Metabolism of endosulfan in the Indian honey bee

Summary Metabolism of topical endosulfan in the Indian honey bee,Apis cerana indica F., was studied. The two endosulfan isomers were found to be interconvertible in bees. The products of metabolism were endosulfan sulfate, diol, ether, hydroxyether, lactone and two unknown compounds. Endosulfan diol, hydroxyether and lactone were found to be conjugated in excreta of the treated bees.Acknowledgment. Authors are grateful to the department of Horticulture and Food Technology for providing gas chromatography facilities.     

The excretion of urea by the larvae ofSpodoptera mauritia boisd. (Noctuidae: Lepidoptera) during development

Summary The concentration of urea in the excreta of the 6th instar larvae ofSpodoptera mauritiavaries from 4.017±0.541 to 31.052±1.193 moles/g dry excreta (mean±SE). The observation confirms that urea excreted is of metabolic origin.     

Purification and properties of glutamine synthetase I fromRhizobium sp. UMKL 20

Summary Glutamine synthetase I was purified fromRhizobium sp. UMKL 20 following polyethylene glycol precipitation. The enzyme had a subunit molecular weight of 58 kd. Apparent K_m values for ammonia and glutamate were 5.6 and 15.2 mM, respectively. Glutamine synthetase I activity was inhibited by several end products of glutamine metabolism. The purified enzyme was highly adenylylated (E _n ^– =8.5).Acknowledgment. I would like to thank Mr J. C. Lai for technical assistance. This work was carried out with the support of Vote F 153/79 from the University of Malaya.     

Partial purification of an endogenous elicitor from suspension-cultured cells of red bean,Vigna angularis

Brief irradiation with a germicidal UV lamp of cells of red bean,Vigna angularis, cultured in suspension in a quartz flask caused the release into the culture medium of an endogenous substance with elicitor activity, as well as the accumulation of isoflavone glucoside stress metabolites in the cells. The active compound was fractionated using phenylalanine ammonia lyase (PAL)-inducing activity in fresh cells as a marker. The elicitor active principle appears to be a low molecular weight (<2000 MW) water-soluble acidic oligosaccharide.     

Regulation of ammonia uptake inAspergillus nidulans

Summary The ammonia uptake inA. nidulans was found to be linear for about 20 min, and was proportional up to 1.5 mg/ml dry cell density. The transport of ammonia does not involve energy. Normal and biotin deficientA. nidulans showed an identical K_m-values of 10.26×10^–5 M ammonia for uptake. The uptake of ammonium ion has been shown to be regulated by the intracellular concentration of ammonia.Acknowledgment. The award of research fellowship by the Govt of India, Department of Atomic Energy, Bombay, to JDD is acknowledged.     

The infectivity of polyhedra of nuclear polyhedrosis virus (N.P.V.) after passage through gut of an insectpredator

Summary Polyhedral inclusion bodies (P.I.B.) of a nuclear polyhedrosis virus ofHeliothis punctigera (Lep.) were found in excreta ofNabis tasmanicus (Het.), a predator ofH. punctigera. An immunofluorescent counting method was used to estimate numbers of P.I.B. Bioassays demonstrated that no loss of infectivity occurred during passage through the gut.Acknowledgment. I am grateful for the comments and criticism offered by Dr D.E. Pinnock, I thank D.J. Cooper for preparation of materials for the immunofluorescent counting method and J. Hardy for supplying me with eggs ofH. punctigera; all of Department of Entomology, Waite Agricultural Research Institute, Glen Osmond, South Australia.     

PcF protein from <Emphasis Type="Italic">Phytophthora cactorum</Emphasis> and its recombinant homologue elicit phenylalanine ammonia lyase activation in tomato

The phytotoxic protein PcF (Phytophthora cactorum-Fragaria) is a 5.6-kDa cysteine-rich, hydroxyproline- containing protein that is secreted in limited amounts by P. cactorum, an oomycete pathogen of tomato, strawberry and other relevant crop plants. Although we have shown that pure PcF triggers plant reactivity, its mechanism of action is not yet understood. Here we show that PcF, like other known fungal protein elicitors involved in pathogen-plant interaction, stimulates the activity of the defense enzyme phenylalanine ammonia a key step in understanding the mechanism of action of PcF at a molecular level is knowledge of its three-dimensional structure, we overexpressed this protein extracellularly in Pichia pastoris. The preliminary structural and functional characterization of a recombinant PcF homologue, N4-rPcF, is reported. Interestingly, although N4-rPcF is devoid of proline hydroxylation and has four additional amino acid residues attached to its N terminus, its secondary structure and biological activity are indistinguishable from wild-type PcF.Received 22 February 2003; received after revision 25 March 2003; accepted 14 April 2003     

Synergism of a natural insect growth inhibitor is mediated by bioactivation

Summary Toxic plant allelochemicals are widespread in nature, but their mechanisms of action are largely unexplored. We report an example of bioactivation of a natural product, -asarone, that is mediated via insect mixed-function oxidases (MFO's), enzymes usually involved in detoxication processes. Bioactivity of -asarone is synergised by menthol, a MFO inducer, and antagonized by the MFO inhibitor piperonyl butoxide. Formation of a bioactive epoxide was confirmed by the identification of asarone epoxide and asarone diol in the insect excreta. These experiments represent the first demonstration of synergism between two natural products (-asarone and menthol) where the mechanism involves induction of enzymes usually involved in detoxication.     

