Acaricidal properties of spinosad against Tetranychus urticae and Panonychus ulmi (Acari: Tetranychidae)

Laboratory bioassays were conducted to characterize the activity of the insecticide spinosad against the twospotted spider mite, Tetranychus urticae Koch, and European red mite, Panonychus ulmi (Koch) (Acari: Tetranychidae). T. urticae females and larvae were individually placed on bean, Phaseolus vulgaris L. (Fabaceae), leaf disks treated with four rates of spinosad (25, 55, 121, and 266 ppm) and a water control. Significantly fewer T. urticae completed development on any spinosad rates (<15%) compared with the control (>85%), whereas spinosad exhibited no significant effects on P. ulmi development; 72.5 and 83.1% of P. ulmi completed development on apple (Malus pumila P. Mill, Rosaceae) leaf disks treated with 75 ppm spinosad and the control, respectively. T. urticae adult females placed on spinosad-treated disks had significantly higher mortality and lower oviposition rates compared with the water control; no significant mortality effects were observed until 3 d after placing adults on leaf disks. In choice tests where half of a bean leaf was treated with 55 ppm spinosad transversally or longitudinally, T. urticae females were repelled by spinosad and largely oviposited and fed on nonspinosad treated areas. Spinosad did not affect the behavior of P. ulmi females. When T. urticae females were released on potted bean plants (two-leaf stage) in which leaves received spinosad sprays on the adaxial or abaxial leaf surfaces, or complete spinosad coverage on one or two of the leaves, mite population increase lagged significantly behind those released on control plants. These results indicate that spinosad has significant acaricidal effects against T. urticae but not P. ulmi.     

Enhanced phosphorylation and enzymatic activity of phosphoglucomutase by the Btk29A tyrosine kinase in Drosophila

The Drosophila Btk29A tyrosine kinase is suggested to be involved in diverse processes, although its target proteins are unknown. In the present study, we investigated substrates of Btk29A tyrosine kinase by expressing a catalytically activated form of Btk29A-P1 (Btk-EG) in Drosophila compound eyes. Expression in eye disks led to the development of the rough-eye phenotype and increased tyrosine phosphorylation of a 65-kDa protein. Partial amino acid sequence analysis of this protein showed that it was phosphoglucomutase. Phosphoglucomutase activity in heads from Btk-EG-expressing flies was higher than that in controls, suggesting that the levels of tyrosine phosphorylation and activity of the enzyme are associated with Btk29A tyrosine kinase activity.     

Metabolic flux analysis of the phenylpropanoid pathway in wound-healing potato tuber tissue using stable isotope-labeled tracer and LC-MS spectroscopy

The metabolic flux of two phenylpropanoid metabolites, N-p-coumaroyloctopamine (p-CO) and chlorogenic acid (CGA), in the wound-healing potato tuber tissue was quantitatively analyzed by a newly developed method based upon the tracer experiment using stable isotope-labeled compounds and LC-MS. Tuber disks were treated with aqueous solution of L-phenyl-d(5)-alanine, and the change in the ratio of stable isotope-labeled compound to non-labeled (isotope abundance) was monitored for p-CO and CGA in the tissue extract by LC-MS. The time-dependent change in the isotope abundance of each metabolite was fitted to an equation that was derived from the formation and conversion kinetics of each compound. Good correlations were obtained between the observed and calculated isotope abundances for both p-CO and CGA. The rates of p-CO formation and conversion (i.e. fluxes) were 1.15 and 0.96 nmol (g FW)(-1) h(-1), respectively, and for CGA, the rates 4.63 and 0.42 nmol (g FW)(-1) h(-1), respectively. This analysis enabled a direct comparison of the biosynthetic activity between these two compounds.     

Range of potential functions of the Drosophila melanogaster hdc gene

The Drosophila melanogaster hdc gene controls trachea branching, which starts during embryo development. Expression in imaginal disks and reproductive organs suggests additional functions for the hdc gene. The gene was demonstrated to have a maternal effect, which was denied previously. Analysis of cell proliferation in imaginal disks with hdc mutations showed that the gene does not possess tumor suppressor properties at the levels of mosaic cuticle clones of adults and transplanted imaginal disks. Transplanted imaginal disks homozygous, but not heterozygous, for an hdc mutation were found to affect oogenesis in the recipient females, implicating the hdc activity in exchanging signals between different organs. Amino acid sequence analysis of the Hdc protein revealed a region homologous to the human HRS proteins, which directly interact with the NF2 tumor suppressor on experimental evidence.     

