bluuebunny/crossref_metadata_2025 huggingface dataset:
Chemical characterization and species specificity of sex pheromones of plusiinae moths in Israel
Authors: Dunkelblum Ezra , Mazor Michal
Published: February 2005

The female sex pheromones of four sympatric Plusiinae species, Autographa gamma, Chrysodeixis chalcites, Cornutiplusia circumflexa, and Trichoplusia ni were investigated in Israel. The research concentrated on the behavioral reproductive isolation regulated by their sex pheromones. The chemical composition of the four sex pheromones was determined by analysis of pheromone gland extracts, and in addition entrained volatiles released from virgin female C. chalcites and T. ni were also analyzed. The pheromones of these latter two species were found to contain several components, among which Z7‐12:Ac was the main one. The pheromone glands of A. gamma and C. circumflexa were found to contain relatively small amounts of material and only Z7–12:Ac and Z7–12:OH could be positively identified. These two pheromone components were present in the two species in roughly inverse rations. Whereas A. gamma utilizes Z7–12:Ac as the main or sole pheromone component, C. circumflexa requires both Z7–12:OH and Z7–12:Ac, with the alcohol being predominant. This is the first reported Plusiinae species using Z7–12:OH as the main pheromone component. Behavioral studies in the wind tunnel and field tests indicate that females of each of the four Plusiinae species utilizes a species‐specific blend of chemicals, consisting of attractive and antagonistic components, to attract conspecific males. Such a complementary strategy provides improved behavioral reproductive isolation among the sympatric species. For example, Z5–12:Ac, which is a specific component of the T. ni sex pheromone, may be redundant as regards the attraction of T. ni males. However, addition of small amounts of this compound to pheromone blends of A. gamma and C. chalcites resulted in complete inhibition of the males' flight behavior and trap catch; thus contributing significantly to the specificity of the T. ni sex pheromone. The relatively high proportion of Z7–12:OH in the sex pheromone of C. circumflexa, essential to the latter, is highly inhinitory to the three other Plusiinae species. The only case of cross‐attraction occurred when T. ni males were attracted to a certain extent to A. gamma lures and females. © 1993 Wiley‐Liss, Inc.
Nonconventional insecticidal effects of pesticides available from the Neem tree, Azadirachta indica
Authors: Ascher K. R. Simon
Published: February 2005

Nonconventional insecticidal effects are exerted by preparations from the Neem tree, also called Indian lilac or Margosa tree (Azadirachta indica A. Juss, syn. Melia azadirachta L. or Antelaea azadirachta L.) (Meliaceae). A closely related species, the chinaberry tree or Persian lilac (Melia azedarch L.), is a source of substances with similar strctures and insecticidal activity. However, the seed kernels, whole fruits or leaves of many M. azedarch chemotypes contain tetranortriterpenoids and other principles highly toxic to mammals (meliatoxins). The most prominent insecticidal constituent of Neem seed kernels is the tetranortriterpenoid azadirachtin and related structures. Lesser concentrations of these active materials are found in the leaves and other parts of the tree. Third World farmers use crude Neem preparations, mainly seed kernel extracts or powders, as insecticides. An industrial Neem product, called Margosan‐O, is in commercial use in the USA. The nonconventional effects of preparations or compounds isolated from Neem may be classified as follows: (i) partial reduction or complete inhibition of fecundity and/or sometimes egg hatchability; (ii) reduction of the life span of adults; (iii) oviposition repellence against females; (iv) direct ovicidal effects; (v) antifeedant effects against larvae (and nymphs) and adults; (vi) formation of permanent larvae; (vii) insect growth regulator effects at molting between larval (or nymphal) instars and especially in the prepupal stage; and (viii) analogous lesions during the emergence of adults. Phenomena (vii) and (viii) give rise to characteristic larval‐pupal, nymphal‐pupal, nymphal‐adult and pupal‐adult intermediates, and to crippled adults. © 1993 Wiley‐Liss, Inc.
Delayed and enhanced biodegradation of soil‐applied diphenamid, carbendazim, and aldicarb
Authors: Aharonson Nadav , Katan Jaacov
Published: February 2005

Recent studies have demonstrated that interaction between various agrochemicals and soil microorganisms may either slow down or enhance processes of degradation. Soil disinfestation is employed for the control of soil‐borne pathogens and weeds. Soil application of such broad‐spectrum biocides, as well as some more selective chemicals, has a strong effect on microbial activity, which may result in drastic reduction in the rate of degradation of pesticides applied to such treated soils. Application of pesticides to previously disinfested soils may extend their biological activity, which in the case of herbicides could cause phytotoxic damage to the next crop. In contrast, repeated application of the same or structurally related pesticides may result in a selective buildup of microbial populations capable of degrading the pesticide at much faster rates. Cases of accelerated degradation were reported for pesticides belonging to various chemical groups. Studies were conducted to evaluate the mechanisms of accelerated degradation. For several pesticides it has been shown that soil fungi are involved in their normal degradation, but not in their accelerated degradation. The shift in the rate of degradation of pesticides such as diphenamid, benomyl, and S‐ethyl dipropylthiocarbamate, in soils that have acquired accelerated degradation, seems to be associated with the buildup of populations of bacterial degraders. Moreover, it has been shown that for the herbicide diphenamid, accelerated degradation is apparently linked to the induction of an oxidative demethylation process in soil bacteria, which might be analogous to the development of resistance in pests. Contrary to studies demonstrating accelerated degradation of the systemic insecticide aldicarb and accumulation of aldicarb sulfoxide in non‐history soils, our work has shown that repeated application of this carbamate at several locations in Israel over a 10‐year period did not induce accelerated degradation. It should be pointed out that in the Israeli soils there was only minimal formation of aldicarb sulfoxide. These studies were performed in soils with a pH ranging from 7.8 to 8.3, which is higher than the pH reported for soils where accelerated degradation was detected. © 1993 Wiley‐Liss, Inc.
Cholinesterase profiles in two species of wild birds exposed to insecticide sprays in their natural habitat
Authors: Yawetz Aminadav , Zook‐Rimon Zohar , Dotan Aharon
Published: February 2005

