γ-Glutamyltransferase (GGT) and γ-glutamylcysteine synthetase (γ-GCS) are two such potential enzymes that are responsible for the production of γ-glutamylpeptides. GGT produces γ-glutamylpeptides by transferring the γ-glutamyl moiety from glutathione to an amino acid or a peptide. γ-GCS normally catalyzes the production of γ-glutamylcysteine from glutamate and cysteine in the glutathione-synthesizing reaction, but other amino acids can also serve as an acceptor of a γ-glutamyl group, thus resulting in the formation of a variety of γ-glutamylpeptides. Based on liquid chromatography–mass spectrometry analyses, we observed differences in the distribution of γ-glutamylpeptides between the liver and kidney and were able to measure the activities of γ-GCS as well as the GGT reactions by quantifying the resulting γ-glutamylpeptides. The enzymatic characterization of γ-GCS in liver homogenates indicated that several γ-glutamylpeptides including γ-glutamyltaurine are actually produced. Cys showed the lowest Km value (0.06 mM) while other amino acids had much higher Km values (ranging from 21 to 1800 mM). The moderate Km values for these amino acids suggest that they were not the preferred amino acids in this conversion but were utilized as acceptor substrates for the production of the corresponding γ-glutamylpeptides by the γ-GCS reaction under Cys-deficient conditions. Thus, the production of these γ-glutamylpeptides by γ-GCS is directly correlated with a low Cys content, suggesting that their measurement in blood plasma could be useful for predicting the presymptomatic disease state of the liver with a defect in GSH redox balance.