In vitro metabolism of the trichothecene 4-monoacetoxyscirpenol by fungus- and non-fungus-feeding insects

Summary The metabolism of the trichothecene 4-monoacetoxyscirpenol by intact gut tissue was determined in the fungus-feeding Nitidulid,Carpophilus hemipterus (L.) and the non-fungus-feeding caterpillars, the fall armyworm,Spodoptera frugiperda (J. E. Smith) and the corn earworm,Heliothis zea (Boddie). The primary metabolite was the hydrolysis product scirpentriol. The amount of metabolism by theC. hemipterus larvae was ca 10 times that of the caterpillars on a per mg protein basis, suggesting metabolic adaptation for feeding on fungi that may contain mycotoxins.Acknowledgment. The authors wish to thank S. Taylor for technical assistance.The mention of firm names or trade products does not imply that they are endorsed or recommended by the U.S. Department of Agriculture over other firms or similar products not mentioned.     

Preliminary observations of a bacteriophage infectingXenorhabdus luminescens (Enterobacteriaceae)

A bacteriophage infective toXenorhabdus luminescens, a bacterial symbiont of heterorhabditid nematodes, was recovered from insects that supported poor nematode development. Plaque tests showed the phage particles to be infective only to primary and not secondary colonies ofX. luminescens. The phage was not infective toX. nenatophilus primaries or secondaries. The bacteriophage particles ranged 80–90 nm in length, with the head ranging from 40 to 50 nm in diameter. Restriction analysis was performed on isolated bacteriophage DNA. This first report of a bacteriophage fromXenorhabdus species has pratical implications since it could be detrimental to cultures ofHeterorhabditis nematodes that are being produced throughout the world for the biological control of insects.     

The evolutionary history ofDrosophila buzzatii. V. Differential survivorship onOpuntia betweenD. buzzatii andD. serido

Summary Survival time ofDrosophila buzzatii adults on anOpuntia (prickly pear) medium was significantly longer than that of its nearest relativeD. serido. A significant difference was also found betweenD. buzzatii adults from two experimental populations, one of them fed onOpuntia rots for more than two years and another one kept on standardDrosophila medium for the same period of time. These results suggest that adult selection may be taking place in cactiphilicDrosophila in their natural habitats and could be responsible for the niche differentiation betweenD. buzzatii andD. serido.This work has been supported by the Comisión Asesora de investigación Científica y Técnica, Spain, by funds to project 4514/79 awarded to the third author.     

Endocrine-dependent expression of circadian clock genes in insects

Current models state that insect peripheral oscillators are directly responsive to light, while mammalian peripheral clock genes are coordinated by a master clock in the brain via intermediate factors, possibly hormonal. We show that the expression levels of two circadian clock genes, period (per) and Par Domain Protein 1 (Pdp1) in the peripheral tissue of an insect model species, the linden bug Pyrrhocoris apterus, are inversely affected by contrasting photoperiods. The effect of photoperiod on per and Pdp1 mRNA levels was found to be mediated by the corpus allatum, an endocrine gland producing juvenile hormone. Our results provide the first experimental evidence for the effect of an endocrine gland on circadian clock gene expression in insects. Received 31 October 2007; received after revision 7 January 2008; accepted 9 January 2008 D. Dolezel, L. Zdechovanova: These authors contributed equally to this work.     

Shoot regeneration fromAgrobacterium tumefaciens-induced tumors of a tropical timber tree,Fagraea fragrans

Summary Agrobacterium tumefaciens A208 with nopaline plasmid pTiT37 was used to obtain stem tumors on plantlets ofFagraea fragrans grown in vitro. Bacterial elimination and tissue proliferation were simultaneously achieved by growing tumors on cefatoxime medium. After some tissue growth the shoots regenerated. An examination of these showed the presence of nopaline, indicating genetic transformation by T-DNA.     