During summer season, a field survey was conducted to assess the extent of stress to which two species of wild birds might have been subjected, due to their dwelling near cotton fields sprayed with insecticides. One of the bird species, the cattle egret Bubulcus ibis, showed no significant changes either in brain acetylcholinesterase (AChE) or in plasma butyrylcholinesterase (BuChE) activities in samples collected before, during, and after the spraying season. With the spur‐wing plover, Hoplopterus spinosus, there was a significant decrease both in brain AChE and plasma BuChE activities in July and August, the period when insecticide spray was most frequent. In laboratory experiments it was found that this species could survive higher doses of parathion, and specimens surviving the treatment showed higher residual plasma BuChE activity and a faster return of that activity to normal compared to Bubulcus specimens. Brain AChE from the Hoplopterus showed a lower Ki value for inhibition by paraoxon or malaoxon compared to the same values recorded for the Bubulcus, indicating that the enzyme from the Hoplopterus brain is less sensitive to inhibition by paraoxon or malaoxon. The finding that the more resistant Hoplopterus showed a decrease in cholinesterase activity during the cotton spraying season may therefore be due to the territorial habits of this species. © 1993 Wiley‐Liss, Inc.
24‐Epi‐makisterone a (not makisterone A) is the major ecdysteroid in the leaf‐cutting ant Acromyrmex octospinosus (reich) (hymenoptera, formicidae: Attini)
Authors: Maurer Patricia , Girault Jean‐Pierre , Larchevêque Marc , Lafont René
Published: February 2005

The chemical nature of the major ecdysteroid from pupae of the leaf‐cutting ant Acromyrmex octospinosus has been reinvestigated. On the basis of reversed phase high performance liquid chromatography and mass spectrometry, this compound was previously identified as makisterone A [Maurer et al., Arch Insect Biochem Physiol 16, 1 (1991)] but a more careful examination has led to the conclusion that it is in fact 24‐epi‐makisterone A. This finding stresses the limits of criteria used for the identification of ecdysteroids and the possible need for reexamination of makisterone A identification in some other species feeding on a 24β‐methyl sterol source. © 1993 Wiley‐Liss, Inc.
Food utilization and antioxidant enzyme activities of black swallowtail in response to plant phototoxins
Authors: Lee Keywan , Berenbaum May R.
Published: February 2005

Phototoxic compounds are widely distributed among plant families; due to their ability to bind covalently to DNA or to react with oxygen and generate toxic oxyradicals, these compounds are toxic to a variety of herbivorous organisms. Black swallowtail (Papilio polyxenes) larvae feed exclusively on phototoxic host plants in the Apiaceae and Rutaceae. In this study, we examined the toxicity of four phototoxins—three furanocoumarins and one β‐carboline alkaloid—to P. polyxenes, as well as the inducibility of antioxidant enzyme defenses in response to these phototoxins. Neither the furanocoumarins nor the β‐carboline alkaloid demonstrated any toxic effect on digestive efficiencies of P. polyxenes in the presence of light; harmine, the alkaloid, did significantly reduce growth and consumption rates. None of the compounds had a significant effect on antioxidant enzyme levels. These findings contrast with those reported in earlier studies for Trichoplusia ni, a generalist noctuid sensitive to both furanocoumarins and β‐carboline alkaloids. Greater detoxicative metabolic capabilities, coupled with substantially higher constitutive levels of antioxidant enzyme activity, likely explain at least in part the absence of induced antioxidant enzyme responses in the specialist feeder on phototoxic plants. © 1993 Wiley‐Liss, Inc.
Characterization of a diuretic hormone receptor from the tobacco hornworm, Manduca sexta
Authors: Regan Jeff D. , Li Jorge P. , Carney Robert L. , Kramer Steven J.
Published: February 2005

We have characterized a diuretic hormone receptor from the tobacco hornworm, Manduca sexta. A single high affinity binding site for the 41 amino acid M. sexta diuretic hormone was found in membranes prepared from Malpighian tubules of fifth stadium larvae. The site has a Kd = 79 pM and Bmax = 3.1 pmol/mg protein. The dissociation rate constant was determined to be 0.11 min−1 with a corresponding half‐life of 6.4 min. Receptor binding of the hormone is inhibited by Ca2+ and Mg2+, while Na+ and K+ inhibit binding to a lesser extent. Truncated diuretic hormone analogs in which up to 20 amino acids were removed from the N‐terminus maintain high affinity for the receptor. A diuretic hormone from Locusta migratoria which has 43% sequence identity with the M. sexta diuretic hormone also possesses a high affinity for the receptor. Conformational analysis of the M. sexta diuretic hormone indicates the core region of the peptide assumes a helical conformation, which may have implications in the binding of the peptide to the receptor. © 1993 Wiley‐Liss. Inc.
Induction of detoxification enzymes in phytophagous insects: Role of insecticide synergists, larval age, and species
Authors: Yu Simon J. , Hsu Err L.
Published: February 2005

