Growth inhibitory,insecticidal and antifeedant effects of some antileukemic and cytotoxic quassinoids on two species of agricultural pests

Summary Several quassinoids, obtained by isolation and derivatization fromSimaba multiflora andSoulamea soulameoides, were evaluated for growth inhibitory and insecticidal effects against the tobacco budworm (Heliothis virescens) and for antifeedant effects againstH. virescens and the fall armyworm (Spodoptera frugiperda). The relative activity of the quassinoids as insect growth inhibitors generally paralleled their known relative potency as antileukemic and cytotoxic agents.Phytochemical aspects of this work were supported by Contract CM-97295 with the Division of Cancer Treatment, National Cancer Institute, National Institutes of Health, Bethesda, Md. Insects were kindly supplied by the agency of the United States Department of Agriculture at Brownsville, TX.     

Beta-carotene affects gene expression in lungs of male and female <Emphasis Type="Italic">Bcmo1</Emphasis><Superscript>−/−</Superscript> mice in opposite directions

Molecular mechanisms triggered by high dietary beta-carotene (BC) intake in lung are largely unknown. We performed microarray gene expression analysis on lung tissue of BC supplemented beta-carotene 15,15′-monooxygenase 1 knockout (Bcmo1 ^−/−) mice, which are—like humans—able to accumulate BC. Our main observation was that the genes were regulated in an opposite direction in male and female Bcmo1 ^−/− mice by BC. The steroid biosynthetic pathway was overrepresented in BC-supplemented male Bcmo1 ^−/− mice. Testosterone levels were higher after BC supplementation only in Bcmo1 ^−/− mice, which had, unlike wild-type (Bcmo1 ^+/+) mice, large variations. We hypothesize that BC possibly affects hormone synthesis or metabolism. Since sex hormones influence lung cancer risk, these data might contribute to an explanation for the previously found increased lung cancer risk after BC supplementation (ATBC and CARET studies). Moreover, effects of BC may depend on the presence of frequent human BCMO1 polymorphisms, since these effects were not found in wild-type mice.     

Effect of acetylcholine on melanophores ofRana tigrina

Summary Acetylcholine produced melanin aggregation and blanching of skin colour inRana tigrina, the common Indian frog. The effects were more prolonged in frogs pretreated with an anticholinesterase agent. Acetylcholine effects were not antagonized by eitherm-cholinolytic (atropine) orn-cholinolytic (pentolinium) agents, but were markedly inhibited by procaine. The results have been discussed in the light of the well-known membrane-stabilizing effect of procaine.     

Selektive insektizide Wirkung des Butazolidins

Summary Butazolidin proved to be insecticidal by ingestion forPediculus humanus and for the first and second instars ofOrnithodorus moubata. Adult specimens ofO. moubata were not affected.     

Disk-elektrophoretische Untersuchungen der Serumproteine europäischer Eschsen der Familien Anguidae und Lacertidae

Summary Serum proteins ofLacerta v. viridis, L. v. meridionalis, L. trilineata, L. agilis, L. vivipara, L. taurica, L. muralis, Ophisaurus apodus, andAnguis fragilis were separated by polyacrylamide disc electrophoresis. Among the lacertids,Lacerta viridis andLacerta trilineata show greater similiarities.Ophisaurus apodus andAnguis fragilis are characterized by praealbumins.     

Intergeneric interactions betweenEimeria separata (Apicomplexa) andNippostrongylus brasiliensis (Nematoda) in the rat

Summary Ova production inNippostrongylus brasiliensis infected rats was significantly greater than in rats singly infected with the helminth whenEimeria separata infections were introduced 4, 6 and 11 days postinoculation withN. brasiliensis. Patent periods were unaltered during concurrent infections. These results suggest that the presence ofE. separata affects helminth fecundity but does not increaseN. brasiliensis longevity as has been shown withE. nieschulzi.This study was supported by NIH MB RS Grant RRO8012-8.     

Chemical and tactile communication between the root aphid parasitoidParalipsis enervis and trophobiotic ants: consequences for parasitoid survival

Females of the aphid parasitoidParalipsis enervis received liquid food by regurgitation (trophallaxis) from workers of the ant speciesLasius niger, but were not fed by workers ofMyrmica laevinodis andTetramorium caespitum. WhileP. enervis was not treated aggressively by workers of any of these species,Lasius flavus workers killed the parasitoid. This different ant behaviour resulted in a different parasitoid longevity. WhileP. enervis survived for only 10 min in the presence ofL. flavus (due to ant aggression) or for approximately one day in the presence ofT. caespitum andM. laevinodis (due to lack of trophallaxis), survival increased significantly to more than five days in the presence ofL. niger, which provided food regularly to the parasitoids. Our study suggests thatP. enervis mimics behavioural signals ofL. niger, as well as odor cues of its host aphidAnoecia corni, to avoid aggression byL. niger.     

