Fungitoxicity ofm-fluorophenylalanine-containing peptides towardsPythium ultimum

Summary The tripeptide L-m-fluorophenylalanyl-L-alanyl-L-alanine was much more fungitoxic towardsPythium ultimum than the dipeptide L-m-fluorophenylalanyl-L-alanine orm-fluorophenylalanine. The fungitoxicity of the tripeptide was reduced by L-alanyl peptides and phenylalanine, but not by other amino acids. In contrast, the fungitoxicity ofm-fluorophenylalanine was unaffected by peptides, and was antagonized by several amino acids. These results suggest the effective delivery ofm-fluorophenylalanine into the cell by a tripeptide carrier.     

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.     

The C4-dicarboxylate transport system ofRhizobium meliloti and its role in nitrogen fixation during symbiosis with alfalfa (Medicago sativa)

TheRhizobium meliloti C₄-dicarboxylate transport (Dct) system is essential for an effective symbiosis with alfalfa plants. C₄-dicarboxylates are the major carbon source taken up by bacteroids. Genetic analysis of Dct^– mutant strains led to the isolation of thedct carrier genedctA and the regulatory genesdctB anddctD. The carrier genedctA is regulated in free-living cells by the alternative sigma factor RpoN and the two-component regulatory system DctB/D. In addition, DctA is involved in its own regulation, possibly by interacting with DctB. In bacteroids, besides the DctB/DctD system an additional symbiotic activator is thought to be involved indctA expression. Further regulation ofdctA in the free-living state is reflected by diauxic growth of rhizobia, with succinate being the preferred carbon source. The tight coupling of C₄-dicarboxylate transport and nitrogen fixation is revealed by a reduced level of C₄-dicarboxylate transport in nitrogenase negative bacteroids.     

Insect photoperiodism: various ways of regulating univoltinism in lacewings (Planipennia: Chrysopidae)

Summary Reproduction inTjederina gracilis adults depends on short-day followed by long-day conditions. InNineta pallida, development of overwintering first-instar larvae is retarded by short days; later on, that of second-and third-instar by long days.Nineta flava andChrysopa perla prepupae automatically enter diapause, but inN. flava, photoperiod regulates diapause eermination, and long days may retard egg laying     

<Emphasis Type="Italic">LAPTM4A</Emphasis> interacts with <Emphasis Type="Italic">hOCT2</Emphasis> and regulates its endocytotic recruitment

Human organic cation transporter 2 (hOCT2) is involved in the transport of endogenous and exogenous organic cations mainly in cells of the kidney and the brain. Here, we focus on the regulation of hOCT2 by direct protein–protein interaction. Screening within a mating-based split-ubiquitin-yeast-two-hybrid system (mBSUS) revealed the lysosomal-associated protein transmembrane 4 alpha (LAPTM4A) as a potential interacting protein. Interaction of LAPTM4A and hOCT2 was confirmed by pulldown assays, FRET microscopy analysis and immunofluorescence microscopy. Functionally, overexpression of LAPTM4A significantly decreased ASP⁺ uptake in HEK293 cells stably transfected with hOCT2, suggesting a negative regulation of hOCT2-mediated transport. Furthermore, overexpression of LAPTM4A leads to a significantly decreased hOCT2 plasma membrane expression in surface biotinylation experiments. In addition, significant expression of LAPTM4A in human kidney was demonstrated by immunoblotting and immunofluorescence.     

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.     

Mating attempts between the Scarlet Tiger Moth,Callimorpha dominula L., and the Cinnabar Moth,Tyria jacobaeae L. (Lepidoptera: Arctiidae), involve a common sex pheromone composition

It has been suggested that a common sex pheromone composition may account for interspecific sexual interactions observed with certain moths in the Arctiidae. In this study, it is demonstrated that the sex pheromones released by females of the Scarlet Tiger Moth,Callimorpha dominula L., and the Cinnabar Moth,Tyria jacobaeae L., have similar activities and elute at the same retention time on analysis by coupled gas chromatography (GC)-electrophysiology with males from each species. Peak enhancement on GC, chiral GC and coupled GC-mass spectrometry using authentic compounds show that the sex pheromone for bothC. dominula andT. jacobaeae is (3Z,6Z,9S,10R)-9,10-epoxyheneicosa-3,6-diene.     

Fungal competition and mycotoxin production on corn

Summary Aflatoxin and secalonic acid D production in corn in laboratory and field by mixed cultures ofPencillium oxalicum andAspergillus flavus orA. parasiticus was lower than production by the pure cultures. However, mixed culture of these molds withFusarium spp. did not affect mycotoxin production.     

Occurrence of a blood group A-like substance in eggs of the prosobranch snailPomacea canaliculata

Summary In the eggs of the prosobranch snailsPomacea canaliculata andPomacea insularum a blood group A-like substance has been detected by anti-A from the snailsHelix pomatia, Helix aspersa andCepaea nemoralis.     