Five insecticide synergists, all of which were either methylenedioxyphenyl compounds or analogs, were compared as to their effect on cytochrome P450 monooxygenase induction caused by an allelochemical in fall armyworm larvae. Feeding the synergists (piperonyl butoxide, safrole, isosafrole, MGK 264, and myristicin) individually to the larvae caused decreases in the microsomal aldrin epoxidase activities ranging from 38% to 74% when compared with controls. Feeding indole‐3‐carbinol resulted in a 4‐fold increase in the microsomal epoxidase activity. However, cotreatment of any of the synergists and the inducer completely eliminated the induction. Sixth instar larvae were more inducible than second instar larvae with respect to microsomal epoxidase and glutathione transferase in the fall armyworm. Enzyme inducibility varied widely among the seven phytophagous Lepidoptera examined. When indole‐3‐carbinol was used as an inducer of microsomal epoxidase, the extent of inducibility of the enzyme was fall armyworm > velvetbean caterpillar > corn earworm > beet armyworm > tobacco budworm > cabbage looper > diamondback moth. When indole‐3‐acetonitrile was used as an inducer, the inducibility of glutathione transferase was fall armyworm > beet armyworm > corn earworm > cabbage looper > velvetbean caterpillar > tobacco budworm > diamondback moth. Inducibility of five microsomal oxidase systems also varied considerably in the corn earworm, indicating the multiplicity of cytochrome P450 in this species. Microsomal epoxidase and glutathione transferase were induced by cruciferous host plants such as cabbage and their allelochemicals in diamondback moth larve. © 1993 Wiley‐Liss, Inc.
Basic isoforms of hemolymph storage proteins expressed during larval metamorphosis
Authors: Jones Grace , Sarkari Neville
Published: February 2005

The hemolymph of metamorphosing, final instar larvae of Trichoplusia ni was analyzed for the presence of basic forms of normally positively charged storage proteins. Basic forms of arylphorin and a normally acidic juvenile hormone suppressible protein were identified. For each of these two proteins, variation was observed in the immunoreactivity of forms with different basic charges, where the antisera had been generated against acidic forms of each protein. A basic protein of high molecular size (ca. M, 150,000) was identified that cross‐reacted specifically with an antiserum raised against a normally basic, M 74,000 juvenile hormone suppressible protein in the hemocyanin superfamily. © 1993 Wiley‐Liss, Inc.
Postendocytic vitellin processing in ovarian follicles of the stick insect Carausius morosus (Br.)
Authors: Giorgi Franco , Masetti Massimo , Ignacchiti Vincenzo , Cecchettini Antonella , Bradley James T.
Published: February 2005

Newly laid eggs of the stick insect Carausius morosus contain two native vitellins (Vit A and Vit B). Under denaturing conditions, these vitellins resolved into 3 (A1, A2, and A3) and 2 (B1 and B2) polypeptides. All of these polypeptides had counterparts in the female hemolymph from which they were shown to be derived by in vivo labelling. During ovarian development, the 2 vitellins changed both in charge and polypeptide composition. In EV and LV follicles, Vit A resolved into 4 distinct vitellin polypeptides (A0, A1, A2 and A3). Using a panel of monoclonal antibodies, polypeptide A0 proved to be immunologically related to polypeptide A2. In follicles about to begin choriongenesis, polypeptide A3 was gradually replaced by a lower Mr polypeptide. Over the same time period, polypeptide B1 changed in charge, but not in Mr. To confirm the existence of a polypeptide processing in C. morosus, ovarian follicles of different developmental stages were exposed in vivo to [35S]‐methionine from 6 to 72 h. Data showed that A0 and B1 were the polypeptides most heavily labelled after short time exposures to the radioisotope. Polypeptides B2 and A3 were also labelled to some extent. With progressively longer exposures, polypeptides A1 and A2 also became labelled. In vivo exposure to [3H]‐GlcNAc caused all vitellin polypeptides to become heavily labelled. Autoradiographic analysis of ovarian follicles labelled this way showed that, during development, radioactivity was gradually transferred from newly formed yolk spheres in the cortical ooplasm to the central ooplasm. Data were interpreted as suggesting a causal relationship between polypeptide processing and progressive yolk sphere fusion to yield the central ooplasm. © 1993 Wiley‐Liss, Inc.
Preparation and partial characterization of amino acid transporting brush border membrane vesicles from the larval midgut of the gypsy moth (Lymantria dispar)
Authors: Wolfersberger Michael G.
Published: February 2005

Brush border membrane vesicles were prepared from both fresh and frozen midguts of Lymantria dispar larvae by Mg/EGTA precipitation and differential centrifugation. The vesicles were enriched 10 to 13‐fold, relative to the homogenate, in aminopeptidase and gamma‐glutamyl transferase activity. No significant difference was found in enzyme enrichment of vesicles prepared from fresh and frozen midguts. Inwardly directed potassium salt gradients resulted in transient accumulation of leucine and lysine but not glutamic acid by the vesicles. © 1993 Wiley‐Liss, Inc.
Purification and characterization of a flavin‐binding storage protein from the hemolymph of Galleria mellonella
Authors: Silhacek Donald L. , Miller Stephen G. , Murphy Curtis L.
Published: February 2005

The 85K storage protein that accumulates in the hemolymph of Galleria mellonella during the final larval instar was isolated and purified from newly molted pupae. The separation of fresh hemolymph proteins from larvae or pupae by different chromatographic and electrophoretic procedures indicated the native protein had a Mr of 170,000 and consisted of two identical 85K subunits. Crosslinking experiments using fresh hemolymph followed by Western blotting also indicated a dimeric structure for the native protein. Analyses of the dimer purified from pupal hemolymph indicated that 85K was a glycoprotein, containing approximately 6.5% neutral sugar and about 1.9% amino sugar. Like other insect flavin‐binding proteins, 85K has a relatively high histidine content but an uncharacteristically high arginine content. The purified 85K dimer did not bind riboflavin, suggesting that the integrity of the molecule had been altered during purification. However, 85K purified in low yield by Affi‐Gel Blue chromatography, did bind riboflavin, indicating that under certain, undefined conditions the functional integrity of the protein could be retained during purification. © 1994 Wiley‐Liss, Inc.