Protein-O-mannosyltransferases in virulence and development

Protein-O-mannosyltransferases (Pmt proteins) catalyse the addition of mannose to serine or threonine residues of secretory proteins. This modification was described first for yeast and later for other fungi, mammals, insects and recently also for bacteria. O-mannosylation depends on specific isoforms of the three Pmt1, 2 and 4 subfamilies. In fungi, O-mannosylation determines the structure and integrity of cell walls, as well as cellular differentiation and virulence. O-mannosylation of specific secretory proteins of the human fungal pathogen Candida albicans and of the bacterial pathogen Mycobacterium tuberculosis contributes significantly to virulence. In mammals and insects, Pmt proteins are essential for cellular differentiation and development, while lack of Pmt activity causes Walker-Warburg syndrome (muscular dystrophy) in humans. The susceptibility of human cells to certain viruses may also depend on O-mannosyl chains. This review focuses on the various roles of Pmt proteins in cellular differentiation, development and virulence. Received 6 September 2007; received after revision 3 October 2007; accepted 5 October 2007     

What leads from <Emphasis Type="Italic">dead-end?</Emphasis>

The 129 mouse strain develops congenital testicular germ cell tumors (TGCTs) at a low frequency. TGCTs in mice resemble the testicular tumors (teratomas) that occur in human infants. The genes that cause these tumors in 129 have not been identified. The defect at the Ter locus increases TGCT incidence such that 94% of 129-Ter/Ter males develop TGCTs. The primary effect of the Ter mutation is progressive loss of primordial germ cells (PGCs) during embryonic development. This results in sterility in adult Ter/Ter mice on all mouse strain backgrounds. However, on the 129 background, Ter causes tumor development in addition to sterility. Therefore, Ter acts as a modifier of 129-derived TGCT susceptibility genes. Ter was identified to be a mutation that inactivates the Dead-end1 (Dnd1) gene. In this perspective, I discuss the possible areas of future investigations to elucidate the mechanism of TGCT development due to Dnd1 inactivation. Received 29 September 2006; received after revision 29 January 2007; accepted 19 February 2007     

Partial sequence of the gene for a serine protease from a halophilic archaeumhaloferax mediterranei R4, and nucleotide sequences of 16S rRNA encoding genes from several halophilic archaea

A part of the gene coding for a halophilic serine protease from a halophilic archaeumHaloferax mediterranei R4 was amplified by PCR and its 672 nucleotide sequence was determined. Tentative translation to the amino acid sequence suggested that the enzyme was quite similar to halolysin produced by another halophilic archaeum strain 172P1. Nucleotide sequences of 16S rRNA encoding genes from 9 halophilic archaea were determined. Alignment of 19 sequences known so far showed that there are more than 20 positions carrying bases or deletions specific for each halobacterial genus:Halobacterium, Haloarcula, Haloferax, andHalococcus.     

Heat shock proteins in three relatedDrosophila species belonging to theobscura group

The effect of heat shock on protein synthesis in three relatedDrosophila species belonging to theobscura group was analyzed on SDS-acrylamide gels. Four major heat shock proteins (hsps) were found in these species, in which synthesis reaches a maximum at 34°C. Although the higher molecular weight proteins are conserved, differences in size were found for the small hsps in these species. By means of in situ hybridization usingD. melanogaster probes for the small hsp genes, it was inferred that the small hsp genes of theobscura group species are clustered at the 27A locus in all three species.     

Effect of tingenone,a quinonoid triterpene,on growth and macromolecule biosynthesis inTrypanosoma cruzi

Summary Tingenone and horminone, two natural quinonoid substances, inhibited the in vitro growth ofTrypanosoma cruzi, 30 M drug concentration producing total inhibition of growth. Tingenone inhibited total uptake and incorporation of [³H]thymidine, [³H]uridine, L-[³H]leucine into parasite macromolecules. Other quinonoids assayed were either less effective (abruquinone A) or even quite inactive (visminone B and ferruginin B). Investigation of several mechanisms for the cytotoxic action of tingenone pointed to the interaction with DNA as the most likely factor involved. Tingenone also inhibited the growth ofCrithidia fasciculata, but the drug was significantly less active on this organism than onT. cruzi.This work was supported by grants of UNDP/World Bank/World Health Organization Special Programme for Research and Training in Tropical Diseases, Organization of American States (Multinational Programme of Biochemistry) and Programa Nacional de Enfermedades Endémicas (SECYT), República Argentina. A preliminary account was given at the Workshop on Oxidative Damage and Related Enzymes, Frascatti (Italy), 1983.     

Spasmin-like proteins in various ciliates revealed by antibody to purified spasmins ofCarchesium polypinum

Summary It was found that some ciliates,Stentor, Spirostomum andBlepharisma, which can contract rapidly like the stalks of Vorticellidae, have Ca²⁺-binding proteins that are very similar to spasmins, in the immunological sense. The presence of spasmins in other Protozoa and in some Metazoa was also investigated.     

Effects of the pyrimidine-containing cytochrome P-450 inhibitor,fenarimol, on the formation of 20-OH ecdysone in flies

Summary The effect of fenarimol, a pyrimidine-containing cytochrome P-450 inhibitor, was tested in vitro on brain-ring gland complexes ofCalliphora vicina (Dipt., Calliphoridae), and on microsomes prepared from the fat body of 0-h wandering stage larvae ofNeobellieria bullata (Dipt., Sacrophagidae). Fenarimol had no influence on the formation of ecdysone, but it was an effective inhibitor of cytochrome P-450-dependent ecdysone 20-monooxygenase.