La formule chromosomique deSorex minutus L. (Mammalia-Insectivora)

Summary The chromosome complement ofSorex minutus L. has been determined. The caryotype is characterized by a diploid number of 42 and by aNF (fundamental number) of 56. The sex-chromosomes are of theX−Y type usual in mammals. The origin and the evolution of the chromosome set ofS. minutus, S. unguiculatus, andS. araneus have been discussed.      

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.     

Sexual behavior mutants revisited: molecular and cellular basis of Drosophila mating

The study of Drosophila melanogaster by a combination of forward genetics with specific mutants, and reverse genetics, in which a given gene is expressed in an appropriate brain area to test its effect on behavior, provides a unique opportunity to explore the causal relationship between a particular gene, its function in the cell and the behavioral outcome at the organismic level. Enhanced male-to-male courtship has been shown to occur as a result of mutations in several different genes. For example, the Voila mutant exhibits intense GAL4 reporter expression in the tarsal gustatory sensilla, suggesting the importance of tapping by a male on the female abdomen with his forelegs. Feminization of parts of the antennal lobe and mushroom body by targeted expression of a female-determining gene transformer ⁺ (tra ⁺) drives the male to court other males. Mutations in the tra target gene fruitless (fru), which is expressed in the antennal lobe as well as the suboesophageal ganglion (the gustatory inputs are processed here), also induce homosexual courtship in males. These results suggest that sensory inputs mediated and/or processed by the tarsal receptors, suboesophageal ganglion, antennal lobe and mushroom body contribute to the regulation of male–female courtship. Mosaic analysis localized the neural center for male courtship behavior to the posterior dorsal brain, in which the sensory information processed by the aforementioned neural structures may be integrated. Another mosaic study mapped the neural center for female sexual behavior, as measured by her receptiveness to copulation, to the anterior dorsal brain. The issue as to how the mutations that reduce female sexual receptiveness, e.g. dissatisfaction (dsf), spinster (spin) and chaste (cht), affect the structure and/or function of this neural center deserves to be addressed urgently. Received 27 April 1999; received after revision 21 June 1999; accepted 8 July 1999     

Un nouveau système polymorphe robertsonien chez une nouvelle espèce de «Leggada» (Mus goundae Petter)

Summary A sample of 6 Leggada from N'Délé (Central African Republic) morphologically different from all these studied up to now, constitue a new species, which will be described by Dr F.Petter asMus goundae Petter. The caryological analysis reveals a new robertsonian polymorphical system. The diploïd numbers are 2N=16, 17, 18 or 19, whereas theN.F. is constant and equal to 30. The 3 first pairs of autosomes, constituted by SM or MC elements are identical in the 4 types, as well as pairs V (MC) and VI (AC) morphologically constant by all individuals. The robertsonian mechanisms, from which polymorphism originates, take place at pairs IV, VII and VIII. The fourth pair, while heterozygote by 2N=19, assembling one MC and two AC elements, is homozygote and MC by 2N=18, 17 and 16; pairs VII and VIII are both constituted by two AC in the cases of 19 and 18 chromosomes; to form caryotypes with 17 and 16 chromosomes these 2 pairs will fusion: by 2N=17 the mutation is heterozygote (1 MC and 2 AC) and it becomes homozygote by 2N=16 giving birth to 2 MC. The sexual chromosomes are from TR Type (X MC andY SM).From these observations arises the problem of low chromosome numbers (16–20) which shall be discussed in a further publication.     

An immune response-dependent mechanism for the vertical transmission of an entomopathogen

Summary An exceptionally efficient mechanism for the vertical transmission of a parasitic gregarine is dependent on the insect host's immune response. Gametocysts ofAscogregarina chagasi on the genital accessory glands of adult female sand flies (Lutzomyia longipalpis) become encapsulated through hemocyte-mediated immune reactions. Oocysts ofA. chagasi, ejected into the lumen of the glands owing to pressure exerted by this capsule, become glued to eggshells and are subsequently ingested by larvae. InL. longipalpis with an experimentally suppressed encapsulation reaction, fewer accessory glands contained oocysts ofA. chagasi.     

Protein O-GlcNAcylation regulates Drosophila growth through the insulin signaling pathway

Modification of nuclear and cytosolic proteins by O-linked N-acetylglucosamine (O-GlcNAcylation) is ubiquitous in cells. The in vivo function of the protein O-GlcNAcylation, however, is not well understood. Here, we manipulated the cellular O-GlcNAcylation level in Drosophila and found that it promotes developmental growth by enhancing insulin signaling. This increase in growth is due mainly to cell growth and not to cell proliferation. Our data suggest that the increase in the insulin signaling activity is mediated, at least in part, through O-GlcNAcylation of Akt. These results indicate that O-GlcNAcylation is one of the crucial mechanisms involved in control of insulin signaling during Drosophila development.     