Endocytosis and inositol hexakisphosphate levels inras transformants ofDictyostelium discoideum amoebae

Summary Fluid-phase pinocytosis kinetics and lysosomal enzyme secretion parameters were measured inDictyostelium discoideum amoebae constructed from strain AX3 by transformation with a multicopy plasmid carrying either a normalras gene (ras-Gly12), a mutatedras gene (ras-Thr12) or by the vector carrying the geneticin resistance gene only (pDNEO2). It was found that the pinocytosis rate and extent as well as the lysosomal enzyme secretion were slightly different in the three strains. These changes, however, were related to minor modifications of the cellular volumes. The overall concentration of inositol hexakisphosphate was similar in the three strains.This work was supported in part by a grant from the Ligue Nationale Française contre le Cancer (n^o 880258) to MS, and by a grant from the Fonds National Suisse de la Recherche Scientifique (No. 3.623-087) to CR.     

Gel-electrophoretic description of European populations ofTerellia virens (Loew) (Diptera,Tephritidae); implications for its use as an agent for the biological control ofCentaurea spp. (Asteraceae) in North America

Allozyme frequencies of 15 enzyme loci, 14 of which were polymorphic, were used to characterize sevenTerellia virens populations originating from three allopatrically distributedCentaurea species. The two populations whose origins were geographically furthest apart, from Israel (onC. iberica) and from Switzerland (onC. vallesiaca), showed relatively high values of genetic distance from the 5 populations sampled in Austria and Hungary (onC. maculosa) (Nei's D>0.07). The latter five displayed a high degree of genetic similarity. No diagnostic (fixed) allelic differences were observed between these three groups ofT. virens populations, but they could be well characterized by significant differences in allelic frequencies at 9 enzyme loci. Independently of this study, the populations from Switzerland (C. vallesiaca) and eastern Austria (C. maculosa) were selected as potential source populations for future introductions into North America for the biological control of introducedC. maculosa andC. diffusa. Based on the observed genetic differences and results from field experiments on the host specificity of these two potential source populations, it is argued that host specificity screening tests should be conducted separately for local (host plant) populations, as such populations might accept a different set of hosts. Biotype mismatch and the risk of spill-overs to native species could thus possibly be reduced.     

Occurrence and biosynthesis of 11 (R)-hydroxy-eicosatetraenoic acid (11-R-HETE) in the Caribbean soft coralPlexaurella dichotoma

Gorgonian soft corals from the Caribbean Sea are known to contain prostaglandin-like compounds as well as other products of arachidonic acid lipoxygenation, and the formation of the latter has been suggested to represent the first step in prostaglandin biosynthesis. Here we present evidence for the presence of 11-R-hydroxy-5Z, 8Z, 12E, 14Z-eicosatetraenoic acid (11-R-HETE), as well as of the enzyme responsible for its biosynthesis, in the Caribbean gorgonianPlexaurella dichotoma. Lipid extracts fromP. dichotoma were purified by conventional SiO₂ column chromatography followed by reversed phase high-pressure liquid chromatography (HPLC). These yielded a component having chromatographic and spectroscopic properties identical to synthetic 11-HETE. Electron impact mass spectrometric analysis of the acetoxy-, methyl ester derivative of the compound confirmed its identity as 11-HETE, while chiral phase HPLC of the methyl ester derivative showed that the stereochemistry of the alcoholic carbon atom wasR. Enzymatically active homogenates fromP. dichotoma were able to convert both unlabelled and [³H] arachidonic acid into 11-HETE. In vitro biosynthesis of the latter metabolite was also observed with homogenates of the Mediterranean gorgonianParamuricea clavata, another non-prostaglandin-containing soft coral, thus suggesting that 11-R-HETE production is not necessarily accompanied by prostaglandin formation in gorgonian corals.     