Effects of ecdysone analogues on development and metabolic activity of wing disks of the fleshfly, Sarcophaga peregrina,in vitro

Effects of ecdysone analogues on development and metabolic activities of Sarcophaga wing disks were studied in cultures. Development of disks was induced by ecdysterone, ponasterone A, and cyasterone in vitro, whereas rubrosterone was quite inactive in inducing development.As well as morphogenetic effects, a proper concentration (3 × 10^?5 M to 3 × 10^?7 M) was required to induce the incorporation of tritiated uridine, thymidine, and leucine into RNA, DNA, and protein, respectively. Higher concentration of the hormone was more favourable to development of disks and enhancement of RNA synthesis. However, the hormone at concentration higher than 2 × 10^?9 M seemed to be rather toxic to both development and metabolic activity.     

Range of potential functions of the Drosophila melanogaster hdc gene

The Drosophila melanogaster hdc gene controls trachea branching, which starts during embryo development. Expression in imaginal disks and reproductive organs suggests additional functions for the hdc gene. The gene was demonstrated to have a maternal effect, which was denied previously. Analysis of cell proliferation in imaginal disks with hdc mutations showed that the gene does not possess tumor suppressor properties at the levels of mosaic cuticle clones of adults and transplanted imaginal disks. Transplanted imaginal disks homozygous, but not heterozygous, for an hdc mutation were found to affect oogenesis in the recipient females, implicating the hdc activity in exchanging signals between different organs. Amino acid sequence analysis of the HDC protein revealed a region homologous to the human HRS proteins, which directly interact with the NF2 tumor suppressor on experimental evidence.     

Formation of amylase in disks of bean hypocotyl

Amylase activity has been studied in disks of hypocotyl of Phaseolus vulgaris L. Although some activity is present in untreated hypocotyls, it is greatly increased when disks are incubated 4 days in water containing kinetin (5 mg/liter). Gibberellic acid and 2,4-dichlorophenoxyacetic acid do not increase this activity appreciably. The increased activity can be shown by subsequently incubating the disks on starch agar plates and testing these with iodine-potassium iodide solution, or by using the original incubation medium and extracts of the disks in reactions with soluble starch. A variety of tests favor the assumption that the enzyme is α-amylase and that it is synthesized during the incubation period.     

Inhibition of ethylene evolution in papaya pulp tissue by benzyl isothiocyanate

Papaya (Carica papaya L.) pulp tissue disks in an incubation medium composed of 0.4 m sucrose evolve ethylene at an optimum pH of 5.25 at 30 C. Disks of young preclimacteric fruit evolve the gas linearly with fruit age until fruit age reaches 4 months. Disks from 5-month-old postclimacteric fruit produce approximately 5-fold more ethylene than disks from 4-month-old fruit. Ethylene evolution by unaged papaya disks is inhibited potently by benzyl isothiocyanate. The compound inhibits production of ethylene by approximately 60% at a concentration of 0.046 mm. However, in aged papaya disks benzyl isothiocyanate causes no inhibition of ethylene production indicating that the compound inhibits the induction of the ethylene-producing system rather than the evolution of the gas per se. Even at a 2-fold higher concentration benzyl isothiocyanate has no effect on respiration of unaged papaya disks. It is proposed that benzyl isothiocyanate may act as an endogenous regulator of ethylene evolution in papaya fruit.     