This article is a US Government work and, as such, is in the public domain in the United States of America.
Toxicokinetics of [3H]‐dihydroazadirachtin in the variegated cutworm Peridroma saucia
Authors: Koul Opender , Isman Murray B. , Arnason John T.
Published: February 2005

The excretion, retention, and tissue distribution of [3H]‐dihydroazadirachtin was investigated in the variegated cutworm, Peridroma saucia (Noctuidae). The candidate compound was rapidly cleared from the hemolymph following either oral exposure or topical administration, with maximum concentrations at 6 h post‐treatment and peak appearance of label in the frass at 12 h. However, approximately 45 and 55% of the labelled material was retained in the body at 72 h in respective treatments. Major depots for retained radioactivity were the gut (24% of the administered oral dose, 18.8% of the administered topical dose) and integument (12.2% of the oral dose and 30.7% of the topical dose). The variation in tissue distribution of dihydroazadirachtin with respect to the mode of application is discussed. A single polar metabolite fraction was obtained from the frass of dihydroazadirachtin‐fed larvae. The physiological and behavioral effects of 22,23‐dihydroazadirachtin and azadirachtin are quantitatively similar. © 1994 Wiley‐Liss, Inc.
Yolk formation in Locusta migratoria and Schistocerca gregaria: Related ligands and oocyte receptors
Authors: Hafer Johannes , Ferenz Hans‐Joerg
Published: February 2005

During vitellogenesis the transport of yolk precursor proteins, the vitellogenins (VTG), from the hemolymph into the oocyte is achieved by receptor‐mediated endocytosis. Recently the receptor for the VTG of Locusta migratoria has been isolated. Now a new protocol has been developed for the purification of the VTG receptor of this locust from ovarian membranes. By CHAPS solubilization of the membranes followed by ion exchange and immunoaffinity chromatography, a 100‐fold purification of the VTG receptor was achieved. The amino acid composition of the receptor protein has been determined. However, first attempts to sequence the receptor failed due to the N‐terminal blocking of the molecule. With the same methods the VTG receptor of another locust, Schistocerca gregaria, has been isolated, purified, and characterized. This receptor has an apparent Mr of 186 kDa under nonreducing conditions. It recognizes L. migratoria VTG and vice versa. However, in cross‐competition experiments in which the Schistocerca VTG competed with Locusta VTG for binding to the Locusta VTG receptor, the Schistocerca VTG was less efficient. Furthermore, the VTG receptor proteins of S. gregaria and L. migratoria are immunologically related as revealed by Western blotting with anti‐Locusta VTG receptor antibodies. It appears that important structural elements required for efficient and specific endocytosis of VTG have been conserved. © 1994 Wiley‐Liss, Inc.
Controlled ecdysteroid accumulation in eggs of the silkworm, Bombyx mori, by an imidazole compound (KK‐42), and embryogenesis in these eggs
Authors: Kadono‐Okuda Keiko , Amornsak Weerawan , Yamashita Okitsugu
Published: February 2005

An imidazole compound (KK‐42), a potent inhibitor of ecdysone synthesis, was applied to the female pharate adult of the silkworm, Bombyx mori, to control ecdysteroid accumulation in developing ovaries and mature eggs. KK‐42 applied on day 2 or later completely suppressed an increase in ecdysteroid content in developing ovaries. The inhibitory action of KK‐42 was restricted to vitellogenic follicles, i.e., those in which active ecdysteroid synthesis is occurring. Ecdysteroid content in the mature eggs of moths remained at the level accumulated in ovaries before KK‐42 application. Thus, KK‐42 was shown to be a novel agent to suppress the ecdysteroid accumulation in eggs. Eggs containing different amounts of ecdysteroids showed different levels of embryonic development. About 80% of the eggs which contained less than 10 ng free ecdysteroids/g eggs were not fertilized. More than 80% of the eggs containing less than 40 ng/g eggs of free ecdysteroids initiated embryogenesis but failed to hatch. Larvae hatched from almost all eggs which accumulated free ecdysteroids of more than 150 ng/g. Thus, maternal ecdysteroids appear to be required at different titers for fertilization, embryogenesis, and hatching of the silkworm larvae. © 1994 Wiley‐Liss, Inc.
Properties and significance of a riboflavin‐binding hexamerin in the hemolymph of Hyalophora cecropia
Authors: Magee John , Kraynack Nathan , Massey Holman C. , Telfer William H.
Published: February 2005

A riboflavin‐binding hexamerin isolated from pupal hemolymph of Hyalophora cecropia has a native Mr of 510,000, subunit Mr of 85,000, and a 5% carbohydrate content. An intrachain cross‐link was confirmed in protease limit digests. Ellman titration confirmed the presence of a sulfhydryl group, which is needed for this linkage. Though Cu2+ is known to promote the linkage, heavy metals were not detected in the isolate. Heat denaturation released ligand with the absorbency, fluorescence spectra, and chromatographic behavior of riboflavin. Binding resulted in substantial quenching of the fluorescence of both the isoalloxazine in riboflavin and of aromatic groups in the apoprotein. Kinetic analysis indicated a KD of 2.5 × 10−7 M for riboflavin, 1.3 × 10−7 M for lumiflavin, and greater than 1 × 10−6 M for FMN and FAD. Over four moles of flavin were bound per mole of hexamerin. The amount of riboflavin in pupal hemolymph is sufficient to occupy only 2–3 of these sites. Riboflavin is also associated with lipophorin and vitellogenin, but the molar ratios after protein isolation were low. On a standard laboratory diet, riboflavin is in great excess, but most of it is apparently excreted before the apoprotein first appears in the hemolymph, just before wandering. The concentration of riboflavin‐binding hexamerin rises to 15–30 mg/ml in pupae; relative to other hexamerins, very little is stored in the fat body. All of the apoprotein and 75% of riboflavin disappear from the hemolymph during adult development. An amount of flavin at least equal to that stored in pupal hemolymph is transferred to the eggs formed during this period. © 1994 Wiley‐Liss, Inc.
Pheromonotropic stimulation of moth pheromone gland cultures in vitro
Authors: Rafaeli Ada
Published: February 2005