Overexpression of SUMO perturbs the growth and development of <Emphasis Type="Italic">Caenorhabditis elegans</Emphasis>

Small ubiquitin-related modifiers (SUMOs) are important regulator proteins. Caenorhabditis elegans contains a single SUMO ortholog, SMO-1, necessary for the reproduction of C. elegans. In this study, we constructed transgenic C. elegans strains expressing human SUMO-1 under the control of pan-neuronal (aex-3) or pan-muscular (myo-4) promoter and SUMO-2 under the control of myo-4 promoter. Interestingly, muscular overexpression of SUMO-1 or -2 resulted in morphological changes of the posterior part of the nematode. Movement, reproduction and aging of C. elegans were perturbed by the overexpression of SUMO-1 or -2. Genome-wide expression analyses revealed that several genes encoding components of SUMOylation pathway and ubiquitin-proteasome system were upregulated in SUMO-overexpressing nematodes. Since muscular overexpression of SMO-1 also brought up reproductive and mobility perturbations, our results imply that the phenotypes were largely due to an excess of SUMO, suggesting that a tight control of SUMO levels is important for the normal development of multicellular organisms.     

DNG1, a Dictyostelium homologue of tumor suppressor ING1 regulates differentiation of Dictyostelium cells

dng1 is a Dictyostelium homologue of the mammalian tumor suppressor ING gene. DNG1 protein localizes in the nucleus, and has a highly conserved PHD finger domain found in chromatin-remodeling proteins. Both dng1 disruption and overexpression impaired cell proliferation. In dng1-null cells, the progression of differentiation was delayed in a cell-density-dependent manner, and many tiny aggregates were formed. Exogenously applied cAMP pulses reversed the inhibitory effect caused by dng1 disruption on the aggregation during early development, but formation of tiny aggregates was not restored. dng1-overexpressing cells acquired the ability to undergo chemotaxis to cAMP earlier and exhibited enhanced differentiation. These phenotypes were found to be coupled with altered expressions of early genes such as cAMP receptor 1 (car1) and contact site A (csA). Furthermore, disordered histone modifications were demonstrated in dng1-null cells. These results suggest a regulatory role of dng1 in the transition of cells from growth to differentiation.Received 29 December 2004; received after revision 24 May 2005; accepted 26 May 2005     

Fluorescence replication banding of frog chromosomes

To identify individual chromosomes of a frog karyotype by their fluorescence banding patterns, chromosomes were stained with actinomycin D and 4,6-diamidino-2-phenylindole (DAPI) after incorporation of BrdU during the late S-phase. The chromosomes of three Rana species which were selected for this study (R. ridibunda, R. lessonae and R. japonica) showed well-defined late replication bands. The fluorescence patterns obtained were the reverse of those produced by a 4Na-EDTA Giemsa-staining technique. Fluorescence patterns of the two water frog species (R. ridibunda and R. lessonae) were similar to each other, except for the different fluorescence of the centromeric heterochromatin, which gave extremely bright signals in R. ridibunda but no signal in R. lessonae. Experiments also showed differences between the fluorescence patterns of R. lessonae chromosome 13 in the Italian and Luxembourgian populations. These results sho w that the fluorescence replication banding using actinomycin D and DAPI is very effective in identifying individual frog chromosomes and detecting their structural changes. Received 7 June 1996; received after revision 23 July 1996; accepted 21 August 1996     

Die α-Glycerophosphat-Oxydation des Heuschreckenbrustmuskels (Locusta migratoria)

Summary The conditions were studied for the glycerophosphate oxidation by homogenate from locust flight muscle, and the O₂-consumption was measured. Maximal oxidation rates were found with 0.087m glycerophosphate, 8 × 10^–6 m cytochromec, 7 × 10^–6 m DPN and pH 7.5. The production of dihydroxyacetone phosphate is followed by further oxidation steps, as could be shown by estimation of the different fractions of acid-soluble phosphate. Comparative studies were made on different insects and vertebrates. The rate of succinate oxidation by insect muscle was found to be ten times higher than that of vertebrate muscle. The relation of glycerophosphate oxidation to succinate oxidation is quite different in insects and vertebrates, but no difference could be detected between the two types of muscle metabolism of insects: the carbohydrate and the fatutilizing type.

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Mit Unterstützung der Deutschen Forschungsgemeinschaft.     

Incorporation of photolabile azido fatty acid probes inNeurospora crassa andSaccharomyces cerevisiae

Summary Photolabile azido fatty acids supported the growth of fatty acid auxotrophs, thecel mutant ofNeurospora crassa and the KD115 mutant ofSaccharomyces cerevisiae. These fatty acids were incorporated into the membrane lipids of thecel and KD115 mutants, but were not used by the wild type strain.This work was presented at the 26th International Congress of Pure and Applied Chemistry, Session 1, Tokyo, September 4–10, 1977, abstracts p. 241.Support was provided by the Govt. of India to B.C. and by NCERT, New Delhi India to D.N.C.To whom requests for reprints should be sent.     