Changes in ribosomal activity during incubation of Daucus carota root disks

Cytoplasmic monoribosomes from freshly cut and ‘aged’ carrot root disks were characterized relative to the Mg²⁺ optima for poly U (polyuridylic acid)-directed phenylalanine incorporation, the ease of dissociation by KCl in the presence of Mg²⁺, the ability to bind ³H-poly U, and acrylamide gel fractionation of the ribosomal proteins. The differences in in vitro amino acid incorporation by ribosomes and supernatant from fresh and ‘aged’ disks were confined to the ribosome fraction. The Mg²⁺ optima for poly U-directed ¹⁴C-phenylalanine incorporation was 16 mM for ribosomes from ‘aged’ disks compared to 20 mM for ribosomes from fresh disks. Monoribosomes from the fresh disks were easily dissociated into subunits (0·2 M KCl in 5 mM Mg²⁺) while the ribosomes from ‘aged’ disks were not completely dissociated even in 0·5 M KCl. Ribosomes from ‘aged’ disks were more effective in binding ³H-poly U than ribosomes from fresh disks. When the disks were subjected to an anaerobic environment prior to ribosome extraction (to strip monoribosomes of peptidyl-t RNA) the above effects of ‘aging’ were reversed. These results suggest that increased monoribosome activity associated with ‘aging’ may be related in part to an increase in the level of peptidyl-tRNA associated with the ribosomes. Acrylamide gel electrophoresis profiles of ribosomal proteins extracted from ribosomes of fresh and ‘aged’ tissue suggest that a change in the protein complement may also be important to the observed changes in ribosomal activity. The ribosomes from ‘aged’ disks contained at least two components not associated with ribosomes from fresh disks.     

Inhibition of the Development of a Cation Accuniulatory System and of Tris-induced Uptake in Storage Tissues by N6-benzyladenine and Kinetin

The effect of N⁶-benzyladenine (BA) and kinetin on cation uptake in disks of beetroot (Beta vulgaris L.) and swede (Brassica napobrassica Mill.) tissue was measured in aerated solutions, containing 1 mM NaCl or 1 mM KCl, and in the presence or absence of trishydroxymethyl ammo methane (tris) buffer at pH 8. This investigation followed a suggestion that the immediate activation of cation uptake in freshly sliced beetroot disks by tris (the tris-effect) may depend on stimulation of hexokinase activity. BA, a competitive inhibitor of hexokinase, caused a complete inhibition of tris stimulated cation uptake in beetroot disks, but the effect was delayed and preceded by a 3–4 hour period of promoted cation uptake. Generally, the effects of BA and kinetin were identical. Use of ¹⁴C-kinetin indicated a rapid incorporation of kinetin into the tissue. BA and kinetin also prevented the development of a cation uptake capability during the aging of the beetroot disks. Swede disks which do not show a tris-effcct and no lag phase in development of cation uptake capability were not affected by BA and kinetin treatments. Mechanisms of tris induced cation uptake are discussed with emphasis on its role as a proton acceptor.     

Inhibitory effect of mispyric acid on mammalian DNA polymerases

Mispyric acid is a novel natural triterpene dicarboxylic acid which has inhibitory activity against DNA polymerase beta (pol beta) isolated from the plant, Mischocarpus pyriformis. In this report, we examine the selectivity of the inhibitory activity against mammalian pols and the mode of inhibition in vitro. Natural mispyric acid (compound 1) inhibited the activities of all the mammalian pols tested (pol alpha, beta, gamma, delta and epsilon) with an IC50 value in the range of 3.6-44.5 microM. The inhibition was strongest for pol gamma among these five pols. The enantiomer of mispyric acid (compound 2, ent-mispyric acid) had similar effects to those of the natural compound. However, derivatives of compounds 1 and 2 with hydroxyl groups instead of carboxyl groups (i.e., compounds 3 and 4, respectively) exhibited no inhibitory effect on mammalian pols. The moiety of two carboxylic acids in mispyric acid was important for the inhibition of pols, and the stereoisomers of mispyric acid had no inhibitory effect.     

Discovery of nosokophic acid,a predicted intermediate of moenomycins,from nosokomycin-producing Streptomyces sp. K04-0144

A new actinomycete metabolite designated nosokophic acid was isolated from the culture broth of nosokomycin-producing Streptomyces sp. K04-0144, and the structure was elucidated by various NMR experiments. Nosokophic acid was found to be 3-phosphoglycosyl-2-sesquiterpenyl dihydroxypropionic acid, a predicted biosynthetic intermediate of nosokomycin-related moenomycins. The compound showed no activity against MRSA, but potentiated imipenem activity against MRSA by 512-fold.     