The direct neurohormonal control of pheromone biosynthesis by pheromone biosynthesis activating neuropeptide (PBAN) was demonstrated in Helicoverpa (Heliothis) spp. using pheromone gland cultures in vitro. Pheromone gland activation involved the de novo production of the main pheromone component (Z)‐11‐hexadecenal as revealed by radio‐TLC, radio‐HPLC, and radio‐GC. Activation was found to be a specific response attributed to pheromone gland cultures alone. Specificity of pheromonotropic activation was demonstrated to be limited to nervous tissue extracts. A sensitive and specific radioimmunoassay was developed using [3H]‐PBAN, and the spatial and temporal distribution of PBAN‐immunore‐activity was studied. PBAN‐immunoreactivity in brain complexes was found throughout the photoperiod and in all ages. From the distribution of PBAN‐immunoreactivity it appears that PBAN release is affected by photoperiod. Pheromone gland cultures were found to be competent to pheromone production irrespective of age and photoperiod. Therefore, the neuroendocrine control of pheromone production operates at the level of neuropeptide synthesis and/or release and not at the level of the target tissue itself. The involvement of cyclic‐AMP as a second messenger system was demonstrated. Brain extracts and PBAN were shown to stimulate dose‐ and time‐dependent changes in intracellular cyclic‐AMP levels. The role of cyclic‐AMP in this mechanism was further verified by the ability of cyclic‐AMP mimetics to mimic the pheromonotropic effect of brain extracts and PBAN. However, dose‐response studies using PBAN and a hexapeptide C‐terminal fragment of PBAN suggested that PBAN induces a two mechanism response, one occurring at low PBAN concentrations (high affinity receptor) and another at higher PBAN concentrations (low affinity receptor). Further evidence indicating a dual receptor system was obtained with the observation that the active phorbol ester (phorbol‐12‐myristate 13‐acetate), the diacyl‐glycerol analog (1,2‐dioleolyl‐sn‐glycerol), and the intracellular calcium ionophore (ionomycin) mimicked the physiological action of PBAN and that lithium chloride had a pheromonostatic effect. The results indicate that pheromone glands also possess receptors that are linked to inositol phosphate hydolysis. © 1994 Wiley‐Liss, Inc.
Effects of allatectomy and ovariectomy on cuticular hydrocarbons inCalliphora vomitoria(diptera)
Authors: Trabalon Marie , Campan Mireille , Hartmann Nadine , Baehr Jean‐Claude , Porcheron Patrick , Clément Jean‐Luc
Published: February 2005

The relationships between female attractiveness, cuticular hydrocarbons, and levels of juvenile hormone and ecdysteroids were studied inCalliphora vomitoria. The experiments were conducted at 48 and 72 h post‐emergence, according to attractiveness appearance and increase. The 48‐h‐old allatectomized females were less attractive than the control females, whereas no changes occurred either in cuticular hydrocarbons total mass production or in the different hydrocarbon families. However, the 72‐h‐old allatectomized females were more attractive than the control females, and, in relative proportions, allatectomy led to an increase in monomethylalkanes and a decrease in n‐alkanes.

Only at 48 h were the ovariectomized females less attractive than the control females and did ovariectomy increase the relative proportions of monomethylalkanes. At 72 h, ovariectomy did not influence female attractiveness, but it decreased the total cuticular hydrocarbon production. Allatectomy and ovariectomy significantly decreased ecdysteroids levels at 48 and 72 h. Ovariectomy did not affect juvenile hormone production.

These results suggest that attractiveness and cuticular hydrocarbon synthesis could be under the direct control of ecdysteroids and the indirect influence of juvenile hormone. © 1994 Wiley‐Liss, Inc.
Cellular defense mechanisms in C. capitata: Recognition and entrapment of E. coli by hemocytes
Authors: Marmaras Vassilis J. , Charalambidis Nektarios D. , Lambropoulou Maria
Published: February 2005

The mechanism of recognition of foreignness and entrapment of invaders by the immune system of insects is unknown. In this report using hemocyte monolayer preparations and biochemical analysis we demonstrate the requirements for recognition of E. coli in vitro, their entrapment by hemocytes, and nodule formation. A model system consisting of an isolated hemocyte protein (47 KDa), isolated hemocyte tyrosinase, isolated hemocytes, tyrosine, and E. coli was used to obtain these results. The 47 kDa polypeptide has the ability to form adducts with tyrosine derivatives generated by the action of tyrosinase and to attach to the E. coli surface. The latter process takes place independently of tyrosinase activity. When the E. coli‐47KDa protein complex was overlaid on hemocyte monolayers followed by tyrosine and tyrosinase or vice versa, the bacteria were entrapped by hemocytes. The same results were obtained when the monolayers were overlaid with 47 KDa protein, followed by E. coli‐47 KDa protein complex and then tyrosine and tyrosinase. The same experimental procedure in test tubes resulted in nodule formation. These results permit us to propose that the most likely explanation for the entrapment of E. coli to hemocytes and the formation of nodules is the production of E. coli‐47 KDa complexes, and their crosslinking through a quinone intermediate generated by the action of tyrosinase on hemocytes. © 1994 Wiley‐Liss, Inc.
Parasitoid‐host interactions: Historical perspectives and current status
Authors: Jones Davy , Strand Michael
Published: February 2005