Possible involvement of the phloem sealing system in the acceptance of a plant as host by an aphid

Possible reasons for the rejection of some lines ofTriticum monococcum (Tm44 and Tm46) by the aphidSitobion avenae were explored. In allT. monococcum lines studied, whether unfavourable (non-host/resistant plant) or favourable (host/susceptible plant), the concentrations of hydroxamic acids, a family of aphid-resistance factors in cereals, were significantly lower than the levels in the favourable host-plantTriticum aestivum cv. Therefore, hydroxamic acids did not account for the host/non-host patterns observed. Phloem sap was collected by stylectomy from young seedlings of favourable and unfavourable plants. In non-aphid-resistant genotypes, the success in stylectomy, the proportion of amputated stylets resulting in long (>1 min) exudations, the average duration of exudation, and the final volume of sap exuded, were higher than in the aphid-resistant genotypes. It is concluded that aphid interference with the phloem sealing system of the plant is a likely mechanism of rejection ofT. monococcum lines Tm44 and Tm46 as hosts byS. avenae.     

Increased susceptibility ofTrypanosoma lewisi infected,or decomplemented rats toSalmonella typhimurium

Summary Rats infected withTrypanosoma lewisi or decomplemented by injection of cobra venom factor or complement activating factor of trypanosomes were found to be more susceptible to infection withSalmonella typhimurium. Decomplemented rats subsequently infected withT. lewisi developed higher blood parasitemia than did normalT. lewisi infected rats.This project is supported by the National Research Council of Canada grant A 0068 and a grant from the International Development Research Center.     

Cytogenetic studies of Hynobiidae (Urodela). IX. Karyological characteristics ofHynobius abei Sato by means of R-and C-banding

Summary Chromosomal characteristics of the salamander speciesHynobius abei, from Ohimya (Kyoto) were revealed by the techniques of R-and C-banding. The karyotype ofH. abei was characterized by the shortness of an R-negative (C-positive) band in the terminal region of the long arm of chromosome 2 and a band encompassing the whole short arm of chromosome 10. These two bands inH. abei were the shortest among those of the variousHynobius species that have been examined. Otherwise no differences could be detected betweenH. abei and seven other pond-type species ofHynobius (2n=56) in terms of the banding patterns of 18 specifically identifiable pairs of their chromosomes.     

On the mechanism of inhibition of p27 degradation by 15-deoxy-Δ12,14-prostaglandin J 2 in lymphoblasts of Alzheimer’s disease patients

It has been proposed that neuroinflammation, among other factors, may trigger an aberrant neuronal cell cycle re-entry leading to neuronal death. Cell cycle disturbances are also detectable in peripheral cells from Alzheimer’s disease (AD) patients. We previously reported that the anti-inflammatory 15- deoxy-Δ^12,14-prostaglandin J ₂ (15d-PGJ ₂) increased the cellular content of the cyclin-dependent kinase inhibitor p27, in lymphoblasts from AD patients. This work aimed at elucidating the mechanisms of 15d-PGJ ₂-induced p27 accumulation. Phosphorylation, half-life, and the nucleo-cytoplasmic traffic of p27 protein were altered by 15d-PGJ2 by mechanisms dependent on PI3K/Akt activity. 15d-PGJ ₂ prevents the calmodulin-dependent Akt overactivation in AD lymphoblasts by blocking its binding to the 85-kDa regulatory subunit of PI3K. These effects of 15d-PGJ ₂ were not mimicked by 9,10-dihydro-15-deoxy-Δ^12,14- prostaglandin J ₂, suggesting that 15d-PGJ ₂ acts independently of peroxisome proliferator-activated receptor γ activation and that the α,β-unsaturated carbonyl group in the cyclopentenone ring of 15d-PGJ ₂ is a requisite for the observed effects. Received 14 July 2008; received after revision 2 September 2008; accepted 12 September 2008     

Satellite DNA sequences and reproductive isolation in theDrosophila willistoni group

Summary Satellite DNAs of theDrosophila willistoni group have been analyzed by Hae III endonuclease digestion. Differences were observed between the 2 speciesD. willistoni andD. paulistorum and also between semispecies ofD. paulistorum and, to a lesser extent, between subspecies ofD. willistoni. Differences did not appear to be consistently greater between strains producing sterile hybrid males than between those producing fertile hybrids.This work was supported by grants 78.01852.65 from the CNR (Rome) and GM 22221 from the PHS (USA).