Induction of Phenylalanine Ammonia-lyase in Strawberry Leaf Disks: Action Spectra and Effects of Wounding, Sucrose, and Light

The increase in phenylalanine ammonia-lyase (PAL) activity in strawberry (Fragaria vesca var. WSU-1232) leaf disks required wounding, sucrose, and light and was cycloheximide-sensitive. In injured leaves and in leaf disks, the highest PAL activity was detected nearest the wounded tissues. Without wounding, no increase in activity was observed when leaves were cultured in sucrose and light.     

Comparison of bioassay techniques for determining baseline susceptibilities to imidacloprid for green apple aphid (Homoptera: Aphididae)

The susceptibility of a clone of green apple aphid, Aphis pomi (De Geer), to the neonicotinyl insecticide imidacloprid was determined by direct and indirect bioassay techniques. Aphid numbers were assessed on potted apple seedlings treated with various concentrations of imidacloprid, adults were dipped in test solutions as per the Food and Agriculture Organization protocol, or nymphs and adults were reared on treated apple leaf disks. Effective concentrations required to kill half of the test population (EC50) varied depending on the bioassay technique, ranging from as low as 0.064 ppm for first instars reared for 3 d on treated leaf disks to 1.79 ppm for adult apterae dipped in solutions of imidacloprid and held for 24 h on clean leaf disks. When imidacloprid was directly applied to aphids, mortality continued to increase over 3 d, but the difference was not statistically significant between day 2 (1.36 ppm) and day 3 (1.19 ppm). Toxicity of neonicotinyls to aphids is expressed rather slowly and primarily after oral ingestion. The effect of imidacloprid on reproduction of green apple aphid was also assessed for adult apterae reared on treated leaf disks. Contrary to previous reports, our results demonstrated that imidacloprid does not have a direct negative effect on the reproductive physiology of this species. Negative effects can mostly be attributed to the antifeedant activity of this compound and the protracted time to death. The results of this study contribute to a better understanding of the most suitable techniques for assessing aphid mortality after exposure to these new insecticides and provides a baseline susceptibility to imidacloprid for green apple aphid.     

L-Phenylalanine ammonia-lyase activity in asparagus spears: Effects of excision and incubation

Incubation of disks sectioned from the basal portion of asparagusspears resulted in a 5-fold increase in L-phenylalanine ammonia-lyase(PAL) (E.C. 4.3.1.5 [EC] ) activity over the level initially presentin the intact tissue. The enhanced activity developed rapidly,with only a slight lag, increasing to a maximum level at 30hr. Thereafter, the level of activity decreased to 50% of maximumactivity and appeared to have attained a new, higher steady-statelevel after 72 hr of incubation. Similar levels of activitydeveloped in basal disks incubated either in buffer solutionor in air, and light had no effect on enzyme activity. The excision-promoted increase in enzyme activity was preventedby cycloheximide (20 ppm) but, unlike some other tissues, delayedaddition of the antibiotic to incubating disks promoted lossof the lyase activity. The phenylpropanoid end products, transcinnamate,p-coumarate and ferulate (at 10^–3 M each), in decreasingorder of effectiveness, also inhibited the excision-promotedincrease in enzyme activity and caused a loss of the enhancedactivity in the incubated disks. The possibility is discussed that the activity initially presentin the spears is under separate control from that activity inducedby excision. (Received April 22, 1972; )     

Induced Resistance Against Plasmopara helianthi in Sunflower Plants by DL-β-Amino-n-butyric acid

Sunflower plants treated with the nonprotein amitio add, DL-β-attiino- n -butyric acid (BABA) were protected against infection with Plasmopara helianthi. Soil drenches at the highest rates (150-250 mg/kg soil), applied one day before the inoculation induced high levels of protection (80-83%) against the disease and more than 90% control was observed when BABA was applied at 300 mg/kg soil. However, at this concentration phytotoxic symptoms were observed. This compound also provided a curative activity when applied one day post-inoculation. BABA had no antifungal activity in vitro against P. helianthi. The effect of BABA on zoosporangia germination was evaluated by treating pre-germinated seeds with the compound solution and the zoosporangia suspension for 3 h. Then, seeds were sown and the percentages of infected plants were determined. The other two aminobutyric acid isomers (a and g) were ineffective against downy mildew. The mechanisms by which DL-β-amino- n -butyric acid protect sunflowers against downy mildew awaits more detailed elucidation.     