The papers contained in this special issue of Archives of Insect Biochemistry and Physiology are published in connection with the symposium entitled “Physiological and Molecular Interactions Between Parasitoids and Their Hosts” held at the XIX International Congress of Entomology, Beijing, China. Speakers in the program discussed advances in the field and provided insight into future directions of study. As a preface to this special issue, we summarize the history of this field as reflected through the continuing series of symposia held in association with the International Congresses.
Biochemical and developmental alterations of Heliothis virescens (F.) (lepidoptera, noctuidae) larvae induced by the endophagous parasitoid Cardiochiles nigriceps viereck (Hymenoptera, braconidae)
Authors: Pennacchio Francesco , Vinson S. Bradleigh , Tremblay Ermenegildo , Tanaka Toshiharu
Published: February 2005

All larval stages of Heliothis virescens (F.) parasitized by the endophagous larval parasitoid Cardiochiles nigriceps Viereck, a braconid species belonging to the subfamily Microgasterinae, exhibit developmental arrest at last instar and fail to pupate. The major part of larval development of the parasitoid is synchronized with the arrested host last larval instar and the parasitoid first molt is never observed before the host attains the late digging stage. At this time, the total ecdysteroid titer of the hemolymph of parasitized hosts is very low and subsequently shows a slow and gradual increase, characterized by a low titer of 20‐hydroxyecdysone (20‐HE) associated with consistent amounts of inactive ecdysteroid polar metabolites. Juvenile hormone esterase (JHE) activity is high in both control and parasitized host larvae at the early digging stage of development, and juvenile hormone analogs (JHA) applied to parasitized host last instar larvae appear to suppress the parasitoid molt. Concurrent with these changes was an increase in the hemolymph titer of proteins which was maintained at a high level in parasitized larvae in contrast to the observed decrease in control larvae at the cell formation stage of development. Neck‐ligation of newly molted host 5th instar parasitized larvae, prior to both JHE release and the increase in protein titers, inhibited growth and molting of the parasitoid. In contrast, ligation after JHE release and with high hemolymph protein titers resulted in parasitoid molting and growth. These data suggest that the host ecdysteroid hormones are not directly involved in the regulation of the parasitoid molt, although high juvenile hormone (JH) levels probably prevent it. More likely, molting is triggered by other biochemical changes, such as proteins or other factors occurring in the hemolymph. Molting of C. nigriceps larvae in vitro into an ecdysone‐free semidefined medium further supported the view that host ecdysone is not necessary for the molt. Teratocytes of C. nigriceps seem to play an important role in the inactivation of 20‐HE through its conversion to inactive polar metabolites, and along with female calyx fluid and venom which depress the secretory activity of the host prothoracic glands, they are the most important sources of host regulatory factors. © 1994 Wiley‐Liss, Inc.
Immunological analysis of lipophorin in the haemolymph, ovaries, and testes of the fall webworm, Hyphantria cunea (Drury)
Authors: Yun Hwa Kyung , Kim Woo Kap , Kim Hak R.
Published: February 2005

Lipophorin (LP) was purified from haemolymph in last instar larvae of Hyphantria cunea (Drury) by KBr density gradient ultracentrifugation and gel filtration. LP is composed of Apo‐LP I and Apo‐LP II with molecular weights of 230 kDa and 80 kDa, respectively.

The level of haemolymph LP in early pupae was somewhat greater than in last instar larvae. In males, this LP concentration is maintained throughout pupal development, whereas the level of haemolymph LP decreases in female pupae beginning at day 7, coincident with the onset of vitellogenesis in the fall webworm. In both male and female adults, haemolymph LP concentrations were dramatically increased in comparison to their pre‐adult levels. Actually, LP was found in the ovary by immunodiffusion, tandem‐crossed immunoelectrophoresis, and Western blotting. Location of LP in the ovary was also traced by immunogold labelling. Also, LP appeared in small amounts in protein yolk bodies of the ovary at an early stage of vitellogenesis, when nurse cells are bigger than the oocyte, but in greater amounts at those stages when the oocyte is larger than nurse cells—that is, when vitellogenesis is actively taking place. This fact clearly reveals that LP is synthesized by fat body and released into the haemolymph, and then taken up by the growing ovary during vitellogenesis. Also, LP was detected in testes by immunological analysis. Western blotting showed that LP was present in testicular fluid but not in the peritoneal sheath and cysts. To test whether LP is also synthesized in testes, testes and fat body tissues were cultured in vitro, indicating that fat body synthsizes LP but testes do not. The result showed that the haemolymph LP itself is taken up into the testes. © 1994 Wiley‐Liss, Inc.
Synthesis and deposition of PCG‐100, the main pupal cuticle glycoprotein of the medfly Ceratitis capitata
Authors: Boccaccio Graciela L. , Quesada‐Allué Luis A.
Published: February 2005