Insulin activates Gαil,2 protein in rat hepatoma (HTC) cell membranes

Insulin action is initiated by binding to its cognate receptor, which then triggers multiple cellular responses by activating different signaling pathways. There is evidence that insulin receptor signaling may involve G protein activation in different target cells. We have studied the activation of G proteins in rat hepatoma (HTC) cells. We found that insulin stimulated binding of guanosine 5′-O-(3-thiotriphosphate) (GTP-γ-³⁵S) to plasma membrane proteins of HTC cells, in a dose-dependent manner. This effect was completely blocked by pertussis toxin treatment of the membranes, suggesting the involvement of G proteins of the Gα _i/Gα _o family. The expression of these Gα proteins was checked by Western blotting. Next, we used blocking antibodies to sort out the specific Gα protein activated by insulin stimulation. Anti-Gα _il,2 antibodies completely prevented insulin-stimulated GTP binding, whereas anti-Gα _o,i3 did not modify this effect of insulin on GTP binding. Moreover, we found physical association of the insulin receptor with Gα _i1,2 by copurification studies. These results further support the involvement of a pertussis toxin-sensitive G protein in insulin receptor signaling and provides some evidence of specific association and activation of Gα _i1,2 protein by insulin. These findings suggest that Gα _i1,2 proteins might be involved in insulin action. Received 23 September 1998; received after revision 23 November 1998; accepted 25 November 1998     

Targeted inhibition of Livin resensitizes renal cancer cells towards apoptosis

Cancer cells are typically characterized by apoptosis deficiency. In order to investigate a possible role for the anti-apoptotic livin gene in renal cell cancer (RCC), we analyzed its expression in tumor tissue samples and in RCC-derived cell lines. In addition, we studied the contribution of livin to the apoptotic resistance of RCC cells by RNA interference (RNAi). Livin gene expression was detected in a significant portion of RCC tumor tissue specimens (13/14, 92.9%) and tumor-derived cell lines (12/15, 80.0%). Moreover, targeted inhibition of livin by RNAi markedly sensitized RCC cells towards proapoptotic stimuli, such as UV irradiation or the chemotherapeutic drugs etoposide, 5-fluorouracil, and vinblastine. These effects were specific for livin expressing tumor cells. We conclude that livin can contribute significantly to the apoptosis resistance of RCC cells. Targeted inhibition of livin could represent a novel therapeutic strategy to increase the sensitivity of renal cancers towards pro-apoptotic agents. Received 30 November 2006; received after revision 22 February 2007; accepted 20 March 2007     

Identification of chicken lysozyme <Emphasis Type="Italic">g</Emphasis>2 and its expression in the intestine

Lysozyme is an important component of the innate immune system, protecting the gastrointestinal tract from infection. The aim of the present study was to determine if lysozyme is expressed in the chicken (Gallus gallus) intestine and to characterise the molecular forms expressed. Immunohistochemical staining localised lysozyme to epithelial cells of the villous epithelium along the length of the small intestine. There was no evidence for lysozyme expression in crypt epithelium and no evidence for Paneth cells. Immunoblots of chicken intestinal protein revealed three proteins: a 14-kDa band consistent with lysozyme c, and two additional bands of approximately 21 and 23 kDa, the latter consistent with lysozyme g. RT-PCR analyses confirmed that lysozyme c mRNA is expressed in 4-day, but not older chicken intestine and lysozyme g in 4- to 35-day chicken intestine. A novel chicken lysozyme g2 gene was identified by in silico analyses and mRNA for this lysozyme g2 was identified in the intestine from chickens of all ages. Chicken lysozyme g2 shows similarity with fish lysozyme g, including the absence of a signal peptide and cysteines involved in disulphide bond formation of the mammalian and bird lysozyme g proteins. Analyses using SecretomeP predict that chicken lysozyme g2 may be secreted by the non-classical secretory pathway. We conclude that lysozyme is expressed in the chicken small intestine by villous enterocytes. Lysozyme c, lysozyme g and g2 may fulfil complimentary roles in protecting the intestine.Received 4 August 2004; received after revision 1 September 2004; accepted 7 September 2004     

Avoidance of competition by spruce bark beetles,Ips typographus andPityogenes chalcographus

Bark beetles,Ips typographus andPityogenes chalcographus, attracted by synthetic or natural pheromone to Norway spruce logs,Picea abies, preferred to colonize uninfested logs rather than logs occupied by these beetles, probably as a means of avoiding intra-and interspecific competition. The aggregation pheromone components ofP. chalcographus, chalcogran and methyl (E, Z)-2,4-decadienoate, inhibited the attraction response ofI. typographus to its pheromone components (methyl butenol andcis-verbenol), while the converse was not true. However, verbenone released from colonized bark inhibited pheromonal response ofP. chalcographus.     

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.