Novel 1,2,4-triazole and imidazole derivatives of L-ascorbic and imino-ascorbic acid: synthesis, anti-HCV and antitumor activity evaluations

Several novel 1,2,4-triazole and imidazole L-ascorbic acid (1, 2, 3, 5, 6 and 9) and imino-ascorbic acid (4, 7 and 8) derivatives were prepared and evaluated for their inhibitory activity against hepatitis C virus (HCV) replication and human tumour cell proliferation. Compounds 6 and 9 exerted the most pronounced cytostatic effects in all tumour cell lines tested, and were highly selective for human T-cell acute lymphoblastic leukaemia cells (CEM/0) with IC(50)s of 10 ± 4 and 7.3 ± 0.1 μM, respectively. Unlike compound 9, compound 6 showed no toxicity in human diploid fibroblasts. One of the possible mechanisms of action of compound 6 accounting for observed cytostatic activity towards haematological malignancies might be inhibition of inosine monophosphate dehydrogenase (IMPDH) activity, a key enzyme of de novo purine nucleotide biosynthesis providing the cells with precursors for DNA and RNA synthesis indispensable for cell growth and division, which has emerged as an important target for antileukemic therapy. In addition, this compound proved to be the most potent inhibitor of the hepatitis C virus replication as well. However, observed antiviral effect was most likely associated with the effect that the compound exerted on the host cell rather than with selective effect on the replication of the virus itself. In conclusion, results of this study put forward compound 6 as a potential novel antitumor agent (IMPDH inhibitor) for treating leukaemia. Its significant biological activity and low toxicity in human diploid fibroblasts encourage further development of this compound as a lead.     

The development of invertase activity in slices of the root of Beta vulgaris L. washed under aseptic conditions

1. When disks of root tissue from sugar or red beet (Beta vulgaris L.) are washed in running aerated tap water the sucrose contained in them disappears and glucose and fructose are formed. 2. Invertase activity in the disks has been measured by a polarimetric method. Freshly cut tissue has a very low activity, but a considerable increase occurs during the first 3–4 days of washing, the final activity being sufficient to hydrolyse the sucrose contained in the disk within a few hours. 3. Disks of red beet have been cut and shaken in water under aseptic conditions. Sucrose breakdown and invertase development still took place. Microbial contamination is therefore not responsible. 4. Trisaccharides that appear in sugar-beet disks during the washing process have been isolated and identified; their formation also suggests that a higher-plant invertase is acting. 5. The significance of these results is discussed in relation to protein synthesis in washed storage-tissue slices, and the occurrence of high invertase activity in growing plant cells.     

Hymenoic acid, a novel specific inhibitor of human DNA polymerase lambda from a fungus of Hymenochaetaceae sp

Hymenoic acid (1) is a natural compound isolated from cultures of a fungus, Hymenochaetaceae sp., and this structure was determined by spectroscopic analyses. Compound 1 is a novel sesquiterpene, trans-4-[(1′E,5′S)-5′-carboxy-1′-methyl-1′-hexenyl]cyclohexanecarboxylic acid. This compound selectively inhibited the activity of human DNA polymerase λ (pol λ) in vitro, and 50% inhibition was observed at a concentration of 91.7 μM. Compound 1 did not influence the activities of the other seven mammalian pols (i.e., pols , γ, δ, ε, η, ι, and κ), but also showed no effect even on the activity of pol β, which is thought to have a very similar three-dimensional structure to the pol β-like region of pol λ. This compound also did not inhibit the activities of prokaryotic pols and other DNA metabolic enzymes tested. These results suggested that compound 1 could be a selective inhibitor of eukaryotic pol λ. This compound had no inhibitory activities against two N-terminal truncated pol λ, del-1 pol λ (lacking nuclear localization signal (NLS), BRCA1 C-terminus (BRCT) domain [residues 133–575]), and del-2 pol λ (lacking NLS, BRCT, domain and proline-rich region [residues 245–575]). The compound 1-induced inhibition of intact pol λ activity was non-competitive with respect to both the DNA template-primer and the dNTP substrate. On the basis of these results, the pol λ inhibitory mechanism of compound 1 is discussed.