We have studied the developmental expression of the main pupal cuticle glycoprotein, PCG‐100, in the Medfly Ceratitis capitata. A polyclonal antiserum was raised against this protein. Western blotting analysis showed that this glycoprotein is integument‐ and stage‐specific. No PCG‐100 or immunologically related polypeptides were detected in other tissues or instars. As studied by microinjection of [35S]methionine in individual flies, in vivo synthesis and deposition of PCG‐100 begins approximately 48 h after the onset of pupariation, shortly after the time of head eversion. Synthesis is maximal at 54–64 h, decreases at 72 h, and practically ceases in fully shaped 4‐day‐old pupae. The time required for PCG‐100 deposition into the cuticle was found to be less than 10 min after its synthesis. This is the first time such in vivo analysis has been performed. © 1994 Wiley‐Liss, Inc.
Purification of a 24 kD parasitism‐specific hemolymph protein from pharate pupae of the caribbean fruit fly, Anastrepha suspensa
Authors: Rolle Rosa S. , Lawrence Pauline O.
Published: February 2005

We report purification of a 24 kD parasitism‐specific protein (24 kD PSP) from pharate pupal hemolymph of the Caribbean fruit fly, Anastrepha suspensa, after parasitization by the braconid wasp, Diachasmimorpha (= Biosteres) longicaudata. We previously utilized isoelectric focusing (IEF) and two‐dimensional (2‐D) gel electrophoresis to demonstrate that the 24 kD PSP consists of two variants with pl 6.7 (more abundant) and pl 6.3. Purification of the more abundant 24 kD PSP variant was accomplished by Concanavalin A (Con A) sepharose B affinity chromatography followed by DEAE column chromatography. A second protocol, utilizing wheat germ agglutinin (WGA) sepharose 6MB affinity chromatography between the ConA and DEAE chromatographic steps, resulted in the purification of a partially deglycosylated form of the 24 kD PSP which retained its immunore‐activity with anti‐PSP serum but which exhibited a greater relative migration in sodium dodecyl sulfate (SDS)‐PAGE than the pl 6.7 24 kD PSP variant.

For structural studies both 24 kD PSP variants were purified from whole hemolymph by flat bed IEF followed by SDS‐PAGE. Peptide cleavage profiles in 1‐D SDS‐PAGE after treatment with BNPS‐skatole, CNBr, and endproteinases Lys‐C and Asp‐N were identical for both 24 kD PSP variants. Primary N‐terminus sequences of at least the first 20 amino acid residues of both variants were identical. A secondary sequence of five amino acids residues was detected in both variants at Thr, the seventh amino acid residue from the N‐terminus of the primary sequence. These data indicate that both 24 kD PSPs are glycoforms of a branched, apparently homogeneous polypeptide. © 1994 Wiley‐Liss, Inc.
Carbohydrate metabolism during the pupal molt of the tobacco hornworm, Manduca sexta
Authors: Siegert Karl J.
Published: February 2005

During the pupal molt of the tobacco hornworm, Manduca sexta, the percentage of active fat body glycogen phosphorylase increased from 5–10 to 20%, but only for a period of 5 h prior to the molt. From the time of the appearance of two sclerotized dorsal bars to the time of the molt, the concentration of total hemolymph carbohydrates doubled to 100 mM trehalose. Initially, the glucose level was high (16 mM) when compared with feeding larvae (approximately 1 mM) but decreased to zero just prior to the molt. The amount of cuticular chitosan decreased from approximately 100 mg to 10 mg at pupation; the exuvia contained approximately 7 mg. While the levels of total lipids in hemolymph were not affected, the lipid content of the fat body decreased significantly prior to the molt but increased sharply thereafter. Fat body glycogen phosphorylase in pharate pupae and pupae of M. sexta was substantially activated by the Manduca adipokinetic peptide hormone, which in pharate pupae, produced the same response at 2 and 20 pmol per insect as in ligated larval abdomens. In pupae the response was clearly reduced. Using chilling to stimulate glycogen phosphorylase, it was found that the enzyme in pharate pupae and pupae responded both in vivo and in vitro as in ligated abdomens of larvae. Thus, a transition to the adult response seems to occur during the pupal and pharate adult development. © 1995 Wiley‐Liss, Inc.
Hypertrehalosemic hormone‐regulated gene expression for cytochrome P4504C1 in the fat body of the cockroach, Blaberus discoidalis
Authors: Lu Kuang‐Hui , Bradfield James Y. , Keeley Larry L.
Published: February 2005

Hypertrehalosemic hormone (HTH) up‐regulates expression of a gene for a cytochrome P450 of family 4 (CYP4C1) in the fat body of adult male B. discoidalis cockroaches. Studies were undertaken to determine the characteristics of HTH‐dependent CYP4C1 expression. A dot‐blot assay was developed for routine measurement of the relative levels of CYP4C1‐mRNA in fat body RNA extracts. A single injection of 10 pmol HTH produced a maximum CYP4C1 response within 8 h. This dose corresponds with the dose needed for a maximum in vivo hypertrehalosemic response to HTH (Keeley et al., 1991). Multiple treatments with HTH at 8 or 24 h intervals were no more effective than a single treatment for producing CYP4C1 expression. These results indicate that CYP4C1 expression is sensitive to physiological doses of HTH and responds rapidly. CYP4C1 expression was suppressed by treatment with α‐amanitin, an inhibitor of RNA polymerase II, but was unaffected by cycloheximide, an inhibitor of protein synthesis. HTH appears to influence CYP4C1 transcription without involvement of intervening regulatory genes. These results suggest that regulation of fat body CYP4C1 expression is a major physiological action of HTH. © 1995 Wiley‐Liss, Inc.
Chitin synthesis in Spodoptera frugiperda wing imaginal discs. III: Role of the peripodial membrane
Authors: Mikołajczyk Piotr , Zimowska Grazyna , Oberlander Herbert , Silhacek Donald L.
Published: February 2005

We determined the contribution of the peripodial membrane to chitin synthesis in cultured wing imaginal discs of Spodoptera frugiperda. This was accomplished by examining chitin synthesis in vitro in intact imaginal discs, in the peripodial membrane, and in imaginal discs in which the peripodial membrane had been injured. Chitin synthesis in peripodial membrane‐deprived imaginal discs, peripodial membrane injured imaginal discs, and peripodial membrane fragments was assessed by measuring incorporation of [14C]GlcNAc after treatment with 20‐hydroxyecdysone in tissue culture. Removing or injuring the peripodial membrane resulted in a marked decrease in ecdysteroid‐dependent chitin synthesis in these wing discs compared with intact wing discs. In addition, a break in the ecdysteroid treatment of 4 h reduced chitin synthesis in the wing discs substantially. These biochemical experiments were supplemented with ultrastructural and immunocytochemical approaches. A wheat germ agglutinin colloidal gold complex was used to visualize the presence of chitin synthesized by wing discs including the peripodial membrane. These experiments confirmed the importance of an intact peripodial membrane for optimal production of cuticle by the wing pouch. Our results demonstrate that for opti‐ma1 production of chitin in tissue culture, wing discs must be treated with 20‐hydroxyecdysone for an uninterrupted period of 48 h, and the peripodial membrane of these imaginal discs must be present and uninjured. © 1995 Wiley‐Liss, Inc.

This article is a US Government work and, as such, is in the public domain in the United States of America.
Purification and immunological characterization of superoxide dismutase of the onion maggot, Delia antiqua
Authors: Ishikawa Yukio
Published: February 2005

Cytosolic superoxide dismutase (SOD) of the onion maggot, Delia antiqua, was purified to apparent homogeneity by ammonium sulfate fractionation followed by anion exchange, hydrophobic interaction, and gel filtration chromatographies. Native molecular mass was estimated as 32,000 daltons. SDS‐PAGE revealed only one subunit of 16,000 daltons, indicating that SOD is a homodimer. Isoelectric focusing revealed 3 charge isomers of pls 5.3, 5.5, and 5.7. The specific activity of purified SOD was 4,250 U/mg protein. A monoclonal antibody (MAb, aSOD2B7) raised against Delia SOD recognized only SOD of the same genus, but another MAb (aSOD1H11) recognized SOD of Drosophila melanogaster as well. © 1995 Wiley‐Liss, Inc.
Separation of multiple forms of acidic glutathione S‐transferase isozymes in a susceptible and a resistant strain of house fly, Musca domestica (L.)
Authors: Chien C. , Motoyama N. , Dauterman W. C.
Published: February 2005

The acidic glutathione S‐transferases from a CSMA (susceptible) strain and a Cornell‐R (resistant) strain of houseflies were purified and separated utilizing affinity chromatography followed by chromatofocusing. Nine fractions were isolated from each house fly strain. Fraction 1 had the highest 1‐chloro‐2,4‐dinitrobenzene vs. 1,2‐dichloro‐4‐nitrobenzene ratio (CDNB/DCNB ratio) in both strains and the ratio of all the other fractions tended to decrease as the isoelectrical points decreased except for fractions 4 and 9. Most fractions from the CSMA strain had higher CDNB conjugation activities than the fractions from the Cornell‐R strain, but all the fractions from the CSMA strain had lower DCNB conjugation activities than fractions from the Cornell‐R strain. Steady‐state kinetics of all the fractions were examined. The Km values obtained from both strains ranged from 0.36 to 1.12 mM, while the Vmax value ranged from 3.0 to 32.6 μmol/min/mg. In the 100,000 g supernatant, the CDNB specific activities in the CSMA strain was about 1/3 of the activity in the Cornell‐R strain but it was about 1.5‐fold following affinity chromatography. The specific activity for DCNB measured in the CSMA strain was only 1/5 of the activities of the Cornell‐R strain in the 100,000 g supernatant, but was about the same after affinity chromatography. The difference was due to the selectivity of the affinity column used in the current study. © 1995 Wiley‐Liss, Inc.
Developmental differences in the sterol composition of Solenopsis invicta
Authors: Ba Amadou S. , Guo De‐An , Norton Robert A. , Phillips Sherman A. , Nes W. David
Published: February 2005

Twenty‐six sterols were isolated from eggs, larvae, workers, and queens of the red imported fire ant, Solenopsis invicta Buren. They were identified by chromatographic (TLC, GLC, and HPLC) and spectral methods (MS and 1H‐NMR). Queens possessed the most varied sterol composition (24 sterols were detected). The major sterols from queens were the doubly bioalkylated 24α‐ethyl cholest‐5‐ and 7‐en‐3β‐ols whereas the major sterol from the other developmental stages was cholesterol, a sterol which lacks a C‐24 alkyl group. From fourth instar larvae were isolated two yeasts, Candida parapsilosis and Yarrowia lipolytica. Both yeasts were found to synthesize similar sterols, primarily ergosterol and zymosterol (90% of the sterol mixture). A minor sterol (approximately 12% of the total sterol mixture) detected in eggs, larvae, and workers was 24‐methyl cholesta‐5,22E‐dien‐3β‐ol (brassicasterol). Brassicasterol may have originated from ergosterol produced by the fungal endosymbiotes. The amount of sterol in each developmental stage was as follows: approximately 24 μg sterol/queen, 3 μg sterol/worker, 2 μg sterol/larvae, and 0.02 μg sterol/egg. The sterol composition of the red imported fire ant differed from that of leaf‐cutting ants previously investigated where 24‐methyl sterols of ectosymbiotic fungal origin were the major sterols detected in soldiers and workers. © 1995 Wiley‐Liss, Inc.